Department of Biotechnology Lund University, Sweden · Biotechnology at a glance The Department of...

Department of BiotechnologyLund University, Sweden

Annual Report 2006

Biotechnology at a glance

The Department of Biotechnology at Lund University was founded in 1985 by Professor Bo Mattiasson and is located at the Center for Chemistry and Chemical Engineering in Lund. During the past 21 years, both the number of research areas and people have grown. Today the department consists of about 60 research-ers, PhD students and staff, and is visited by around 20 master students and 40 international guest researchers every year. The research activities at the department are focused on various as-pects of environmental-, analytical-, food- medical- and industrial biotechnology and consist of both basic and applied research. The main research areas are protein structure and function, biodi-versity, green chemicals and materials, bioseparation, new ma-terials for biotechnical applications, bioanalysis, functional food, bioremediation and renewable energy. Every year around 10 PhD students graduate from the department. The department is re-sponsible for 8 undergraduate courses within the Biotechnology programme at LTH, two free-standing courses and a course pro-gram for PhD students and takes active part in the International Masters programme in Bio- and Food technology at LTH, Lund Institute of Technology.

Foreword

The need to inform about activities in academic research laboratories is escalating. There is a need felt from the society in general, from the funding bodies and from present and future collaborators. With this in mind, we have since two years back published an annual reeport compiling the activities within the department of Biotechnology.As you will see from the text, the activities in the department cover a range of fields, from the very basic issues to some very applied. It is indeed a privilege to be active in a field with so close contact between basic science and applications, and at the same time in fields where the development is very fast.We hope that the information presented in this report will stimulate new thoughts and a wish to know more about some specific issues. Please use the contacts given to the individual scientists, or contact the editors of this document.

COVER PHOTOS. Top from left to right: HPLC analysis of volatile fatty acids, Chemo-enzymatic epoxidation of vegetable oil, Hot spring in Bolivia, Activated sludge process in course lab, Hemp as energy crop, Collection of sea weed for biogas production

Dept. of Biotechnology Annual Report 2006 �

CONTENTS

Head of Department´s report 4External communication 5Services 6 Research activities 8 Protein structure and function 9 Biodiversity 10 Green chemicals and materials 11 Bioseparation 12 New materials 13 Bioanalysis 14 Functional Foods 15 Bioremediation 16 Renewable energy 17 Education 18 Courses 18 Masters education in Bio- and Food technology 19 Master theses 19Staff 20 PhD students 21Visiting researchers and students 22International collaborations 24Funding and grants 26Publications 27 Journal articles 27 Doctoral degrees and theses 30 Licentiate degrees and theses 30 Conferences 30

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Dept. of Biotechnology Annual Report 2006�

activities that have come a long way. There are indi-cations that these activities will be strengthened and proliferated during the coming years. The research program Greenchem with a focus on production of “green” chemicals was granted continued funding for Phase 2 comprising 4 years.

Biotechnology for medical/pharmaceutical applica-tions is developing into an interesting field at the department. New technology for separation of mam-malian cells has been developed. Likewise, systems for cultivation of mammalian cells in reactors as well as implantable materials have been studied. Studies have been ongoing for the development of process analytical tools (PAT) for monitoring of products and impuri-ties during production of biopharmaceuticals. There have also been projects in the area of food biotechnol-ogy such as those dealing with antioxidants and lipid modification.

During 2006, the department has been successful in acquiring grants from the Research Council to a larger

Head of Department´s reportAlthough a broad spectrum of research activities are pursued at the department, some focus areas and even some new directions have emerged. It became quite

extent than ever before, and at the same time increasing interactions with industries have also led to funding of projects to a larger extent. We are also proud to have

“2006 - A successful year on the whole and also a year of transition for Department of Biotechnology”

clear that our work within the area of in-dustrial biotechnology is very much in line with the global trends towards envi-ronmental sustainability. Biotechnologi-cal means of production of energy, chemi-cals and materials from renewable, surplus biomass or wastes, and waste-water treat-ment have evolved as the strong research themes, and have further involved exten-sive contacts with industry and society. Production of chemicals using biocataly-sis, screening and development of bio-catalysts from nature´s diversity and by protein engineering, production of biode-gradable plastics by fermentation, process intensification of biogas production from various wastes, and development of tech-nology for enrichment and treatment of environmental pollutants are some of the

been a birthplace for some entrepreneurial ventures.

The Department has also been heavily involved in undergraduate courses both for the Master of Science in Engineering Programme and International Master Programme. At the graduate level, six PhDs passed out from the department during 2006 and a few students were registered for their PhD training. In a number of research projects and also in education, Department of Biotechnology has collaborated with some other de-partments at Lund University, and other universities in Sweden and abroad. We hope to continue this trend.

This introduction is a collaborative effort of the new and the former head of department. Bo Mattiasson has left the chairmanship after almost 22 years and Rajni Hatti-Kaul is now in charge.

Lund, April 2007

Bo MattiassonRajni Hatti-Kaul

Present and former Head of Department, Rajni Hatti-Kaul and Bo Mat-tiasson (photo Marika Murto)


External communication

Today, the scientific institutions traditionally known for their research and education, are being transformed by expanding interactions with the surrounding soci-ety. This is also the case with Department of Biotech-nology. We consider engagement with business and industry as well as interactions with the wider commu-nity as very important in being able to meet the needs and expectations from the surroundings. Besides the academic staff, the department has two technology co-ordinators with the task of being ambassadors for en-trepreneurial activities at the department and encour-aging the interactions between academia and industry. We actively make contacts with industries and other potential collaborators outside the university. Among the activities we engage ourselves in are collective pro-grammes such as Greenchem, bilateral agreements including contract research, training of company per-sonnel by e.g. industrial PhD students or specialized courses and scientific collaborations of different kind. The Department of Biotechnology recognises the importance of allowing valuable research ideas be-ing tested for their commercial importance and sup-ports the start-up of new businesses. One example of this is the company “Bioprocess control” which was established as a result of collaboration between Dept. Biotechnology (Jing Liu and Bo Mattiasson) and Industrial Electrical Engineering and Automa-tion (Gustaf Olsson). During 2006 the company, which is run by Jing Liu, research associate at Dept. Biotechnology, received 250 000 SEK by winning the competition Vinn Nu. Besides this their busi-ness idea ended up at third place in Venture Cup. Jing Liu was noticed for his combined research and entre-preneurial assignments in “Process Control”, 8/06.

During the year, a large number of activities have been carried out to spread information about our research in food, medical, industrial and environmental biotech-nology. In our research programme Greenchem, the external communication has been quite extensive during 2006. Through several scientific papers, one doctoral thesis, four Greenchem conferences/semi-nars, several lectures at external conferences/semi-nars, eight public articles and one radio program, the Greenchem newsletter and the Greenchem

homepage (www.greenchem.lu.se), knowledge about industrial biotechnology has been spread to scientists, politicians, business people and the general public. The Biogas area continued to attract attention during 2006. The research station Anneberg was vis-ited by several groups of people, showing the large in-terest in the Biogas research the department is involved in. Within the programme BiogasÖresund, which is a network for matchmaking, knowledge exchange and cooperation in the field of biogas in the Öresund re-gion, several workshops and meetings were held. The project is managed by the Department of Biotechnol-ogy and the Institute of Environment & Resources at the Technical University of Denmark. The goal is to promote and develop the production and use of biogas in the Öresund region. In June a workshop “Biogas process optimization, manure treatment and nutrient recovery” was held in Denmark to inform about and discuss the latest research results in the biogas area.

Participants at the seminar “Applied biogas research in the Nordic countries – the role of energy crops in agricultural biogas production” (Photo Marika Murto)

In October another public seminar about “Applied bi-ogas research in the Nordic countries – the role of en-ergy crops in agricultural biogas production” was held in Malmö.

Several of our researchers have been invited to give lectures at different seminars, conferences and meet-ings as well as industries, both in Sweden and abroad.

The webpage for the department of Biotechnol-ogy was during 2006 visited by almost 10.000 visitors, and is expected to be upgraded with a new design and more information during year 2007.

The Department of Biotechnology is a scientific institution providing an inspiring environment where sci-entific work and education goes hand in hand with interactions with society. During 2006 we continued our mission of enhancing our contacts with the surrounding society.

Dept. of Biotechnology Annual Report 20066

Services for external customersThe department offers services to other academic institutions and industry in different areas where the department has expertise and possesses special equipment or research facilities. We have the possibility to run pilot scale fermen-tations upto 400 litres scale, design and perform biogas production, assist in running mass spectrometry experiments on our LC-MS/MS equipment or perform other type of evaulations and optimizations of biotechnological processes or technologies.

The contact persons are the following:Pilot plant fermentations: Prof. Olle Holst: [email protected], phone: 046-222 98 44LC-MS/MS: Martin Hedström: [email protected], phone: 046-222 08 82DSC: Eva Nordberg Karlsson: [email protected], phone: 046-222 46 26All other services: Maria Andersson, technology coordinator. [email protected], phone: 046-222 75 78 or Josefin Ahlqvist, technology coordinator. [email protected], phone: 046-222 48 38

LC-MS/MS Mass spectrometer (Q-Star)At the department we have a mass spectrometer of the type Quadrupole-Time Of Flight, Q-TOF (Q-Star, Applied Biosystems), which is con-nected to a LC-system. The mass spectrometer is equipped with an electrospray (Turbospray) ionization source and a nanospray system. The nanospray system can be connected to syringe pumps in order to get a continuous nano-system equipped with capillary columns for separation on a column packed with C18 or other packing material of interest. It also has a Maldi ioniza-tion interface.

Victor Ibrahim, a student at the department running an MS experiment (photo Martin Hedström)

DSC Differential Scanning Calorimeter

The department of Biotechnology owns a differential scanning calorimeter (VP-DSC). This is a sensitive, easy to use differential scanning calorimeter, with an active cell volume of approximately 0.5 ml used for studying samples in solution. The operating temper-ature of the instrument is in the range of –10ºC to +130ºC, and the instrument has a selectable scan rate in the range of 0ºC to 90ºC per hour (upscan), allow-ing studies of fast or slow transition processes.

It can be used to measure the intramolecular stability of a broad spectrum of biomolecules, including pro-teins, nucleic acids, lipids and detergent micellar sys-tems. The VP-DSC provides fast, accurate transition midpoint (Tm) determination, as well as a thermody-namic profile that can provide insight into the factors that affect conformation and stability.

Christina Wennerberg working at the DSC (photo Eva Nordberg-Karlsson)


The department offers as a service evaluation and optimization of processes and technologies in the field of biotechnology based on the genuine knowledge in fermentation, protein purification, immobilization, enzyme ca-talysis, bioremediation, biogas production, bioanalysis and other areas within biotechnology. During 2006 several projects were initiated together with industry for optimization of processes or evaluation of technologies.

Evaluations and optimizations

Pilot plant fermentor

The station for Biogas oriented research at Anneberg is situated in close connection to a crop farm. It is an impor-tant site for demonstration and for getting increased acceptance of farm based biogas technology.

The research station consists of a wide range of test reactors in 1-350 m3 scale which are useful as test rigs in the

The fermentation pilot plant was renovated in 2004 and is located in close connection to the department´s course lab. The new lab facility contains bright, espe-cially ventilated rooms with course lab, chemical stor-age room, washing room, sterile room and office. The pilot plant is equipped with fermentors (15, 75 and 400 liters scale), equipment for downstream process-ing of microorganisms (filters and cell disintegrator), as well as for downstream processing of biological mol-ecules. The whole facility is approved for work with class 2 organisms.

Research station for biogas oriented research

ongoing biogas research. The set-up allows many types of process design to be investi-gated and new methods for on-line process monitoring to be evaluated.

For a successful project close co-opera-tion between research and agriculture is necessary. The project is managed by Prof. Bo Mattiasson and Dr. Lovisa Björnsson at the Department of Biotechnology, Lund University together with Agricultural Tech-nologist Kjell Christensson. The studies are carried out at Lund University and at the Research Station Anneberg situated in the municipality of Svalöv.


Research activities

Towards a sustainable developmentBiotechnology for a sustainable development is coming in focus at the same time as the awareness of climate change and shortage of water for a growing population are becoming focal points among politicians around the world. We feel that during year 2006 the research activities within our department became increasingly valuable and important for the surrounding society, and in the light of this development, it appears that earlier deci-sions to direct the research towards the development of environmentally friendly processes have been correct.

Over the years the research activities within the department of Biotechnology have evolved towards a sustainabili-ty thinking and research projects within this field have emerged as an integrated part of the scientific programme at the department and led the development one step further. During the past years, the research has been focused within the fields of ENVIRONMENTAL biotechnology, MEDICAL biotechnology and FOOD biotechnology. INDUSTRIAL biotechnology or “white” biotechnology is seen as another important field to position our research in, especially since we believe that biotechnology will play an important role in the future when it comes to replac-ing materials based on fossile carbon with renewable feed stocks.

The wide knowledge base at the department within bioseparation, cell cultivation techniques and enzyme technol-ogy makes us well equipped for research within diverse areas such as energy production, waste water treatment, bioremediation, conversion of renewable feed stocks into value added products (green chemicals and materials), tissue engineering, production of food components with improved properties, improvement of process analytical technology and production of pharmaceutical proteins.

Food biotechnology

Industrial biotechnology

Environmental biotechnology

Enzyme technology

Medical biotechnology

Bioseparation Cell cultivation techniques

Bioorganic synthesis

Enzyme stability and specificity

Protein engineering

Affinity interaction

Smart polymers

Cryogels

Simulated moving bed technology

Aqueous two-phase systems

Renewable energy

Bioremediation

Lipids

Antioxi-dants

Bacteriocins

Feed additives

Polysaccharides

Green chemicals

Biomaterials

Chiral products

Biodiversity

Extremophiles and extremozymes

Recombinant protein production

Tissue engineeringPharmaceutical

proteins

Plasmid DNA

Process analytical technology

PAT

Illustration: Overview of the research activities at the Department of Biotechnology


Protein Structure and Function

Eva Nordberg KarlssonAssociate Professor

Gashaw MamoResearch engineer

Creating functionality and stabilityStable and selective catalysts are prerequisites for efficient biocatalytic proc-esses, and development of novel enzymes paired with a good understanding of the target proteins is thus an area of exquisite importance for creation of future biocatalysts.

ExtremozymesAt the dept of Biotechnology, we have specialized in research on enzymes from extremophiles, i.e. microorganisms adapted to extreme ecological con-ditions (heat or cold, high or low pH, or high salt conditions). Our research is based on use of recombinant enzymes, thus allowing genetic development of the selected target molecules, selected based on proper-ties like, stability, reaction specificity or production yield. Our aim is de-

Projects:- Fundamental studies of glycosidases from extreme environments- DATAGENOM- A thermostable carbohydrate binding module as a general scaffold for diversity and specificity - High pH catalytic adaptation of extremozymes - Enhancing the thermal stability of alkaline active enzymes

PhD students:Pernilla Turner

Figure. Front cover of the Green chemistry issue 8, 2006 (Turner et al, 2006). Selected thermophilic β-glucosidases used in combination with sub-critical water extraction for extraction and modification of anti-oxidants from onion waste.

Figure (top). Substrate bound structure of the cellulase Cel12A from Rhodothermus marinus (Solved in collaboration with Dr S.J. Crennell at the University of Bath, UK [Crennell et al, 2006])

velopment of biocatalysts for environmental friendly processing, e.g. for the production of green chemicals, with the focus on carbohy-drate modifying enzymes.

Carbohydrate modifying enzymesCarbohydrates are essential components in life, and biomass (estimated produced quan-tity is about 60 Gt/year) contains an array of structural and storage polysaccharides (like starch, cellulose and hemicellulose). These polysaccharides are richly available renewable rawmaterial resources, many of which are im-portant in industrial applications. Glycoside hydrolases (GHs) are enzymes which hydrolyse glycosidic bonds between two or more carbohydrates or between a carbohy-drate and a non-carbohydrate moiety. These

enzymes are subject of growing interest due to their wide range of possible applications contributing to environmental-friendly processes. Creation of more efficient biocatalysts with good stability is however crucial to make biocatalytic processes a viable alternative. We employ both rational and random engineering strategies in studies aiming at transforming a number of GHs into more perfect biocatalysts, and via collaborations utilize struc-tural biology resources to understand the molecular reasons for the changed properties that improve their performance in applications.

Enzyme productionThe possibility to produce the recombinant protein is a necessity for use and characterization of created variants. To achieve these practical produc-tion levels, usage of genetically modified microorganisms is essential. As a natural complement to enzyme development, batch and fed-batch strate-gies for their heterologous production are utilized and developed.


Biodiversity

Rajni Hatti-KaulProfessor

Gashaw MamoResearch engineer

Bo MattiassonProfessor

Olle HolstProfessor

Projects- Screening for thermostable alcohol dehydrogenases- Biodiversity of alkaline and saline environments- Isolation and characterization of extremophiles from highly alkaline environments.- Anaerobic microorganisms for biotechnological applications

PhD studentsDaniel Guzman Laura MendozaCarla CrespoRosa Aragao

Post DocJorge Quillaguaman

Nature - an infinite source of activitiesMicrobial diversity existing in nature is extremely vast. Only a small frac-tion of the natural microorganisms have so far been identified. Most of the biotechnological applications have been made possible with the help of microbial cells or their products. Microorganisms inhabiting extreme envi-ronments, the so called extremophiles, have been of interest both for un-derstanding the mechanisms used by them for survival as well as for exploit-ing their potential for applications requiring extreme conditions. Over the years, a diverse group of extremophiles have been isolated at Department of Biotechnology from samples of alkaline soda lakes in East African Rift Valley, hot springs in Iceland, and saline environments of Altiplano region of Bolivia. A number of novel microorganisms belonging to genera Bacillus, Dietzia, Brevibacterium, Halomonas, Amphibacillus etc have been identified. The studies on biodiversity have been performed in collaboration with uni-versities in the respective countries. Some interesting hydrolytic enzymes from extremophiles have been investigated. One of the halophilic bacteria is being used for the production of biopolyester, polyhydroxybutyrate and compatible solutes.

lows many more organisms to be isolated and identified. Sulphate reducing bacteria are important in the sulphur cycle and also with regard to heavy metal ions in environment. Several new species have been isolated e.g. some new Clostridium species.

Degradation of aromatic structures is often complicated for microorgan-isms. A group of very capable organisms is that of white rot fungi. Isolation and characterization of such organisms from the tropical regions of Bolivia has given some very interesting producers of oxidative enzymes including peroxidases and laccase.

Anaerobic microor-ganisms comprise a group that to a large extent has been over-looked. These organ-isms play an impor-tant role in many cycles of elements in nature, however they are poorly understood. During the year the work has continued to isolate new organisms with special enzyme producing properties. The isolation is based on the microdroplet technique which al-

Not yet identified halophilic strains isolated from the saline lake ‘Laguna Verde’ in Bolivia (Photo: Jorge Quillaguaman)

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Green Chemicals and Materials

Greenchem – Speciality chemicals from re-newable resourcesIndustrial biotechnology has a promising role as an enabling technology for production of chemicals, materials and energy from bio-based renew-able raw materials. The production processes are based on fermentation or biocatalysis using whole cells or enzymes, and are characterized by reduced energy consumption and waste generation, and biodegradable products.At Department of Biotechnology, a research program Greenchem focuses on the production of “green” chemicals. The programme is interdiscipli-nary involving cooperation between biotechnology, environment systems analysis and technology innovation systems analysis, and also with indus-tries representing an integrated value chain from raw materials to the users of industrial chemicals. The products studied within Greenchem are tar-geted for applications in environment-friendly surface coatings, lubricants, consumer care and cleaning.

Coating and lubricant productsRapeseed methyl ester, tall oil fatty acids, linseed oil and allyl ethers have been epoxidized using a chemo-enzymatic process. Acrylation of epoxidized oil has also been performed. A few different polyester acrylates have been prepared by combining chemical synthesis of the polyester with subsequent enzymatic acrylation. Epoxidized oil and acrylated product samples have been evaluated for wood coating. The results indicated several good proper-ties. Biolubricants based on vegetable oils are generally used to replace min-eral oil, but they are unsuitable for use at extreme temperatures. Branched fatty acids with potentially superior properties than vegetable oils have been synthesized and evaluated.

Biosurfactant productsSugar based surfactants generally have low toxicity, low irritability and good biodegradability. Synthesis of one group of sugar based surfactants, alkyl glycosides, by reversed hydrolysis reactions between glucose and alcohols (hexanol and octanol) was catalysed efficiently by β-glucosidase B from Thermotoga neapolitana. Another group of environment-friendly surfactants with a broad range of applications are alkanolamides. Using a solvent free enzymatic process a yield of 95% was obtained within a few hours. The product was equivalent to the highest quality products currently available.

BiopolyestersPolyhydroxyalkanoates (PHAs) are polymers produced and accumulated in-tracellularly by several bacteria as energy reservoirs. The properties of these polymers are similar to polyethylene and polypropylene, but differ in being biodegradable. We have isolated a number of novel moderately halophilic bacteria from saline lakes in Bolivia showing the production of PHAs. In 2006, polyhydroxybutyrate (PHB) production was achieved using glucose, sucrose and hydrolysed wheat bran respectively as carbon source. Recovery of the polymer in high yields was achieved using environmentally benign conditions.

Dietlind AdlercreutzReseach associate

Patrick AdlercreutzProfessor

Eva Nordberg KarlssonAssociate Professor

Rajni Hatti-KaulProfessor


Projects- Coating and lubricant products - Biosurfactant products- Biopolyesters- Biocatalyst screening and develop-ment- Enzyme production

PhD students:Thuoc Doan, Daniel Guzman, Hector Guzmán Suarez , Marlene Munoz , Mathias Nordblad , Anna Petersson, David Svensson, Pär Tufvesson, Pernilla Turner, Ulrika Törnvall

Post DocsJorge Quillaguaman, Piyush Lathi, Sangita Kasture

Olle HolstProfessor


Bioseparation

Igor GalaevAssociate professor

Alexander IvanovAssociate professor


Sorting molecules and cellsResearch in the area of bioseparation has traditionally dealt with develop-ment of procedures for isolation of specific biomolecules, or for develop-ment of new separation media. The development today is at a breakthrough point in the sense that several new trends can be identified that are growing and soon will constitute separate research areas:

materials for cell separation/ cultivation and their application in medi-cal technology

• environmental separation• process analytical technology (PAT) for downstream processing

The activities during the last year can be divided into the following areas:Development and applications of cryogels

new types of superporous gels cryogels for operation in stirred tank reactorsdesigned cryogels for enzyme/cell immobilizationcryogels as scaffolds for cell cultivationnew technology for affinity separation of cellscryogels for applications in environmental separation

Development and applications of smart polymers and smart surfacesusing smart surfaces with boronates for cell colonizationsmart boronate polymers in biosensors for glucose

Simulated moving bed technology for efficient protein purificationlarge scale processes for bulk proteinsSMB and affinity chromatography

Use of molecular imprinting technology combined with suitable process technology for design of processes for removal of pollutants present in waste water at low concentrations

enrichment for later destruction of endocrine disruptorscapture of pharmaceuticals from waste water

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Projects:Smart surfaces

Carbohydrate-specific synthetic poly-mers as components of mucoadhesive gels and stimuli-responsive supports for cell separation and analysis

Smart Polymers for selective recogni-tion of biomolecules

Simulated moving bed technology for efficient protein purification

PhD students: Jonatan Andersson, Wim Noppe

Researchers: Dr. Fatima Plieva, Dr. Maria Dainiak, Dr. Irina Savina, Deepti Sahoo

Visiting researchers: Dr. Ashok Ku-mar (India), Suthasenee Nilsang (Thai-land), Homayon Ahmad Panahi (Iran).

Photo (top) Scanning electron micrographs of HCT 116 human colon cancer cell line grown for 6 days in cryogels with RGD (Arg-Gly-Asp) mimic (left) and collagen (right) coupled to the pore surface. (Photo Maria Dainiak)

A pulse of whole blood through a cryogel column indicating than no cells are trapped inside the column.

Dept. of Biotechnology Annual Report 2006 1�

New Materials

Igor GalaevAssociate professor

Alexander IvanovAssociate professor

Polymers in the service of Biotechnology

By exploring the interface between polymer technology and biotechnology, we have developed a range of new and interesting materials for use in areas such as bioseparation, medical biotechnology and bioanalysis.

Cryogels We produce macroporous gels, so called cryogels, by polymerization in a partially frozen state when the ice crystals perform as porogens. After com-pleting the polymerization and melting ice crystals a system of large and inteconnected pores is formed. The high mechanical strength along with unique tissue-like elasticity and uniform porous structure with large pores make cryogels very promising materials for bioseparation of nanoparticles, as scaffolds for the cultivation of mammalian cells and implants in tissue

Projects:

Smart surfaces

Carbohydrate-specific synthetic polymers as components of mucoadhesive gels and stimuli-responsive supports for cell separation and analysis

Biomaterials for wound healing and tissue regeneration

Disposable bioreactors for therapeutic protein production

Researchers: Prof. Bo Mattiasson, Dr. Fatima Plieva, Dr. Maria Dainiak, Dr. Irina Savina

Visiting researchers: Dr. Ashok Kumar (India), Marina Kuzimenkova (Russia), Natalia Zhurav-leva (Russia), Chongdee Thammakhet (Thailand), Homayon Ahmad Panahi (Iran), Suthasenee Nilsang (Thailand), Vishal Nehru (India)

Picture 3D reconstruction of micro-computed tomography (µ-CT) images of the HEMA-DMAAm-cryogel

engineering. In 2006, the main emphasis in the project was on studying mechanical properties of cryogels as well as the factors controlling the pore structure in cryogels and on developing new methods of modifica-tion of pore surface e.g. atom transferred radical graft polymerization (in collaboration with Prof. F. Du Prez, University of Gent, Belgium). Cryogels with differ-ent chemistries of pore surface modification, includ-ing newly developed mimic of cell-recognition motif RGD (Arg-Gly-Asp), were studied in 96-well format as scaffolds for the cultivation of adherent and non-adherent mammalian cells. Besides, the work has been initiated on using cryogels in biomedical area as scaf-folds in tissue engineering. Preliminary data indicate that cryogels are well tolerated when implanted in the body. This work is done in collaboration with Prof. E. Piskin in Hacettepe University in Ankara, Turkey .

Smart surfaces and smart gelsThe surfaces with grafted smart polymers, so called smart surfaces re-spond in a strong and predefined way to small changes in environment like temperature, pH, ionic strength, the presence of specific chemicals. A new grafting chemistry has been developed to control the wettability of the surface in response to temperature or in response to the presence of a certain chemical, e.g. sugar. We have developed smart surfaces with boronate ligands using graft-copolymerization of N,N-dimethylacrylamide (DMAA) and N-acryloyl-m-phenylboronic acid (NAAPBA). Smart semi-transparent boronate-containing gels were developed for optical detection of saccharides. The polymer grafts were characterized with time-of-flight secondary ion mass-spectrometry (ToF SIMS) and contact angle measure-ments. Cancer cells of different cell lines have different ability to attach to the DMAA-NAAPBA copolymer-grafted glass slides. The adhered cells are efficiently detached from the grafted slides with fructose. Smart surfaces could be used for cell-based toxicity tests performed on the transparent supports like glass slides.


Bioanalysis

Maria AnderssonTechnology coordinator

Martin HedströmResearch engineer

Selective and sensitive tools for bioprocess monitoring and controlThe bioanalytical research at the department is focused on using bio-logical molecules as recognition elements or catalysts in analytical equip-ment such as biosensors and in diagnostic tests. With the increased need for the identification of biological molecules and reaction products from different enzymatically catalysed reactions, much of the activities also fo-cus on developing methods for HPLC, mass spectrometry, light-scatter-ing and differential scanning calorimetry for the analysis and charac-terization of small molecules, peptides, proteins and other biopolymers. Within the department we started early to develop biosensors for process monitoring. These were used to monitor down stream processing, both target molecules and impurities. With the programme initiated by the U.S. Food and Drug Administration (FDA) to primarily promote the devel-opment of process analytical technology (PAT) as a concept within pharma-ceutical manufacturing, our earlier activities suddenly came very much into

Projects:

Development of analytical systems for the trace-amount detection of bacterial toxins.

A process analytical technology (PAT) approach for the concomitant on-line monitoring of target proteins and bacterial contaminants present during purification.

Monitoring and control of host cell proteins (HCPs) during recombinant production using a capacitive biosen-sor.

Monitoring and control of recombinant human growth hormone during fer-mentation and downstream processing using capillary-LC/MS.

Sol-gel encapsulation of biomolecules for the use in diagnostic tests

Scientist involved:Prof. Bo Mattiasson

Visiting Researchers:Mahmoud LabibMohammad Mazlomi Kosin Teeparuksapun

Capacitive biosensor analytical system (Photo Kosin Teeparuksapun)

focus. Several initia-tives for collaborations with “big pharma” have now been taken. The objective of PAT is a full exploitation of the benefits of an in-tegrated process ana-lytical control where real-time information on the product quality can be obtained along the production line.One example of our efforts involves an ultra-sensitive capacitive biosensor, utilized to monitor trace amounts of toxic bacteria-related contaminants. The transducer surface is covered with a self-assembled monolayer (SAM) of alkylthiols. Biomolecules, e.g. anti-bodies directed towards the contaminant, are immobilized on this SAM. A capacitance decrease is then registered and measured when target molecules bind selectively to the sensor surface. The sensor is mounted in a flow injec-tion system, and a continuous stream of liquid is passed over the sensing surface. After binding and washing, the registration takes place prior to the forced dissociation of the affinity complex by a regeneration solution and the system is reconditioned for still another assay. The assays are sensitive. In several cases a detection limit in the order of 10-18 moles per liter has been achieved, and quantification to at least 10-17. The types of analytes that have been monitored by this technique are e.g. endotoxin, enterotoxins, host cell proteins from the expression host when producing transgenic proteins, plas-mids and protein A. The biosensor system can also be used in continuous monitoring and are hence of interest for further development as a PAT-tool.


Functional foods

Patrick AdlercreutzProfessor

Biotechnology for production of healthy food

Recent advances in biotechnology have increased the possibilities to make food components with improved properties. Enzymatic methods are used to tailor-make lipids containing health-promoting fatty acids, such as ome-ga-3 fatty acids. During 2006 enrichment of these fatty acids from different marine oils has been carried out. The principle behind the enrichment is that many lipases convert these fatty acids much slower than other fatty acids. It was thus possible to remove a large portion of the other fatty acids from the triglyceride molecules and keep EPA and DHA in the glyceride fraction. After removal of the free fatty acids a concentrate of EPA and/or DHA was obtained. This forms an interesting ingredient in functional food products.

ProjectsSafe food products containing long-chain omega-3 fatty acids

Health promoting lipids containing alfa-linolenic acid

Efficiency of antioxidants in the protec-tion of vital biomolecules

PhD studentsCarl Elovson GreyAnn-Marie LybergJulia Larsson

“The development of more healthy food will be essen-tial for combating life style

related diseases, such as diabetes and heart disease”

All aerobic organsims are ex-posed to reactive oxygen species (hydrogen peroxide, superoxide radicals, hydroxyl radicals, etc.), which can damage vital bio-molecules, such as DNA, lipids and proteins. In order to cope with this, the organisms protect themselves in different ways, and one important way is by low molecular weight antioxidants, which are often obtained from the diet. It has been shown that fruits and vegetables promote human health and antioxidants are supposed to play an important role. In this project the ability of antioxidants to protect important biomol-ecules is evaluated. Effects on DNA and its building blocks have been stud-ied earlier in the project and during 2006 the work focused on protection of proteins, especially the enzyme chloroperoxidase. This is an enzyme that catalyses a wide range of interesting reactions, but its practical utility is lim-ited by its poor operational stability. It was discovered that antioxidants ef-ficiently protected the enzyme and thus prolonged its lifetime and increased the amounts of reaction products obtained. The most efficient antioxidant was caffeic acid, which is found in relatively large amounts in coffee.

COOH

COOH

DHA

EPA

EPA and DHA are long chain omega-3 fatty acids of importance for building up and maintaining the brain and nervous system. Fat fish is the main source of these fatty acids in the human diet. We are investigating possibilities to incorporate the enriched products into functional food products. Such products can become a good alternative for those who do not eat fish often enough! Because of the many double bonds in the molecules, they are quite sensitive, and the processing is therefore carried out by mild enzymatic methods.


Bioremediation

Benoit GuieysseResearch Associate


Marika MurtoResearch engineer

Degrading the “undegradable”Persistent organic pollutants (POPs) include a large variety of substances such as pesticides, surfactants or wood preservatives (i.e. creosote mixture) as well as products released from industrial activities (i.e. dioxins, polyaro-matic hydrocarbons). Heavy metals is another group of compounds caus-ing severe environmental problems. A new group of potentially hazardous chemicals have come into focus in recent years: pharmaceuticals and mol-ecules with endocrine disrupting activity.

Projects- Removal of heavy metal ions using sulphate reducing bacteria- Enrichment of heavy metals on chelating matrices before release and precipitation- Enrichment and destruction of endo crine disruptors in wastewater- Development of separation tools and systems specially adapted to waste water treatment- Degradation of textile dyes in waste water from textile industries- Fungal bioreactors for degradation of POPs- Degradation of chlorophenols in water- Microbial ecology studies of chloro- phenol degrading consortia- Microbial degradation of EDTA- Algal-bacterial photobioreactors- Photochemical oxidation of organic pollutants as a pre-step to biodegradation- Treatment of wastewater from a coke factory in Egypt- Biosensor for environmental analysis

Basin for residual water from textile industry in southern India (Photo: Bo Mattiasson)

Deposit of silt from precipitation step dur-ing textile processing (Photo: Bo Mattiasson)

The department has focused much attention on de-veloping biotech-nological meth-ods, either alone or in combina-tion with physical treatment, to de-grade or immobi-lize these hazard-ous compounds. Sometimes the concentration at which the hazard-ous compound is present is very low, and then an enrichment step is used before treatment. This is used especially for endocrine disruptors and phar-maceuticals.

“Access to clean water is a key issue for a peaceful development in the world. Efficient treatment of polluted water is one important step to meet this

demand”

Photo (top): Identifica-tion of polychloroph-enol degraders using fluorescence in situ hybridization (Photo: Hamid Zilouei)

PhD students: Hamid Zilouei, Mathieu Le Noir, Britt-Marie Pott, Maryam Lati-fian, Gerly Morades

Researchers: Paqui Blánquez, Paula Fernández Álvarez, Marja Palmroth, Maria Jonstrup, Laura Montelongo, To-bias Hey, Nararat Thongsrinoon


Renewable energy

Lovisa BjörnssonAssociate professor

Jing LiuResearch associate


Marika MurtoResearch engineer

A sustainable bioenergy system should 1. be resource efficient, 2. have a high energy efficiency, 3. have maximized environmental benefits and 4. be cost efficient. These four issues are addressed in the bioenergy research at Department of Biotechnology, with focus on the biogas production proc-ess.

Projects- Improved hydrolysis of residual municipal waste- Improvement of biodegradation of manure with biotechnological solu tions- Sewage sludge pre-treatment for biodegradability improvement-Optimising bioenergy production from annual crops - Manure, energy crops and crop residues for biogas production in the Nordic countries- Evaluation of the biogas potential as a renewable energy source in Chile- Biogas production from seaweed us ing a two-stage process - Removal of heavy metals for im proved fertilizer quality using an eco logical kidney- Advanced process monitoring and control of the anaerobic digestion process- Molecular microbiology and statistics- Bioenergy production with extremo philes - Microbial fuel cell for electricity production

PhD students: Irene Bohn, Carla Cre-spo, Emma Kreuger

Visiting researchers: Federico Esco-bar, Ivo Achu Nges, Valentine Nkemka Nkongndem, Robert Selling, Yeroolt Tangad, Zhidan Liu

system, one alternative is to utilize energy crops. Here, cost efficiency is a great challenge, which in our research is addressed by:

- Improved energy efficiency, giving higher energy output. This is achieved by optimized process design, improved process control, or pretreatment of substrates for higher conversion rates.

- Quantification and optimisation of the environmental benefits that are inherent in the process.

Photo (top) Energy crops for biogas production (Photo: Lovisa Björnsson)

To produce biogas from residuals is an established concept, but chal-lenging research issues are how to improve microbial and operational efficiency of the systems. To further increase the potential contribution of biogas into our energy supply

Microbial fuel cells with computer based data acquisition system. (Photo: Jing Liu)

“Biotechnical solutions will reduce our fossil

fuel dependence”

Cactus - Opuntia Ficus-Indica - energy crop for biogas production.

- Biorefinery concepts, where higher value produced, com-bined with biogas production from the residuals to ensure sus-tainability.

Anaerobic digestion reactors. (Photo: Jing Liu)


Education: courses

Course Code Credits No. of stu-dents

Responsible teacher

Programme related courses

Biotechnology KKK070 12 65 Olle Holst

Process Engineering for the Biotechnical and Food Industry

BLT010 4 50 Olle Holst

Bioanalytical chemistry KBT050 5 30 Maria Andersson

Biotechnology KBT070 5 20 Olle Holst

Enzyme technology KBK031 5 37 Patrick Adlercreutz

Biotechnological separation processes KBT060 5 27 Igor Galaev

Environmental biotechnology KBT080 5 32 Marika Murto

Biotechnology, Process and Plant Design KBT041 10 12 Olle Holst

Free-standing courses

Entrepreneurship in Biotechnology TNK021 5 19 Maria Andersson

Green chemistry and Biotechnology TNK280 5 20 Eva Nordberg Karlsson

PhD course

Mass spectrometry 2 4 Martin Hedström

The department is responsible for giving eight courses at Master´s level and three courses belonging to the LTH free-standing course program. During the past year the responsible teachers have been involved in the transition towards a new Masters education system according to the Bologna process, and more transitional work is ex-pected during the coming year. The number of students have increased over the past years which has resulted in a slightly higher work load for responsible teachers but also created a larger pool of students available for master thesis projects and as possible recruits for PhD positions. We continued to take active part in the International Master´s programme in Bio- and Food Technology which every year accepts around 25 international students. In total 24 students from Sweden and abroad chose to do their master thesis projects at the department during 2006. Courses offered during 2006 are listed below.

Environmental Biotechnology (Photo: Erik Andersson) Process Engineering for the Biotechnical and Food Industry (Photo:Erik Andersson)


BENJAMIN ALAMU OLATUNJI, Physical and chemical pre-treatment of sewage sludge for biodegradability Imrove-ment

AGATA ANDERSSON, Deammonification in a moving bed biofilm process

VALERY BISONG AGBOR, Enhanced solubilisation of the residual fraction of municipal solid waste by two-stage digestion and enzyme addition

MANDAVA CHANDRA SEKHAR, Overexpression and puri-fication of ribosomal proteins (L7/L12&L10) and elonga-tion factor G

YAN CUI, Site-directed mutagenesis of a cyclomaltodextri-nase of thermophilic origin, and evaluation of the effects on hydrolytic and transglycosylating activities

MARIA DALEKE, Detection and quantification of the heavy metal efflux pump genes czcA and copA in polluted ma-rine Sediments by Real Time PCR

ENONGENE EKWE SYLVANUS, Site-directed mutagen-esis of beta-gucosidase A from Thermotoga neapolitana (TnBgl1A) for improved alkylglycoside synthesis

DENIS EPIE EBONG, Partial characterization of xylanases produced by two newly isolated alkalophilic anaerobes from Kenyan soda lakes

CHENG FANG, Microbial fuel cell used as biosensor for on-line monitoring anaerobic digestion processes

MIMMI GERSBRO, Optimization & transcriptional analysis of fed-satch culture during plasmid production in DH5-alfa host cells

MARTIN GRÄBER, A novel direct screening method for eval-uation of alkyl glucoside producing Beta-glucosidases

EMMANUEL IKECHUKWU AMADI, Optimization of chemo-enzymatic epoxidation of allyl ethers

PABLO LORENZO AMIGO, Activity characterization of ADHs from thermophilic bacteria towards different sub-strates and pHs.

SARA MARTINS BADENES, Chemo-enzymatic epoxidation of oils

SANAM S. MONAVARI, Development of water quality test

kit based on substrate utilization and toxicity resistance in river microbial communities

ACHU NGES IVO, Process optimisation of sewage sludge volume reduction through anaerobic degradation

BEATRICE NILSSON, Analysis of off-flavour compounds-In-vestigations of the analytical techniques dynamic head-space and Solid Phase Micro Extraction in combination with GC/MS

PATRIK NILSSON, Use of boronate-containing copolymers for glucose sensing at neutral pH

ANNA PERSSON, Biological degradation of EDTA in Nordic paper Seffle pulp mill effluents

EMIL POHL AND MAGNUS PRAHL Enzymatic production of polyester acrylates

FABIAN RUNDBÄCK, Reaction conditions for production of alkyl glucosides by reversed hydrolysis using beta-glu-cosidase B from Thermotoga neapolitana in a two-phase system

VALGERDUR TOMASDOTTIR, Genetic and biochemical characterization of the novel short-chain alcohol dehy-drogenase ADH101 from a thermophilic Bacillus strain from Iceland

YIZHE ZHANG, Development of economical and simple en-zymatic diagnostic kit

The broad area of biotechnology continued to attract Master students, both from Sweden and abroad. In all, 24 students chose to do their master thesis work at the department, on such diverse topics as enzyme modification and characterisation, development of bioanalytical kits and improvement of sewage sludge degradation.

Education: Master thesis projects

During 2006 the Department of Biotechnology continued to take active part in teaching and supervising stu-dents on the International Master’s programme in Bio- and Food Technology. The objective of this programme is to offer courses focused on modern methods of Food Science and Biotechnology. Prof. Olle Holst is the coordi-nator of the programme which in 2006 accepted around 25 students.

Education: International Master’s programme in Bio- and Food Technology


Visiting researchers and students

Name University Country

Muhammad Al-Najjar Philadelphia University, Amman Jordania

Rosa Aragao Peralta Universidad Mayor de San Simon, Cochabamba La Paz Bolivia

Khalid Atia Metwally Nuclear Research Center, Cairo, Egypt

Mahmoud Aziz Labib Cairo University Egypt

Paqui Blanquez Universitat Autònoma de Barcelona Spain

Thalea Coitlan University of Braunsweig Germany

Carla Crespo Universidad Mayor de San Andrés La Paz Bolivia

Maria Del Mar University of Almería Spain

Elena DeSeta Faculty of Pharmacy, University of Naples FedericoII Italy

Tarik Diab Philadelphia University, Amman Jordania

Sinan Egri Hacettepe University, Ankara Turkey

Aly El Tayeb Cairo University, Cairo Egypt

Reza Faryar Shabestar Azad University Iran

Paula Fernández Álvarez University of Santiago de Compostela Spain

Victor Ibrahim Holy Spirit University, Jounieh Lebanon

Sangita Kasture National Chemical Laboratory, Pune India

Harald Kirsebom Lund University Sweden

Marina Kuzimenkova MIFCT Moscow Academy of Fine Chemical Technology Russia

Piyush Lahti University Institute of Chemical Technology (UICT) Mumbai India

Maryam Latifian Shiraz University, Shiraz Iran

Zhidan Liu Institute of Process Engineering,Chinese Academy of Sciences, Beijing China

Nadhem Machta National Institute of Applied Sciences and Technology (INSAT), Tunis Tunisia

Laura Montelongo National Autonomus University of Mexico (UNAM), Mexico City Mexico

Isabella Musolino Faculty of Science, University of Rome “Tor Vergata” Italy

Dept. of Biotechnology Annual Report 2006 21

Vishal Nehru Indian Institute of Technology, Kanpur India

Suthasinee Nilsang Asian Institute of Technology, Bangkok Thailand

Marja Palmroth Tampere University (TUT) Finland

Dino Pignetti “Leonardo da Vinci Program Trainee”(Unipharma Graduated project) Italy

Irina Savina A.N.Nesmeyanov Institute of Organoelement Compounds, Moscow Russia

Yumei Sun College of Bio & Food Engineering, Dalian Institute of Light Industry, China

Kosin (Jan) Teeparuksapun Prince of Songkhla University, Hat Yai Thailand

Chongdee (Cook) Tham-makhet

Prince of Songkhla University, Hat Yai Thailand

Nararat (Noi) Thongsrinoon Prince of Songkhla University, Hat Yai Thailand

Nicolas Viudez University of Montpellier France

Natalia Zhuravleva Far East State University in Vladivostok Russia

Some of our guest researchers in action on one of the warmest days in summer 2006 From left: Thuy, Laura, Deepti, Sangita, Suthanisee and Cheng (Photo: Bo Mattiasson).

Excursion to Rövarkulan in June 2006 (Photo: Siv Holmqvist)Christmas dinner in December 2006


International collaborations

North, Central and South America1. USAUniversity of Houston, Houston

2. MEXICOUniversidad Autonomia Metropolitana, Mexico City.

�. NICARAGUAUniversidad Nacional Autonoma de Nica-ragua, Managua.

�. BOLIVIAUniversidad Mayor de San Andres, La PazUniversidad Mayor de San Simón, Cochabamba.

�. CHILEFundacion Chile, Santiago

Europe6. ICELANDProkaria Ltd. Reykjavik.

�. FINLAND Department of Biology and Environmen-tal Science in Jyväskylä University

�. GREAT BRITAINUniversity of Newcastle, Newcastle upon Tyne

University of Bath.

�. FRANCEForeign Affairs Ministry, Paris

Veolia Water, Paris,

Anjou recherche

Veolia Environnement, Maisons-Laffitte

10. DENMARKTechnical University of Denmark, Kgs. Lyngby.

11. GERMANYTechnical University, Braunschweig

Christian-Albrechts University, Kiel

12. AUSTRIATechnical University, Graz

1�. PORTUGALMinho University, Braga

1�. SPAINValladolid University, Valladolid.

1�. ITALYPolitecnico di Milano

16. RUSSIARussian Academy of Sciences, Moscow.

Africa1�. EGYPTSuez Canal University, Ismailia

Cairo University, Cairo

Beni-Sueif University

Nuclear Research Centre, Cairo

1�. KENYAUniversity of Nairobi, Nairobi.

1�. TANZANIAUniversity of Dar es Salaam

20. SOUTH AFRICA

During 2006, the department collaborated with universities, institutes and companies in more than 35 countries worldwide. The collaboration included both minor initiatives such as student exchange as well as large research programmes spanning over several years. The collaboration covered all major research areas at the department.

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Durban Institute of Technology University of Limpopo.

21. TUNISIANational Institute of Applied Science and Technology (INSAT)

Asia

22. TURKEYHacettepe University, Ankara.

2�. ISRAELBar-Ilan University, Ramat Gan.

2�. GEORGIATbilisi State University, Tbilisi.

2�. JORDAN Philadelphia University, Amman

26. AZERBADIJANInstitute of Polymer Materials, Baku.

2�. IRANAzad University, Tehran

Isfahan University of technology, Isfahan.

2�. INDIAIndian Institute of Technology, Delhi

Indian Institute of Technology, Kanpur

Agharkar research institute, Pune

Bhabha Atomic Research Centre, Mumbai

University of Calcutta, Calcutta

Central Food Technology Research Insti-tute, Mysore

2�. SRI LANKAUniversity of Jaffna, Jaffna.

�0. THAILANDAsian Institute of Technology, Bangkok

Prince of Songkhla University, Hat Yai.

�1. VIETNAMHanoi University of Education, Hanoi.

�2. PHILIPPINESDe La Salle University, Manila.

��. CHINAInstitute of process engineering, Chinese Academy of Sciences, Bejing.

��. JAPANMiyazaki University, Miyazaki.

Australia

��. AUSTRALIAUniversity of Queensland, Brisbane.

Sampling of microorganisms in Sajama, Bolivia (Photo: Rajni Hatti-Kaul)

Visit at the Suez Canal University, Egypt, (Photo: Rajni Hatti-Kaul)

Evalution of the biogas potential in Chile. Biogas course for people from Fundacion Chile.


Funding and grants

As many other departments and institutions at the university, the Dept. of Biotechnology is increasingly dependent on external sources for carrying out the research. Foundations such as Mistra (The Founda-tion for Strategic Environmental Research) and public authorities such as SIDA (The Swedish International Development Cooperation Agency) are major finan-cial contributors. During 2006 the contribution from education and industry increased, mirroring the in-creased activities in teaching and industrial collabora-tion. The total funding for 2006 was approximately 32 MSEK (3.3 MEuro). The distribution of sources is shown below.

Source Funds (kSEK)

Faculty 6 466

Education 3 505

Research councils 2 558

EU 334

Public authorities 8 843

Foundations 8 469

Industry 1 070

Others 715

Total �1 600

Foundations

EU

Research councils

Faculty

Education

Public authorities

IndustryOthers

29%

26%

1%

8%

11%

20%

2%3%


J. Ahlqvist, A. Kumar, H. Sundström, E. Ledung, G. Hörnsten, S-O. Enfors and B. Mattiasson, Affinity bind-ing of inclusion bodies on supermacroporous monolithic cryogels using labeling with specific antibodies. J. Biotech-nol., 122, 216-225 (2006)

J. Ahlqvist, M. Dainiak, A. Kumar, H. Sundström, E. Ledung, E. G. Hörnsten, S-O. Enfors, I. Yu. Galaev and B. Mattiasson, Monitoring the production of inclusion bodies during fermentation and ELISA analysis of intact inclusion bodies using 96-minicolumn cryogel microplates. Anal. Biochem., 354, 229-237 (2006)

M. T. Alvarez, T. Pozzo and B. Mattiasson, Enhance-ment of sulphide production in anaerobic packed bed bench-scale biofilm reactors by sulphate reducing bacteria. Biotechnol. Lett., 28, 175-181 (2006)

J. Andersson and B. Mattiasson, Simulated moving bed technology with a simplified aproach for protein purifica-tion: Separation of lactoperoxidase and lactoferrin from whey protein concentrate (WPC). J. Chrom. A. 1107, 88-95 (2006)

V. Bansal, P. K. Roychoudhury, B. Mattiasson and A. Kumar, Recovery of urokinase from integrated mammalian cell culture cryogel bioreactor and purification of the en-zyme using p-aminobenzamidine affinity chromatography. J. Mol. Rec., 19, 332-339 (2006)

L. Cicortas Gunnarsson, L. Dexlin, E. Nordberg Karls-son, O. Holst and M. Ohlin, Evolution of a carbohydrate binding module into a human IgG4-specific protein binder. Biomol. Eng., 23, 111-117 [2] (2006)

L. Cicortas Gunnarsson, Q. Zhou, C. Montanier, E. Nor-dberg Karlsson, H. Brumer III, and M. Ohlin, Engineered xyloglucan specificity in a carbohydrate-binding module. Glycobiology, 16, 1171-1180 (2006)

L. Cicortas Gunnarsson, E. Nordberg Karlsson, M. Andersson, O. Holst and M. Ohlin, Molecular engineer-ing of a thermostable carbohydrate binding module. Biocat. Biotrans., 24, 31-37 (2006)

D. G. Cirne, L. Björnsson, M. Alves, and B. Mattiasson, Effects of bioaugmentation by an anaerobic lipolytic bac-terium on anaerobic digestion of lipid-rich waste. J. Chem. Technol. Biotechnol., 81, 1745-1752 (2006)

D. G. Cirne, O. D. Delgado, S. Marichamy and B. Mat-tiasson, Clostridium lundense sp nov., a novel anaerobic lipolytic bacterium isolated from bovine rumen. Int. J. Syst. Evol. Micr., 56, 625-628 (2006)

S. J. Crennell, D. Cook, A. Minns, D. Svergun, R. L. Andersen and E. Nordberg Karlsson, Dimerisation and

an increase in active site aromatic groups as adaptations to high temperatures: X-ray solution scattering and substrate-bound crystal structures of Rhodothermus marinus endog-lucanase Cel12A. J. Mol. Biol., 356, 57-71 (2006)

G. Crocetti, M. Murto and L. Björnsson, An update and optimisation of oligonucleotide probes targeting methano-genic Archaea for use in fluorescence in situ hybridisation (FISH). Journal of Microbiol. Meth., 65(1), 194-201 (2006)

M. Dainiak, A. Kumar, I. Yu. Galaev and B. Mattias-son, Detachment of affinity captured bioparticles by elastic deformation of macroporous hydrogel. Proc. Natl. Acad. Sci. (USA), 103, 849-854 (2006)

M. B. Dainiak, I. Yu. Galaev and B. Mattiasson, Affinity cryogel monoliths for screening for optimal separation conditions, chromatographic separation of cells J. Chrom., 1123, 145-150 (2006)

O. Delgado, J. Quillaguaman, S. Bakhtiar, B. Mattias-son, A. Gessesse, R. Hatti-Kaul, Nesterenkonia aethiopica sp. nov., an alkaliphilic, moderate halophile isolated from an Ethiopian soda lake. International J. Syst. Evol. Micro-biol., 56(6), 1229-1232 (2006)

E. N. Efremenko, Y. A. Votchitseva, F. M. Plieva, I. Yu. Galaev and B. Mattiasson, Purification of His

6-organo-

phosphate hydrolase using monolithic supermacroporous polyacrylamide cryogels developed for immobilized metal affinity chromatography. Appl. Microbiol. Biotechnol., 70, 558-563 (2006)

C. Elovson Grey and P. Adlercreutz, Time and concentra-tion dependence of Fenton induced oxidation of dG. Nucle-osides Nucleotides Nucleic Acids, 25, 259-278 (2006)

C. Elovson Grey and P. Adlercreutz, Evaluation of mul-tiple oxidation products for monitoring effects of antioxi-dants in fenton oxidation of 2´deoxyguanosine. J. Agric. Food Chem., 54, 2350-2358 (2006)

T. Essam, A. Amin Magdy, O. El Tayeb, B. Mattiasson and B. Guieysse, Biological treatment of industrial wastes in a photobioreactor. Wat. Sci. Technol., 53, 117-125 (2006)

T. Essam, H. Zilouei, A. Amin Magdy, O. El Tayeb, B. Mattiasson and B. Guieysse, Sequential UV–biological degradation of chlorophenols. Chemosphere 63, 277-284 (2006)

A. Hanora, I. Savina, F. M. Plieva, V. A. Izumrudov, I. Yu. Galaev and B. Mattiasson, Direct capture of bacterial plasmid DNA from non-clarified cell lysate using monolith columns from macroporous gel grafted with polycations. J. Biotechnol., 123, 343-355 (2006)

During 2006, 66 research papers were published in peer reviewed journals and another 4 papers got available on-line. For reprints of papers contact Siv Holmqvist, [email protected]

Publications

Journal articles


R. Hatti-Kaul, H.O. Birgisson and B. Mattiasson, Cold active enzymes in food processing. In Food Biotechnology, 2nd Edition Eds. K. Shetty, G. Paliyath, A. Pometto and R.E. Levin, CRC, Taylor & Francis, Boca Raton, 1631-1653 (2006)

A. E. Ivanov, J. Ekeroth, L. Nilsson, I. Yu. Galaev, B. Bergenståhl and B. Mattiasson, Variations in Wettability and protein adsorption on solid siliceous carriers grafted with poly(N-isopropylacrylamide). J. Coll. Interph. Sci., 296, 538-544 (2006)

A. Ivanov, I. Yu. Galaev and B. Mattiasson, Interaction of sugars, polysaccharides and cells with boronate-contain-ing copolymers: from solution to opolymer brushes. J. Mol Rec., 19, 322-331 (2006)

A. E. Ivanov, H. A. Panahi, M. V. Kusimenkova, H. S. Vaquif, M. Jahanshahi, I. Yu. Galaev and B. Mattias-son, Affinity adhesion of carbohydrate particles and yeast cells to boronate-containing polymer brushes grafted to siliceous supports. Chemistry – A European Journal, 12, 7204-7214 (2006)

A. E. Ivanov, K. Shiomori, Y. Kawano, I. Yu. Galaev and B. Mattiasson, Effects of polyols, sccharides and glycopro-teins on thermoprecipitation of phenylboronate-containing copolymers. Biomacromolecules, 7, 1014-1024 (2006)

A. A. Khalil, S. S. Mohamed, F. S. Taha and E. Nordberg Karlsson, Production of functional protein hydrolysates from Egyptian breeds of soybean and lupin seeds. Afr. J. Biotechnol., 5, 907-916 (2006)

M. A. Kumar, R. S. Chouhan, M. S. Thakur, B. E. Amita Rani, Bo Mattiasson and N. G. Karanth, Automated flow enzyme-linked immunosorbent assay (ELISA) system for analysis of methyl parathion. Anal. Chim. Acta, 560, 30-34 (2006)

A. Kumar, V. Bansal, K. S. Nandakumar, I. Yu. Galaev, P. K. Roychoudhury, R. Holmdahl and B. Mattiasson, Integrated bioprocess for the production and isolation of urokinase from animal cell culture using supermacroporous cryogels. Biotechnol. Bioeng., 93, 636-646 (2006)

A. Kumar, V. Bansal, J. Andersson, P. K. Roychoudhury and B. Mattiasson, Supermacroporous cryogel matrix for integrated protein isolation: IMAC purification of uroki-nase from cell culture broth of a human kidney cell line. J. Chrom. A, 1103, 35-42 (2006)

M. R. Lacayo, E. Terrazas, B. van Bavel and B. Mattias-son, Degradation of toxaphene by Bjerkandera sp strain BOL12 using waste biomass as a cosubstrate. Appl. Micro-biol. Biotechnol. 71, 549-554 (2006)

P. S. Lathi and B. Mattiasson, Green approach for the preparation of biodegradable lubricant base stock from epoxidized vegetable oil. Appl. Cat. B: Env., 69, 207-212 (2006)

A. Lehtomaki and L. Björnsson, Two-stage anaerobic digestion of energy crops: Methane production, nitrogen mineralisation and heavy metal mobilisation. 27(2), 209-218 (2006)

M. Le Noir, B. Guieysse and B. Mattiasson, Removal of trace contaminants using molecularly imprinted polymers. Wat. Sci. Technol., 53, 205-212 (2006)

W. Limbut, P. Kanatharnan, B. Mattiasson, P. Asa-watreratanaku, P. Thavarungkul, A comparative study of capacitive immunosensors based on self-assembled mon-olayers formed from thiourea, thiotic acid and 3-mercapto-propionic acid. Biosens. Bioel., 22, 233-240 (2006)

W. Limbut, P. Kanatharnan, B. Mattiasson, P. Asa-watreratanaku, P. Thavarungkul, A reusable capacitive immunosensor for carcinoembryonic antigen (CEA) detec-tion using thiourea modified gold electrode. Anal. Chim. Acta, 561, 55 – 61 (2006)

J. Liu, G. Olsson and B. Mattiasson, Extremum-seeking with variable gain control for intensifying biogas produc-tion in anaerobic fermentation. Wat.Sci. Technol., 53(4-5), 35-44 (2006) J. Liu, Intensified anaerobic digestion processes, In “Focus on Biotechnology Research”, Ed. Edwin C. Hearns, ISBN: 1-59454-863-3, Nova Science Publishers, Inc, New York, USA. 179-212 (2006)

A-M. Lyberg and P. Adlercreutz, Monitoring monohy-droperoxides in docosahexaenoic acid using high-perform-ance liquid chromatography. Lipids, 41, 67-76 (2006)

G. Mamo, R. Hatti-Kaul and B. Mattiasson, Fusion of carbohydrate binding modules from Thermotoga neapoli-tana with a family 10 xylanase from Bacillus halodurans S7. Extremophiles, 11, 169-177.

G. Mamo, R. Hatti-Kaul and B. Mattiasson, A ther-mostable alkaline active endo-β-1-4-xylanase from Bacil-lus halodurans S7: Purification and characterization. Enz. Microb. Technol., 39(7), 1492-1498 (2006)

G. Mamo, O. Delgado, A. Martinez, B. Mattiasson and R. Hatti-Kaul, Cloning, sequence analysis, and expression of a gene encoding an endoxylanase from Bacillus halo-durans S7. Mol. Biotechnol., 33(2), 149-160 (2006)

A. Mshandete, L. Björnsson, A. K. Kivaisi, S. T. Rubind-amayugi and B. Mattiasson, Effect of particle size on biogas yield from sisal fibre waste. Ren. En., 31, 2385-2392 (2006)

A. Msandete, L. Björnsson, A. Kivaisi, M. Rubind-amayugi and B. Mattiasson, Performance of a fixed bed anaerobic digester filled with sisal fibres for biogas produc-tion from semi solid sisal pulp waste. Tanz. J. Sci., 31 (2) 41-52 (2006)

R. Munoz, M. T. Alvarez, A. Munoz, E. Terrazas, B. Guieysse and B. Mattiasson, Sequential removal of heavy metal ions and organic pollutants using an algal-bacterial consortium. Chemosphere, 63, 903-911 (2006)

C. Nilsson, F. Nilsson, P. Turner, M. Sixtensson, E. Nordberg Karlsson, O. Holst, A. Cohen and L. Gorton, Characterization of two novel cyclodextrinases using micro-dialysis sampling on-line with HPAEC-PAD. Anal. Bioanal.


Chem., 385, 1421-1429 (2006)

S. Nitsawang, R. Hatti-Kaul and P. Kanasawud, Purifica-tion of papain from Carica papaya latex: Aqueous two-phase extraction versus two-step salt precipitation. Enz. Microb. Technol., 39(5), 1103-1107 (2006)

W. Noppe, F. M. Plieva, I. Yu. Galaev, K. Vanhoorel-beke, B. Mattiasson and H. Deckmyn, Phages as affinity ligands for direct purification of lactoferrin from defatted milk. J. Chrom. A., 1101, 79-85 (2006)

K. R. Oluoch, U. Welander, M. M. Andersson, F. J. Mu-laa, B. Mattiasson and R. Hatti-Kaul, Hydrogen peroxide degradation by immobilized cells of alkaliphilic Bacillus halodurans. Biocatal. Biotrans. 24(3), 215-222 (2006)

W. Parawira, M. Murto, R. Zvauya and B. Mattias-son, Comparative performance of a UASB reactor and an anaerobic packed-bed reactor when treating potato waste leachate. Ren. En., 31(6), 893-903 (2006)

F. M. Plieva, M. Karlsson, M-R. Aguilar David Gomez, S. Mikhalovsky, I. Yu. Galaev and B. Mattiasson, Pore structure of macroporous monolithic cryogels prepared from poly(vinyl alcohol). J. Appl. Pol. Sci., 100, 1057-1066 (2006)

F. Plieva, A. Oknianska, E. Degerman, I. Yu. Galaev and B. Mattiasson, Novel supermacroporous dextran gels. J. Biomat. Sci., Polymer Edition, 17, 1075-1092 (2006)

F. Plieva, B. Bober, M. Dainiak, I. Yu. Galaev and B. Mattiasson, Macroporous polyacrylamide monolithic gels with immobilized metal affinity ligands. The effect of porous structure and ligand coupling chemistry on protein binding. J. Mol. Recogn., 19, 305-312 (2006)

F. Plieva, X. Huiting I. Yu. Galaev , B. Bergenståhl and B. Mattiasson, Macroporous elastic polyacrylamide gels prepared at subzero temperatures: control of porous struc-ture. J. Mat. Chem., 16, 4065-4073 (2006)

J. Quillaguaman, O. Delgado, B. Mattiasson and R. Hatti-Kaul, Poly(ββ-hydroxybutyrate) production by a mod-erate halophile, Halomonas boliviensis LC1. Enz. Microb. Technol., 38(1-2), 148-154 (2006)

F. Rahimpour, F. Feyzi,S. Maghsoudi and R. Hatti-Kaul, Purification of plasmid DNA with polymer-salt aqueous two-phase system: optimization using response surface methodology. Biotechnol. Bioeng., 95(4), 627-637 (2006)

S.O. Ramchuran, V.A. Vargas, R. Hatti-Kaul and E. Nor-dberg Karlsson, Production of a lipolytic enzyme originat-ing from Bacillus halodurans LBB2 in the methylotrophic yeast Pichia pastoris. Appl. Microbiol. Biotechnol., 71(4), 463-472 (2006)

P. K. Roychoudhury, S. S. Khaparde, B. Mattiasson and A. Kumar, Synthesis, regulation and production of urokinase using mammalian cell culture: A comprehensive review. Biotechnol. Adv., 24, 514-528 (2006)

I. N. Savina, I. Yu. Galaev and B. Mattiasson, Ion-ex-change macroporous hydrophilic gel monolith with grafted

polymer brushes. J. Mol Rec., 19, 313-321 (2006)

I. N. Savina, I. Yu. Galaev, B. Mattiasson, Graft polym-erization of vinyl monomers inside macroporous polyacryla-mide gel, cryogel, in aqueous and aqueous-organic media initiated by diperiodatocuprate(III) complexes. J. Pol. Sci., 44, 1952-1963 (2006)

A. Soares, M. Murto, B. Guieysse and B. Mattiasson, Biodegradation of nonylphenol in a continuous bioreactor at low temperatures and effects on the microbial popula-tion. Appl. Microbiol. Biotechnol., 69(5), 597-606 (2006)

A. Soares, B. Guieysse and B. Mattiasson, Influence of agitation on the removal of nonylphenol by the white-rot fungi Trametes versicolor and Bjerkandera sp. BOL 13. Bio-technol. Lett., 28, 139-143 (2006)

L. M. Svensson, L. Björnsson, and B. Mattiasson, Straw bed priming enhances the methane yield and speeds up the start-up of single-stage, high-solids anaerobic reactors treating plant biomass. J. Chem. Technol. Biotechnol., 81, 1729-1735 (2006)

C. Turner, P. Turner,G. Jacobson, M. Waldebäck, P. Sjöberg, E. Nordberg Karlsson, and K. Markides, Subcritical water extraction and beta-glucosidase-catalyzed hydrolysis of quercetin in onion waste. Green Chem., 8, 949-959 (2006)

H. Zilouei, B. Guieysse and B. Mattiasson, Biological degradation of chlorophenols in packed-bed bioreactors using mixed bacterial consortia. Proc. Biochem. 41, 1083-1089 (2006)

H. Zilouei, A. Soares, M. Murto, B. Guieysse and B. Mattiasson, Influence of temperature on process efficiency and microbial community response during the biological removal of chlorophenols in a packed-bed bioreactor. Appl. Microbiol. Biotechnol., 72(3), 591-599 (2006)

Papers available online 2006

M. B. Dainiak, I. Yu. Galaev and B. Mattiasson, Ma-croporous monolithic hydrogels in a 96-minicolumn plate format for cell surface-analysis and integrated binding/quantification of cells . Enz. Microb. Technol. Available on-line 6 June 2006.

S. F. Deraz, F. Plieva, I. Yu. Galaev, E. Nordberg Karls-son and B. Mattiasson, Capture of bacteriocins directly from non-clarified fermentation broth using macroporous monolithic cryogels with phenyl ligands. Enz. Microb. Tech-nol. Available on-line 1 August (2006)

S. F. Deraz, M. Hedström, E. Nordberg Karlsson, S. Linse, A. Khalil and B. Mattiasson, Production and physicochemical characterization of acidocin D20079, a bacteriocin produced by Lactobacillus acidophilus DSM 20079. World J. Microbiol. Biotechnol. Available on-line 5 December (2006)

U. Törnvall, C. Orellana-Coca, R. Hatti-Kaul and D. Adlercreutz, Stability of immobilized Candida antarctica lipase B during chemo-enzymatic epoxidation of fatty acids. Enz. Microb. Technol. Available on-line 12 July (2006)


Doctoral degrees and theses

Irene Bohn (photo Erik Anders-son)

Martin Hedström (photo Erik Andersson)

Conferences

Monolith Summer School on Biochromatogra-phy, Bioconversion and Solid Phase Synthesis, Portoroz, Slovenia, 28-31 May.

CBioSep-WGDSP Meeting in Lund, Sweden, 19-20 June.

International Congress on Biocatalysis, Ham-burg, Germany, 3-7 September.

IWA World Water Congress, Beijing, China, 10-14 September.

The 2nd International Symposium towards En-vironmentally Harmonized Energy- Conversion, Miyazaki-Japan, 14-15 September

Extremophiles, Brest, France, 17-21 September

Below are listed the national and international conferences and symposia attended by biotechnology co-workers during 2006.

Purification of Biological Products, 2nd Annual Meeting, Thousand Oaks, CA, USA, 18-20 Sep-tember 18-20.

20-th European Conference on Biomaterials, Nantes, France, 27 September - 1 October.

2nd STIPOMAT WORKSHOP, Seggau, Austria, 23-25 October.

BIND-06: International Conference on Design of Biomaterials, Indian Institute of Technology, Kanpur-India 8-11 December

Environmental Biocatalysis: From Remediation with Enzymes to Novel Green Processes, Cor-doba, Spain, 23-26 April.

Licentiate degrees and theses

2� JANUARYKevin OluochAlkaline active enzymes from Bacillus halodurans LBK261 with potential application in textile industry.

12 JUNEJosefin AhlqvistUsing monolithic cryogels for direct capture of inclu-sion bodies during fermentation- a way to monitor the process.

2 OCTOBERTamer EssamSolar-based physicochemical-biological processes for the treatment of toxic and recalcitrant effluents.

20 DECEMBERCarl Elovson GreyProtection of biomolecules by antioxidants-Mechanisms and applications.

1� JANUARYGashaw Mamo Basazenen An alkaline active xylanase from Bacillus halodurans S7 Molecular and structural aspects.

20 JANUARYCecilia Orellana Coca Chemo-enzymatic epoxidation of unsaturated fatty acids.

1� FEBRUARYDores Gurgo e CirneEvaluation of bio-logical strategies to enhance hydrolysis during anaerobic digestion of complex waste.

� JUNEIrene Bohn Anaerobic digestion of crop residues at low temperatures.

1� JUNEMartin HedströmProcess analytical tools for the monitoring and char-acterization of bacterial toxins.

Dept. of Biotechnology Annual Report 2006�0

Department of BiotechnologyCenter for Chemistry and Chemical Engineering

Lund UniversityP.O. Box 124

SE-221 00 LundSWEDEN

www.biotek.lu.se

Email addresses Personal addresses are constructed by

name and surname as follows:[email protected]

Annual report 2006

PHOTOS: AGRIGAS (TOP), BO MATTIASSON, EVA NORDBERG-KARLSSON, MARIKA MURTO

Publisher: Rajni Hatti-Kaul. Editors: Josefin Ahlqvist, Maria Andersson, Marika Murto

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