Page i

A Project Proposal

on

Biofortification of Selenium in Edible Mushrooms from Se-rich Crop

Residues

Submitted to

Department of Biotechnology

Ministry of Science and Technology

New Delhi

by

Dr. N.Tejo Prakash

Associate Professor

Department of Biotechnology and Environmental Sciences

Thapar University, Patiala

Patiala

&

Dr. Swaranjit Singh Cameotra

Scientist F

Institute of Microbial Technology

Chandigarh

Page ii

Project Summary

Title : Biofortification of Selenium in Edible Mushrooms from Se-rich Crop Residues

1. Name of the Institute/University/Organization/Industry submitting the Project Proposal:

Thapar University, Patiala

2. State: Punjab

3. Is the project Single Institutional or Multiple-Institutional? Multi-Institutional

Principal Investigator: Dr. N.Tejo Prakash, Associate Professor, Thapar Univ., Patiala

Co-Investigator(s) : Dr S.S. Cameotra, Scientist F, IMTECH, Chandigarh

Dr Ranjana Prakash, Associate Professor, Thapar Univ., Patiala

4. Status of the Institution (PI): Deemed University

5. Name and Designation of the Executive Authority of the Institution forwarding the application:

The Dean, Research and Sponsored Projects

Thapar University, Patiala – 147004

6. Rationale / Objective of the project

Agriculturally rich state like Punjab, with major produce in the form of wheat and rice,

generates 23 million tonnes of rice straw and 17 million tonnes of wheat straw, annually. The

seleniferous region of Punjab bordering the districts of Nawanshahr and Hoshiarpur has

approximately 1200 hectares under cultivation. The conventional practice of discarding the post-

harvest residue is dominantly through burning of residue in field, with very marginal use as fodder.

The agri-waste residues contain significantly high levels of selenium accumulated in crop produce

as well as post-harvest residues as detected by the investigators. With extensive burning of these

residues being a major environmental concern in North-western region of India, any approach that

would facilitate re-use of post-harvest agri-residue for extraction of selenium (Se) would help

reducing the atmospheric pollution and in particular provide appropriate outlet for revenue

generation especially in the Se-impacted region.

Mushrooms are thought of as healthy foods with relatively few calories and fat but rich in

vegetable proteins, chitin, vitamins and minerals. Several agricultural residues have been used to

produce the edible mushrooms belonging to genera such as Pleurotus (oyster mushrooms),

Agaricus (button mushrooms), Boletus (Porcini), Ustilago (Huitlacoche), Ganoderma, etc. With

strong evidences indicating bioaccumulation of selenium in crop plant parts, the proposed project

envisages mobilization and fortification of selenium from agri-residues generated during the harvest

of wheat, rice and other cereals as growth-substrates for mushroom cultivation. Till date there is

no evidence on Se uptake in mushrooms cultivated on agri-residue based substrates naturally

enriched with selenium in either commercial or experimental culture conditions.

Objectives

Demonstrate the potential of Se-fortified mushrooms in term of bioactive properteisl

Examine potential of Se-fortified mushrooms as a functional food supplements/products.

Page iii

7. Total Project Costs: Rs. 34.8 lakhs; Duration: 2 Years

Non-Recurring: Rs. 13,00,000/-

Recurring:

Manpower Rs. 4,50,000/-

Consumables Rs. 5,00,000/-

Travel Rs. 1,50,000/-

Contingency (Incl. analytical services) Rs. 5,00,000/-

Overheads @ 20% Rs. 5,80,000/-

Grand Total Rs. 34,80,000/- (Rs. 34.80 lakhs)

Page 1

PROFORMA – I

PROFORMA FOR SUBMISSION OF PROJECT PROPOSALS ON RESEARCH AND

DEVELOPMENT, PROGRAMME SUPPORT

PART I: GENERAL INFORMATION

1. Name of the Institute/University/Organisation

submitting the Project Proposal : Thapar University

2. State: Punjab 3. Status of the Institute: University

4. Name and designation of the Executive Authority of the Institute/University forwarding the

application : The Dean, Research and Sponsored Projects, Thapar University, Patiala

5. Project Title : Mobilization and Biofortification of Selenium to Edible Mushrooms from Se Rich

Crop Residues

6. Category of the Project (Please tick) :R&D / Programme Support:

7. Specific Area (Please see Annexure - II): 8.6 – Mineral Biofortification

8. Duration : Two Years (24 Months)

9. Total Cost (` in lakhs) : 34.80

10. Is the project Single Institutional or Multiple-Institutional (S/M) ? : Multi-Institutional

11. If the project is multi-institutional, please furnish the following :

Name of Project Coordinator : Dr N.Tejo Prakash

Affiliation : Department of Biotechology and Environmental Sciences, Thapar University, Patiala,

India.

Address : Associate Professor, Department of Biotechology and Environmental Sciences, Thapar

University, Patiala, India.

12. Scope of application indicating anticipated product and processes

This study is aimed to demonstrate cultivation of selenium fortified mushrooms using Se rich

crop residues for their use as selenium food supplements.

13. Project Summary (Not to exceed one page. Please use separate sheet). Attached

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PART II: PARTICULARS OF INVESTIGATORS

Principal Investigator:

14. Name: N.Tejo Prakash ; Date of Birth: 27-07-1965; Sex (M/F): Male

Designation: Associate Professor

Department: Department of Biotechnology and Environmental Sciences

Institute/University: Thapar University, Patiala

Address: Department of Biotechnology and Environmental Sciences, Thapar University, Patiala,

147004, India.

Telephone: 0175-2393319; Fax:. 0175-2364498; E-mail:.ntejoprakash@thapar.edu

Number of research projects being handled at present:. 02 (Two) & 02 (as CoPI)

Co-Investigator:

15. Name: Swaranjit Singh Cameotra; Date of Birth : 30.01.1959; Sex : Male

Designation : Scientist F ; Department : Microbial Type Culture Collection;

Institute: Institute of Microbial Technology

Address : Sector 39 A, Chandigarh, 160036, India.

Telephone: 0172-665223/224, 09888146750 Fax No.: 1722690632

E-mail : ssc@imtech.res.in

Number of Research projects being handled at present: 03 (Three)

Co-Investigator:

16. Name : Ranjana Prakash; Date of Birth: 19-11-1968; Sex(M/F) : Female

Designation : Associate Professor

Address : School of Chemistry and Biochemistry, Thapar University, Patiala ; PIN : 147004

Telephone : 0175-2393119; Fax: 0175-2364498; Email: rprakash@thapar.edu

Number of Research projects being handled at present: 02 (PI) & 02 (Co-PI)

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PART III : TECHNICAL DETAILS OF PROJECT

17. Introduction

17.1 Origin of the proposal

As the concept of fortification through nutrition vis-à-vis crop which have augmented capacity to accumulate selenium hold a market value in terms of use as Se supplements, they can be commercially exploited in the specific geographical regions which are naturally deficient for Se. The proposal aims at cultivating mushrooms on Se rich crop residues harvested from seleniferous belt of Punjab, India for exploring and exploiting the bioactive and health related properties of Se enriched mushrooms.

17.2 (a) Rationale of the study supported by cited literature

Agriculturally rich state like Punjab, as major producer of wheat and rice, generates 23 million tonnes of rice straw and 17 million tonnes of wheat straw, annually. The seleniferous region of Punjab bordering the districts of Nawanshahr and Hoshiarpur has approximately 1200 hectares under cultivation. The conventional practice of discarding the post-harvest residue is dominantly through burning of residue in field, with very marginal use as fodder. These agri-waste residues contain significantly high levels of selenium as detected by the investigators. 1,2

Mushrooms constitute an increasing share as healthy foods with relatively few calories and fat but rich in proteins, chitin, vitamins and minerals.3 Several agricultural residues have been used to produce the edible mushrooms belonging to genera such as Pleurotus (oyster), Agaricus (button), Lentinula (Shitake), Ganoderma (Reishi), etc. With strong evidences indicating bioaccumulation of selenium in crop plant parts as reported by other researchers 4 and the investigating research group, the proposed project envisages mobilization and fortification of selenium from agri-residues generated during the harvest of wheat, rice and other cereals as growth-substrates for mushroom cultivation. Till date, there is no report on Se uptake in mushrooms cultivated on agri-residue based substrates naturally enriched with selenium in either commercial or experimental culture conditions. In addition, there is limited information on the biological and health related properties accrued by Se enriched mushrooms when compared to commonly (non-Se) grown mushrooms.

(b) Hypothesis

Selenium enrichment in mushrooms growing on natural Se rich crop residue would facilitate enhance bioactivity over normal mushrooms; and therefore can be cultivated for nutritional and pharmacological applications.

(c) Key questions

What would be the extent of selenium accumulation in fruiting bodies of mushrooms during the process of cultivation?

What are the biological activities of mushrooms that get modulated due to selenium enrichment?

What is the feasibility of commercial production of Se enrich mushrooms as neutraceuticals?

1 Sharma et al., 2009. J.Radioanal. Nucl. Chem., 281: 59-62.

2.Cubadda et al., 2010, J. Agri. Food Chem., 58: 2295-2301.

3 Lindequist et al., 2005, e CAM., 2: 285-299.

4 Dhillon and Dhillon, 2003, Adv. Agron., 79: 119-184.

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17.5 Current status of research and development in the subject

The concept of using plants in place of chemical or nutrient factories to supply the direly needed macro and micro-elements is gaining significant attention and constitutes the first step towards biotechnology-based, nutritionally fortified foods.1 Although, it has been known since late 1800s that edible biota such as mushrooms can accumulate extraordinary levels of metals, yet the use of these systems as nutrient-supplements has been very limited. Edible mushrooms can accumulate elements such as selenium which can act as essential nutrients, and therefore can provide natural source of such elements to both animals and human beings.

Selenium deficiency and suboptimality are manifested in populations as increased rates of thyroid dysfunction, cancer, severe viral diseases, cardiovascular disease and various inflammatory conditions. Se deficiency probably affects at least a billion individuals in Europe and Scandanavian countries.1 The strategies that applied to overcome these deficiencies include increase consumption of higher-Se foods, individual supplementation, food fortification, etc.2

The seleniferous region of Punjab bordering the districts of Nawanshahr and Hoshiarpur having approximately 1200 hectares under cultivation generates agri-waste residues that contain significant levels of selenium accumulated in plant parts.3 These post-harvest agricultural residues can effectively be used to produce both edible and medicinal mushrooms belonging to variety of genera such as Pleurotus, Agaricus, Ganoderma, etc. With strong evidences indicating bioaccumulation of selenium in agri-residues, the proposed project envisages mobilization and fortification of selenium from these substrates to mushrooms. Although, limited evidences have appreared in recent past on selenium uptake in commercial mushrooms such as Pleurotus, Ganoderma, etc., under experimental conditions,4,5 there is no evidence on Se uptake in mushrooms with Se rich substrates and their bioactivity.

17.6 The relevance and expected outcome of the proposed study

This study is expected to demonstrate biological potential of selenium fortified mushrooms for their use as selenium enriched neutraceuticals. In addition to the determination of health related properties the project would facilitate re-use of post-harvest agri-residue as an alternative to the burning of agri-waste, a practice prevalent in the region. Thus, reducing the atmospheric pollution and provide appropriate outlet for revenue generation.

17.7 Preliminary work done so far

The research group of PI has been associated with sponsored research projects, since year 2004, on selenium removal and its accumulation in biological systems such as bacteria and crop plants cultivated in seleniferous region of Punjab. The initial focus of the group was to exploit the potential of selenium tolerant bacterial isolates to mobilize selenium from soils and to understand the mechanisms of biological transformations (CSIR project 2003-07). Further studies were carried to characterize the biotransformed forms of selenium (DRDO project 2008-09). Current on-going projects, are aimed to understand speciation in environmental and biological samples from seleniferous region of Punjab. As a part of the study, the selenium levels have been quantified in crop produce such as rice, wheat, maize and mustard; and their post harvest residues. In addition, detailed speciation studies have been carried out in few grain varieties and their processed food materials. Presently, cultivation of Pleurotus sp. (oyster mushrooms) and Agaricus sp. (button mushrooms) have been carried out on the Se rich agri-residues. Preliminary quantification of selenium in Se fortified mushrooms indicate significant Se uptake. Studies are in progress to examine the bioactivity of the test mushrooms.6,7

1 Lyons et al., 2003, Nutr. Res. Rev., 16:45-60

2 Combs, 2001, Br. J. Nutr., 81:517-547

3 Dhillon and Dhillon, 2003, Adv. Agron., 79: 119-184

4 Slejkovec et al., 2000, Biol. Trace Elem. Res., 75: 139-155.

5 Estrada et al., 2009, World J. Microbiol. Biotechnol., 25:1597-1607

6 Bhatia et al., 2010, in. Proc. Natl. Conf. Emerging Trends in Biopharmaceuticals, Thapar University, Patiala, pp. 221

7 Bhatia et al., 2011.. in. International Conf. Trace Elements in Foods, Aberdeen, UK.(forthcoming)

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18. Specific objectives

Demonstrate the potential of Se-fortified mushrooms in terms of bioactive

properties

The studies under this objective would broadly aimed

(a) quantification and speciation of selenium in edible and medicinal mushrooms cultivated

on Se rich substrates using florescence spectrometry and further validation using NAA and/or

ICP-MS ;

(b) determining the modulations in bioactivity of Se enriched mushrooms in comparison the

commonly (non-Se) cultivated mushrooms, focusing on various nutritional (protein, vitamin,

anti-oxidant levels, etc) and medicinal (antibiotic, fibriolytic, anti-cancer, etc) properties using

standard protocols and in vitro models wherever applicable.

Examine potential of Se-fortified mushrooms as a functional food

supplements/products.

This component of the study would be focused to examine the quality and techno-economic

feasibility of scale-up cultivation, processing and marketability of Se-fortified edible and

medicinal mushroom products for neutraceutical and pharmacological requirements.

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19. Work Plan

19.1 Work plan & 19.2 Connectivity of the participating institutions and investigators

a) Selenium quantification in agricultural residues

Agricultural (post-harvest straw) residues from wheat, rice and other crops will be collected following seasonal and cropping patterns in seleniferous belt of of Hoshiarpur- Nawashahr regions during respective harvest periods. Samples will be collected from different pockets of seleniferous along with non-seleniferous sites. Total selenium will be estimated in residues using florescence spectrometer followed by validation using neutron activation analysis (NAA).

To be carried out at Thapar University, Patiala & utilization of BARC facility

b) Mushroom cultivation on Agricultural Residues

The cultivation of selected species of edible and medicinal mushrooms belonging to genera Pleurotus, Agaricus, Ganoderma, Lentinula, Flammulina and Volvariella will be established using agriculture and plantation based substrates collected from field under study, following conventional protocols already standardized by PAU, Ludhiana and NRCM, Solan.

To be carried out at Thapar University, Patiala

c) Examine selenium uptake by mushroom from Se-rich growth substrates and its bioaccessibility

The uptake profile and localization of selenium would be examined from fruit body stage, across the duration of cultivation, so as to determine the fate and speciation of selenium over different flushes. Quantification of Se would be carried out using florescence spectrometry followed by NAA using research reactors at BARC. Speciation of Se would be carried out by separating appropriate fractions using aqueous (water soluble), acid (inorganic) and in vitro (gastric/ gastrointestinal) digestion methods followed by INAA based quantification of Se in each of the fractions.

To be carried out at Thapar University, Patiala and utilization of BARC facility

d) Examine bioactive properties of Se-fortified mushrooms

The modulations in the nutritional (protein content, mineral and vitamin profile, anti-oxidant effect, etc.) parameters of fruiting bodies, as induced by selenium accumulation, would be examined using standard methods and in vitro digestion protocols along with the Se quantification. The extracts obtained from edible portions of the test mushrooms will be examined for various bioactive and health related properties such as anti-microbial activity, anti-oxidant activity, anti-inflammatory response, etc. The studies would dominantly be carried out using in vitro models. Attempt would also be made to replicate the studies under in vivo conditions with animal models.

Based on the evidence of various bioactive properties, further studies will be carried out at University, Patiala & IMTECH, Chandigarh along with utilization analytical service/commercial facilities available at NIPER, SAIF and other organizations.

e) Scale-up production of mushrooms

The feasibility of the scale up production of Se rich mushroom products will be examined for use as functional foods and raw material for neutraceutical preparations

To be carried out at Thapar University, Patiala

19.3 Alternate strategies

Focus would be diverted towards isolation and characterization of bioactive molecules and/or metabolites generated by Se enriched mushrooms.

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20. Timelines

Period of study Achievable targets

6 Months

Establishment of mushroom cultivation facility

Quantification of selenium in agricultural and other substrates to

be used for cultivation of mushrooms

12 Months Procurement and cultivation of selected species of edible and

medicinal mushrooms in phased manner

Mushroom cultivation at culture facility

Quantification of Se uptake and bioaccessibility following various

protocols

18 Months Determination of nutritive and bioactive properties of Se

fortified mushrooms using standard protocols and in vitro

models respectively

24 Months Scale up cultivation and examination of quality parameters

associated with commercial production of Se fortified

mushrooms

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PART IV: BUDGET PARTICULARS Budget (In Rs)

A. Non-Recurring (e.g. equipments, accessories, etc.)

S.

No.

Item Year 1 Yr.2 Yr.3 Total

1

Mushroom cultivation facility

(Infrastructural requirement include establishment of

small scale mushroom culture facility with

temperature and humidity controlled systems to

facilitate 365 day cultivation)

12,00,000/- 13,00,000/-

2 UPS and A/C facility of florescence spectrometry and

analytical lab

1,00,000/-

Sub-Total (A)

B. Recurring

B.1 Manpower (See guidelines at Annexure-III)

S. No. Position

No.

Consolidated Emolument Year 1 Year 2 Total

1 JRF (1)

Rs. 14,000/- + 15% HRA + MA 1,94,000 1,94,000 3,88,000

2 Field Assistant On daily wage basis based on

prevailing rates

62,000

Manpower would include recruitment of a research fellow (1) and a field assistant (1) for the

duration of 2 years.

Sub-Total (B.1) = 4,50,0000

B.2 Consumables

S. No. Item

Quantity Year 1 Year 2 Total

1 Glassware, labware and

standards

As per the

requirement

2,50,000 2,50,000 5,00,000

Consumables and labware are required for mushroom cultivation and lab scale analysis of routine

parameters

Sub-Total (B.2) = 5,00,000

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Other items Year 1 Year 2 Total

B.3 Travel

75,000 75,000 1,50,000

B.4 Contingency

2,50,000 2,50,000 5,00,000

Travel includes transport of raw material for from field to TU-Patiala; and mutual

interim visits of investigators at TU-Patiala and IMTECH-Chandigarh, NABI-Mohali,

and various analytical facilities in the region as and when required.

Contingency expenditure and expenditure towards analytical services would

payment of service charges for carrying analysis of nutrient and bioactive

properties using facilities at NIPER, SAIF-PU and IISER, Mohali or other facilities

on paid basis.

B.5 Overheads (@20%) 5,80,000

Sub-total of B (B.1+B.2+B.3+B.4+B.5) 16,00,000

Grand Total (A + B) 34,80,000

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PART V : EXISTING FACILITIES

Infrastructure facilities related to project activity available at PI’s institute (Thapar Univ.)

1. Atomic Absorption Spectrophotometer with hydride generator/GF accessory

5 Temperature controlled room & cold chamber/sub-zero cabinets

2. DNA & Protein Electrophoresis 6. Autoclaves

3. Refrigerated Centrifuges 7. Laminar air flows

4. Innova refrigerated orbital shakers 8. Microfuges

In addition, Neutron activation analysis, X-Ray Florescence spectrometer, and other nuclear analytical facilities are available to PI through collaborative programme with BARC, Mumbai

Infrastructure facilities related to project activity available at CI’s institute (IMTECH)

1. Confocal Microscope 5 HPLC

2. Animal House 6. GC

3. Culture Collection 7. SEM

4. AAS 8. TEM

Page 11

Page 12

PART VII: PROFORMA FOR BIOGRAPHICAL SKETCH OF INVESTIGATORS

A. Principal Investigator

Name : N.Tejo Prakash

Designation : Associate Professor

Department/Institute/University : Department of Biotechnology and Environmental Sciences,

Thapar University, Patiala 147004, India

Date of Birth : 27.07.1965; Sex (M/F) : Male; SC/ST : Not applicable

Education (Post-Graduation onwards & Professional Career)

S. No. Institution/ Place Degree Awarded Year Field of Study

1 S.V.University, Tirupathi M.Sc 1987 Zoology

2 Pondicherry University, Pondicherry Ph.D 1994 --

A. Position and Honors

Position and Employment (Starting with the most recent employment)

S. No. Institution/ Place Position From (Date) To (date)

1 Thapar University, Patiala Associate

Professor

September 2010 Till date

2 Thapar University, Patiala Assistant

Professor

February 2003 August 2010

3 Thapar Centre for Industrial Research

& Development, Patiala

Scientist July 1997 January 2003

Honors/Awards

2009 – Thermo‐Scientific Award for best poster presentation – 3rd International Conference on Trace Element in Foods, Rome, Italy

2008 ‐ Erasmus Mundus MESPOM Scholarship as Visiting Faculty at University of Manchester, UK 2008 ‐ Kirkhouse Trust Travel Award, Kirkhouse Trust, Institute of Advanced Study, Oxford

University, UK 2003 ‐ JSTP – Post Doctoral Fellowship, Japan Society for Promotion of Science, Japan (Could not

be availed) 1996 ‐ SETAC – Summer School Visitorship, Society of Environmental Toxicology and Chemistry

(SETAC) – Italy, Europe 1995 ‐ MASHAV Postdoctoral Visitorship, Centre for International Cooperation, Israel 1996 ‐ DRDO Research Associateship, Defence Research and Development Organization (DRDO),

India

Professional Experience and Training relevant to the Project

The research group of PI has been associated with sponsored research projects, since year 2004, on

selenium removal and its accumulation in biological systems such as bacteria and crop plants

cultivated in seleniferous region of Punjab. The initial focus of the group was to exploit the potential

of selenium tolerant bacterial isolates to mobilize selenium from soils and to understand the

mechanisms of biological transformations (CSIR project 2003-07). Further studies were carried to

characterize the biotransformed forms of selenium (DRDO project 2008-09). Current on-going

projects, are aimed to understand speciation in environmental and biological samples from

Page 13

seleniferous region of Punjab (BRNS 2008-11; UGC-SAP; 2007-11). Presently, cultivation of Pleurotus

sp. (oyster mushrooms) and Agaricus sp. (button mushrooms) have been carried out on the Se rich

agri-residues. Preliminary quantification of selenium in Se fortified mushrooms indicate significant

Se uptake. Studies are in progress to examine the bioactivity of the test mushrooms.

B. Publications

Books/Book Chapters : 05; Research Papers: 28; Patents filed: 02;

Others: 01 (http://umbbd.ahc.umn.edu/bzs/ bzs_map.html)

Selected peer-reviewed publications

1. Singh M, Khanna S, Tejo Prakash N, 2010. Influence of cellulolytic bacteria augmentation on

organic carbon and available phosphorus in sandy loam soil under cultivation. Journal of

Agricultural Science, 2: 137-145.

2. Tejo Prakash N, Sharma N, Prakash R, Acharya R, 2010. Removal of selenium from Se enriched

natural soils by a consortium of Bacillus isolates. Bulletin of Environmental Contamination and

Toxicology, 84: 214-218.

3. Cubadda F, Aureli A, Ciardullo S, D’Amato M, Raggi A, Acharya R, Reddy AVR, Tejo Prakash N,

2010. Changes in selenium speciation associated with increasing tissue concentration of

selenium in wheat grain. Journal of Agricultural and Food Chemistry, 58: 2295-2301.

4. Tejo Prakash N, Sharma N, Prakash R, Nathaniel N, Acharya R, Reddy AVR, 2010. Selenium

fortification and pro/anti-oxidant responses in Allium cepa (onion plant) cultivated in Se

supplemented soils. Experimental Agriculture, 46: 531-540.

5. Gupta S, Ranjana Prakash, Pearce CI, Pattrick RAD, Hery M, Lloyd JR, Tejo Prakash N, 2010.

Selenium mobilization by Pseudomonas aeruginosa (SNT-SG1) isolated from seleniferous soils

from India, Geomicrobiology Journal, 27: 35-42.

6. Tejo Prakash N, Sharma N, Ranjana Prakash, Raina KK, Fellowes J, Pearce CI, Lloyd JR, Pattrick

RAD, 2009. Aerobic microbial manufacture of nanoscale selenium: Exploiting nature’s

bionanomineralization potential. Biotechnology Letters, 31: 1857-1862.

7. Sharma N, Prakash, R, Srivastava A, Sadana US, Acharya R, Tejo Prakash N, Reddy AVR, 2009.

Profile of selenium in soil and crops in seleniferous area of Punjab, India by neutron activation

analysis. Journal of Radioanalytical and Nuclear Chemistry, 281: 59-62.

8. Pearce CI, Coker VC, Cutting R, Pattrick RAD, Lloyd JR, Tejo Prakash N, 2008, Bio-nano-

engineering: From cleanup to catalysis, Geochimica et Cosmochimica Acta (Suppl. P) 72: 730-

731.

9. Arun Kapoor, Ranjana Prakash, Tejo Prakash N, 2003. Predictive Biotransformation of Potential

Toxic Chemicals, Chemicke Listy, 97: 472-473.

10. Tejo Prakash N, Rao KSJ, 1995. Modulations in anti-oxidant enzymes in different tissues of

marine bivalve Perna viridis during heavy metal exposure. Molecular and Cellular Biochemistry,

146: 107-113.

List maximum of five recent publications* relevant to the proposed area of work.

i. Cubadda F, Aureli A, Ciardullo S, D’Amato M, Raggi A, Acharya R, Reddy AVR, Tejo Prakash N,

2010. J. Agri. Food Chem., 58: 2295-2301.

ii. Tejo Prakash N, Sharma N, Prakash R, Nathaniel N, Acharya R, Reddy AVR, 2010. Exper. Agri.,

46: 531-540.

iii. Tejo Prakash N, Sharma N, Prakash R, Acharya R, 2010. Bull. Environ. Contam. Toxicol.,, 84:

214-218.

Page 14

iv. Gupta S, Ranjana Prakash, Pearce CI, Pattrick RAD, Hery M, Lloyd JR, Tejo Prakash N, 2010.

Geomicrobiol. J., 27: 35-42.

v. Sharma N, Prakash, R, Srivastava A, Sadana US, Acharya R, Tejo Prakash N, Reddy AVR, 2009.

J. Radioanal. Nucl. Chem., 281: 59-62.

*Details of the publications given in the list of the best publications

C. Research Support

Ongoing Research Projects

Sl

No.

Title of Project Funding

Agency

Amount Date of sanction

and Duration

1 Studies on Se and its speciation ….biological samples from seleniferous region of Punjab

BRNS, Govt.

of India

11 Lakhs May 2008 ; 3 yrs

2 Selenium mobilization ….. seleniferous Regions

UGC-SAP 13 Lakhs

(Thrust Area II)

Oct. 2007; 4 yrs

Completed Research Projects

S. No. Title of Project Funding Agency Amount Date of

completion

1 Role of plant microbe

interactions …… soils

CSIR 9.94 April 2007

2 The microbial controls on

selenium ….. in the environment

Univ. Manchester –

ESRF, Grenoble

Synchrotron

facility and

travel

support

July 2009

3 Biogenesis of Se nanoparticles DRDO 6.80 November 2009

Place : Patiala

Date : 25/01/2011

Page 15

B. Curriculum vitae (CV) of Co-Investigator (CI)

Name & Designation: Dr. Swaranjit Singh Cameotra Scientist F, Institute of Microbial Technology Date & Place of Birth: 30 Jan 1959 Nationality: Indian Present post: Scientist E II (wef 1.1.2002) Institution with address: IMTECH, Sector 39 A, Chandigarh 160036 Telephone No. (with STD code):0172-665223/224, 09888146750; Fax No.: 0172- 2690632 E-mail: ssc@imtech.res.in Qualifications: Ph. D. Biochemistry, RRL Jorhat (Gauhati University 1986, M. Sc. Biochemistry BHU 1980 EXPERIENCE: ENVIRONMENTAL/INDUSTRIAL APPLIED BIOTECHNOLOGY, BIODIVERSITY: 22 years after Ph. D & as Scientist. Presently a Scientist F. Attended a fortnight long IPR training course in IPMD, CSIR in 2006. Completed a 45 days electronic course of WIPO on different aspects of IPR/Patents on the 15th November 2006 with excellent grade awarded by WIPO. IPR papers: (i) Preservation of microorganisms as ‘deposits’ in Patent Applications. Swaranjit S. Cameotra BBRC 353(4):849-50. Feb 23, 2007. (ii) IPR Protection of Geographical Indicators: communicated 2007. (iii) Lesser Known IPR Protection of New Varieties of Plants. Plant Breeding org. February 5, 2007. (iv) A database on Microbial Surfactants 2007 has been submitted for publication. The following projects are ongoing: 1. Bioremediation of Heavy Metals. 2. MTCC & Gene Bank Teamwork has made MTCC an International Depository (IDA). 3. National Project on Biodiversity, and 4. Production and applications of Lipases and Biosurfactants as adjuvants. During my Ph. D. work (1981-1986) I elucidated the mechanism of Petroleum Hydrocarbons uptake / utilization by bacterial and yeast strains. Investigation and study made on the "Mechanism of Hydrocarbon Utilization by Microorganisms" using Yeast and Bacterial sp. Participated in an oceanographic cruise onboard R.V. Gaveshani organized by Department of Ocean Development (DOD) from October 4-20, 1986. Work in the Bacterial and Plasmid Wing of the Microbial Type Culture Collection & Gene Bank (MTCC) since October 1987. MTCC is an ongoing National Project/Facility set up by CSIR and DBT jointly. I am involved in preservation, maintenance, distribution and identification of bacterial strains. The strains are preserved by

freeze drying, storage in 10% glycerol in liquid nitrogen and at -70 C. Viability record of the lyophilized cultures is maintained. Strain data computerization and data files maintenance was started in 1987. I am involved in the identification of bacterial cultures. MTCC is now an International Depository Authority (IDA) and the strains for patent filing can be deposited in the IDA in our country. I take care of the Liquid Nitrogen Plant for smooth supply of LN2 that is vital for the long-term preservation of cell lines and microbial cultures since 1987. These are very important microbiology activities for the conservation of the rich microbial diversity of India. I successfully completed very interesting and applied projects in IMTECH: Production of Biosurfactants by Microorganisms and Microbial degradation of Petroleum Sludge and Pesticides. I guided 5 Ph. D. students (Ph. D. degrees awarded). Presently I am supervising 4 Ph. D. students. The results and data were consistently published in high impact journals like AEM, BBRC, TIBTECH, COM, Biotechnol. Bioeng., JIM, Biotechnology Techniques, CRC Rev.

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Biotechnol. etc. ~45 Publications-The citation index (CI) of my publications is 800. h- Index is 13. Basing on my contributions to Microbiology I am a Task Group Member of two WFCC. I am a Member of the Board of Studies of the Microbiology Wing of the Punjab University Chandigarh, GNDU Amritsar and the HAU, Hisar. Use of biosurfactants in the proliferation of T and B cells/immune response is in progress. The findings will be of therapeutic value and are currently being carried out in mice. In June 2007 I delivered an invited talk on “Bioremediation of Xenobiotics” in the Asia Pacific Conference on Plant Tissue Culture and Agribiotechnology (APaCPA) 2007, Kuala Lumpur, Malaysia. On 16th October 2007, I delivered an invited talk on “Conservation of Microbial Diversity” in the G. B. Pant Institute Of Himalayan Environment & Development and conducted Hands-on-Training on this aspect. Delivered an invited lecture in the IBS 2008 in Dalian, China.I wrote many popular articles that were enjoyed by people. Dr. Swaranjit Singh is a Member of Biotechnology & Biosafety of the National Biodiversity Authority, Chenna and a Consortium Member of the ICAR NAIP Project- Development of non-destructive systems for evaluation of microbial and physico-chemical quality parameters of mango. Wrote many popular science articles in magazines and papers.

Awards & Fellowships: Fellow Association of Microbiologists, India (FAMI). Member Nat.Aca. of Sciences(M FNA Sc.) Member WFCC Task Groups. Member Society of Scientific Values(SSV) and Member SIS. Successfully completed the CSIR Leadership Development Programme LDP08-02. Received 1st Prize for my essay on “Team CSIR in the labs and fields” organized by CSIR by the DGSIR in Lucknow on the 20th February 2005. Post Doctoral Research in Pennsylvania State University, USA 1999. Kobe University, Japan 2005. The citation index (CI) of the publications is 800. h- Index is 13. Publications during the last 5 years/or relevant to the project

1. Potential of Microbial Surfactants in Biomedical Sciences. Pooja Singh and Swaranjit Singh

Cameotra, TIBTECH 22(3): 142 - 146 (2004). 2. Recent applications of biosurfactants as biological and immunological molecules. Swaranjit

Singh Cameotra and Randhir S. Makkar. Current Opinion in Microbiology, 7(3): 262-266, 2004.

3. Isolation and characterization of a bacterial strain capable of degrading Atrazine. Pooja Singh, C. Raman Suri and Swaranjit Singh Cameotra. BBRC: 317(3): 697-702, 2004.

4. Preservation of microorganisms as deposits in Patent Applications. Swaranjit Singh Cameotra. BBRC 353(4):849-50. Feb 23, 2007.

5. Lesser known IPR Protection of New Plant Varieties. Swaranjit Singh Cameotra. Plant Breeding February 5, 2007. http://plantbreeding.org/ article.php

6. New Definition of Xenobiotics. Wikipedia. Swaranjit Singh Cameotra. February 15, 2007. http://www.katrinapetsneedhelp.com/pets/Xenobiotics

7. Surfactin description in Wikipedia. http://en.wikipedia.org/wiki/Surfactin 8. Atrazine uptake by Acinetobacter radioresistens using a flourometric method. Accepted for

Publication 2008. 9. Mode of Hexadecane Uptake and Rhamnolipid Synthesis by P.aeruginosa. Accepted for

Publication (In Press) 2008. 10. Bioremediation of oil sludge using crude biosurfactants. Int. Biodeterio. Biodegradation

2008 (Online/In Press). 11. Priyanka Lal, Vishnu Agarwal, Parul Pruthi, Vikas Pruthi and Swaranjit Singh Cameotra. 2008,

Biofilm Formation by Microorganisms Isolated from Paper Mill Effluent and its Public Health Implications. Clean: soil, air, water 36 (12): 927 – 929, 2008.

12. Deepak Sharma, Vikas Pruthi and Swaranjit Singh Cameotra 2008, Characterization and industrial usage of biosurfactant produced by a hydrocarbon degrading Bacillus pumilus strain. Applied Biochemistry and Biotechnology (Submitted for publication)

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13. Silk-fiber immobilized lipase-catalysed hydrolysis of emulsified sunflower oil. Sushovan Chatterjee, Lepakshi Barbora, Swaranjit Singh Cameotra and Pranab Goswami. In Press 2008.

14. Biosurfactant-enhanced bioremediation of hydrophobic pollutants. Swaranjit Singh Cameotra. J. Biotechnol. 136, 678, 2008.

15. Aerobic biogenesis of selenium nanospheres by Bacillus cereus isolated from coalmine soil, Soniya Danjal and SS Camotra, Microbial Cell Factories, 9: 52-59.

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C. Curriculum vitae (CV) of Co-Investigator (CI)

Name : Dr. Ranjana Prakash

Date of Birth : 19-11-1968

Designation : Assistant Professor

Educational Qualifications

Degrees Discipline Year University

B.Sc Biological Sciences 1987 Kanpur University

M.Sc Chemistry 1990 Kanpur University

Ph.D Applied Chemistry 1999 Kanpur University

Teaching and Research Experience

Experience (Years) : Teaching – 10 Years

Research – 8 Years (post-Ph.D)

Thesis guidance (postgraduate & Ph.D.) : Ph.D: 04 (on-going)

M.Phil: 01 (Completed)

PG: 10 (completed)

01 (on-going)

Area of Specialization : Biotransformations and biodegradation

Sponsored Research (ongoing) :

1. Strain improvement of Aspergillus sp. (RBD01) for enhanced tranesterification of waste-

cooking and non-

BRNS (2011-2014)

2. Fungus mediated biodiesel generation from waste edible oils.

CSIR (2012-2015) List of Publications 1. Jaiswal S, Prakash R, Acharya R, Reddy AVR, Tejo Prakash N, 2012, Selenium content in seed, oil

and oil cake of Se hyperaccumulated Brassica juncea (Indian Mustard) cultivated in seleniferous

region of India. Food Chemistry, 134: 401-404.

2. Jaiswal S, Prakash R, Acharya R, Nathaniel TN, Reddy AVR, Tejo Prakash N, 2012, Bioaccessibility

of selenium from Se rich food grains of seleniferous region of Punjab, India as analyzed by

Instrumental and Chemical NAA. CyTA Journal of Food, 10: 160-164.

3. Prakash R, Aulakh SS. and Kalra R. (2010). Effect of frying time on free fatty acid generation and

esterification rate in Aspergillus sp. catalysed transesterification of cottonseed oil. Biocatalysis

and Biotransformation, 28: 403-407

4. Aulakh SS, Agarwal S, Chhibber M, Ranjana Prakash, 2010. Lipase catalyzed transesterification

of cottonseed oil. Journal of Pure and Applied Microbiology, 4: 367-372.

5. Mathur, C., Prakash, R., Ali, A., Kaur, J., Cameotra, S.S., Tejo Prakash, N., 2010. Emulsification

and hydrolysis of oil by a soil borne fungus, Syncephalastrum racemosum. Defence Science

Journal, 60: 251-254

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6. Aulakh SS, Ranjana Prakash, 2010. Optimization of medium and process parameters for

production of lipase from an oil tolerant Aspergillus sp. Journal of Basic Microbiology, 50: 37-42.

7. Prakash NT, Sharma N., Ranjana Prakash, Nathaniel N, Acharya R, Reddy AVR, 2010, Selenium

fortification and pro/anti-oxidant responses in Allium cepa (onion plant) cultivated in Se

supplemented soils. Experimental Agriculture, 46: 531-540

8. Gupta, S., Ranjana Prakash, Carolyn I. Pearce, Richard A.D. Pattrick, M. Hery, Jonathan R. Lloyd,

Prakash NT, 2010, Selenium mobilization by Pseudomonas aeruginosa (SNT-SG1) isolated from

seleniferous soils from India, Geomicrobiology Journal, 27: 35-42

9. Prakash NT, Sharma N, Ranjana Prakash, Raina KK, Fellowes J, Pearce CI, Lloyd JR, Pattrick RAD,

2009, Aerobic microbial manufacture of nanoscale selenium: Exploiting nature’s

bionanomineralization potential. Biotechnology Letters. 31: 1857-1862

10. Sharma N, Ranjana Prakash, Srivastava A, Sadana US, Acharya R., Prakash NT., Reddy AVR,

2009, Profile of selenium in soil and crops in seleniferous area of Punjab, India by neutron

activation analysis. Journal of Radioanalytical and Nuclear Chemistry 281: 59-62

11. Yadav V, Sharma N, Ranjana Prakash, Raina, KK, Bharadwaj LM, Prakash NT, 2008, Generation

of selenium containing nano-structures by soil bacterium, Pseudomonas aeruginosa.

Biotechnology, 7: 299-304

12. Yadav SS, Garg MK, Ranjana Prakash, Prakash NT, 2007, Selenium induced reduction in pyrene

toxicity in bacteria, Environ. Inform. Arch., 5: 690-693

13. Shikha Yadav, Gupta S, Ranjana Prakash, Prakash N.T, Spallholz J.E., 2007, Selenium uptake by

Allium cepa in Se-spiked soils, Am. Eur. J. Agri. Environ. Sci. 6: 81-84

14. Sharma N, Kumar A, Ranjana Prakash, Prakash NT, 2007, Selenium accumulation and Se-

induced anti-oxidant activity in Allium cepa. Environ. Inform. Arch., 5: 328-336

15. Arun Kapoor, Ranjana Prakash, Prakash NT, 2003, Predictive Biotransformation of Potential

Toxic Chemicals, Chemicke Listy, 97: 472-473

16. Srivastava AP, Ranjana Bajpai, Srivastava RB, 1998, Anti-termite treatment of rubber cups used

in underground mines, Materials und Organismen, P-1: 1-6

C. Full paper publications in Books/Proceedings

1. Srivastava A, Prakash N. T., Prakash R, 2006, Study of selenium in soils and plants using NAA,

Proc. of DAE-BRNS Meet on Current Trends and Perspectives of Neutron Activation Analysis, held

at Bhabha Atomic Res. Centre (BARC), Mumbai, India – November 2006

2. Gupta S, Kaur H, Ranjana Prakash, Ganguli A, Prakash NT, 2004, Bioreduction and crystallization

of selenium oxyanions by aerobic bacterium, Bacillus cereus. In: Biotechnological Approaches in

Sustainable Development (Eds. Reddy MSR, Khanna SK), Allied Publishers, New Delhi, Pp. 323-33

3. Ranjana Prakash, Prakash NT, 2003, Biocorrosion potential of fungi isolated from

biocontaminated fuel, Proceedings of the National Conference on Materials and Related

Technologies held at TIET, Patiala – September 2003

4. Prakash NT, Ranjana Prakash, Srivastava RB, 2002, A method for testing material susceptibility to

biocorrosion in oil-water interface. In: Biodeterioration of Materials Vol. 2 (Eds. Srivastava RB,

Mathur GN, Agarwal OP), INTACH-DMSRDE Publication, Lucknow, Pp. 42-45

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