PART I: GENERAL INFORMATION Name of the Institute...

PART I: GENERAL INFORMATION

1. Name of the Institute/University/Organization Submitting the Project

Proposal:

Department of Microbiology

University of Pune

Ganeshkhind,

Pune: 411007

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

4. Name and Designation of the Executive Authority of the University

Forwarding the Application: Dr M. L. Jadhav

Registrar, University of Pune

5. Project Title: “Development of an Efficient Microbial System for the Production of

Polyhydroxyalkanoate”

6. Category of the Project: R&D/ Programme Support

7. Specific Area: 7.1- Microbiology/Strain Improvement

7.2- Bio Process Optimization and Scale Up

8. Duration: Three Years

9. Total Cost (Rs.): 32,35,000/-

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

11. If the Project is Multi-institutional, please furnish the following: Not

Applicable

Prof. R.L.Deopurkar, Department of Microbiology, University of Pune

12. Scope of Application Indicating Anticipated Product and Processes:

The project aims at developing an efficient microbial system for

production of polyhydroxybutyrate (PHB), a well recognized candidate for

biodegradable plastic. Extensive use of polypropylene based plastic (PP plastic)

all over the world poses a serious environmental hazard. Biodegradable,

biocompatible and ecofriendly substitutes to PP plastic can alleviate

environmental problems arising out of PP plastics. However, the present

production cost of PHB is prohibitory. Newer microorganisms capable of

producing PHB using cheap indigenous raw materials, mutations to enhance

PHB productivity of culture, and careful investigation of environmental factors

affecting growth and production of PHB would make PHB production

economically competitive. Understanding the physiology, biosynthesis and

mechanism of accumulation of polyhydroxybutyrate is extremely important in

developing microbial system and the improved process of producing PHB.

Production of PHB and a mixed copolymer using cheap agricultural

waste/residues available in our country, using high yielding strains is of great

importance in promoting the usage of PHA based plastics.

The work would result into development of indigenous microbial

cultures giving greater yields (70% wt/wt) of PHB and/ its copolymer using

agricultural residues available in our country.


13. Project Summary

The current global plastic industry turnover is about $1 trillion/year and

represents 3.7% of the world Gross Domestic Product (GDP) [4]. The extensive

use of polypropylene based plastic (PP plastic) all over the world is of serious

concern because

PP plastics are inherently resistant to biodegradation.

These non biodegradable plastics accumulate in the environment at a rate of

millions of tons per year [4].

Chemical degradation of PP plastics often generates toxic substances [10].

Production of polypropylene plastic is closely linked with petrochemical

resources which are fast depleting. Production of one ton of plastic needs as

large as one ton of fossil fuel and the present estimated amount of PP plastic in

the world is 150 million tons [26].

Polyhydroxyalkanoates in general and PHB-co-PHV in particular

are well accepted substitutes to PP plastics. Because of the biological origin,

PHA based plastics are biodegradable and consequently nonpolluting and

ecofriendly[7]. Since polyhydroxyalkanoates in general are immunologically

inert and slowly degraded in human tissue, they have immense applications as

biocompatible medical/surgical implants [30].

The estimated cost of synthesis and subsequent extraction of PHA

from bacteria is about 3-4 US$/Kg, which is 5-10 times higher than that of

polypropylene [26]. The enormous cost of production prohibits the widespread

use of PHB.

Systematic studies of PHB production at low cost indeed are

necessary to make PHB production economical.

The production cost of PHB can substantially be reduced by

a) Screening and isolation of microorganisms producing greater amount of PHB.

b) Optimizing the conditions to improve the yields of PHB.

c) Use of cheap raw materials for the growth and production of PHB

d) Improving the potential of culture.


PART II: PARTICULARS OF INVESTIGATORS

Principal Investigator:

14. Name: Dr. R. L. Deopurkar

Date of Birth: August 15, 1953 Sex (M/F): M

Designation: Former Head and Professor.

Department: Department of Microbiology.

Institute/University: University of Pune.

Address: Department of Microbiology, University of Pune.

Ganeshkhind, Pune. PIN: 411007.

Telephone: 020-25690643 E-mail: [email protected]

Number of research projects being handled at present: Three

15. Co-Investigator: Nil


PART III: TECHNICAL DETAILS OF PROJECT

16. Introduction

16.1 Origin of the Proposal

The massive usage of PP plastics poses serious threats to the environment. The

accumulation of conventional polypropylene based plastic goods widely used in

day-to-day life, has become a major environmental concern. They are inherently

resistant to biodegradation and upon chemical degradation lead to production of

toxic substances. Also only a few and not all the plastics are recyclable and

reusable.

Moreover, production of conventional plastic is closely linked with

petrochemical resources which are fast depleting. The world currently consumes

about 140 million tons of plastic per annum processing of which uses 150 million

tons of fossil fuel [26].

Polyhydroxyalkanoates (PHA) in general and Polyhydroxybutyrate

(PHB) in particular have physical properties very close to those of PP plastics

[Appendix 1]. The copolymers of PHB and PHV are best suited as biodegradable

plastics, because polymers of PHB alone are little harder and brittle. PHB based

plastics are nontoxic, biodegradable and biocompatible [27]. PHBs are water

insoluble and they also sink in water, and hence can be easily disposed of by

putting them in sea beds where they get anaerobically degraded.

The major hurdle in widespread usage of PHB-PHV based plastics is

the higher cost of production of PHB-PHV; the production of pp is 5 – 10 times

cheaper.

16.2 Rationale of the Study Supported by Cited Literature

Enough literature has been published on PHB as future biodegradable plastics [3,

26, 27 and 30]. Despite the studies on genetics, biochemistry and physiology of

synthesis and accumulation of PHB by microorganisms, the use PHB as plastic

source has still remained impracticable largely due to high cost of production.[2,

5, 25 and 28].

The search for newer microbial strains specially suited to utilize

inexpensive carbon sources, improvement in strains, use of cheap alternative


substrates for growth of microorganisms to produce PHB will make PHB

production economically competitive and commercially more attractive.

16.3 Hypothesis

Development of microbial strains giving higher yields of PHA, use of less

expensive substrates for microbial growth and production of PHAs and well

optimized production process would make it possible to have intracellular

accumulation as high as 60 to 70% of the biomass, leading to economically

cheaper production of PHAs

16.4 Key Questions

The production cost of PHB plastics should be comparable to that of PP plastics.

Currently the PP plastics are 5-10 times cheaper than PHB plastics.

Studies on production of PHB should address the following questions.

• Can we substantially decrease the cost of production of PHB by using better

strains?

• Can we use cheap alternative raw materials like molasses, whey, paper pulp

sludge and sugarcane liquor as cheap carbon substrate for the production of PHB?

• Can we increase the PHB production and specific accumulation per unit biomass by

mutation in cultures?

• Can we get good copolymer of PHB and PHV?

16.5 Current Status of Research and Development in the Subject

(Both International and National Status)

International Status:

It is also generally accepted fact that it is still costly and inefficient to use PHB

as plastic source [5, 7]. Biopol, commercially available biodegradable

thermoplastic, was produced by fed batch fermentation of Ralstonia eutropha

using glucose and propionate as substrate. A major factor in the prohibitively

high price of Biopol was that the propionate is considerably expensive than

glucose [2].

Genetics, biochemistry and physiology of synthesis and accumulation of

PHB by microorganisms have been studied in great details. The increased


production of PHB by Azospirillum brasilense was shown to be paralleled with

increased specific activities of hydroxybutyrate dehydrogenase, β ketothiolase,

and thiophorase [28].

The genes for PHB production have been well studied in few

organisms like Alcaligenes eutrophus and Azospirillum brasilense. Recombinant

E.coli has been constructed by transferring genes of PHB production from

Alcaligenes eutrophus to E.coli [25].

Choi et al has documented production of P(3HB-co-3HV) by

recombinant E.coli and the production was 10 times higher than that of natural

PHB producer [6].

National Status:

In our country too, the importance of PHB as potential candidate for

biodegradable plastic has been well recognized by many of laboratories. A few

laboratories have focused their attention on exploring biodiversity with the aim to

get better PHB producers.

There are reports on production of PHB (46-65% dry wt of cell mass)

using jack fruit seed powder (Ramadas et al from Kerala) [22, 23], pea shell

(Kumar at al) [13], cheese whey (Nath et al from M.S. Baroda) [19] and phenols

(Nair et al from Kerala) [18] as raw material. Aarthi et al isolated Bacillus

mycoides and Bacillus cereus from garden soil and showed the intracellular

accumulation of PHB to the extent of 12.18% to 57.2% by dry wt [1]. Also leather

industry effluent was the source of Rhodobacter capsulatus and was capable of

producing PHB [16]. Halomonas campisalis isolated from Lonar lake showed

production of PHB-co-PHV in the range of 45-81% [12].

Since PHB lacks sufficient flexibility for the effective use as plastic,

Otari et al from Shivaji University, Kolhapur studied the production of copolymer

hydroxybutyrate and hydroxyvalerate [21].

The studies by Kumaravel et al (from Tamilnadu) on degradation of

polyhydroxybutyrate and polyhydroxyvalerate by fungal isolates are important in

understanding of “non persistent” nature of PHA based plastics [14].

Sardesai et al (from University of Delhi) studied the relationship between

accumulation of PHB and cold tolerance in Rhizobium [24].


16.6 The Relevance and Expected Outcome of the Proposed Study

Relevance to the present day problems and needs of the society and the

country:

The threat of accumulation of polypropylene plastic has led to the intensive search

of 'bioplastic' made up of biopolymer of PHB/PHV. Currently, the main limitation

for bulk production of PHB/PHV is its high production and recovery cost [14].

Production of PHB and a mixed copolymer using cheap agricultural

waste/residues available in our country, using high yielding strains is of great

importance in promoting the usage of PHA based plastics.

The work would result into development of indigenous microbial

cultures giving greater yields (70% wt/wt) of PHB and/ its copolymer using

agricultural residues available in our country.

16.7 Preliminary work done so far

After extensive screening of PHB producing bacteria using E2 mineral

medium followed by staining with Nile blue stain [11], thirty cultures have

been already isolated and characterized.

Ten of these cultures show significant accumulation of PHB as seen under

light microscope after sudan black staining.

Quantitation of intracellular PHB in terms of dry wt basis for these cultures is

under progress.

The PI has done exploratory interactions with Vasantdada Sugar Institute and

Katraj dairy to supply sugar cane liquor and whey.

17. Specific objectives

Though PHAs are acceptable biodegradable, ecofriendly substitute to pp plastics,

enormous production cost of it at present prohibits its widespread use as plastic.


This project therefore specifically aims at economizing the production of PHB:

search for high yielding microorganisms, use of cheap raw materials, strain

improvement through mutagenesis are important steps in achieving the objective.

Specific objectives of the project are as follows:

1) To identify cultures:

Methods to be used-

• Biochemical tests (Phenotype Diagnostic Tests)

• 16S rDNA sequencing of the isolates

• BLAST (Basic Local Alignment Search Tool) analysis

Verifiable indicators – Taxonomically identified better PHA producers at hand.

2) To optimize parameters for PHA production:

• Shake flake studies to optimize PHA production with respect to pH, temperature,

salts, and nitrogen as well as phosphorus source

• Experimental design and analysis of data – by Response surface methodology

(23)

Verifiable indicators – Standardized/optimized process for best production of PHA

(70% wt/wt).

3) To produce PHA using cheap carbon substrates such as molasses, whey, sugar

cane liquor, industrial waste, and paper pulp sludge:

• PHA production using cheap carbon substrate in laboratory fermenter (5-10L).

Verifiable indicators – Data on production of PHA using molasses, whey and

sugar cane liquor at hand.

4) To improve strain through mutations:

• Mutagens to be used- UV, acriflavin and 5 bromouracil.

• Screening methods – a) 3-HB dehydrogenase deficient mutants [20]

b) Staining with nile blue [9]

Verifiable indicators – Mutants accumulating PHB as high as 70% of biomass.

5) To characterize PHA produced by the selected culture:

PHA produced will be characterized (12) using techniques such as


• Gas chromatography to determine constituent monomers present in the

polymer.

• Gel permeation chromatography to determine molecular weight spectrum of

the PHA produced.

• Differential scanning calorimetry to determine melting temperature and degree

of crystallinity of the polymer.

Verifiable indicators – Well characterized PHA especially with respect to

copolymer (PHB-co-PHV) composition.

18. Work Plan:

18.1 Work Plan (Methodology/Experimental design)

18.1.1 Identification of PHB producing microorganisms isolated from diverse

environments and study of the type of PHAs (such as PHB

(polyhydroxybutyrate), PHV (polyhydroxyvalerate) and PHB-co-HV (copolymer

of PHB and PHV) produced –

The cultures will be identified using nucleic acid based methods viz 16S rDNA

sequencing and blast analysis [5]. Pure isolates will be identified by sequencing

partial sequences of their 16S rDNA. The partial 16S rDNA will be amplified

using universal primers. Purified DNA from the PCR will be then sequenced by

using DNA sequencer. The sequences of partial 16S rDNA will be compared

with the 16S rDNA sequences available in the public nucleotide databases

(NCBI) by using their World Wide Web site and the BLAST algorithm [5].

The PHA isolation and purification method involves digestion of the

cell pellet using sodium hypochlorite, followed by washing of residue with

distilled water, acetone and finally with diethyl ether. The residue is dried and

polymer is extracted into boiling chloroform. After filtration polymer is

reprecipitated by addition of chloroform solution and cooling to -200C [15, 31].

For spectrophotometric assay chloroform is evaporated and polymer

is dissolved in concentrated H2SO4 and is heated for 10 mins at 1000C in water

bath. After cooling, absorbance at 235nm is measured [15].

Identification of types of PHAs would be done by GC MS

chromatography [12].


18.1.2 Shake flask studies – Using shake flask cultures, various parameters (pH,

temperature, salts, nitrogen as well as phosphate limitation) will be determined for

maximal production of PHB [8]. PHB production using cheap carbon substrates

(molasses, whey, sugar cane liquor, industrial waste, and paper pulp sludge) will be

investigated.

18.1.3 Fermentation studies – Two-stage fed-batch process (rich medium for growth

and nitrogen limiting medium for PHB production) will be investigated for

fermentative production of PHB [17, 22, and 29]. The optimized conditions (pH,

temperature, and nitrogen and phosphorus limitation) of PHB production as revealed

by shake flask studies will be used in fermentation studies. Cheap carbon substrates

such as molasses, whey, sugar cane liquor, industrial waste, wafer residue, rice chaff

and paper pulp sludge will be used.

18.1.4 Mutation studies - Physical as well as chemical mutagens such as UV,

acriflavin and 5 bromouracil will be used to produce mutants [9, 20]. Two strategies

will be used for screening of mutants:

Selection of mutants:

a) 3 HB dehydrogenase deficient mutants - These mutants are known to accumulate

more PHB. They grow on glucose containing minimal medium but not on 3-HB

containing minimal medium [20].

b) Cells producing more of PHB give rise to colonies that are intensely fluorescent

after staining with nile blue [9].

18.1.5 Characterization of PHA produced by selected culture –

1] Gas chromatographic (GC) analysis: GC analysis of methyl esters of PHA, PHB,

and PHB-co-PHV will be performed by methanolysis.

Experimental conditions will be: oven temperature – 900C, injector temperature –

1700C and detector temperature – 1900C, extra pure grade nitrogen gas as a carrier gas

with flow rate 35.5 ml/min. Standard methyl esters of 3-hydroxybutyric acid and stan-

dard methyl esters of 3- hydroxyvaleric acid will be used as standard [12].

2] Gel Permeation Chromatography: Molecular weight / molecular weight distribu-

tion of the PHA formed will be determined by Gel Permeation Chromatography.

Chloroform will be used as mobile phase and polystyrene as a molecular weight

standard. Molecular weight analysis of standard PHB and standard PHB-co-PHV

will be used for comparison [12].


3] Differential Scanning Calorimetric analysis (DSC): DSC of purified PHA sam-

ple standard PHB and standard PHB-co-PHV will be carried out. Approximately

5 mg of purified PHA sample will be encapsulated in aluminum pans and heated

in a temperature range from 50 to 2000C at the rate of 100C/min using Differen-

tial Scanning Calorimeter. Melting temperature will be recorded at the peak of

melting endotherm. Enthalpy of fusion will be determined from the graph and

percentage crystallinity will be calculated. The melting temperature of purified

PHA will be compared with standard PHB and standard PHB-co-PHV (9:1) [12].

18.2 Connectivity of the participating institutions and investigators: Not applicable

18. 3 Alternate strategies: Not applicable

19. Timelines:


Achievable targets Period of

studyIdentification of isolated bacteria using microbiological techniques and

molecular biology techniques, such as 16S rDNA gene sequencing

and Selection of bacterial isolates producing high amount of PHB

and/ its copolymer.

6 Months

Identification of types of PHAs using GC MS chromatography. 3 Months

Optimization of the parameters for high PHB producing strain (such as

temp., pH, inexpensive substrates) in shake flask experiments using

standard strain as well as best new isolate.

6 Months

Fermentation studies to optimize parameters for high PHB production

using best new isolate/mutant strain.

6 Months

Development of mutant strain 6 Months

Characterization of PHA produced by the selected culture using Gas

chromatography, Gel permeation chromatography and NMR

spectroscopy.

6 Months

Documentation, compilation of data, publication and patenting. 3 Months


Identification and selection of best isolate

Identification of types of PHAs Shake flask

studiesFermentation studies

First Year Third Year Second Year

Development of mutant strain

Characterization of PHA produced by selected isolate

Documentation

20. Name and address of five experts in the field


Sr. No. Name Designation Address1 Prof. Anjana Desai Professor Maharaja Sayajirao University of

Baroda

Baroda – 390 002

Gujarat, India.

Mob. No. - 098792844492 Dr. P.P. Kanekar Scientist Agharkar Research Institute

Gopal Ganesh Agarkar Road,

Pune – 411 004, India

Ph. No. - 020 – 256536803 Prof. S.N. Bhosale Professor Goa University

Taleigao Plateau,

Goa – 403206, India

Mob. No. - 097639299114 Dr. Anuradha Nerulkar Associate

Professor

Maharaja Sayajirao University of

Baroda

Baroda – 390 002

Gujarat, India.

Mob. No. - 094263843655 Prof. S. Mohan Professor Swami Ramanand Teerth

Marathwada University

Vishnupuri, Nanded – 431 606,

Maharashtra, India.

Mob. No. - 09421839512

PART IV: BUDGET PARTICULARS

Budget (In Rupees)

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

Sr.

No.

Item I Year

(Rs)

II Year

(Rs)

IIIYear

(Rs)

Total

(Rs)

1

2

Equipments

Columns for Gas Chromatography

CP Wax 52 CB (25m X 0.32mm)

CP Wax 58 FFAP (50m X 0.32mm)

Fermenter

1,00,000

15,00,000

1,00,000

15,00,000

Sub-Total (A) = Rs.16, 00,000

B. Recurring

B.1 Manpower

Sr.

No.

Position

No.

I Year

(Rs)

II Year

(Rs)

III Year

(Rs)

Total

(Rs)

1 JRF (Junior Research

Fellow for two years and

SRF for one year)

1,92,000 1,92,000 2,16,000 6,00,000

Sub-Total (B.1) = Rs. 6,00,000


B.2 Consumables

Sr.

No.

Item Year 1

(Rs)

Year 2

(Rs)

Year 3

(Rs)

Total

(Rs)1 Bacteriological media 60,000 50,000 40,000 1,50,000

Basic inorganic chemicals60,000 50,000 40,000 1,50,000

2Fine chemicals (DNA isolation and PCR chemicals, SDS PAGE chemicals, Standard PHB,Nile Blue sulphate, GC-MS analysis , Acriflavin, Ethyl methane sulphonate)

1,50,000 1,00,000 50,000 3,00,000

6Glassware

70,000 70,000 40,000 1,80,000

Sub-Total (B.2) = Rs. 7,80,000

Other items Consolidated

Emolument

Year 1

(Rs)

Year 2

(Rs)

Year 3

(Rs)

Total

(Rs)

B.3 Travel 10,000 10,000 10,000 30,000

B.4 Contingency 50,000 50,000 50,000 1,50,000

B.5 Overhead 25,000 25,000 25,000 75,000

Sub-total of B

(B.1+B.2+B.3+B.4+B.5)

16,35,000

Grand Total (A + B) 32,35,000


Financial Year: April - March

PART V: EXISTING FACILITIES

Resources and additional information

1. Laboratory:

Present building has about 250 sq. meter laboratory space available.

Construction of new departmental laboratories (Approx. 2180 sq.m) is nearly

completed and laboratory will be ready to use latest by January 2012. It houses

four research laboratories, three temperature control rooms and one cold room.

a. Manpower

Lab assistants, Store keeper

PI has eight Ph.D. students, one UGC Post Doctoral Fellow (D.S. Kothari Post

Doctoral Fellowship).

b. Equipments

Equipment and accessories available within the Investigator’s group/Dept. which can be utilized for the project.

Sr. No.

Name of Equipment/

Accessories

Make (No.) Model Year of Procurement

1. Autoclave Fabwell Engineering, Mumbai

Horizontal 2002

2. Incubator Remi Instruments, Mumbai (2)

- 1987

3. Cooling Incubator Remi, Mumbai C1-125 1995

4. Deep freezer Angeltonii, Italy Polar 110,H 340,V

2004, 2007

5. Freeze Drier Martin Christ - 2010

6. Gel Documentation System

Alfa Innotech Corporation California, USA.

Alpha Imager

2200

2001

7. High speed Centrifuge

Remi Instruments, Mumbai

Kubota (2)

Plastocraft

C24

-

Rota4R

2002

2007, 2010

2009

8. Laminar Air Flow System

Microfilt India, Pune (2)



-

-

-

1984

1995

2006

9. PCR Gradient Eppendorf 2007

10. Phase Contrast Nikon-Japan - 1985


Sr. No.

Name of Equipment/

Accessories

Make (No.) Model Year of Procurement

Microscope

11. Rotary Incubator Shaker

Steelmet Industries, Pune.

Jeobiotech

-

-

2004

2007

12. Rotary Shaker Steelmet Industries, Pune Two tier 2010

13. Tensiometer Angeltoni Dataphysics, 2004

14. UV spectrophotometer

Schimadzu Corporation, Japan.

UV-1601 1995

15. Gas Chromatograph 2014 ATF with FID and TCD

Schimadzu Corporation, Japan

2014 ATF 2011

16. Lyophilizer Martin Christ CB18-40 2010

17. Ice flaker Manitowoc RF0266A 2010

. Other Infrastructural facilities:

Sr. No.

Item Name Yes/No/NR*

Sr. No.

Item Name Yes/No/NR*

1. Workshop Yes 7. Telecommunication Yes2. Water & Electricity Yes 8. Transportation Yes3. Standby power supply Yes 9. Administrative l support Yes4. Laboratory space & furniture Yes 10. Library facilities Yes5. AC room for equipment Yes 11. Computational facilities Yes6. Refrigerators Yes 12. Animal/Glass house No

NR*: Not Required

2. Other resources such as clinical material, animal house facility, glass house.

Experimental garden, pilot plant facility etc. Not Applicable


PART VI: DECLARATION/CERTIFICATION

It is certified that

a) the research work proposed in the scheme/project does not in any way duplicate

the work already done or being carried out elsewhere on the subject.

b) the same project proposal has not been submitted to any other agency for financial

support.

c) the emoluments for the manpower proposed are those admissible to persons of

corresponding status employed in the institute/university or as per the Ministry of

Science & Technology guidelines (Annexure-III)

d) necessary provision for the scheme/project will be made in the

Institute/University/State budget in anticipation of the sanction of the

scheme/project.

e) if the project involves the utilization of genetically engineered organisms, we

agree to submit an application through our Institutional Biosafety Committee. We

also declare that while conducting experiments, the Biosafety Guidelines of the

Department of Biotechnology would be followed in toto.

f) if the project involves field trials/experiments/exchange of specimens, etc. we will

ensure that ethical clearances would be taken from concerned ethical

Committees/Competent authorities and the same would be conveyed to the

Department of Biotechnology before implementing the project.

g) it is agreed that any research outcome or intellectual property right(s) on the

invention(s) arising out of the project shall be taken in accordance with the

instructions issued with the approval of the Ministry of Finance, Department of

Expenditure, as contained in Annexure-V.

h) we agree to accept the terms and conditions as enclosed in Annexure-IV. The

same is signed and enclosed.


i) the institute/university agrees that the equipment, other basic facilities and such

other administrative facilities as per terms and conditions of the grant will be

extended to investigator(s) throughout the duration of the project.

j) the Institute assumes to undertake the financial and other management

responsibilities of the project.

Signature of Principal Investigator: Signature of Executive Authority

of Institute/University with seal

Date: Date:


PART VII: PROFORMA FOR BIOGRAPHICAL SKETCH OF

INVESTIGATORS

Name: Prof. R.L.Deopurkar

Designation: Professor

Department/Institute/University: Department of Microbiology, University of Pune

Date of Birth: August 15, 1953 Sex (M/F) M SC/ST: Not Applicable

Education (Post-Graduation onwards & Professional Career)

Sr. No. Institution

Place

Degree

Awarded

Year Field of Study

1 Indian Institute of

Science, Bangalore

Ph.D 1990 Protoplasts of fungi

2 M.S. University of

Baroda

M.Sc 1976 Microbiology

A. Position and Honors

Position and Employment

Sr. No.Institution

PlacePosition From To

1Department of Microbiology,

University of PuneProfessor 2006 Contd.


University of PuneReader 1992 2006


University of PuneLecturer 1979 1992

4

BAIF-BRIAH, Wagholi

Foot and Mouth Viral vaccine

manufactures

Quality Control

Officer1977 1979

5GITS Food production, Ltd.

Hadapsar

Laboratory

Incharge1976 1977


Honors/Awards

1) Functioned as the ‘Joint Organizing Secretary’ of the 50th Annual Conference

of the ‘Association of Microbiologists of India’ [15-18 December` 2009]

2) Elected as the vice-president of the ‘Association of Microbiologists of India’

[Pune Unit] on 14 June, 2010.

3) Felicitated by Haffkine Biopharmaceutical, Corporation Ltd. On the occasion

of 35th foundation day[1 Sep, 2009]

4) Secretary, Marathi Vidnyan Mahasangh, Pune, for last 10 years.

5) Nominated as the member of the experts in the preparation of ‘Krishi Dnyan

Kosh’, ‘Khand 4&5’ [published] and for ‘Khand 6’ [in preparation], published

by ‘Bhartiya Sanskritikosh Mandal

6) Invited as an author for the 10th standard biology book [published in 2007] of

the Maharashtra State Board of Higher Secondary Education.

7) Member of the Board of studies in Microbiology, Goa University.

8) Member of the Board of studies in Para-Clinical subjects, Pravara Institute of

Medical Sciences, Loni

9) Best teacher award, from Uttar Bhartiya Sangh, Pune

10) Nominated as the editor, AMI, Pune Unit Newsletter, for Sep 2007 - 2010

Professional Experience and Training relevant to the Project

B. Publications (Numbers only)

Books: 02 Research Papers, Reports: 17 General articles:

Patents: Others (Please specify):

Selected peer-reviewed publications

1. Ahire K.C., G.J. Kulkarni, Y. S. Shouche, B.P. Kapadnis, and R.L. Deop-

urkar (2011) Biodegradation of tributyl phosphate by novel bacteria isolated

from enrichment culture technique. Biodegrdation. (Accepted and In Press,

DOI 10.1007/s 10532-011-9496-7).

2. Salvi, N., A. Waghmare, R.L. Deopurkar, M. Khadilkar, M. Kalolikar,

and S.K. Gade (2010) Validation of indirect ELISA for quantitative testing of

rabies antibodies during production of anti-rabies serum using equines. Pro-

ceedia in Vaccinology 2: 3-11.


3. Waghmare, A., R. L. Deopurkar., N. Salvi, M. Khadilkar, M. Kalolikar,

and S.K. Gade (2009) Comparison of montanide adjuvants, IMS 3012

(Nanoparticle), ISA 206 and ISA 35 (Emulsion based) along with incomplete

freund’s adjuvant for hyperimmunization of equines used for production of

polyvalent snake antivenom. Vaccine 27: 1067-1072.

4. Atre, A. N., S. V. Surve, Y. S. Shouche, J. Joseph, M. S. Patole, and R. L.

Deopurkar (2009) Association of small Rho GTPases and actin ring

formation in epithelial cells during the invasion by Candida albicans. FEMS

Immunology & Medical Microbiology 55(1): 74-84.

5. Kalantar, E. and R. L. Deopurkar (2007). Application of factorial design

for the optimized production of antistaphylococcal metabolite by Aureobasidi-

um pullulans. Jundishapur J. Natural Pharma. Products 2 (2): 69-77.

6. Kalantar, E., R. L. Deopurkar, and B. P. Kapadnis (2005). Antistaphylo-

coccal metabolites from A. pullulans: Production and Characterization.

African Journal of Clinical and Experimental Microbiology 6: 177-182.

7. Kalantar, N. E., R. L. Deopurkar. and B.P. Kapadnis (2005) Antimicrobial

activity of indigenous strains of Aureobasidium isolated from Santalum album

leaves. Iranian J. Pharma. Res. 1: 59-64.

8. Deshpande, M.S., R. L. Deopurkar, and V.B. Rale (1989). Beta – galactosi-

dase from Aureobasidium pullulans. Letters in Applied Microbiology 9: 21-

24.

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

- Nil.


C. Research Support

Ongoing Research Projects:-

Sr.

No.Title of the Project

Funding

Agency

Amount

(Rs.)

Date of sanction

and Duration

1 Biochemical investigation of

thermophilic aerobic digestion:

Pretreatment process for

anaerobic digestion

Dept. of Atomic

Energy, Govt.

of India, BRNS

24,85,000 15/02/2011

Three years

(2011 – 2014)

2 Molecular mechanism of

Quorum sensing (Q.S.) and

viable but non culturable state

in food borne pathogens

BARC UoP joint

research project

- Three years

(2010 – 2013)

3 Study of Uricase and its

PEGylated form in treatment of

hyperuricemia and associated

diseases

UGC - 26/08/2011

Three years

(2011-2014)

Completed Research projects:

Sr.

No.Title of the Project

Funding

Agency

Amount

(Rs.)

Date of

completion

1 Kinetics and catalytic

properties of amylase from

Aureobasidium pullulans

BCUD,

University of

Pune

2,00,000 2006

2 Purification and

characterization of

beta-galactosidase from A.

Pullulans

University of Pune,

under Quality

improvement

Programme

3,00,000 2008


3 ‘Restoration of soil and water

polluted with pesticide

residues using indigenous

microorganisms

Dept. of Atomic

Energy, Govt.

of India, BRNS

14,19,200 2007

4 Studies on the production of

biofuel/bioethanol using

alcohol tolerant Yeast

BCUD,

University of

Pune

85,000 2009

Appendix 1:

Table 1: Properties of PHAs and polypropylene (PP) (Verlinden et al. 2007)

Different Types of PHAs

Sr.

NoParameter PHB PHBV PHB4B PHBHx PP

1Melting Point

(0C)177 145 150 127 176

2Glass transition

temp (0C)2 -1 -7 -1 -10

3 Crystallinity (%) 60 56 45 34 50-70

4Tensile strength

(Mpa)43 20 26 21 38

5Extension to

break (%)5 50 444 400 400

Appendix 2:

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Place: Signature of Investigator

Date:


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