N N Sharma CV, 2012
Dr. Nitya Nand Sharma
Ph D in Biotechnology
Division of Plant Protection
Central Potato Research Institute,
Shimla, India, PIN-171 001
[email protected], [email protected]
Mobile: +91 9418189407
Thrust area
Plant immune receptors, transgenic development against biotic and abiotic stress.
Education
Degrees
University/Department
Subjects/Topic
Ph D in Biotechnology
2010
Department of Biotechnology,
Himachal Pradesh University,
Shimla, India
Department of Biotechnology,
Himachal Pradesh University,
Shimla, India
Presidency College, University
of Calcutta, India
Hyperinduction of nitrilase in Nocardia globerula
NHB-2 and its application in biotransformation of
N-substituted aromatic nitriles.
Biotechnology
M Sc Biotechnology
2003 (First division)
B Sc (Botany Hons)
2001 (First division)
Botany (Hons), Zoology, Chemistry
Current position (March 2009 to till date)
Research Associate, under supervision of Dr S K Chakrabarti, Head, Division of Plant Protection, Central Potato
Research Institute, Shimla 171001.
Engineering late blight resistance in susceptible commercial Indian potato cultivars (ABSP II)
Major tasks completed
 Generation of transgenic potato cultivars of India with RB gene (from Solanum bulbocastanum) through
Agrobacterium mediated gene transfer.
 Expression profiling of RB gene in transgenic potato lines through qRT PCR.
 Sequencing of whole RB gene (9.3 kb)from pCLD04541.
 Cloning and sequencing of RB transcript (3kb).
Additional tasks
 Sequencing of potato BAC clones from PGSC using Roche GSFLX Titanium sequencer.
 Development of immunosorbant dip sticks for detection of PVX, PVY, and PVA.
Major achievements of Ph D
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Development of bioprocess for the production of nicotinic acid and isonicotinic acid from their
corresponding substrate (3-cyanopyridine/4-cyanopyridine) at fastest rate using nitrilase from Nocardia
globerula NHB-2.
The nitrilase from N. globerula NHB-2 has been purified and biochemical characteristics have been
studied.
The nitrilase of this organism have also been studied at molecular level through PCR amplification and
sequencing.
Academic achievements
DST International Travel Grant for attending XIIIth International Biotechnology Symposium and Exhibition,
Oct 12-17, Dalian, China.
 CSIR-UGC (JRF) - Dec 2002 and June 2005 (ranked in top 20% candidates).
 National Level Entrance Test conducted by Jawaharlal Nehru University, New Delhi for admission to DBT
Govt. of India Sponsored M. Sc. Biotechnology programme of Indian Universities.
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Scholarships and awards
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National scholarship during M Sc from Department of Biotechnology, Govt. of India
Junior Research Fellowship from Council of Scientific and Industrial Research, New Delhi during Ph D
programme (Sept 2003-Aug 2005)
 Senior Research Fellowship from Council of Scientific and Industrial Research, New Delhi during Ph D
programme (Sept 2005-Septemer 2008)
Page 1 of 5
N N Sharma CV, 2012

DST International Travel Grant for attending XIIIth International Biotechnology Symposium and Exhibition,
Oct 12-17, Dalian, China.
Publications
Research Papers
1.
2.
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4.
5.
Sharma M, Sharma NN, and Bhalla TC (2012) Biotransformation of acetamide to acetohydroxamic acid at
bench scale using acyl transferase activity of amidase of Geobacillus pallidus BTP-5x MTCC 9225. Indian
Journal of Microbiology, 52, 76-82.
Sharma NN, Sharma M and Bhalla TC (2011) An improved nitrilase mediated bioprocess for synthesis of
nicotinic acid from 3-cyanopyridine with hyperinduced Nocardia globerula NHB-2. Journal of Industrial
Microbiology and Biotechnology, 38, 1235-1243.
Raj J, S Prasad, Sharma NN and Bhalla TC (2010) Bioconversion of acrylonitrile to acrylamide using
polyacrylamide entrapped cells of Rhodococcus rhodochrous PA-34. Folia Microbiologica, 55, 442-446.
Raj J, Sharma NN, S. Prasad, and Bhalla TC (2008) Acrylamide synthesis using agar entrapped cells of
Rhodococcus rhodochrous PA-34 in a partioned fed batch reactor. Journal of Industrial Microbiology and
Biotechnology, 35, 35-40.
Sharma NN, Sharma M, Kumar H and Bhalla TC (2006) Nocardia globerula NHB-2: Bench scale production of
nicotinic acid. Process Biochemistry, 41, 2078-2081.
Reviews
1. Sharma M, Sharma NN, and Bhalla TC (2005) Hydroxynitrile lyases: At the interface of biology and
chemistry. Enzyme and Microbial Technology, 37, 279-294.
2. Sharma M, Sharma NN, and Bhalla TC (2009) Amidases: versatile enzymes in nature. Reviews in
Environmental Science and Biotechnology, 8,343-366.
Manuscripts in preparation
1. Shandil RK, Sharma NN, Sharma S, Thakur G, Tomar G, Kaushik SK, Singh BP and Chakrabarti SK (2012)
Introgression of durable late blight resistance gene (RB) in susceptible commercial Indian potato cultivars (Kufri
Jyoti and Kufri Bahar) evaluation via crossing of with RB-transgenic Katahdin (SP951) as male parentSharma
NN, Shandil RK, Sharma S, Thakur G, Tomar G, Kaushik SK, Singh BP and Chakrabarti SK (2012) Impact of
genotypic backgrounds on RB (rpi-blb1) mediated late blight resistance in potato and tomato.
2. Sharma NN, Tomar G, Shandil RK, Sharma S, Thakur G, Singh BP and Chakrabarti SK (2012) Presence of
multigene family of elongation factor alpha (EF1-α) in the genome of Indian potato cultivars similar to
Arabidopsis.
Abstracts/Posters
1.
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Sharma NN, Nayak S, Sharma M, Kaur IJ, Bhalla TC (2007) Optimization of culture parameters for apple like
fruity fragrance production by yeast isolate GF-4. In: International Conference on the Biology of Yeasts and
Filamentous Fungi, Feb 15-17, 2007 at National Chemical Laboratory, Pune INDIA, pp 99
Sharma NN, Sharma M and Bhalla TC (2008) Hyper induction of nitrilase in Nocardia globerula NHB-2:
Growth and expression kinetics in the presence of isobutyronitrile. In: XIII th International Biotechnology
Symposium and Exhibition, Oct 12-17, Dalian, China. Journal of Biotechnology, 136, pp 298-299.
Sharma M, Sharma NN and Bhalla TC (2008) A new thermophilic amidase of Bacillus sp BTP-5x isolated
from Tatapani thermal spring of Himachal Pradesh, India. In: XIIIth International Biotechnology Symposium and
Exhibition, Oct 12-17, Dalian, China. Journal of Biotechnology, 136, pp 342.
Bhalla TC, Mehta PK, Sharma NN and Bhatia S.K (2009) Production of isonicotinic acid using agar entrapped
whole cells of Nocardia globerula NHB-2. XVIIth International Conference on Bioencapsulation, Groningen,
Netherlands ; September 24-26, 2009.
Sharma S, Jeevalatha A, Sagar V, Sharma NN, Chakrabarti SK and Singh BP (2010) Detection of latent
infection of Phytophthora infestans in potatio seed tubers. In: Annual meeting of Indian Phytopathological
Society (NZ) & Symposium on Emerging Plant Diseases in North India: Status and Management Strategies,
CCS Haryana Agricultural University, Hisar (IHaryana), INDIA, October 28-29, 2010, pp 35.
Jeevalatha A., Kaundal P, Sharma NN and Chakrabarti SK (2010) Expression of recombinant coat protein of
Apical Leaf Curl Virus in Escherichia coli. In: Perspective in the Plant Health Management, Dec 14-16, 2010 at
Anand Agricultural University, Anand, Gujrat. INDIA.
Sharma NN, Shandil RK, Sharma S, Thakur G, Tomar G, Kaushik SK, Singh BP and Chakrabarti SK (2012)
Impact of genotypic backgrounds on RB (rpi-blb1) mediated late blight resistance in potato and tomato.
International Conference on Plant Biotechnology for Food Security: New Frontiers, February 21-24, 2012 at
National Agricultural Science Centre, Pusa, New Delhi, India. pp 86
Thakur G, Sharma S, Tomar G, Shandil RK, Sharma NN, Kaushik SK, Singh BP and Chakrabarti SK (2012)
Molecular mapping of late blight horizontal resistance in Solanum chacoense. International Conference on Plant
Biotechnology for Food Security: New Frontiers, February 21-24, 2012 at National Agricultural Science Centre,
Pusa, New Delhi, India. pp 87
Page 2 of 5
N N Sharma CV, 2012
9.
Tomar G, Jeevalatha A, Thakur G, Sharma NN, Shandil RK, Azmi W,, Singh BP and Chakrabarti SK (2012)
RNAi technology for resistance development against Apical Leaf Curl Virus in potato. International Conference on
Plant Biotechnology for Food Security: New Frontiers, February 21-24, 2012 at National Agricultural Science
Centre, Pusa, New Delhi, India. pp 145
Chapter in Books
1.
2.
3.
Bhalla TC, Sharma NN and Sharma M (2007) Food And Industrial Microbiology, In e-books for NSDL,
National Institute of Science Communication and Information Resources, CSIR- New Delhi.
http://nsdl.niscair.res.in/handle/123456789/129
Bhalla TC, Sharma M and Sharma NN (2007) Applied Microbiology, In e-books for NSDL, National Institute of
Science
Communication
and
Information
Resources,
CSIRNew
Delhi.
http://nsdl.niscair.res.in/handle/123456789/648
Bhalla TC, Sharma M and Sharma NN (2008) Nitrile metabolizing yeasts. In: Yeast Biotechnology: Diversity
and Applications, eds T. Satyanarayana and Kunze, Springer New York Inc. pp 696-715.
Technical expertise
Genetic analyzers
ABI 310, ABI 3500, Agilent Bioanalyzer 2100
Whole genome sequencing
Roche GSFLX Titanium Sequencer
Real time PCR
Applied Biosystems StepOne Plus and 7900HT, Roche LightCycler 480 II
Analytical instruments
UV-VIS Spectrophotometer, High Performance Liquid Chromatography, Gas
Chromatography
Microbiology
Isolation, characterization and maintenance of bacterial cultures, identification of
bacteria, screening of microbes for industrial enzymes, strain preservation
(lyophilization)
Protein purification
Akta Prime Fast Flow Liquid Chromatography, SDS & native PAGE
Biochemical engineering
Laboratory fermenter (2 and 15 L), Ultrafilteration
Recombinant DNA
technology
Transformation of E. coli, genomic DNA isolation from bacteria, plant & blood,
plasmid isolation, restriction digestion & ligation of DNA, DNA amplification by
PCR, Southern Blotting.
Enzyme technology
Assay of various enzyme, immobilization of enzymes, biotransformation of
various nitriles to amides and acids using nitrilase
Computers/Bioinformatics
Internet, Microsoft Word, Excel, Power Point, Publisher, Adobe Photoshop,
SigmaPlot, ChemSketch, FORTRAN 77, Exome, BLAST, ClustalW, ORF
searching, Rasmol
Plant Biotechnology
Maintenance of potato lines through tissue culture, potato transgenics
generation by Agrobacterium mediated gene transfer.
Workshops and training
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International Conference on Plant Biotechnology for Food Security: New Frontiers, February 21-24,
2012 at National Agricultural Science Centre, Pusa, New Delhi, India.
XIIIth International Biotechnology Symposium and Exhibition, Oct 12-17, 2008, Dalian, China.
International Conference on the Biology of Yeats and Filamentous Fungi, Feb 15-17, 2007 at National
Chemical Laboratory, Pune (INDIA)
National Roving Seminar on Patenting in Biotechnology, November 21, 2005, organized by Himachal
Pradesh University, Summer Hill, Shimla-171 005.
Biotechnological Methods in Pollution Control, August 3, 2004, All India Council for Technical Education
sponsored workshop, organized by N. C. College of Engineering, Israna, Panipat, India
Trends in Genomics and Proteomics, October 20-22, 2003, Department of Biotechnology, Govt. of India
sponsored Workshop-cum-Training Programme, organized by Bioinformatics Center, Himachal Pradesh
University, Summer Hill, Shimla-171 005
Knowledge Millennium III: The Business of Biotechnology, March 21-23, 2003, at New Delhi organized by
The Associated Chambers of Commerce and Industry of India.
Memberships/Affiliations
Lifetime member of Association of Microbiologists of India (Since 2005)
Referees
Page 3 of 5
N N Sharma CV, 2012
 Prof. T C Bhalla, Chairman, Department of Biotechnology, Himachal Pradesh University, Summer Hill,
Shimla 171005, HP (India) email: [email protected]
 Dr. S K Chakrabarti, Head, Division of Crop Protection, Central Potato Research Institute, Shimla 171001,
HP (India) email: [email protected]
 Dr. B P Singh, Director, Central Potato Research Institute, Shimla 171001, HP (India) email:
[email protected]
 Dr. Sanjeev Sharma, Senior Scientist, Division of Crop Protection, Central Potato Research Institute,
Shimla 171001, HP (India) email: [email protected]
 Prof. S S Kanwar, Department of Biotechnology, Himachal Pradesh University, Summer Hill, Shimla
171005, HP (India) email: [email protected]
 Prof. Reena Gupta, Department of Biotechnology, Himachal Pradesh University, Summer Hill, Shimla
171005, HP (India) email: [email protected]
Personal Details
Date of Birth
Sex
Category
Marital status
Nationality
Languages Known
Permanent address
May 25, 1980
Male
General
Married
Indian
English, Hindi, Bengali and German (Certificate course)
Village: Jura Korari, PO: Minwa, Tehsl: Sahajanwa, Distt: Gorakhpur, Uttar
Pradesh, PIN 273209, INDIA
Declaration
I hereby declare that all the information given above is true to the best of my knowledge.
Shimla
March 21, 2011
(Nitya Nand Sharma)
Page 4 of 5
N N Sharma CV, 2012
Abstracts of Research Work
Doctor of Philosophy in Biotechnology (2011)
Title: Hyper induction of nitrilase in Nocardia globerula NHB-2 and its application in biotransformation of Nsubstituted aromatic nitriles
Abstract:
Nocardia globerula NHB-2 harbours an inducible nitrilase specific for N-substituted aromatic nitriles (3cynaopyridine, 4-cyanopyridine). The growth of N. globerula NHB-2 and expression kinetics of nitrilase has been
studied to explore the key role of inducer (isobutyronitrile). The depletion of inducer in production medium resulted in
decrease of nitrilase activity after 18 h of incubation The exponential increase of concentration of isobutyronitrile in
the growth medium through multiple feeding at an interval of 6 h for 24 h (0.08% to 1.28% v/v isobutyronitrile in five
feeding) resulted in hyper induction (seven fold increase) of nitrilase in N. globerula NHB-2 (4.93 U/mg dcw) after
30 h of induction. The presence of isobutyronitrile has inhibitory effect on growth and glucose utilization of N.
globerula NHB-2.
The nitrilase of N. globerula NHB-2 had higher specific activity towards aromatic heterocyclic nitriles
(benzonitrile>3-cyanopyridine>4-cyanopyridine>m-tolunitrile>p-tolunitrile). The substrate specificity pattern
suggests that π-electrons of >C=C< in acrylonitrile and aromatic nitriles might play some role in the positioning of –
CN group for interaction with the cystine residue of the catalytic site. The reaction pH variation showed that nitrilase
in the free cells of N. globerula NHB-2 was active from 4.0-10.5 and maximum activity was recorded in 0.1 M citrate
buffer (pH 6.0). The nitrilase of N. globerula NHB-2 showed highest activity at 55°C and above this a sharp decline
in activity was observed with a complete loss at 80°C. The nitrilase of this organism was highly labile at higher
temperature and 100% activity was lost in 6 h at 55°C. The nitrilase of this organism was stable at 35°C.
The nitrilase of N. globerula has been purified having molecular weight of 40 kD (monomer) and exists as
dimmer (70 kD) in native form. In presence of substrate (3-cyanopyridine, benzonitrile) formation of multimeric
aggregate (>220kD, 6-8 mer) have been confirmed by native-PAGE analysis. Further, the gene encoding nitrilase
has been amplified through PCR techniques. The amino acid sequence derived from the nucleotide sequence of the
forward primer shows homology with nitrilases of R.rhodochrous tg1-A6, R.rhodochrous J1 and R.rhodochrous PA34. Similar to the nitrilases of the above microorganisms, nitrilase of N. globerula efficiently hydrolyses benzonitrile.
The substitution of leucine (125) with glycine near the catalytic triad of the nitrilase might be responsible for higher 3cyanopyridine/4-cyanopyridine hydrolysing activity in comparison to R. rhodochrous J1 nitrilase. Though at position
142 tyrosine was present in the nitrilase of N. globerula NHB-2 its affinity towards aliphatic nitriles was low. The
presence of glycine at 125 position might be the possible reason for such behaviour of the enzyme and at the same
time could have improved the affinity for 3-cyanopyridine and 4-cyanopyridine.
In a one liter reaction performed at 35°C containing hyper induced resting cells of N. globerula NHB-2
equivalent to 10 U/ml nitrilase activity in one liter reaction mixture a total of one 1 mole of nicotinic and 0.7 mole of
isonicotinic acid has been synthesized from their respective substrates in 200 min and 140 min, respectively. This is
the highest ever conversion rate (37 g acid/h) achieved for the synthesis of nicotinic acid and isonicotinic acid
through biotransformation route.
Project work for partial fulfillment of M Sc Biotechnology


Bioconversion of 3-cyanopyridine to nicotinic acid using resting cells of Rhodococcus rhodochrous
NHB-2 (Major research project: July 2002-June 2003)
Abstract: The intracellular nitrilase of Nocardia globerula NHB-2 induced by propionitrile catalyses the
hydrolysis of 3-cyanopyridine to nicotinic acid without detectable formation of nicotinamide. N. globerula NHB2 showed highest conversion rate at 40 mM 3-cyanopyridine, in 0.1 M NaH2PO4/Na2HPO4 buffer (pH 7.5)
containing 5 mM 2-mercaptoethanol. In 1 l fed-batch reaction, containing 0.1 M NaH2PO4/Na2HPO4 buffer (pH
7.5), 4.2 g resting cells (dry weight), at 35°C, 400 rpm, a total of 1 mol of 3-cyanopyridine added at feed rate
of 0.12 mol h-1 was converted to nicotinic acid (98.6%) in 9 h. The rate of nicotinic acid production was 26
mmol h-1g-1 DW.
Molecular genetic analysis of Rajputs of Himachal Pradesh using microsatellite Markers (Short
research training: Jan-Feb, 2002)
Abstract: DNA isolation from the blood samples collected from 97 individuals belonging to Rajput community
inhabiting in Shimla district of Himachal Pradesh (INDIA) was performed and six STR loci D31S1358(3p),
D16S539(16q24-qter), THO1(11p11.5), TPOX(2p23-2pter, CSF1PO(5q33.3-34), D7S820(7q11.21-22) were
amplified through multiplex PCR using AmpFlSTR CofilerTM loci kit. The genetic analysis of the amplified
products was performed using ABI 377 sequencer. The sampled population was in Hardy-Weinberg
equilibrium (HWE) for all the six analysed markers in the Cofiler system, with p>0.05, for the Exact, Chisquare tests. The observed heterozygosity varied from 0.587 in TPOX to 0.804 for the D31S1358 loci. The
power of discrimination varied from 0.696 for TPOX to 0.801 for D7S820. The neighbour joining tree prepared
between Rajputs of Himachal Pradesh and other Kshtriya population of Indian Subcontinent (Rajputs of Bihar,
Thakurs of Uttar Pradesh and Khandayats of Orissa) showed clear proximity between Rajputs of Himachal
Pradesh and Bihar.
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