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Hire Dr. Amrit K.

Saint Martin (French Part)

Profile Summary

• Current Research Interest at Genozymes Network (http://www.fungalgenomics.ca/wiki/Main_Page) Various microbial enzymes participating in lignocellulose decomposition has major role as renewable energy resource and bio-based products. To better understand the perplexing diversity of these enzymes and their roles in degradation of complex polysacchride substrates, detailed characterization of their molecular function and specificity is needed. The Genozymes project has used genomic and ancillary approaches to identify, analyze and develop potential fungal enzymes as catalysts for bio-processing. In this regard, thousand fold of enzymes with potential lignocellulolytic activity were identified and their functional domains were cloned and expressed in E. coli and Aspergillus niger. These proteins were functionally characterized as well as tried for crystallization. I was involved in selecting target for structure determination, domain designing and structural characterization of selected targets in terms of active site, catalytic mechanism and substrate specificity. I have structurally characterized xyloglucan specific glycosyl hydrolases such as GH12, GH10, GH62 and also carbohydrate esterases such as CE16 and CE3 class of enzymes. I have further investigated the GH62 and CE16 enzyme active sites and substrate specificities by structure guided site directed mutagenesis. I have used approaches like domain design to optimize expression of fungal proteins and have purified several low expression proteins. I have further performed enzymatic assays, kinetics study and effect of inhibitors. I am trained Molecular Microbiologist and in this project I have learned to analyze the structure function relationship of microbial enzymes. Further, wherever possible I have co-related the obtained data with native gene expression in Aspergillus and synergism exhibited among these enzymes in vivo and in vitro applications. • Previous work in Ph.D. and Postdoctorate at MPI for Chemical Energy Conversion PhD Dissertation title ‘‘Genetic and Molecular Characterizations of Hydrogenases and Sulfate metabolism proteins of Desulfovibrio vulgaris Miyazaki F’’ Desulfovibrio vulgaris Miyazaki F (DvMF) is an H2S producing strictly anaerobic bacterium that exhibit high level of expressions of several metal containing enzymes. Periplasmic heterodimeric [NiFe] and [NiFeSe]- hydrogenases are two such examples that contains iron sulfur clusters and have oxygen sensitive catalytic activities. Although the yields of such proteins from native DvMF are high, the functional and genetic manipulations were a challenging task due to high antibiotic resistance of DvMF as well as non-availability genomic sequences at that time. My initial goal was to establish an expression system that could enable expression of this periplasmic protein such into E. coli or into a hydrogenase knock out mutant of original strain. By preparing and probing cosmid library by southern hybridization, I sequenced two additional hydrogenases two subunit [NiFeSe] hydrogenase, a six subunit Ech hydrogenase as well as five maturation (post-translational modification) genes and genes coding for complexes 5’-adenylyl sulfate reductase (aprA and aprB) and desulfoviridin (dsvA, dsvB, dsvD and dsvC). I also participated in protein purification of these native proteins, some of which were crystallized. The co-expression hydrogenases and their maturation genes was achieved, however the resultant hydrogenases produced in E.coli were not functionally active. The post-translational modifications of microbial hydrogenases is a complex theme for investigation and in this regard I made some interesting advances for [NiFe] and [NiFeSe]- hydrogenases of anaerobic DvMF. • Other relevant projects Research work ‘Transcriptional fusions of esterase and lipases of Pseudomonas aeruginosa with GFP (green fluorescent protein) and its unstable variant.’ Several enzymes including the lipolytic enzymes of P. aeruginosa play roles in biofilm formation of P. aeruginosa. In this project I cloned the regulatory sequences of lipases (lipA, lipC) and esterases (estA) in transcription fusions with GFP and an unstable variant of GFP. Using conjugation, these transcription fusions were transformed into wild and mutant strains of P. aeruginosa which were chromosomally tagged with a red fluorescent protein. These constructs were further used to visualize in vivo stable and transient expression in Pseudomonas biofilms and future goals were set to study relative architecture of biofilms in regard to the enzyme studied.

Subject Matter Expertise

• ☆ Separation & Purification
• ☆ Microbiology
• ☆ Gram-Negative Bacteria
• ☆ Structural Biology
• ☆ Proteomics
• ☆ Genome Sequencing
• ☆ Biotechnology & Bioengineering
• ☆ Microbial Biotechnology
• ☆ Cluster Analysis
• ☆ Protein/Enzyme Engineering
• ☆ Antibiotic Development
• ☆ Audio Transcription

Services

• Writing
• Consulting
• Data & AI
• Product Development

Work Experience

Postdoctoral Fellow

University of Toronto

November 2011 - January 2015

PostDoc Fellow

University of Toronto

November 2011 - January 2015

Postdoctoral Fellow

Max Planck Institute of Chemical Engergy Conversion

February 2009 - March 2010

Post-Doktoranden

Max-Planck-Institut für chemische Energiekonversion

February 2009 - February 2010

Research Fellow

Max Planck Institute of Chemical Engergy Conversion

July 2004 - January 2009

Research Fellow

Institute of Enzyme Technology, Dusseldorf University

April 2003 - June 2004

Research Fellow

University Clinics Heidelberg

September 2002 - December 2002

Research Fellow

University Clinics, Essen-Duisburg University

April 2002 - August 2002

Education

Ph.D. Biological Sciences

Dusseldorf University

July 2004 - January 2009

PhD Degree/Dr. rer. nat.

Max-Planck-Institut für chemische Energiekonversion

July 2004 - January 2009

M.Sc. Biotechnology

G.N.D University

July 1999 - August 2001

B.Sc. Industrial Microbiology

G.N.D University

June 1996 - June 1999

Certifications

• Certification details not provided.

Publications

JOURNAL ARTICLE

Amrit Pal Kaur, Boguslaw P. Nocek, Xiaohui Xu, Michael J. Lowden, Juan Francisco Leyva, Peter J. Stogios, Hong Cui, Rosa Di Leo, Justin Powlowski, Adrian Tsang, et al.(2014). Functional and structural diversity in {GH}62 $\upalpha$-L-arabinofuranosidases from the thermophilic fungus S cytalidium thermophilum . Microbial Biotechnology. p. n/a--n/a. Wiley-Blackwell

W. Wang, G. Mai-Gisondi, P. J. Stogios, A. Kaur, X. Xu, H. Cui, O. Turunen, A. Savchenko, E. R. Master(2014). Elucidation of the Molecular Basis for Arabinoxylan-Debranching Activity of a Thermostable Family {GH}62 ~-L-Arabinofuranosidase from Streptomyces thermoviolaceus . Applied and Environmental Microbiology. 80. (17). p. 5317--5329. American Society for Microbiology

Hideaki Ogata, Yasuhito Shomura, Aruna Goenka Agrawal, Amrit Pal Kaur, Wolfgang Gärtner, Yoshiki Higuchi, Wolfgang Lubitz(2010). Purification, crystallization and preliminary X-ray analysis of the dissimilatory sulfite reductase from Desulfovibrio vulgaris Miyazaki F . Acta Cryst F. 66. (11). p. 1470--1472. International Union of Crystallography ({IUCr})

Hideaki Ogata, Aruna Goenka Agrawal, Amrit Pal Kaur, Richard Goddard, Wolfgang Gärtner, Wolfgang Lubitz(2008). Purification, crystallization and preliminary X-ray analysis of adenylylsulfate reductase from Desulfovibrio vulgaris Miyazaki F . Acta Cryst F. 64. (11). p. 1010--1012. International Union of Crystallography ({IUCr})