Department of Biosciences and Bioengineering : Indian Institute of Technology Guwahati

Arun Goyal

Professor

arungoyl[AT*]iitg.ac.in

Research Areas:

Molecular Biology, Protein Engineering, Structural and Functional Proteomics of Carbohydrate active enzymes and other industrially important microbial enzymes

Web page: https://kaziranga.iitg.ac.in/~biotech/faculty_profile.php?fname=Arun&lname=Goyal&iitg=1115&mail=arungoyl@iitg.ac.in

Area of Research:

Molecular Biology, Protein Engineering, Structural and Functional Proteomics of Carbohydrate active enzymes and other industrially important microbial enzymes.

I) Ongoing Research:

A. Microbial Diversity for industrially useful microorganisms from North-East India.

Identification and functional analysis of glucansucrase and fructansurase producing Leuconostoc mesenteroides, Pediococcus pentosaceus, Lactobacillus plantarum, Lactobacillus fermentum, Weisella confusa and Weisella cibaria.
Optimization of conditions for culture for maximum secretion of enzyme using Batch fermentation.
Purification of enzyme by fractionation and size exclusion chromatography. Identification of enzyme by activity staining and molecular size analysis by SDS-PAGE.
Linkage analysis of glucans by ¹H & ¹³C NMR and FTIR techniques.
Production and structure characterization of glucans from novel microbes as agents for anticancer, biomedical, functional food and cereal applications.
Isolation and characterization of probiotic microbes Lactobacillus plantarum & Weisella cibaria for functional food applications.
Production and purification of Prebiotic Isomalto-, Gluco-, Gentio-, Manno- and Xylo-oligosaccharides by enzymes.
Glucan production from fruit waste by Lactic acid bacteria for food applications.

B. Molecular, structural, functional and biochemical analysis of Carbohydrate Active Enzymes.

Hemicellulases (L-?-arabinofurnasidase, a family 43 glycoside hydrolase (GH43), glucoronoxylan-xylanohydrolase, (GH30) and ?-mannanase (GH26) from Clostridium thermocellum.
Polysaccharide Lyases (Pectate lyases, PL1A, PL1B, PL9 and Rhamnogalacturonan lyase, PL11) from Clostridium thermocellum.
Pectic methyl esterase of family 8 Carbohydrate esterase (CE8) from Clostridium thermocellum.
Xylanase (GH11) from Pedobacter saltans and xylanase (GH10) from Clostridium thermocellum.
Carbohydrate binding modules (CBM6 and CBM35) of xylanase, arabinofuranosidase and polysaccharide Lyases.
Constructing mutants of carbohydrate binding modules (CBMs) from Clostridium thermocellum by in vitro site-directed mutagenesis to identify amino acids involved in binding the polysaccharides.
Protein engineering of carbohydrate enzymes (GH5, GH43, Chimeric GH5-GH43) for developing novel biocatalysts.
Crystallization and 3D structure analyses of Xyn30A, GH43, GH81, GH26 and PL1B from Clostridium thermocellum
Molecular cloning, expression and biochemical characterization of Chondroitin ABC lyase (PL8a) from Pedobacter saltans.
Structure and functional analysis of a novel Laminarinase (GH81) with multi-substrate specificity from Clostridium thermocellum.

C. Microbial conversion of cellulosic agrowaste to bioethnol using microbial cultures, natural and recombinant cellulases and hemicellulases for bioethanol production.

Selection of agrowaste on the basis of their structural carbohydrate composition: Estimation of cellulose, hemicellulose and lignin of different lignocellulosic leafy biomasses viz., jamun, neem, asoka, bamboo, poplar, wild grass, thatch grass, eucalyptus, mango, water hyacinth, corn cob, sugarcane bagasse and whitetop weed.
Identification of efficient pretreatment process aiding in improved saccharification: Confirmation of structural destabilization with porosity increment and lignin breakdown by field emission scanning electron microscopy (FESEM) and Fourier transform infrared (FTIR) spectroscopic analyses.
Efficiency of fermentation mode in terms of ethanol yield in shake flask: Separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF) involving various enzymes viz. T. reesei cellulase, B. subtilis, B. amylofaciens cellulase, recombinant Clostridium thermocellum cellulase, hemicellulase, xylanase and fermentative microbes viz. Zymomonas mobilis, Saccharomyces cerevisiae and Candida shehatae. Selection of fermentation by single enzyme-single culture and mixed enzyme-mixed culture system.
Optimization of fermentation process parameters at shake flask level: Standardization of fermentation process parameters viz. concentrations of substrate and enzyme, microbial inoculua, pH and temperature for higher ethanol titre and yield.
Scale-up at bioreactor level and subsequent product recovery with purification: Shake flask fermentation with increased substrate concentration. Scale-up in 7.0 L bioreactor with optimization of process parameters.
Recovery and purification of bioethanol: By distillation, pervaporation, vacuum evaporation. Purification by semi-permeable membrane, molecular sieve and pressure reduction.

II) At Faculty of Veterinary Medicine, Lisbon, Portugal I was involved in various projects:

Genetic control of cellulase and xylanase expression from Pseudomonas fluorescens and Celvibrio mixtus. The project involved cloning and expression of a regulatory protein involved in controlling cellulase and xylanase synthesis. Both, the full-length regulatory protein RegA (50 kDa) and a truncated form, DNA binding domain RDNAF (14 kDa) from Cellvibrio mixtus were cloned and expressed.
Cloning, expression and determining substrate and ligand specificity of catalytic modules GH5 and GH26 and carbohydrate binding module, CBM11 of bifunctional cellulase H from Clostridium thermocellum, cloned separately and together and elucidating mechanisms and role of these modules.
Cloning expression and elucidation of role of carbohydrate binding module, CBM22 of Xylanase from Clostridium thermocellum.
Cloning, expression and biochemical characterization of acetyl xylan esterase (carbohydrate esterase family 2, CE2) of cellulase, Cel 5D from Clostridium thermocellum.
Determining ligand specificity of carbohydrate binding module of family 6 (CBM6) from Celvibrio mixtus.
The first crystals of family 11, carbohydrate binding module 11 (CBM11) and of the catalytic module, glycoside hydrolase 26 (GH26), both being the modules of Cellulase H (Cel H) were produced. The crystal structures of both have been solved.

III) At BISR, Jaipur, I worked on a project on Microbial Biodiversity of saline lakes and desert regions of Rajasthan, India. The aim was to selectively screen the enzymes of bioprocess importance such as hydrolases (lipase, protease, xylanases and cyclodextrin glycosyltransferase) from the isolates. We isolated few strains from soil samples of Asia's largest salt lake (Sambhar Lake), capable of growing at high pH and at saturating concentrations of salt.

IV) I worked on Molecular Genetics of RNA at WSU, Detroit. I was involved in determination of structural and functional relationship of the 790 loop and 1400-1500 regions of 16S ribosomal RNA of E. coli. The 790 loop is a conserved hairpin located between positions 786 and 796 of 16S rRNA and is required for ribosomal function. With the development of a method for `instant evolution' in which random mutagenesis and in vitro selections were used to identify both structural and functional features of 790 loop and 1400-1500 bp regions. I performed the random mutations at 1401, 1404, 1497 and 1500 positions of 16S rRNA in pWK1 plasmid separately and then random mutations at all four positions together in pRNA123 plasmid. I accomplished the transfer of nine single 790-loop mutants of 16S rRNA from plasmid pRNA122 to pRNA123, to study the functional aspects of ribosome.

V) At OSU, Columbus, I worked on purification of a newly identified methyl branched chain fatty acid synthesising enzyme from Mycobacterium tuberculosis H37Rv. Mycobacterial cell wall contains a variety of lipids with many types of multiple methyl-branched fatty acids. The aim was to elucidate the mechanisms involved in the biosynthesis of these lipids and to test their possible functions in the host-pathogen interaction. I was involved in the purification of a mycocerosic acid synthase, identification of products generated, seeking the identity of the gene that encodes the synthase in the genomic data bank using the protein sequence and in determining the biochemical and functional consequences of knocking out of this gene. The results would help identify the cell wall lipids critical for pathogenesis that might be suitable targets for new antimicrobial drugs.

VI) At UCD, Dublin, I investigated co-operative behaviour of hybrid hexamers of different mutant forms of glutamate dehydrogenase (GDH) from Clostridium symbiosum. Hybrid hexamers were constructed by in vitro subunit hybridisation. Clostridial GDH is an allosteric enzyme and is hexameric and has only one type of subunits. This implies that allosteric effects are mediated by interactions among six identical subunits. To study this defined inter-subunit hybrids were constructed in which some of the subunits were functionally disabled with respect to substrate binding or catalysis. The results demonstrated that it is possible to dissect the basis of allosteric interaction in GDH by constructing hybrid hexamer approach.

VII) In doctoral research at IIT Kanpur, I worked on dextransucrase from Leuconostoc mesenteroides NRRL B-512F. I discovered a new medium for the maintenance of culture and a new medium for higher production of enzyme. I developed a simple and effective method for purification by fractionation using polyethylene glycol 400. The total amino acid composition of dextransucrase was determined. I established for the first time by chemical modification studies using pyridoxal 5'-phosphate, o-phthalaldehyde and 2,4,6-trinitrobenzenesulphonic acid that a lysine residue present at the active site is essential for enzyme activity.

VIII) In M.Tech. project at IIT Delhi, I worked on isolation and purification of coenzymes NADH and ATP from Saccharomyces cerevisiae. I developed a method to isolate and purify these adenine coenzymes. The cells were permeabilised by ether for the recovery of coenzymes. The purification was carried out by affinity-ultrafiltration technique using alcohol dehydrogenase and hexokinase for NADH and ATP respectively, as affinity molecules.

1995 Ph.D. Chemistry (8.33/10.0) (worked on Microbiology and Enzymology), Indian Institute of Technology Kanpur, Kanpur, India.

1989 M.Tech. Biochemical Engineering and Biotechnology (6.96/10.0), Indian Institute of Technology Delhi, New Delhi.

1988 M.Sc. Chemistry, (6.82/10.0), Indian Institute of Technology Delhi, New Delhi.

1986 B.Sc. (Honours) Chemistry (70.6%) Hindu College, University of Delhi, Delhi.

1983 12th Class, Delhi Senior School Cetificate Examination, CBSE New Delhi (75.0%) Ramjas Senior Secondary School No. 5, New Delhi.

1981 10th Class, Delhi Secondary School Examination, CBSE New Delhi (72.6%), Ramjas Senior Secondary School No. 5, New Delhi.

Feb 2017- Mar 2017 Visiting Fellow (UGC), Department of Microbiology, Panjab University, Chandigarh for 2 weeks (Feb-Mar, 2017)
Dec 2015 – contd. Professor HAG (Higher Administrative Grade), Department of Biosciences & Bioengineering, Indian Institute of Technology Guwahati, Assam
Jun 2015 - Jul 2015 Visiting Professor, Department of Animal Production, Faculty of Veterinary Medicine, University of Lisbon, Portugal.
May 2015- May 2015 Visiting Professor, Department of Food and Environmental Science, University of Helsinki, Finland.
Dec 2014 - Dec 2014 Visiting Professor, Department of Food and Environmental Science, University of Helsinki, Finland.
Jun 2014 - Jul 2014 Visiting Professor, Department of Animal Production, Faculty of Veterinary Medicine, University of Lisbon, Portugal.
Jun 2014 - Jun 2014 Visiting Professor, Department of Food and Environmental Science, University of Helsinki, Finland.
Mar 2013 - Nov 2013 Visiting Professor, Department of Animal Production, Faculty of Veterinary Medicine, University of Lisbon, Portugal.
Dec 2009 - Dec 2015 Professor, Department of Biotechnology, Indian Institute of Technology Guwahati, Assam, India.
Oct 2012 - Oct 2012 Visiting Professor, Department of Food and Environmental Science, University of Helsinki, Finland.
May 2011 - Jun 2011 Visiting Professor, Department of Animal Production, Faculty of Veterinary Medicine , University of Lisbon, Portugal.
Jun 2010 - Jul 2010 Visiting Professor, Department of Biochemistry, Plovdiv University, Bulgaria.
Apr 2009 - Feb 2013 Head, Department of Biotechnology, Indian Institute of Technology Guwahati, Assam, India.
May 2004 - Nov 2009 Associate Professor, Department of Biotechnology, Indian Institute of Technology Guwahati, Assam, India.
May 2005 - Apr 2006 Vice-Chairman, Graduate Aptitude Test in Engineering (GATE) 2006.
Aug 2003 - May 2004 Visiting Associate Professor, Department of Biotechnology, Indian Institute of Technology Guwahati, Assam, India
Sep 2001 - Aug 2003 Post-doctoral Scholar at Faculty of Veterinary Medicine (FMV), Technical University of Lisbon (UTL), Lisbon,
Portugal, sponsored through i) 6 year (2001-2007) scholarship awarded by Foundation of Science and Technology
(FCT), Portugal and ii) Interdisciplinary Center for Investigation in Animal Health (CIISA) at FMV, Lisbon.
Sep 2000 - Aug 2001 Post-doctoral Fellow, Faculty of Veterinary Medicine (FMV), Technical University of Lisbon (UTL), Lisbon, Portugal.
Feb 2000 - Jun 2000 Scientist, Department of Biotechnology, Birla Institute of Scientific Research (BISR), Jaipur, India.
Feb 1999 - Sep 1999 Post-doctoral Fellow, Department of Biological Sciences, Wayne State University (WSU), Detroit, Michigan, USA.
Nov 1998 - Feb 1999 Visiting Scholar, Neurobiotechnology Center, Ohio State University (OSU), Columbus, Ohio, USA.
Jan 1996 - Oct 1998 Post-doctoral Research Associate, Department of Biochemistry, University College Dublin (UCD), Dublin, Ireland.

Involved in teaching courses at IIT Guwahati.

Ph.D. courses (number of times taught)

BT 602 Basic Biotechnology (Biochemical Engineering, Downstream Processing) (1)
BT 607 Plant Biotechnology (Plant cell culture) (1)
BT 608 Microbial Biotechnology (3)
BT 630 Protein Structure and Function (1)

M.Tech. courses

BT 504 Biomolecular and Cellular Process Engineering (3)
BT 520 Applied Biology and Bio-engineering Laboratory (2)

B.Tech. courses (number of times taught)

BT 101 Modern Biology (2)
BT 204 Plant Biotechnology (Plant cell culture) (1)
BT 220 Microbiology Laboratory (4)
BT 202 Microbiology (2)
BT 240 Plant Biotechnology Laboratory (Molecular Biology techniques) (1)
BT 305 Industrial Microbiology (8)
BT 380 Molecular Biotechnology Laboratory (1)
BT 426 Enzyme Structure and Reaction Mechanism (Elective) (1)

Journals

1. Kedar Sharma, Carlos M.G.A. Fontes, Shabir Nazmuddin and Arun Goyal "Molecular organization and protein stability of Clostridium thermocellum glucuronoxylan endo-?-1,4-xylanase of family 30 glycoside hydrolase in solution. ", Journal of Structural Biology.. DOI., Vol.206, , P.P 335-344 , (2019)

2. Kedar Sharma, Vikky Rajulapati and Arun Goyal "Green synthesis of arabinoxyloglucan coated antimicrobial copper nanoparticles. ", Trends in Carbohydrate Research. DOI., Vol.11(1), , P.P 22-30 , (2019)

3. Priyanka Nath, Arun Dhillon, Krishan Kumar, Kedar Sharma, Sumitha Banu Jamaldheen, Vijayanand Suryakant Moholkar and Arun Goyal "Development of bi-functional chimeric enzyme (CtGH1-L1-CtGH5-F194A) from endoglucanase (CtGH5) mutant F194A and ?-1,4-glucosidase (CtGH1) from Clostridium thermocellum with enhanced activity and structural integrity. ", Bioresource Technology. . DOI., Vol.282, , P.P 494-501 , (2019)

4. Ritesh S. Malani, Sachin B. Umriwad, Arun Goyal and Vijayanand S. Moholkar "Ultrasound- assisted enzymatic biodiesel production using blended feedstock of non-edible oils and thermodynamic analysis.", Energy Conversion and Management. DOI., Vol.188, , P.P 142-150 , (2019)

5. Aruna Rani, Vikky Rajulapat and Arun Goyal "Antitumor effect of chondroitin AC lyase (PsPL8A) from Pedobacter saltans on melanoma and fibrosarcoma cell lines by in vitro analysis.", Pharmacological Reports.. DOI., Vol. 71, , P.P 167-174 , (2019)

Conference Publication

1. Arun Goyal "Green process of degumming of jute fiber and bioscouring of cotton fabric by alkaline pectinases from Clostridium thermocellum", National Conference on Recent Trends and Advancements in Chemical Sciences, University of Delhi, March 29-31, 2019 (Invited Talk). Vol., , P.P - , (2019)

2. Arun Goyal "Chimera construction from cellulose hydrolysing enzymes by protein engineering for enhancing biomass saccharification", 8th International Forum on Industrial Bioprocessing (IBA-IFIBiop 2019) “Bridging Sustainability and Industrial Revolution through Green Bioprocessing”, 1-5 May, 2019, Imperial Hotel, Miri, Sarawak, Malaysia (Invited Talk). Vol., , P.P - , (2019)

3. Vikky Rajulapati, Arun Dhillon and Arun Goyal "Green process of degumming of jute fiber and bioscouring of cotton fabric by alkaline pectinases from Clostridium thermocellum", National Conference on Recent Trends and Advancements in Chemical Sciences, University of Delhi, March 29-31. Vol., , P.P - , (2019)

4. Shweta Singh, Priyanka Nath, Krishan Kumar and Arun Goyal "Analysis of mechanism for enhanced catalytic efficiency of CMCase from Bacillus amyloliquefaciens SS35 UV2 mutant strain.", 8th International Forum on Industrial Bioprocessing (IBA-IFIBiop 2019) “Bridging Sustainability and Industrial Revolution through Green Bioprocessing”, 1-5 May, 2019, Imperial Hotel, Miri, Sarawak, Malaysia.. Vol., , P.P - , (2019)

5. Mohanapriya.N*, Shweta Singh, Priyanka Nath, Sumitha Banu and Arun Goyal "Saccharification of Sorghum durra by chimeric enzyme (?-glucosidase and endo ?-1,4 glucanase, CtGH1-L1-CtGH5-F194A) and cellobiohydrolase (CtCBH5A) from Clostridium thermocellum for bioethanol production", 8th International Forum on Industrial Bioprocessing (IBA-IFIBiop 2019) “Bridging Sustainability and Industrial Revolution through Green Bioprocessing”, 1-5 May, 2019, Imperial Hotel, Miri, Sarawak, Malaysia. Vol., , P.P - , (2019)

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