At present, research in the laboratory focuses on enzymes used by major national industries. Current interest involves lignocellulose degrading enzymes, amylolytic enzymes and lipases/esterases as well as some other specialized enzymes of biotechnological value. Enzymes capable of working under extreme conditions applicable for industrial processes are also of special interest. Our work in the laboratory currently involves three aspects of research:
The aim is to identify enzymes with desirable properties by two approaches:
(a) Culture-dependent approach: This approach is employed to identify potent enzymes from cultured organisms such as fungi and bacteria. At present more than 37,000 microbial isolates at the BIOTEC Culture Collection provide a promising resource for enzyme screening.
Enzyme purification by using AKTA Explorer FPLC system
Quantitatively assays for various industrial enzymes
Culture-dependent enzyme discovery research
(b) Culture-independent approach: This approach is employed to identify enzymes from microorganisms in the environment without the necessity of culturing. This approach is very useful to find a broad range of novel enzymes since only approximately 1% of microbes naturally residing in the environment can be grown in normal laboratory conditions. Advanced DNA technology is used to construct so-called “metagenomic libraries”, representing the genomes of all microbes from a particular environment. Then, genes encoding enzymes with desirable properties can be identified using several approaches, including activity-based screening, sequence-based screening and pyrosequencing. Metagenomic libraries have been constructed from various sources, including unique or extreme ecosystems e.g. hot springs, peat swamp forest, sugarcane bagasse compost and lignocellulolytic microbial consortium as well as particular biological niches including termite gut and cattle digestive tracts. The two approaches enable us to obtain enzymes from both cultured and uncultured microorganisms with high activity and functional stability suitable for biotechnological application. The work also attributes to the awareness for conservation and sustainable use of the
Enzyme discovery from metagenomes (Modified from K. J. Shelswell. 2009. Metagenomics: The science of biological diversity. (www.scq.ubc.ca/metagenomics-the-science-of-biological-diversity/)
Optimization of Large-Scale Enzyme Production:
In close collaboration with the Fermentation and Biochemical Engineering Laboratory, BIOTEC, the research includes identification of appropriate conditions for enzyme production in large scale, so that particular enzymes can be produced in sufficient quantity by bioreactor to be used in industrial process optimisation and field trial. This includes both enzymes produced by native fungal strains and by recombinant systems. Various types of agricultural wastes are tested for their possible
use in culturing media for high production of enzymes.
Enzyme Application in Industry:
Current research and development projects with industrial sectors include application of enzymes in three major industries, including animal feed industry, biorefinery, and pulp & paper processing. These include the use of non-starch polysaccharide hydrolyzing enzymes and phytases in the animal feed industry, the application of lignocellulolytic enzymes in biomass conversion process of agro-industrial by-products to value-added products, including biofuels, bio-plastics and chemicals and also application of cellulolytic/hemicellulolytic enzymes for the pulp and paper industry.
Current collaborative researches with academic institutions and industrial sectors include:
Development of biomass pretreatment process for agro-industrial by-products in collaboration with Advanced Industrial Science and Technology (AIST), Japan
Collaboration with the Joint Graduate School of Energy and Environment (JGSEE) for production of alternative biofuels, including biodiesel and liquid alkane biofuel by thermocatalytic and biocatalytic processes
Development of a non-thermal cassava feedstock saccharification process using fungal multi-enzyme for bioethanol production with Kasetsart Agricultural and Agro-Industrial Product Improvement Institute (KAPI), Kasetsart University
Collaboration with the University of Tokyo, Japan to establish active lignocellulolytic microbial consortium with structural and functional stability for application in biomass degradation and discovery of lignocellulolytic enzymes
Collaboration with Mahidol University and KMITL (King Mongkut’s Institute of Technology Ladkrabang) to identify enzymes with special characteristics or enzymes suitable to be used in feedstuff
Research collaboration with National Institute of Advanced Industrial Science and Technology (AIST), Japan on production of bio-based plastic/monomer
Collaboration with Betagro Science Center, Asia Star Animal Health Co. Ltd. and Sunfeed on screening and production of enzymes for animal feed industry
Collaboration with SCG Paper, Public Co. Ltd. to develop enzymes for pulp and paper production process
Contract research with Electricity Generating Authority of Thailand (EGAT) in collaboration with King Mongkut’s University of Technology North Bangkok
A database describing non-starch polysaccharide hydrolyzing enzymes and amylases obtained from screening fungi in the BIOTEC Culture Collection with high potential for various industries is available in the form of “Enzyme Catalogue” for interested private sectors and researchers. Enzyme Technology Laboratory also provides enzyme activity analysis service and consulting service for enzyme application in industries. In addition, several products aiding enzyme analysis, namely Enzhance Overlay Enzyme Detection Kit, Enzhance Microbial Cell Lysis Buffer and Enzyme Assay Strip Test have been developed. These are considered important steps for the efficient advancement of enzyme application in Thailand.
ENZhance Complete Kit for screening of enzyme activity from bacterial samples
ENZhance Cell Lysis Buffer for active protein extraction from bacteria, yeast and fungi
ENZstrip test strips for enzyme activity detection in commercial products