Ikram ul Haq
GC University, Pakistan
Title: Cloning, characterization and saccharification analysis of GH12 endo-1,4-β-glucanase from Thermotoga petrophila in a mesophilic host
Biography
Biography: Ikram ul Haq
Abstract
The increasing demand of energy has strongly stimulated the research on conversion of lignocellulosic plant biomass by the action of cellulases enzymes into reducing sugars, for the subsequent production of bioethanol. Endoglucanases are mainly responsible for hydrolyzing the internal glycosidic bond to decrease the length of the cellulose chains. Obtaining efficient and thermostable endoglucanase has become the goal of much research worldwide. Therefore, our research work was focus to search for new resources of endoglucanases, which was thermostable and with high catalytic efficiency. The article focuses on the thermotolerant endo-1,4-β-glucanasegene, of Thermotoga petrophila RKU-1, was cloned and over-expressed in E. coli strain BL21 CodonPlus for its potential usage for the hydrolysis of lignocellulosic biomass and in different industrial applications. Thermostable endoglucanase can be used simultaneously and directly in the saccharification procedure without a pre-cooling process of biomass. Purified enzyme was optimally active with 530 Umg-1 of specific activity against CMC at pH 6.0 and 95°C, which has exhibited a half- life (t1/2) of 6.6 min even at temperature as high as 97°C and stable upto 8h at 80°C. The recombinant enzyme saccharified pre-treated wheat straw and baggase to 3.32% and 3.2%, respectively after 6h incubation at 85°C. Its thermostability, resistance to heavy metal ions and high specific activity make endoglucanase a potential and promising candidate for various industrial applications such as in textile industry (in biostoning and biofinishing), in animal feed production, in processing of beer and fruit juice, in biomass hydrolysis (bioethanol production) and paper industry.