HIGHLY POTENT SACCHARIFICATION OF ARTHROSPIRA MAXIMA GLYCOGEN BY FUNGAL AMYLOLYTIC ENZYME FROM TRICHODERMA SPECIES J113

Abstract

The first generation of biofuels was mostly generated from plant materials that competed directly with food and feeds for human and animals leading to commodity market instability and negative impact on global food prices. The use of cellulose and lignin, requires pretreatment before these materials can be subjected to fermentation for bioethanol production. Recently, photosynthetic microorganisms, particularly cyanobacteria, are taking interest as an alternative to plant-based biomass for renewable energy production. Our goal was to identify a new source of enzymes with improved amylolytic efficiency in cyanobacterial glycogen hydrolysis. We isolated a new Trichoderma species J113 strain from the coastal terrains of Korea, and then determined that the fungus has a high amylolytic enzyme activity. We cultured the fungus on wheat bran to stimulate enzyme production, and the crude extract was subsequently purified through filtrations, precipitation, and chromatography. We observed that J113 enzyme consists of two putative major amylases, Ayt40 and Ayt70, that were determined as an α-amylase and a glucoamylase, respectively. While these two amylases exhibited different pH and temperature requirements for optimum performance, collectively J113 enzyme showed the highest activity at pH 4 and 60°C. In addition, we were able to drastically enhance the amylolytic capacity of Ayt70 gluco-amylase by 291% with 5 mM Mn2+ amendmose and lignin, requires pretreatment before these materials can be subjected to fermentation for bioethanol production. Recently, photosynthetic microorganisms, particularly cyanobacteria, are taking interest as an alternative to plant-based biomass for renewable energy production. Our goal was to identify a new source of enzymes with improved amylolytic efficiency in cyanobacterial glycogen hydrolysis. We isolated a new Trichoderma species J113 strain from the coastal terrains of Korea, and then determined that the fungus has a high amylolytic enzyme activity. We cultured the fungus on wheat bran to stimulate enzyme production, and the crude extract was subsequently purified through filtrations, precipitation, and chromatography. We observed that J113 enzyme consists of two putative major amylases, Ayt40 and Ayt70, that were determined as an α-amylase and a glucoamylase, respectively. While these two amylases exhibited different pH and temperature requirements for optimum performance, collectively J113 enzyme showed the highest activity at pH 4 and 60°C. In addition, we were able to drastically enhance the amylolytic capacity of Ayt70 gluco-amylase by 291% with 5 mM Mn2+ amendm