Separate expression and co-expression of MTSase and MTHase from Arthrobacter ramosus S34 in Escherichia coli BL21(DE3)
Systems Microbiology and Biomanufacturing(2023)
Abstract
Maltooligosyl trehalose synthase (MTSase) and maltooligosyl trehalose hydrolase (MTHase) are used to produce trehalose, a disaccharide of interest to many different industries, from starch. MTSase and MTHase from Arthrobacter ramosus S34 were first produced using separate Escherichia coli BL21(DE3) strains. The activities obtained in a 3-L fermenter under optimized conditions were 1608.3 U mL −1 and 8766.2 U mL −1 , respectively. Then, MTSase and MTHase were co-produced in E. coli BL21(DE3) using a co-expression construct. After optimizing induction conditions, the MTSase and MTHase activities produced by the superior strain reached 1827.4 U mL −1 and 2944.9 U mL −1 , respectively. When the co-produced enzymes were used to synthesize trehalose from starch, a conversion rate identical to that achieved using separately produced enzymes (about 67%) was obtained. This is the first describing the co-production of the MTSase and MTHase in a 3-L fermentor. The results represented the highest MTSase production level reported to date, and the MTHase activity from co-production was sufficient for trehalose synthesis. Using co-produced enzymes during trehalose synthesis would lower costs without sacrificing yield. Therefore, this study provided a foundation for the industrial synthesis of trehalose using co-produced enzymes.
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Key words
Maltooligosyl trehalose synthase,Maltooligosyl trehalose hydrolase,Escherichiacoli,Co-expression,Fermentation optimization
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