Article ID Journal Published Year Pages File Type
16823 Enzyme and Microbial Technology 2016 8 Pages PDF
Abstract

•A novel PTS related gene cluster with overlapped gene was found from metagenomic library of termite hindgut.•There was an interaction between the PTS components Glu1392 and Glu1923, and the co-expression of these two components was responsible for the utilization of the glucoside substrates in E. coli.•The glucosidase Glu1392 had a very high tolerance to glucose with a Ki value of 2.25 M.

Functional screening of a metagenomic library of termite hindgut identified an overlapping gene cluster encoding the phosphotransferase system (PTS) components, which consisted of a glucoside specific PTS enzyme II gene (glu1923) and a glycoside hydrolase gene (glu1392). Hydrolytic experiments revealed that the combined effect of Glu1923 and Glu1392 was responsible for the utilization of glucosidic substrates in recombinant Escherichia coli (E. coli) strains. Yeast two hybrid analysis suggested that there was an interaction between Glu1923 and Glu1392, and the domain EIIA of Glu1923 played an important role for the interaction. In addition, the hydrolase Glu1392 displayed hydrolysis ability toward cellobiose and maltose, and had a very high tolerance to glucose with a Ki value of 2.25 M. These properties make Glu1392 an interesting candidate in biotechnology applications after further study.

Related Topics
Physical Sciences and Engineering Chemical Engineering Bioengineering
Authors
, , , , , , ,