Functional Characterization of Trehalose Biosynthesis Genes from E. coli: An Osmolyte Involved in Stress Tolerance |
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Authors: | Toms C. Joseph Lawrance Anbu Rajan Nirmala Thampuran Roswin James |
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Affiliation: | (1) Microbiology, Fermentation and Biotechnology Division, Central Institute of Fisheries Technology, Matsyapuri P.O., Cochin, 682 029, Kerala, India |
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Abstract: | Trehalose (1-α-d-glucopyranosyl-1-α-d-glucopyranoside), a non-reducing disaccharide is a major compatible solute, which maintains fluidity of membranes and protects the biological structure of organisms under stress. In this study, trehalose-6-phosphate synthase (otsA) and trehalose-6-phosphate phosphatase (otsB) genes encoding for trehalose biosynthesis from Escherichia coli was cloned as an operon and expressed in E. coli M15(pREP4). The recombinant E. coli strain showed a threefold increase in the activity of otsBA pathway enzymes, compared to the control strain. The transgenic E. coli accumulated up to 0.86 mg/l of trehalose. The sequence of otsA and otsB genes reported in this study contains several base substitutions with that of reported sequences in GenBank, resulting in the altered amino acid sequences of the translated proteins. |
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