The tresI gene of Myxococcus sp. strain V11 was cloned, and found to encode a trehalose synthase comprising 551 amino acids. The deduced molecular weight of the encoded TreS I… Click to show full abstract
The tresI gene of Myxococcus sp. strain V11 was cloned, and found to encode a trehalose synthase comprising 551 amino acids. The deduced molecular weight of the encoded TreS I protein 64.7 kDa and the isoelectric point (pI) was predicted to be 5.6. The catalytic cleft consists of the Asp202-Glu244-Asp310 catalytic triad and additional conserved residues. The recombinant (His)6-tag enzyme was expressed in Escherichia coli BL21(DE3) and purified by Ni2+-affinity chromatography, resulting in a specific activity of up to 172.7 U/mg. TLC and HPLC results confirmed that rTreS I can convert maltose into trehalose, with a yield of 61%. The KM and Vmax values of recombinant TreS I for maltose were 0.62 mM and 25.5 mM·min-1 mg-1 protein, respectively. TreS I was optimally active at 35° and stable at temperatures of <25°C. TreS I was stable within a narrow range of pH values, from 6.0 to 7.0. The enzymatic activity was slightly stimulated by Mg2+ and strongly inhibited by Fe3+, Co2+ and Cu2+. TreS I was also strongly inhibited by SDS and weakly by EDTA and TritonX-100.
               
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