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The pulA gene enconding a thermostable pullulanase of 844 amino acids was cloned by PCR amplification from the genomic DNA of Thermotoga maritima MSB8. We successfully expressed the pulA gene in a protease-deficient strain Bacillus subtilis WB600, and the expressed pullulanase was secreted into the culture broth. Preliminary results showed that the pullulanase exhibited optimal activity at 90°C, and...
Three recombinant Escherichia coli strains were constructed to produce guanosine 5'-diphosphate (GDP)-mannose, donor of GDP-fucose, which is an essential substrate for synthesis of fucosyloligosaccharides. Glucokinase (glk), phosphomannomutase (manB), and mannose-1-phosphate guanylytransferase (manC), are three crucial enzymes for the de novo GDP-mannose biosynthesis, were overexpressed in recombinant...
Arthrobacter simplex is extensively used in pharmaceutical industry to introduce C1-C2 double bond into steroid nucleus. This dehydrogenation reaction is known characterized by its 3-ketosteroid-Δ1-dehydrogenase(KSDH), which is encoded by ksdD gene. In this research, we report the expression of KSDH in Escherichia coli. The activity of the enzyme has been determined by spectrophotometric method. We...
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