초록
<P>Glycerol dehydratase (GDHt) is a key and rate‐limiting enzyme in the biological process of producing 1, 3‐propanediol from glycerol. Using a uniform design to get the optimal system of amplifying <I>dha</I>B<SUB>1</SUB>B<SUB>2</SUB> by PCR, a new GDHt and its reactivase were obtained from <I>Clostridia butyricum</I>. SDS‐PAGE showed that the relative molecular weights of the <I>dha</I>B<SUB>1</SUB>B<SUB>2</SUB> expression products were about 88 and 35 kDa, which confirmed the coexpression of <I>dha</I>B<SUB>1</SUB>B<SUB>2</SUB> in <I>Escherichia coli</I> BL21. The best inducing conditions, 0.1 mM IPTG, 28°C, and 11‐h induction time, were obtained by the uniform design and regression analysis. By anaerobic inducible expression in <I>E. coli</I> BL21, the activity of GDHt activity was six times higher than in <I>C. butyricum</I> (2.37 U/mL), and its specific activity was 36.3 U/mg. These results and methods would be useful for effective PCR amplification on a long gene fragment, high‐efficiency expression of the recombinant enzyme, and enzymatic production of 1, 3‐propanediol.</P>