Abstract: 1,3-propenediol oxidoreductase isoenzyme is one of the key enzymes in biotransformation systhesis of 1,3-propenediol，and is encoded by the yqhD gene. YqhD gene was amplified from E. coli by polymerase chain reaction (PCR). The fragment was inserted into plasmid pET28 cut by two restriction enzymes BamHI and NheI. The yqhD gene after recombinant and transformation was induced and expressed by IPTG in E. coli at 25 ℃ for 12 h. The expression product was checked by SDS-PAGE. The cloned yqhD gene was obtained and protein expression was realized. The catalytic activity of 1,3-propenediol oxidoreductase isoenzyme was improved by 88 fold. The yield of 1,3-propenediol in whole cell fermentation with recombined strain could reach 65%，higher than wild strain (yield 9.5%).

摘要： 大肠杆菌1,3-丙二醇氧化还原酶同工酶被yqhD的基因编码，能催化羟基丙醛生成1,3-丙二醇。采用PCR方法从大肠杆菌中扩增到yqhD，测序结果显示该片段大小为1164 bp。将目标片段克隆到原核表达载体pET28（α+）中并转化到大肠杆菌Novablue，经IPTG诱导后，SDS-PAGE 电泳鉴定，结果表明克隆得到的yqhD基因能在大肠杆菌Novablue-pET28中实现蛋白表达；在25 ℃条件下，IPTG诱导12 h后，1,3-丙二醇氧化还原酶同工酶的酶活力达到124 U/mg，而野生型的酶活力仅为1.4 U/mg；重组菌全细胞发酵结果显示，产物1,3-丙二醇的产率能达到65%，而野生型仅为9.5%。