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利用温控载体pHsh将编码甘油脱水酶的基因dhaB和编码1,3-丙二醇氧化还原酶同工酶的基因yqhD串联构建重组质粒pHsh-dhaB-yqhD,将其转化大肠杆菌得到新型产1,3-丙二醇重组大肠杆菌JM109(pHsh-dhaB-yqhD),并对影响该重组菌发酵的营养因子进行研究.实验结果表明:该重组菌转化甘油合成1,3-丙二醇的适宜培养基组成为甘油60g·L-1、酵母膏5·0g·L-1、维生素B120·05g·L-1以及KH2PO47·5g·L-1;以此培养基在5L发酵罐上进行放大,1,3-丙二醇产量、转化率和生产能力分别达到43·26g·L-1、72·2%和1·55g·L-1·h-1.
The pHha-dhaB-yqhD recombinant plasmid pHsh-dhaB-yqhD was constructed by tandemly linking the gene dhaB encoding glycerol dehydratase and the gene yqhD encoding 1,3-propanediol oxidoreductase isozyme using temperature-controlled vector pHsh to obtain novel 1,3 - propylene glycol recombinant Escherichia coli JM109 (pHsh-dhaB-yqhD), and the factors that affect the fermentation of the recombinant bacteria were studied.The experimental results show that: the suitable medium for the synthesis of 1,3-propanediol from glycerol was 60g L-1, yeast extract 5.0g · L-1, vitamin B120 · 05g · L-1 and KH2PO47 · 5g · L-1. The medium was amplified on a 5L fermentor and the yield of 1,3-propanediol , Conversion rate and production capacity reached 43.26g · L-1,72 · 2% and 1.55g · L-1 · h-1, respectively.