Construction of a novel recombinant Escherichia coli strain capable of producing 1,3–propanediol and optimization of fermentation parameters by statistical design
Construction of a novel recombinant Escherichia coli strain capable of producing 1,3–propanediol and optimization of fermentation parameters by statistical design
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DOI：
10.1007/s11274-006-9139-z
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发表时间：
2006-03
影响因子：
4.1
通讯作者：
Xiaomei Zhang;Yin Li;B. Zhuge;Xueming Tang;W. Shen;Z. Rao;H. Fang;J. Zhuge
中科院分区：
文献类型：
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作者：
Xiaomei Zhang;Yin Li;B. Zhuge;Xueming Tang;W. Shen;Z. Rao;H. Fang;J. Zhuge
采用PCR方法扩增了野生型大肠杆菌编码1,3-丙二醇氧化还原酶同工酶的结构基因yqhD和弗氏柠檬酸杆菌编码甘油脱水酶的结构基因haB。将携带theyqhD和dhaB基因的温控表达载体pHsh转化至大肠杆菌中。 coliJM109 产生重组菌株E。大肠杆菌JM109（pHsh-dhaB-yqhD）。然后应用响应面法(RSM)进一步优化重组菌株的发酵条件。然后开发了一个数学模型来显示每种培养基成分及其相互作用对重组菌株 E 生产 1,3-丙二醇的影响。大肠杆菌JM109。该模型估计，当甘油、酵母提取物和维生素 B12 的浓度分别设置为 61.8 g/l、6.2 g/l 和 49 mg/l 时，可以获得 1,3-丙二醇的最大产量（43.86 g/l）；发酵时间为30小时。这些预测值也通过验证实验得到验证。与实验设计中其他运行获得的值相比，优化后的培养基导致1,3-丙二醇的产率显着增加。在最佳参数和工艺下，产量和生产率可达43.1 g/l和1.54 g/l/h。使用优化培养基在 5 升发酵罐中实现了 41.1 克/升的最大 1,3-丙二醇产量。这使得该工程菌株在工业规模的甘油转化为1,3-丙二醇方面具有潜在的应用前景。
The structural geneyqhD from a wild-typeEscherichia coliencoding 1,3-propanediol oxidoreductase isoenzyme and the structural genedhaB fromCitrobacter freundiiencoding glycerol dehydratase were amplified by using the PCR method. The temperature control expression vector pHsh harboring theyqhD anddhaB genes was transformed intoE. coliJM109 to yield the recombinant strainE. coliJM109 (pHsh-dhaB-yqhD). The response surface method (RSM) was then applied to further optimize the fermentation condition of the recombinant strain. A mathematical model was then developed to show the effect of each medium composition and their interactions on the production of 1,3-propanediol by recombinant strainE. coliJM109. The model estimated that a maximal yield of 1,3-propanediol (43.86 g/l) could be obtained when the concentrations of glycerol, yeast extract and vitamin B12were set at 61.8 g/l, 6.2 g/l and 49 mg/l, respectively; and the fermentation time was 30 h. These predicted values were also verified by validation experiments. Compared with the values obtained by other runs in the experimental design, the optimized medium resulted in a significant increase in the yield of 1,3-propanediol. The yield and productivity under the optimal parameters and process can reach 43.1 g/l and 1.54 g/l/h. Maximum 1,3-propanediol yield of 41.1 g/l was achieved in a 5-l fermenter using the optimized medium. This makes the engineered strain have potential application in the conversion of glycerol to 1,3-propanediol on an industrial scale.