GENETIC-IMPROVEMENT OF ESCHERICHIA-COLI FOR ETHANOL-PRODUCTION - CHROMOSOMAL INTEGRATION OF ZYMOMONAS-MOBILIS GENES ENCODING PYRUVATE DECARBOXYLASE AND ALCOHOL DEHYDROGENASE-II
GENETIC-IMPROVEMENT OF ESCHERICHIA-COLI FOR ETHANOL-PRODUCTION - CHROMOSOMAL INTEGRATION OF ZYMOMONAS-MOBILIS GENES ENCODING PYRUVATE DECARBOXYLASE AND ALCOHOL DEHYDROGENASE-II
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DOI：
10.1128/aem.57.4.893-900.1991
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发表时间：
1991-04-01
影响因子：
4.4
通讯作者：
INGRAM, LO
中科院分区：
文献类型：
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作者：
OHTA, K;BEALL, DS;INGRAM, LO
将丙酮酸脱羧酶 (pdc) 和乙醇脱氢酶 II (adhB) 的运动发酵单胞菌基因整合到丙酮酸甲酸裂解酶基因 (pfl) 内或附近的大肠杆菌染色体中。 整合提高了运动发酵单胞菌基因在大肠杆菌中的稳定性，但需要进一步选择以增加表达。 选择对高水平氯霉素具有抗性的自发突变体，该突变体也表达高水平运动发酵单胞菌基因。 选择类似的突变体以增加醛指示板上乙醇脱氢酶的表达。 这些突变体在功能上与之前用于将木糖和葡萄糖发酵成乙醇的基于质粒的菌株相同。 从 10% 葡萄糖和 8% 木糖中分别获得 54.4 克/升和 41.6 克/升的乙醇浓度。 由于复杂营养素的分解代谢额外产生了乙醇，因此基于添加糖的转化效率超出了理论极限（0.51 克乙醇/克糖）。 引入进一步的突变来灭活琥珀酸产生（frd）并阻止同源重组（recA）。
Zymomonas mobilis genes for pyruvate decarboxylase (pdc) and alcohol dehydrogenase II (adhB) were integrated into the Escherichia coli chromosome within or near the pyruvate formate-lyase gene (pfl). Integration improved the stability of the Z. mobilis genes in E. coli, but further selection was required to increase expression. Spontaneous mutants were selected for resistance to high levels of chloramphenicol that also expressed high levels of the Z. mobilis genes. Analogous mutants were selected for increased expression of alcohol dehydrogenase on aldehyde indicator plates. These mutants were functionally equivalent to the previous plasmid-based strains for the fermentation of xylose and glucose to ethanol. Ethanol concentrations of 54.4 and 41.6 g/liter were obtained from 10% glucose and 8% xylose, respectively. The efficiency of conversion exceeded theoretical limits (0.51 g of ethanol/g of sugar) on the basis of added sugars because of the additional production of ethanol from the catabolism of complex nutrients. Further mutations were introduced to inactivate succinate production (frd) and to block homologous recombination (recA).