在大肠杆菌 B (BL21) 的 ackA (-) pta (-) poxB (-) 三重突变体中，通过替代代谢途径积累醋酸盐。
Acetate accumulation through alternative metabolic pathways in ackA (-) pta (-) poxB (-) triple mutant in E. coli B (BL21).
机构信息
Biotechnology Core Laboratory, NIDDK NIH Bethesda, Bldg 14A Room 173, Bethesda, MD 20892, USA.
出版信息
Biotechnol Lett. 2010 Dec;32(12):1897-903. doi: 10.1007/s10529-010-0369-7. Epub 2010 Aug 12.
Abstract
Individual deletions of acs and aceA genes in E. coli B (BL21) showed little difference in the metabolite accumulation patterns but deletion of the ackA gene alone or together with pta showed acetic acid gradually accumulated to 3.1 and 1.7 g/l, respectively, with a minimal extended lag in bacterial growth and a higher pyruvate formation. Single poxB deletion in E. coli B (BL21) or additional poxB deletion in the ackA-pta mutants did not change the acetate accumulation pattern. When the acetate production genes (ackA-pta-poxB) were deleted in E. coli B (BL21) acetate still accumulated. This may be an indication that perhaps acetate is not only a by-product of carbon metabolism; it is possible that acetate plays also a role in other cellular metabolite pathways. It is likely that there are alternative acetate production pathways.
摘要
单个缺失 E. coli B (BL21) 中的 acs 和 aceA 基因在代谢产物积累模式上几乎没有差异，但单独缺失 ackA 基因或同时缺失 ackA 和 pta 基因会导致乙酸逐渐积累到 3.1 和 1.7 g/L，细菌生长的延滞期最小，丙酮酸的形成量更高。在 E. coli B (BL21) 中单独缺失 poxB 基因或在 ackA-pta 突变体中进一步缺失 poxB 基因不会改变乙酸的积累模式。当 E. coli B (BL21) 中的乙酸生成基因 (ackA-pta-poxB) 被缺失时，乙酸仍然会积累。这可能表明，乙酸不仅是碳代谢的副产物；它也可能在其他细胞代谢物途径中发挥作用。很可能存在替代的乙酸生成途径。
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本文引用的文献
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Acetate consumption activity directly determines the level of acetate accumulation during Escherichia coli W3110 growth.
J Microbiol Biotechnol. 2009 Oct;19(10):1127-34. doi: 10.4014/jmb.0902.0097.
2
Engineering Escherichia coli for an efficient aerobic fermentation platform.
J Biotechnol. 2009 Oct 12;144(1):58-63. doi: 10.1016/j.jbiotec.2009.06.021. Epub 2009 Jun 27.
3
Minimizing acetate formation in E. coli fermentations.
J Ind Microbiol Biotechnol. 2007 Nov;34(11):689-700. doi: 10.1007/s10295-007-0244-2. Epub 2007 Aug 1.
4
Free CoA-mediated regulation of intermediary and central metabolism: an hypothesis which accounts for the excretion of alpha-ketoglutarate during aerobic growth of Escherichia coli on acetate.
Res Microbiol. 2005 Sep;156(8):874-9. doi: 10.1016/j.resmic.2005.04.008. Epub 2005 Jun 16.
5
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Biotechnol Prog. 2005 Jul-Aug;21(4):1062-7. doi: 10.1021/bp050073s.
6
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Biotechnol Bioeng. 2005 Jun 30;90(7):805-20. doi: 10.1002/bit.20478.
7
Pyruvate oxidase contributes to the aerobic growth efficiency of Escherichia coli.
Microbiology (Reading). 2001 Jun;147(Pt 6):1483-1498. doi: 10.1099/00221287-147-6-1483.
8
Metabolic flux analysis of Escherichia coli deficient in the acetate production pathway and expressing the Bacillus subtilis acetolactate synthase.
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9
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