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FAS 延伸循环基因表达与脂肪酸生成的相关性研究,以提高大肠杆菌中长链脂肪酸的产量。

Correlations between FAS elongation cycle genes expression and fatty acid production for improvement of long-chain fatty acids in Escherichia coli.

机构信息

Department of Chemical and Biomolecular Engineering, Sogang University, Seoul 121-742, Republic of Korea.

出版信息

Appl Biochem Biotechnol. 2013 Mar;169(5):1606-19. doi: 10.1007/s12010-012-0088-8. Epub 2013 Jan 17.

Abstract

Microorganisms have been used for biodiesel (fatty acid methyl ester) production due to their significant environmental and economic benefits. The aim of the present research was to develop new strains of Escherichia coli K-12 MG1655 and to increase the content of long-chain fatty acids by overexpressing essential enzymes that are involved in the fatty acid synthase elongation cycle. In addition, the relationship of β-ketoacyl-acyl carrier protein (ACP) synthase (fabH), β-ketoacyl-ACP reductase (fabG), β-hydroxyacyl-ACP dehydrase (fabZ), and β-enoyl-ACP reductase (fabI) with respect to fatty acid production was investigated. The four enzymes play a unique role in fatty acid biosynthesis and elongation processes. We report the generation of recombinant E. coli strains that produced long-chain fatty acids to amounts twofold over wild type. To verify the results, NAD(+)/NADH ratios and glucose analyses were performed. We also confirmed that FabZ plays an important role in producing unsaturated fatty acids (UFAs) as E. coli SGJS25 (overexpressing the fabZ gene) produced the highest percentage of UFAs (35 % of total long-chain fatty acids), over wild type and other recombinants. Indeed, cis-9-hexadecenoic acid, a major UFA in E. coli SGJS25, was produced at levels 20-fold higher than in wild type after 20 h in culture. The biochemically engineered E. coli presented in this study is expected to be more economical for producing long-chain fatty acids in quality biodiesel production processes.

摘要

由于微生物具有显著的环境和经济效益,因此被用于生物柴油(脂肪酸甲酯)的生产。本研究的目的是开发新的大肠杆菌 K-12 MG1655 菌株,并通过过表达参与脂肪酸合成延伸循环的必需酶来提高长链脂肪酸的含量。此外,还研究了β-酮酰-酰基载体蛋白(ACP)合酶(fabH)、β-酮酰-ACP 还原酶(fabG)、β-羟基酰-ACP 脱水酶(fabZ)和β-烯酰-ACP 还原酶(fabI)与脂肪酸生成的关系。这四种酶在脂肪酸生物合成和延伸过程中发挥着独特的作用。我们报告了产生能够产生两倍于野生型量的长链脂肪酸的重组大肠杆菌菌株。为了验证结果,进行了 NAD(+) / NADH 比值和葡萄糖分析。我们还证实 FabZ 在产生不饱和脂肪酸(UFAs)方面起着重要作用,因为大肠杆菌 SGJS25(过表达 fabZ 基因)产生的 UFAs 百分比最高(占总长链脂肪酸的 35%),高于野生型和其他重组体。事实上,在培养 20 小时后,大肠杆菌 SGJS25 中主要的 UFA 顺-9-十六烯酸的产量比野生型高 20 倍。本研究中生物化学工程改造的大肠杆菌有望在质量更好的生物柴油生产过程中更经济地生产长链脂肪酸。

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