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参与红平红球菌N9T-4贫营养生长的甲基营养型氧化还原酶的基因表达分析

Gene expression analysis of methylotrophic oxidoreductases involved in the oligotrophic growth of Rhodococcus erythropolis N9T-4.

作者信息

Yoshida Nobuyuki, Hayasaki Takuya, Takagi Hiroshi

机构信息

Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma 630-0192, Japan.

出版信息

Biosci Biotechnol Biochem. 2011;75(1):123-7. doi: 10.1271/bbb.100700. Epub 2011 Jan 7.

Abstract

Rhodococcus erythropolis N9T-4 shows extremely oligotrophic growth requiring atmospheric CO₂ without any additional carbon or energy source. We performed a gene expression analysis of the oxidoreductases, which are involved in methanol metabolism, under various growth and induction conditions in N9T-4. A real-time PCR analysis revealed that the genes encoding NAD-dependent formaldehyde dehydrogenase (nFADH) and N,N'-dimethyl-4-nitrosoaniline-dependent methanol dehydrogenase (MDH) were strongly expressed under the oligotrophic conditions at levels of 20-100 fold those under heterotrophic conditions, in which n-tetradecane was used as the sole carbon source, while glucose did not affect the gene expression pattern in a minimum medium. The genes encoding mycothiol-dependent formaldehyde dehydrogenase (mFADH) and formate dehydrogenase were not induced under oligotrophic conditions, although mFADH expression was observed when formaldehyde was added to the induction medium. These results suggest that N9T-4 had three distinct formaldehyde oxidation systems, and that MDH and nFADH were the key enzymes in its oligotrophic growth.

摘要

红平红球菌N9T-4表现出极低营养生长特性,需要大气中的二氧化碳,无需任何额外的碳源或能源。我们在N9T-4的各种生长和诱导条件下,对参与甲醇代谢的氧化还原酶进行了基因表达分析。实时PCR分析表明,编码NAD依赖性甲醛脱氢酶(nFADH)和N,N'-二甲基-4-亚硝基苯胺依赖性甲醇脱氢酶(MDH)的基因在贫营养条件下强烈表达,其表达水平是在以正十四烷作为唯一碳源的异养条件下的20至100倍,而葡萄糖在基本培养基中不影响基因表达模式。编码巯基乙醇依赖性甲醛脱氢酶(mFADH)和甲酸脱氢酶的基因在贫营养条件下未被诱导,尽管当甲醛添加到诱导培养基中时观察到mFADH表达。这些结果表明,N9T-4具有三种不同的甲醛氧化系统,并且MDH和nFADH是其贫营养生长中的关键酶。

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