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破伤风梭状芽孢杆菌丙酮解偶联丁醇生产的发酵及基因组分析

Fermentation and genomic analysis of acetone-uncoupled butanol production by Clostridium tetanomorphum.

作者信息

Gong Fuyu, Bao Guanhui, Zhao Chunhua, Zhang Yanping, Li Yin, Dong Hongjun

机构信息

Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, No.1 West Beichen Road, Chaoyang District, Beijing, China.

University of the Chinese Academy of Sciences, Beijing, China.

出版信息

Appl Microbiol Biotechnol. 2016 Feb;100(3):1523-1529. doi: 10.1007/s00253-015-7121-0. Epub 2015 Nov 17.

Abstract

In typical acetone-butanol-ethanol (ABE) fermentation, acetone is the main by-product (50 % of butanol mass) of butanol production, resulting in a low yield of butanol. It is known that some Clostridium tetanomorphum strains are able to produce butanol without acetone in nature. Here, we described that C. tetanomorphum strain DSM665 can produce 4.16 g/L butanol and 4.98 g/L ethanol at pH 6.0, and 9.81 g/L butanol and 1.01 g/L ethanol when adding 1 mM methyl viologen. Butyrate and acetate could be reassimilated and no acetone was produced. Further analysis indicated that the activity of the acetate/butyrate:acetoacetyl-CoA transferase responsible for acetone production is lost in C. tetanomorphum DSM665. The genome of C. tetanomorphum DSM665 was sequenced and deposited in DDBJ, EMBL, and GenBank under the accession no. APJS00000000. Sequence analysis indicated that there are no typical genes (ctfA/B and adc) that are typically parts of an acetone synthesis pathway in C. tetanomorphum DSM665. This work provides new insights in the mechanism of clostridial butanol production and should prove useful for the design of a high-butanol-producing strain.

摘要

在典型的丙酮-丁醇-乙醇(ABE)发酵中,丙酮是丁醇生产的主要副产物(占丁醇质量的50%),导致丁醇产量较低。已知一些破伤风梭状芽孢杆菌菌株在自然条件下能够生产丁醇而不产生丙酮。在此,我们描述了破伤风梭状芽孢杆菌DSM665菌株在pH 6.0时可产生4.16 g/L丁醇和4.98 g/L乙醇,添加1 mM甲基紫精时可产生9.81 g/L丁醇和1.01 g/L乙醇。丁酸和乙酸可被重新同化,且不产生丙酮。进一步分析表明,负责丙酮生产的乙酸/丁酸:乙酰乙酰辅酶A转移酶在破伤风梭状芽孢杆菌DSM665中失去活性。对破伤风梭状芽孢杆菌DSM665的基因组进行了测序,并以登录号APJS00000000保存在DDBJ、EMBL和GenBank中。序列分析表明,在破伤风梭状芽孢杆菌DSM665中没有典型的基因(ctfA/B和adc),这些基因通常是丙酮合成途径的组成部分。这项工作为梭菌丁醇生产机制提供了新的见解,对高产丁醇菌株的设计应具有重要意义。

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