Engineering alternative butanol production platforms in heterologous bacteria
Engineering alternative butanol production platforms in heterologous bacteria
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DOI：
10.1016/j.ymben.2009.05.003
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发表时间：
2009-07-01
影响因子：
8.4
通讯作者：
Prather, Kristala L. Jones
中科院分区：
文献类型：
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作者：
Nielsen, David R.;Leonard, Effendi;Prather, Kristala L. Jones
替代微生物宿主已被设计为丁醇生物合成的生物催化剂。首先在大肠杆菌中重建了乙酰丁酸梭菌的丁醇合成途径，为与其他宿主进行比较奠定了基础。途径基因的多顺反子表达导致了34 mg/L丁醇的产生，而途径基因的单独表达使效价提高到200 mg/L，通过共表达酿酒酵母甲酸脱氢酶提高了效价，而过表达大肠杆菌甘油醛3-磷酸脱氢酶提高了糖酵解通量，使效价提高到580 mg/L，恶臭假单胞菌和枯草杆菌也被探索作为替代生产宿主。丁醇生物合成基因的多顺反子表达使恶臭假单胞菌和枯草杆菌的丁醇滴度分别达到120和24 mg/L。专性好氧菌恶臭假单胞菌的产量依赖于BCD-etfAB的表达。这些结果证明了在各种异源微生物中工程合成丁醇的潜力，包括那些在好氧条件下培养的微生物。(C)2009 Elsevier Inc.保留所有权利。
Alternative microbial hosts have been engineered as biocatalysts for butanol biosynthesis. The butanol synthetic pathway of Clostridium acetobutylicum was first re-constructed in Escherichia coli to establish a baseline for comparison to other hosts. Whereas polycistronic expression of the pathway genes resulted in the production of 34 mg/L butanol, individual expression of pathway genes elevated titers to 200 mg/L. Improved titers were achieved by co-expression of Saccharomyces cerevisiae formate dehydrogenase while overexpression of E. coli glyceraldehyde 3-phosphate dehydrogenase to elevate glycolytic flux improved titers to 580 mg/L. Pseudomonas putida and Bacillus subtilis were also explored as alternative production hosts. Polycistronic expression of butanol biosynthetic genes yielded butanol titers of 120 and 24 mg/L from P. putida and B. subtilis, respectively. Production in the obligate aerobe P. putida was dependent upon expression of bcd-etfAB. These results demonstrate the potential of engineering butanol biosynthesis in a variety of heterologous microorganisms, including those cultivated aerobically. (C) 2009 Elsevier Inc. All rights reserved.