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异丙醇生产 天然质粒pCA复制子

isopropanol production native plasmid pCA replicon.

作者信息

Nogle Robert, Nagaraju Shilpa, Utturkar Sagar M, Giannone Richard J, Reynoso Vinicio, Leang Ching, Hettich Robert L, Mitchell Wayne P, Simpson Sean D, Jewett Michael C, Köpke Michael, Brown Steven D

机构信息

LanzaTech Inc., Skokie, IL, United States.

Center for Cancer Research, Purdue University, West Lafayette, IN, United States.

出版信息

Front Bioeng Biotechnol. 2022 Aug 5;10:932363. doi: 10.3389/fbioe.2022.932363. eCollection 2022.

Abstract

is a model gas-fermenting acetogen for commercial ethanol production. It is also a platform organism being developed for the carbon-negative production of acetone and isopropanol by gas fermentation. We have assembled a 5.5 kb pCA plasmid for type strain DSM10061 (JA1-1) using three genome sequence datasets. pCA is predicted to encode seven open-reading frames and estimated to be a low-copy number plasmid present at approximately 12 copies per chromosome. RNA-seq analyses indicate that pCA genes are transcribed at low levels and two proteins, CAETHG_05090 (putative replication protein) and CAETHG_05115 (hypothetical, a possible Mob protein), were detected at low levels during batch gas fermentations. Thiolase (), CoA-transferase (), and acetoacetate decarboxylase () genes were introduced into a vector for isopropanol production in using the native plasmid origin of replication. The availability of the pCA sequence will facilitate studies into its physiological role and could form the basis for genetic tool optimization.

摘要

是用于商业乙醇生产的典型气体发酵产乙酸菌。它也是一种正在开发的平台生物,用于通过气体发酵以碳负方式生产丙酮和异丙醇。我们利用三个基因组序列数据集为模式菌株DSM10061(JA1-1)组装了一个5.5 kb的pCA质粒。pCA预计编码七个开放阅读框,估计是一种低拷贝数质粒,每条染色体上约有12个拷贝。RNA测序分析表明,pCA基因转录水平较低,在分批气体发酵过程中检测到两种蛋白质,CAETHG_05090(假定的复制蛋白)和CAETHG_05115(假定的,可能是Mob蛋白)水平较低。硫解酶()、辅酶A转移酶()和乙酰乙酸脱羧酶()基因被引入到一个载体中,用于在利用天然质粒复制起点生产异丙醇。pCA序列的可用性将有助于对其生理作用的研究,并可为遗传工具优化奠定基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc1a/9413188/0dc8ccd3b991/fbioe-10-932363-g001.jpg

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