Identification of a sensor histidine kinase (BfcK) controlling biofilm formation in Clostridium acetobutylicum

doi:10.1016/j.cjche.2021.04.024

Chinese Journal of Chemical Engineering ›› 2022, Vol. 46 ›› Issue (6): 84-93.DOI: 10.1016/j.cjche.2021.04.024

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Identification of a sensor histidine kinase (BfcK) controlling biofilm formation in Clostridium acetobutylicum

Dong Liu^1,2, Shikai Ge¹, Zhenyu Wang¹, Mengting Li², Wei Zhuang¹, Pengpeng Yang¹, Yong Chen^1,2, Hanjie Ying^1,2

1 State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211816, China;
2 School of Chemical Engineering and Energy, Zhengzhou University, Zhengzhou 450001, China

Received:2021-01-27 Revised:2021-03-23 Online:2022-07-20 Published:2022-06-28
Contact: Yong Chen,E-mail:chenyong1982@njtech.edu.cn;Hanjie Ying,E-mail:Yinghanjie@njtech.edu.cn
Supported by:
The authors thank Prof. Nigel P. Minton from the University of Nottingham for kindly providing the ClosTron plasmids, and Prof. Sheng Yang from Shanghai Institutes for Biological Sciences for kindly providing pSY8 plasmid. This work was supported by the Key Program of the National Natural Science Foundation of China (Grant No. 21636003), the Outstanding Youth Foundation of Jiangsu (Grant No. SBK2017010373), the National Key Research and Development Program of China (Grant No. 2019YFD1101204), and the Jiangsu Synergetic Innovation Center for Advanced Bio-Manufacture. Dong Liu is supported by the Jiangsu Qinglan Talent Program.

Identification of a sensor histidine kinase (BfcK) controlling biofilm formation in Clostridium acetobutylicum

Dong Liu^1,2, Shikai Ge¹, Zhenyu Wang¹, Mengting Li², Wei Zhuang¹, Pengpeng Yang¹, Yong Chen^1,2, Hanjie Ying^1,2

1 State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211816, China;
2 School of Chemical Engineering and Energy, Zhengzhou University, Zhengzhou 450001, China

通讯作者: Yong Chen,E-mail:chenyong1982@njtech.edu.cn;Hanjie Ying,E-mail:Yinghanjie@njtech.edu.cn
基金资助:
The authors thank Prof. Nigel P. Minton from the University of Nottingham for kindly providing the ClosTron plasmids, and Prof. Sheng Yang from Shanghai Institutes for Biological Sciences for kindly providing pSY8 plasmid. This work was supported by the Key Program of the National Natural Science Foundation of China (Grant No. 21636003), the Outstanding Youth Foundation of Jiangsu (Grant No. SBK2017010373), the National Key Research and Development Program of China (Grant No. 2019YFD1101204), and the Jiangsu Synergetic Innovation Center for Advanced Bio-Manufacture. Dong Liu is supported by the Jiangsu Qinglan Talent Program.

Abstract

Abstract: Clostridium acetobutylicum has been extensively exploited to produce biofuels and solvents and its biofilm could dramatically improve the productivities. However, genetic control of C. acetobutylicum biofilm has not been dissected so far. Here, to identify potential genes controlling C. acetobutylicum biofilm formation, over 40 gene candidates associated with extracellular matrix, cell surface, cell signaling or gene transcription, were tried to be disrupted to examine their individual impact. A total of 25 disruptants were finally obtained over years of attempts, for which biofilm and relevant phenotypes were characterized. Most of these disruptants formed robust biofilm still, or suffered both growth and biofilm defect. Only a strain with a disrupted histidine kinase gene (CA_C2730, designated bfcK in this study) abolished biofilm formation without impairing cell growth or solvent production. Further analysis revealed that bfcK could control flagellar biogenesis and cell motility at protein levels. The bfcK also appeared to repress the phosphorylation of a serine/threonine protein kinase (encoded by CA_C0404) that might negatively regulate biofilm formation. Based on these findings, possible bfcK-mediated mechanisms for biofilm formation were proposed. This is a big step toward understanding the biofilm formation in C. acetobutylicum and will help further engineering of its biofilm-based industrial processes.

Key words: Clostridium acetobutylicum, Biofilm, Histidine kinases, CA_C2730, Phosphoproteomics, Flagellar motility

摘要： Clostridium acetobutylicum has been extensively exploited to produce biofuels and solvents and its biofilm could dramatically improve the productivities. However, genetic control of C. acetobutylicum biofilm has not been dissected so far. Here, to identify potential genes controlling C. acetobutylicum biofilm formation, over 40 gene candidates associated with extracellular matrix, cell surface, cell signaling or gene transcription, were tried to be disrupted to examine their individual impact. A total of 25 disruptants were finally obtained over years of attempts, for which biofilm and relevant phenotypes were characterized. Most of these disruptants formed robust biofilm still, or suffered both growth and biofilm defect. Only a strain with a disrupted histidine kinase gene (CA_C2730, designated bfcK in this study) abolished biofilm formation without impairing cell growth or solvent production. Further analysis revealed that bfcK could control flagellar biogenesis and cell motility at protein levels. The bfcK also appeared to repress the phosphorylation of a serine/threonine protein kinase (encoded by CA_C0404) that might negatively regulate biofilm formation. Based on these findings, possible bfcK-mediated mechanisms for biofilm formation were proposed. This is a big step toward understanding the biofilm formation in C. acetobutylicum and will help further engineering of its biofilm-based industrial processes.

关键词: Clostridium acetobutylicum, Biofilm, Histidine kinases, CA_C2730, Phosphoproteomics, Flagellar motility

Dong Liu, Shikai Ge, Zhenyu Wang, Mengting Li, Wei Zhuang, Pengpeng Yang, Yong Chen, Hanjie Ying. Identification of a sensor histidine kinase (BfcK) controlling biofilm formation in Clostridium acetobutylicum[J]. Chinese Journal of Chemical Engineering, 2022, 46(6): 84-93.

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Identification of a sensor histidine kinase (BfcK) controlling biofilm formation in Clostridium acetobutylicum

Identification of a sensor histidine kinase (BfcK) controlling biofilm formation in Clostridium acetobutylicum

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