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Function Enhancement of a Metabolic Module via Endogenous Promoter Replacement for Pseudomonas sp. JY-Q to Degrade Nicotine in Tobacco Waste Treatment

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Abstract

Nicotine-degrading Pseudomonas sp. JY-Q is a preferred strain utilized in reconstituted tobacco process for tobacco waste treatment. However, its efficiency of nicotine metabolism still requires to be improved via genomic technology such as promoter engineering based on genomic information. Concerning upstream module of nicotine metabolic pathway, we found that two homologous genes of nicotine dehydrogenase (nicA2 and nox) coexisted in strain JY-Q. However, the transcriptional amount of nox was 20-fold higher than that of nicA2. Thus, the nicA2 expression required improvement. Combinatorial displacement was accomplished for two predicted endogenous promoters, named as PnicA2 and Pnox for nicA2 and nox, respectively. The mutant with Pnox as the promoters for both nicA2 and nox exhibited the best nicotine metabolic capacity which increased by 66% compared to the wild type. These results suggested that endogenous promoter replacement is also feasible for function improvement of metabolic modules and strain enhancement of biodegradation capacity to meet real environment demand.

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Availability of Data and Materials

GenBank accession number of strain JY-Q is CP011525. Data and materials supporting this study have been provided in the text.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Funding

This work was financially supported by grants from the National Natural Science Foundation of China (31970104, 31800118, 21938012, 31670115).

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Author notes

  1. Jun Li and Fengmei Yi contributed equally to this work.

Authors and Affiliations

  1. College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, 310032, China

    Jun Li, Fengmei Yi, Guoqing Chen, Zeyu Chen, Zeling Zhang, Xiaotong Mei & Weihong Zhong

  2. Technology Center, China Tobacco Zhejiang Industrial Co., Ltd., Hangzhou, 310009, China

    Fanda Pan, Yang Yang & Ming Shu

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  1. Jun Li
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Contributions

Jun Li: Conceptualization, resources, data curation, software, formal analysis, investigation, methodology, writing—original draft, project administration, writing—review and editing. Fengmei Yi: Data curation, software, formal analysis, writing—original draft, visualization, methodology. Guoqing Chen, Zeyu Chen, Zeling Zhang, Xiaotong Mei: Software, investigation. Fanda Pan, Yang Yang, Ming Shu: Investigation, methodology. Weihong Zhong: Conceptualization, resources, supervision, funding acquisition, investigation, methodology, project administration, writing—review and editing.

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Correspondence to Yang Yang or Weihong Zhong.

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Li, J., Yi, F., Chen, G. et al. Function Enhancement of a Metabolic Module via Endogenous Promoter Replacement for Pseudomonas sp. JY-Q to Degrade Nicotine in Tobacco Waste Treatment. Appl Biochem Biotechnol 193, 2793–2805 (2021). https://doi.org/10.1007/s12010-021-03566-0

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