Engineering the TetR-family transcriptional regulator XNR_0706 to enhance heterologous spinosad production in Streptomyces albus B4 chassis

The TetR family of regulators are an important group of transcription regulators that regulate diverse cellular processes in prokaryotes. In this study, we found that XNR_0706, a TetR family regulator, controlled the expression of XNR_0345, XNR_0454, XNR_0513 and XNR_1438 putatively involved in fatt...

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Main Authors: Xingjun Cui, Hao Tang, Wenzong Wang, Wenping Wei, Jing Wu, Bang-Ce Ye
Format: Article
Language:English
Published: KeAi Communications Co., Ltd. 2025-03-01
Series:Synthetic and Systems Biotechnology
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Online Access:http://www.sciencedirect.com/science/article/pii/S2405805X24001194
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author Xingjun Cui
Hao Tang
Wenzong Wang
Wenping Wei
Jing Wu
Bang-Ce Ye
author_facet Xingjun Cui
Hao Tang
Wenzong Wang
Wenping Wei
Jing Wu
Bang-Ce Ye
author_sort Xingjun Cui
collection DOAJ
description The TetR family of regulators are an important group of transcription regulators that regulate diverse cellular processes in prokaryotes. In this study, we found that XNR_0706, a TetR family regulator, controlled the expression of XNR_0345, XNR_0454, XNR_0513 and XNR_1438 putatively involved in fatty acid β-oxidation by interacting with the promoter regions in Streptomyces albus B4. The transcription level of these four genes was downregulated in XNR_0706 deletion strain (ΔXNR_0706) and restored by XNR_0706 complementation in Δ0706/pIB-0706, demonstrating that XNR_0706 was a positive transcriptional regulator of the genes. With toxic long-chain fatty acids addition in TSB media, deletion of XNR_0706 caused significantly poor growth, whereas XNR_0706 complementation increased the utilization of additional fatty acids, resulting in restored growth. Fatty acid β-oxidation is one source of acetyl- and malonyl-CoA precursors for polyketides biosynthesis in actinobacteria. Overexpression of XNR_0706 in B4/spnNEW, a spinosad heterologous expression strain derived from S. albus B4, increased spinosad yield by 20.6 %. Additionally, supplement of 0.3 g/L fatty acids resulted in a further 42.4 % increase in spinosad yield. Our study reveals a regulatory mechanism in long-chain fatty acids metabolism in S. albus and these insights into the molecular regulation of β-oxidation by XNR_0706 are instrumental for increasing secondary metabolites in actinobacteria.
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spelling doaj-art-417adf8da46d4c0396733f66bcb4586c2025-08-20T02:12:03ZengKeAi Communications Co., Ltd.Synthetic and Systems Biotechnology2405-805X2025-03-0110121822510.1016/j.synbio.2024.08.008Engineering the TetR-family transcriptional regulator XNR_0706 to enhance heterologous spinosad production in Streptomyces albus B4 chassisXingjun Cui0Hao Tang1Wenzong Wang2Wenping Wei3Jing Wu4Bang-Ce Ye5Institute of Engineering Biology and Health, Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, 310014, ChinaInstitute of Engineering Biology and Health, Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, 310014, ChinaInstitute of Engineering Biology and Health, Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, 310014, ChinaInstitute of Engineering Biology and Health, Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, 310014, ChinaInstitute of Engineering Biology and Health, Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, 310014, ChinaInstitute of Engineering Biology and Health, Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, 310014, China; State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China; Corresponding author. Institute of Engineering Biology and Health, Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, 310014, China.The TetR family of regulators are an important group of transcription regulators that regulate diverse cellular processes in prokaryotes. In this study, we found that XNR_0706, a TetR family regulator, controlled the expression of XNR_0345, XNR_0454, XNR_0513 and XNR_1438 putatively involved in fatty acid β-oxidation by interacting with the promoter regions in Streptomyces albus B4. The transcription level of these four genes was downregulated in XNR_0706 deletion strain (ΔXNR_0706) and restored by XNR_0706 complementation in Δ0706/pIB-0706, demonstrating that XNR_0706 was a positive transcriptional regulator of the genes. With toxic long-chain fatty acids addition in TSB media, deletion of XNR_0706 caused significantly poor growth, whereas XNR_0706 complementation increased the utilization of additional fatty acids, resulting in restored growth. Fatty acid β-oxidation is one source of acetyl- and malonyl-CoA precursors for polyketides biosynthesis in actinobacteria. Overexpression of XNR_0706 in B4/spnNEW, a spinosad heterologous expression strain derived from S. albus B4, increased spinosad yield by 20.6 %. Additionally, supplement of 0.3 g/L fatty acids resulted in a further 42.4 % increase in spinosad yield. Our study reveals a regulatory mechanism in long-chain fatty acids metabolism in S. albus and these insights into the molecular regulation of β-oxidation by XNR_0706 are instrumental for increasing secondary metabolites in actinobacteria.http://www.sciencedirect.com/science/article/pii/S2405805X24001194Streptomyces albus B4TetR family transcriptional regulatorSpinosadPrecursor supplyFatty acids metabolismHeterologous expression
spellingShingle Xingjun Cui
Hao Tang
Wenzong Wang
Wenping Wei
Jing Wu
Bang-Ce Ye
Engineering the TetR-family transcriptional regulator XNR_0706 to enhance heterologous spinosad production in Streptomyces albus B4 chassis
Synthetic and Systems Biotechnology
Streptomyces albus B4
TetR family transcriptional regulator
Spinosad
Precursor supply
Fatty acids metabolism
Heterologous expression
title Engineering the TetR-family transcriptional regulator XNR_0706 to enhance heterologous spinosad production in Streptomyces albus B4 chassis
title_full Engineering the TetR-family transcriptional regulator XNR_0706 to enhance heterologous spinosad production in Streptomyces albus B4 chassis
title_fullStr Engineering the TetR-family transcriptional regulator XNR_0706 to enhance heterologous spinosad production in Streptomyces albus B4 chassis
title_full_unstemmed Engineering the TetR-family transcriptional regulator XNR_0706 to enhance heterologous spinosad production in Streptomyces albus B4 chassis
title_short Engineering the TetR-family transcriptional regulator XNR_0706 to enhance heterologous spinosad production in Streptomyces albus B4 chassis
title_sort engineering the tetr family transcriptional regulator xnr 0706 to enhance heterologous spinosad production in streptomyces albus b4 chassis
topic Streptomyces albus B4
TetR family transcriptional regulator
Spinosad
Precursor supply
Fatty acids metabolism
Heterologous expression
url http://www.sciencedirect.com/science/article/pii/S2405805X24001194
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