Development of a highly efficient p-coumaric acid-responsive biosensor in Saccharomyces cerevisiae

Developing biosensors to monitor and regulate intracellular biosynthesis pathways can significantly enhance natural product yields in microbial cell factories. This study created a novel biosensor in Saccharomyces cerevisiae to respond to p-coumaric acid, a critical precursor in the biosynthesis of...

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Main Authors: Xueqing Pang, Jingyuan Zhu, Yuqing Li, Jing Xiao, Xinyu Zhang, Depeng Ren, Pingping Zhou
Format: Article
Language:English
Published: KeAi Communications Co., Ltd. 2025-12-01
Series:Synthetic and Systems Biotechnology
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Online Access:http://www.sciencedirect.com/science/article/pii/S2405805X25001048
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author Xueqing Pang
Jingyuan Zhu
Yuqing Li
Jing Xiao
Xinyu Zhang
Depeng Ren
Pingping Zhou
author_facet Xueqing Pang
Jingyuan Zhu
Yuqing Li
Jing Xiao
Xinyu Zhang
Depeng Ren
Pingping Zhou
author_sort Xueqing Pang
collection DOAJ
description Developing biosensors to monitor and regulate intracellular biosynthesis pathways can significantly enhance natural product yields in microbial cell factories. This study created a novel biosensor in Saccharomyces cerevisiae to respond to p-coumaric acid, a critical precursor in the biosynthesis of polyphenols and flavonoids. This biosensor was constructed by expressing the BsPadR repressor from Bacillus subtilis and engineering hybrid promoters. Notably, the PBS1-CCW12 hybrid promoter exhibited tight regulation by BsPadR and enhanced activity in response to p-coumaric acid. However, excessive BsPadR expression negatively impacted yeast growth, which was mitigated by using weaker promoters, PBST1 and PERG9. Furthermore, the impact of nuclear localization signal (SV40-NLS) positioning on BsPadR functionality was explored, revealing that fusion of an SV40-NLS at the C-terminus of BsPadR enhanced the biosensor's performance. To validate its utility, we applied this system to dynamically regulate CrtE (geranylgeranyl pyrophosphate synthase), a key enzyme in lycopene biosynthesis. By coupling p-coumaric acid production with lycopene biosynthesis, we enabled high-throughput colorimetric screening for enzyme evolution and strain selection. This novel biosensor serves as a valuable tool for future studies aimed at optimizing the production of p-coumaric acid and its derivatives in S. cerevisiae, thereby advancing the efficiency of biosynthetic processes in microbial cell factories.
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spelling doaj-art-4bd972570bc147869056f276c8efc81d2025-08-20T03:59:37ZengKeAi Communications Co., Ltd.Synthetic and Systems Biotechnology2405-805X2025-12-011041284129310.1016/j.synbio.2025.07.007Development of a highly efficient p-coumaric acid-responsive biosensor in Saccharomyces cerevisiaeXueqing Pang0Jingyuan Zhu1Yuqing Li2Jing Xiao3Xinyu Zhang4Depeng Ren5Pingping Zhou6College of Bioscience and Biotechnology, Yangzhou University, Yangzhou, 225009, PR ChinaCollege of Bioscience and Biotechnology, Yangzhou University, Yangzhou, 225009, PR ChinaCollege of Bioscience and Biotechnology, Yangzhou University, Yangzhou, 225009, PR ChinaCollege of Bioscience and Biotechnology, Yangzhou University, Yangzhou, 225009, PR ChinaCollege of Bioscience and Biotechnology, Yangzhou University, Yangzhou, 225009, PR ChinaCollege of Bioscience and Biotechnology, Yangzhou University, Yangzhou, 225009, PR ChinaCorresponding author.; College of Bioscience and Biotechnology, Yangzhou University, Yangzhou, 225009, PR ChinaDeveloping biosensors to monitor and regulate intracellular biosynthesis pathways can significantly enhance natural product yields in microbial cell factories. This study created a novel biosensor in Saccharomyces cerevisiae to respond to p-coumaric acid, a critical precursor in the biosynthesis of polyphenols and flavonoids. This biosensor was constructed by expressing the BsPadR repressor from Bacillus subtilis and engineering hybrid promoters. Notably, the PBS1-CCW12 hybrid promoter exhibited tight regulation by BsPadR and enhanced activity in response to p-coumaric acid. However, excessive BsPadR expression negatively impacted yeast growth, which was mitigated by using weaker promoters, PBST1 and PERG9. Furthermore, the impact of nuclear localization signal (SV40-NLS) positioning on BsPadR functionality was explored, revealing that fusion of an SV40-NLS at the C-terminus of BsPadR enhanced the biosensor's performance. To validate its utility, we applied this system to dynamically regulate CrtE (geranylgeranyl pyrophosphate synthase), a key enzyme in lycopene biosynthesis. By coupling p-coumaric acid production with lycopene biosynthesis, we enabled high-throughput colorimetric screening for enzyme evolution and strain selection. This novel biosensor serves as a valuable tool for future studies aimed at optimizing the production of p-coumaric acid and its derivatives in S. cerevisiae, thereby advancing the efficiency of biosynthetic processes in microbial cell factories.http://www.sciencedirect.com/science/article/pii/S2405805X25001048BiosensorSaccharomyces cerevisiaep-Coumaric acidBsPadR transcriptional repressorHybrid promoter
spellingShingle Xueqing Pang
Jingyuan Zhu
Yuqing Li
Jing Xiao
Xinyu Zhang
Depeng Ren
Pingping Zhou
Development of a highly efficient p-coumaric acid-responsive biosensor in Saccharomyces cerevisiae
Synthetic and Systems Biotechnology
Biosensor
Saccharomyces cerevisiae
p-Coumaric acid
BsPadR transcriptional repressor
Hybrid promoter
title Development of a highly efficient p-coumaric acid-responsive biosensor in Saccharomyces cerevisiae
title_full Development of a highly efficient p-coumaric acid-responsive biosensor in Saccharomyces cerevisiae
title_fullStr Development of a highly efficient p-coumaric acid-responsive biosensor in Saccharomyces cerevisiae
title_full_unstemmed Development of a highly efficient p-coumaric acid-responsive biosensor in Saccharomyces cerevisiae
title_short Development of a highly efficient p-coumaric acid-responsive biosensor in Saccharomyces cerevisiae
title_sort development of a highly efficient p coumaric acid responsive biosensor in saccharomyces cerevisiae
topic Biosensor
Saccharomyces cerevisiae
p-Coumaric acid
BsPadR transcriptional repressor
Hybrid promoter
url http://www.sciencedirect.com/science/article/pii/S2405805X25001048
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AT jingxiao developmentofahighlyefficientpcoumaricacidresponsivebiosensorinsaccharomycescerevisiae
AT xinyuzhang developmentofahighlyefficientpcoumaricacidresponsivebiosensorinsaccharomycescerevisiae
AT depengren developmentofahighlyefficientpcoumaricacidresponsivebiosensorinsaccharomycescerevisiae
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