Corynebacterium glutamicum for Microbial Production of Chitin Oligosaccharides Using Modular Engineering

ABSTRACT Microbial fermentation is a potent strategy for eco‐friendly and sustainable chitin oligosaccharide (CHOS) production. Nonetheless, hurdles (e.g., imbalanced metabolic flow and the need for uridine diphosphate (UDP)‐sugar donor consumption in CHOS synthesis) hinder enhanced and efficient pr...

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Main Authors: Chen Deng, Ruijie Xin, Xingjian Li, Liqiang Fan, Yongjun Qiu, Liming Zhao
Format: Article
Language:English
Published: Wiley 2025-06-01
Series:Food Bioengineering
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Online Access:https://doi.org/10.1002/fbe2.70012
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author Chen Deng
Ruijie Xin
Xingjian Li
Liqiang Fan
Yongjun Qiu
Liming Zhao
author_facet Chen Deng
Ruijie Xin
Xingjian Li
Liqiang Fan
Yongjun Qiu
Liming Zhao
author_sort Chen Deng
collection DOAJ
description ABSTRACT Microbial fermentation is a potent strategy for eco‐friendly and sustainable chitin oligosaccharide (CHOS) production. Nonetheless, hurdles (e.g., imbalanced metabolic flow and the need for uridine diphosphate (UDP)‐sugar donor consumption in CHOS synthesis) hinder enhanced and efficient production. In this study, we aimed to use Corynebacterium glutamicum as the foundational organism for de novo CHOS synthesis. Initially, we developed the CHOS synthesis pathway in C. glutamicum, attaining a CHOS titer of 113.34 mg/L. Furthermore, we fortified the uridine 5′‐diphospho‐N‐acetylglucosamine (UDP‐GlcNAc) synthesis module, vital for CHOS and other functional sugar synthesis, and developed a system for regenerating uridine triphosphate (UTP) precursors. Finally, we performed C. glutamicum‐mediated scale‐up CHOS production in a 5‐L bioreactor yielding a titer of 5.08 g/L. The CHOS chassis strain provides a robust foundation for mass CHOS production via metabolic engineering. Altering the intracellular UDP‐sugar donor creation pathway could reportedly significantly enhance CHOS production. We attained the peak concentration of 829.33 mg/L with the heightened expression of glmM, glmU, and the metabolic equilibrium of PCM1 and AmgK. Bacterial growth remained unaffected by the excessive gene expressions or external gene incorporations. In addition, the swift growth and C. glutamicum accumulation in the fermenter led to increased CHOS production, reaching a titer of 5.08 g/L from the recombinant strain CGSL63, being 4.43 times higher than in the case of shake flask fermentation. The engineering strategies used in this study might be helpful for the C. glutamicum‐mediated microbial synthesis of functional sugars. The methods applied in this study are broadly applicable for boosting the microbial generation of other valuable functional sugars.
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spelling doaj-art-91dd01b7c28148a1a152c78aedb1d0c32025-08-20T03:31:31ZengWileyFood Bioengineering2770-20812025-06-014212714110.1002/fbe2.70012Corynebacterium glutamicum for Microbial Production of Chitin Oligosaccharides Using Modular EngineeringChen Deng0Ruijie Xin1Xingjian Li2Liqiang Fan3Yongjun Qiu4Liming Zhao5State Key Laboratory of Bioreactor Engineering East China University of Science and Technology Shanghai ChinaState Key Laboratory of Bioreactor Engineering East China University of Science and Technology Shanghai ChinaState Key Laboratory of Bioreactor Engineering East China University of Science and Technology Shanghai ChinaState Key Laboratory of Bioreactor Engineering East China University of Science and Technology Shanghai ChinaState Key Laboratory of Bioreactor Engineering East China University of Science and Technology Shanghai ChinaState Key Laboratory of Bioreactor Engineering East China University of Science and Technology Shanghai ChinaABSTRACT Microbial fermentation is a potent strategy for eco‐friendly and sustainable chitin oligosaccharide (CHOS) production. Nonetheless, hurdles (e.g., imbalanced metabolic flow and the need for uridine diphosphate (UDP)‐sugar donor consumption in CHOS synthesis) hinder enhanced and efficient production. In this study, we aimed to use Corynebacterium glutamicum as the foundational organism for de novo CHOS synthesis. Initially, we developed the CHOS synthesis pathway in C. glutamicum, attaining a CHOS titer of 113.34 mg/L. Furthermore, we fortified the uridine 5′‐diphospho‐N‐acetylglucosamine (UDP‐GlcNAc) synthesis module, vital for CHOS and other functional sugar synthesis, and developed a system for regenerating uridine triphosphate (UTP) precursors. Finally, we performed C. glutamicum‐mediated scale‐up CHOS production in a 5‐L bioreactor yielding a titer of 5.08 g/L. The CHOS chassis strain provides a robust foundation for mass CHOS production via metabolic engineering. Altering the intracellular UDP‐sugar donor creation pathway could reportedly significantly enhance CHOS production. We attained the peak concentration of 829.33 mg/L with the heightened expression of glmM, glmU, and the metabolic equilibrium of PCM1 and AmgK. Bacterial growth remained unaffected by the excessive gene expressions or external gene incorporations. In addition, the swift growth and C. glutamicum accumulation in the fermenter led to increased CHOS production, reaching a titer of 5.08 g/L from the recombinant strain CGSL63, being 4.43 times higher than in the case of shake flask fermentation. The engineering strategies used in this study might be helpful for the C. glutamicum‐mediated microbial synthesis of functional sugars. The methods applied in this study are broadly applicable for boosting the microbial generation of other valuable functional sugars.https://doi.org/10.1002/fbe2.70012chitin oligosaccharidesCorynebacterium glutamicummetabolic engineeringsynthetic biologyUDP‐sugar donor
spellingShingle Chen Deng
Ruijie Xin
Xingjian Li
Liqiang Fan
Yongjun Qiu
Liming Zhao
Corynebacterium glutamicum for Microbial Production of Chitin Oligosaccharides Using Modular Engineering
Food Bioengineering
chitin oligosaccharides
Corynebacterium glutamicum
metabolic engineering
synthetic biology
UDP‐sugar donor
title Corynebacterium glutamicum for Microbial Production of Chitin Oligosaccharides Using Modular Engineering
title_full Corynebacterium glutamicum for Microbial Production of Chitin Oligosaccharides Using Modular Engineering
title_fullStr Corynebacterium glutamicum for Microbial Production of Chitin Oligosaccharides Using Modular Engineering
title_full_unstemmed Corynebacterium glutamicum for Microbial Production of Chitin Oligosaccharides Using Modular Engineering
title_short Corynebacterium glutamicum for Microbial Production of Chitin Oligosaccharides Using Modular Engineering
title_sort corynebacterium glutamicum for microbial production of chitin oligosaccharides using modular engineering
topic chitin oligosaccharides
Corynebacterium glutamicum
metabolic engineering
synthetic biology
UDP‐sugar donor
url https://doi.org/10.1002/fbe2.70012
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