Research advances in biosynthesis of chondroitin sulfate and its oligosaccharides

doi:10.12211/2096-8280.2025-051

Synthetic Biology Journal

Research advances in biosynthesis of chondroitin sulfate and its oligosaccharides

ZHANG Jin¹^,²^,³, ZHANG Weijiao¹^,²^,³, XIONG Haibo¹^,²^,³, XIE Zhuan¹^,²^,³, XU Ruirui¹^,²^,³, KANG Zhen¹^,²^,³^,⁴

^1.Key Laboratory of Carbohydrate Chemistry and Biotechnology，Ministry of Education，School of Biotechnology，Jiangnan University，Wuxi 214122，Jiangsu，China
^2.Science Center for Future Foods，Jiangnan University，Wuxi 214122，Jiangsu，China
^3.Key Laboratory of Industrial Biotechnology，Ministry of Education，School of Biotechnology，Jiangnan University，Wuxi 214122，Jiangsu，China
^4.Jiangsu Province Basic Research Center for Synthetic Biology，Jiangnan University，Wuxi 214122，Jiangsu，China

Received:2025-05-29 Revised:2025-08-13 Published:2025-08-16
Contact: KANG Zhen

硫酸软骨素及其寡聚糖的生物合成进展

张瑾¹^,²^,³, 张维娇¹^,²^,³, 熊海波¹^,²^,³, 谢专¹^,²^,³, 胥睿睿¹^,²^,³, 康振¹^,²^,³^,⁴

^1.江南大学生物工程学院，糖化学与生物技术教育部重点实验室，江苏无锡 214122
^2.江南大学，未来食品科学中心，江苏无锡 214122
^3.江南大学生物工程学院，工业生物技术教育部重点实验室，江苏无锡 2141223
^4.江苏省合成生物基础研究中心，江苏无锡 214122

通讯作者: 康振
作者简介:张瑾（2002—），女，硕士研究生。研究方向为硫酸软骨素的生物合成与酶工程改造应用。E-mail：6240201068@stu.jiangnan.edu.cn
康振（1982—），男，博士，教授，博士生导师，研究方向为微生物合成生物学与生物制造研究。E-mail：zkang@jiangnan.edu.cn
基金资助:
国家重点研发计划项目(2024YFF1106300);国家自然科学基金项目(32400056);江苏省合成生物基础研究中心资助项目(BK20233003);江苏省基础研究计划自然科学基金-青年基金项目(BK20241616);国家资助博士后研究人员计划项目(GZC20240607);江南大学至善青年(JUSRP622003)

Abstract

Abstract:

Chondroitin sulfate (CS) is a type of glycosaminoglycan widely distributed in animal connective tissues, characterized by diverse biological activities including anti-inflammatory, moisturizing, and antioxidant effects. Owing to these beneficial properties, CS has been extensively utilized in joint health supplements, cosmetics, and pharmaceuticals. Traditionally, CS is extracted from animal tissues. However, this conventional extraction process presents several significant challenges, including heterogeneity in molecular weight, potential risks of pathogen contamination, long production cycles, and environmental pollution caused by chemical reagents and waste disposal. Such issues limit the ability of animal-derived CS to meet the stringent purity, safety, and quality standards required in high-end applications, especially in precision medicine and advanced biomaterials.With the rapid advancement of synthetic biology, the green and sustainable production of CS using engineered microbial cell factories has emerged as a key area of research. By reconstructing the CS precursor synthesis pathway and constructing an efficient sulfation modification system in microbial cell factories, the controlled biological synthesis of CS has been initially achieved, and the preparation of CS oligosaccharides has been realized by introducing CS degrading enzymes. This microbial synthesis approach not only addresses safety and environmental concerns but also offers advantages in scalability, product consistency, and cost-effectiveness.This review article provides a comprehensive summary of recent progress in biosynthesis of CS. It focuses on analyzing the optimization strategies for the synthesis pathways of chondroitin precursors and the design and engineering of key enzyme components. It delves deeply into the construction of the CS biosynthetic system and the multi-level optimization approaches. Meanwhile, the paper also provides a detailed introduction to the precise preparation processes of CS oligosaccharides, laying a solid foundation for achieving biosynthesized CS with highly consistent structural and functional properties. Based on these latest research advancements, this paper thoroughly analyzes the main challenges faced in the biosynthesis of CS and its oligosaccharides, including inadequate supply of precursors, insufficient catalytic activity and stability of sulfotransferase, unclear spatiotemporal coordination regulation of chondroitin sulfate glycan chain polymerization and sulfation modification, as well as low catalytic efficiency of hydrolysis enzymes in oligosaccharide preparation. It also provides a perspective on the future development direction of CS biosynthesis research: future studies should focus on achieving green and efficient large-scale production of CS and promoting its innovative applications in multiple fields through dynamic metabolic regulation, artificial intelligence-assisted enzyme modification, and the deep integration of synthetic biology and enzyme engineering.

Key words: chondroitin sulfate, chondroitin, sulfonate donor, sulfotransferases, metabolic engineering, synthetic biology

摘要：

硫酸软骨素（chondroitin sulfate， CS）是一类广泛存在于动物结缔组织中的糖胺聚糖，具有良好的抗炎、保水、抗氧化等多种生物活性，广泛应用于关节保健品、化妆品及医药等领域。然而，传统的动物组织提取工艺面临分子量不均一、潜在病原体风险、工艺周期长、环境污染等问题，难以满足高标准应用领域对产品纯度与安全性的要求。随着合成生物学的发展，利用工程化微生物实现CS的绿色制造已经成为研究热点。通过在微生物细胞工厂中重构CS前体合成路径和构建高效硫酸化修饰体系，已初步实现CS的可控生物合成，并通过引入CS降解酶实现了CS寡聚糖的制备。本综述系统梳理了CS的生物合成研究进展，重点讨论了前体软骨素合成途径优化及关键酶元件设计优化、CS的生物合成体系构建与优化及寡聚糖的精准制备技术。基于这些最新的研究进展，本文深入分析了CS及其寡聚糖生物合成面临的主要挑战，包括前体供应不足、硫酸基转移酶催化活性及稳定性差、软骨素糖链聚合与硫酸化修饰的时空协同调控不明确，以及CS寡聚糖制备中水解酶催化效率低下等问题，并对CS及其寡聚糖生物合成研究的未来发展方向进行了展望：未来研究应着力通过动态代谢调控、人工智能辅助酶改造以及合成生物学与酶工程的深度融合，实现CS绿色、高效的规模化生产及推进其在多领域的创新应用。

关键词: 硫酸软骨素, 软骨素, 硫酸基供体, 硫酸基转移酶, 代谢工程, 合成生物学

CLC Number:

Q815

ZHANG Jin, ZHANG Weijiao, XIONG Haibo, XIE Zhuan, XU Ruirui, KANG Zhen. Research advances in biosynthesis of chondroitin sulfate and its oligosaccharides[J]. Synthetic Biology Journal, DOI: 10.12211/2096-8280.2025-051.

张瑾, 张维娇, 熊海波, 谢专, 胥睿睿, 康振. 硫酸软骨素及其寡聚糖的生物合成进展[J]. 合成生物学, DOI: 10.12211/2096-8280.2025-051.

Figures/Tables 7

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底盘细胞	生物法合成方式	PAPS来源	产量/转化率/硫酸化度	来源
E. coli Origami B (DE3)	体外酶法	直接添加	CSA产量：- CSE转化率：50.00%	[54]
Bacillus subtilis 168/ E. coli BL21(DE3)/ Komagataella phaffii GS115	体外酶法	依赖AST IV再生	CSO产量：7.15 g/L CSA转化率：98.00% CSC转化率：96.00%	[55]
Bacillus subtilis 168/ E. coli BL21(DE3)/ Komagataella phaffii GS115	体外酶法	依赖AST IV再生	CSO产量：- CSA转化率：98.00%	[56]
Komagataella phaffii GS115/ E. coli BL21(DE3)	体外酶法	基于ATP再生系统合成PAPS	CSO产量：2.60 g/L CSA硫酸化度：40.00%	[57]
Corynebacterium glutamicum/ E. coli BL21(DE3)/ Komagataella phaffii GS115	体外酶法	基于ATP再生系统合成PAPS	CSO产量：- CSA硫酸化度：97.00%	[58]
E. coli Rosetta (DE3)	全细胞催化法	基于ATP再生系统与依赖AST IV再生	CSO产量：- CSA转化率：89.50%	[59]
E. coli Rosetta (DE3)	全细胞催化法	基于ATP再生系统与依赖AST IV再生	CSO产量：- CSE转化率：72.20%	[60]
E. coli MG1655	微生物一步发酵法	胞内PAPS天然合成	CSO产量：- CSA产量：27.00 μg/g DCW CSA硫酸化度：96.12%	[61]
Komagataella phaffii GS115	微生物一步发酵法	胞内PAPS天然合成	CSO产量：189.80 mg/L CSA产量：2.10 g/L CSA硫酸化度：4.00%	[62]
E. coli BL21 STAR (DE3)	微生物一步发酵法	基于AMP-PAPS循环再生途径	CSO产量：2.95 g/L CSA产量：1.89 g/L CSA硫酸化度：76.00%	[63-64]
Komagataella phaffii GS115	微生物一步发酵法	基于ATP-PAPS循环再生途径	CSA产量：1.15 g/L CSA硫酸化度：96.00%	[65]

底盘细胞	生物法合成方式	PAPS来源	产量/转化率/硫酸化度	来源
E. coli Origami B (DE3)	体外酶法	直接添加	CSA产量：- CSE转化率：50.00%	[54]
Bacillus subtilis 168/ E. coli BL21(DE3)/ Komagataella phaffii GS115	体外酶法	依赖AST IV再生	CSO产量：7.15 g/L CSA转化率：98.00% CSC转化率：96.00%	[55]
Bacillus subtilis 168/ E. coli BL21(DE3)/ Komagataella phaffii GS115	体外酶法	依赖AST IV再生	CSO产量：- CSA转化率：98.00%	[56]
Komagataella phaffii GS115/ E. coli BL21(DE3)	体外酶法	基于ATP再生系统合成PAPS	CSO产量：2.60 g/L CSA硫酸化度：40.00%	[57]
Corynebacterium glutamicum/ E. coli BL21(DE3)/ Komagataella phaffii GS115	体外酶法	基于ATP再生系统合成PAPS	CSO产量：- CSA硫酸化度：97.00%	[58]
E. coli Rosetta (DE3)	全细胞催化法	基于ATP再生系统与依赖AST IV再生	CSO产量：- CSA转化率：89.50%	[59]
E. coli Rosetta (DE3)	全细胞催化法	基于ATP再生系统与依赖AST IV再生	CSO产量：- CSE转化率：72.20%	[60]
E. coli MG1655	微生物一步发酵法	胞内PAPS天然合成	CSO产量：- CSA产量：27.00 μg/g DCW CSA硫酸化度：96.12%	[61]
Komagataella phaffii GS115	微生物一步发酵法	胞内PAPS天然合成	CSO产量：189.80 mg/L CSA产量：2.10 g/L CSA硫酸化度：4.00%	[62]
E. coli BL21 STAR (DE3)	微生物一步发酵法	基于AMP-PAPS循环再生途径	CSO产量：2.95 g/L CSA产量：1.89 g/L CSA硫酸化度：76.00%	[63-64]
Komagataella phaffii GS115	微生物一步发酵法	基于ATP-PAPS循环再生途径	CSA产量：1.15 g/L CSA硫酸化度：96.00%	[65]

Research advances in biosynthesis of chondroitin sulfate and its oligosaccharides

硫酸软骨素及其寡聚糖的生物合成进展

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