重组胶原蛋白的绿色生物制造及其应用

doi:10.16085/j.issn.1000-6613.2020-2109

摘要/Abstract

摘要：

胶原蛋白存在于各个组织器官，与动物胶原蛋白相比，重组胶原蛋白组分单一、安全性高、生产过程可控。本篇综述简述了重组胶原蛋白不同表达体系的构建，包括动植物以及微生物表达体系，比较了不同体系的优缺点。着重介绍了微生物体系中影响产物表达的不同发酵参数的调控，产物的分离纯化工艺以及重组胶原蛋白在医学领域的应用。提出微生物发酵体系较动植物体系成本低，操作简单，易于扩大生产；温度、pH、溶解氧、葡萄糖、乙酸浓度等影响大肠杆菌发酵中的蛋白表达量；酵母发酵中，甲醇添加量、温度、pH和溶解氧是主要影响参数；微生物发酵体系均需通过不同的粗纯及精纯技术获得纯度较高的产物。同时，重组胶原蛋白在生物医学领域发挥着重要作用。

关键词: 重组胶原蛋白, 大肠杆菌, 酵母, 发酵, 纯化, 应用

Abstract:

Collagen exists in all tissues and organs. Compared with animal collagen, recombinant collagen has a more single component, higher safety, and more controllable production process. This review briefly introduces the construction of different expression systems of recombinant collagen, including the expression systems of animals, plants, and microorganisms, and their advantages and disadvantages are compared. The regulation of different fermentation parameters affecting the expression of the product in the microbial system, the separation and purification process of the product, and the application of recombinant collagen in the medical field were introduced emphatically. It is pointed out that the microbial fermentation system has a lower cost, simple operation, and easier to expand production than the animal and plant system. Temperature, pH, dissolved oxygen, glucose, and acetic acid concentration affect the protein expression in Escherichia coli fermentation. In yeast fermentation, the addition amount of methanol, temperature, pH, and dissolved oxygen are the main influencing parameters. Microbial fermentation systems need to obtain high purity products through different crude and precision technologies. Meanwhile, recombinant collagen plays an important role in the field of biomedicine.

Key words: recombinant collagen, Escherichia coli, yeast, fermentation, purification, application

中图分类号:

Q815

李阳, 朱晨辉, 范代娣. 重组胶原蛋白的绿色生物制造及其应用[J]. 化工进展, 2021, 40(3): 1262-1275.

LI Yang, ZHU Chenhui, FAN Daidi. Green biological manufacture and application of recombinant collagen[J]. Chemical Industry and Engineering Progress, 2021, 40(3): 1262-1275.

图/表 12

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项目	天然胶原蛋白	重组类人胶原蛋白
与人体亲和性	异体胶原、弱人体亲和性	同质胶原、高人体亲和性
生产工艺	化学提取、可控性差	基因工程、品质可控
安全性	易携带动物病毒（疯牛病等）	酵母发酵无病毒
致敏性	异源蛋白、易过敏	同质蛋白、不易过敏
生物活性	无胶原空间结构、无生物活性	有胶原空间结构、保留生物活性
纯度	混合胶原，成分复杂	单一胶原，成分固定，纯度达95%
保湿性	弱	高

项目	天然胶原蛋白	重组类人胶原蛋白
与人体亲和性	异体胶原、弱人体亲和性	同质胶原、高人体亲和性
生产工艺	化学提取、可控性差	基因工程、品质可控
安全性	易携带动物病毒（疯牛病等）	酵母发酵无病毒
致敏性	异源蛋白、易过敏	同质蛋白、不易过敏
生物活性	无胶原空间结构、无生物活性	有胶原空间结构、保留生物活性
纯度	混合胶原，成分复杂	单一胶原，成分固定，纯度达95%
保湿性	弱	高

表达体系	优点	缺点
动物	蛋白活性高，更接近于天然蛋白；正确的高级结构（如糖基化）；适合表达完整的大分子蛋白	培养基成本极高，培养周期长；培养较困难，表达量较低；操作技术要求高、产率低；不易大规模生产
植物	植物来源经济；安全性高	发酵纯化成本较高；产量低；不易于产业化生产
微生物	成本低；遗传背景清晰；生长繁殖快，培养周期短；可操作性强，易于产业化	大肠杆菌体系，纯化困难，产物活性低；毕赤酵母体系，甲醇为潜在风险

表达体系	优点	缺点
动物	蛋白活性高，更接近于天然蛋白；正确的高级结构（如糖基化）；适合表达完整的大分子蛋白	培养基成本极高，培养周期长；培养较困难，表达量较低；操作技术要求高、产率低；不易大规模生产
植物	植物来源经济；安全性高	发酵纯化成本较高；产量低；不易于产业化生产
微生物	成本低；遗传背景清晰；生长繁殖快，培养周期短；可操作性强，易于产业化	大肠杆菌体系，纯化困难，产物活性低；毕赤酵母体系，甲醇为潜在风险

微生物	发酵	纯化
原核	受pH、温度、溶解氧、乙酸浓度、碳源氮源等影响	大部分为胞内表达，需先释放胞内物质；涉及沉淀、超滤、层析等粗纯和精纯
真核	部分需要甲醇诱导受pH、温度以及甲醇含量的影响	既能胞内分泌，亦可胞外分泌，胞内释放时细胞破碎难度大，胞外释放不需细胞破碎；均涉及沉淀、超滤、层析等粗纯和精纯