维生素A生物合成的研究进展

doi:10.11949/0438-1157.20220838

摘要/Abstract

摘要：

维生素A是维持人体代谢的必需维生素，在食品、药品及护肤品等多个领域占有重要地位，是三大维生素支柱产品之一，应用前景广阔。市场上的维生素A主要来源于化学合成和天然提取。近年来，随着绿色生物制造的发展，维生素A的生物合成研究取得了重大的进展。总结了维生素A生物合成的研究现状，并对维生素A生物合成的优化改造方案进行了分析归纳，概述了维生素A发酵生产以及产品组分调控和储存策略，最后对生物合成维生素A的现状进行总结与展望。

关键词: 维生素A, 细胞工厂, 合成生物学, 视黄醇, 酶, 代谢

Abstract:

Vitamin A is an essential vitamin to maintain human metabolism. It plays an important role in many fields such as food, medicine and skin care products, and is one of the three pillar vitamins with broad application prospects. Vitamin A on the market mainly comes from chemical synthesis and natural extraction. In recent years, with the development of green biomanufacturing, the biosynthesis of vitamin A has made great progress. This review firstly summarized the research status of vitamin A biosynthesis, analyzed and summarized the optimization of vitamin A biosynthesis, then summarized the fermentation production of vitamin A, regulation of product components and storage strategies, and finally summarized and prospected the status quo of vitamin A biosynthesis.

Key words: vitamin A, cell factory, synthetic biology, retinol, enzyme, metabolize

中图分类号:

Q 74

王欣慧, 王颖, 姚明东, 肖文海. 维生素A生物合成的研究进展[J]. 化工学报, 2022, 73(10): 4311-4323.

Xinhui WANG, Ying WANG, Mingdong YAO, Wenhai XIAO. Research progress of vitamin A biosynthesis[J]. CIESC Journal, 2022, 73(10): 4311-4323.

图/表 10

参考文献 72

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公司	产地	上游中间体	生产工艺	产能/(t/a)
新和成	中国	自配	Roche^[7]	10000
帝斯曼	荷兰等	外购	Roche	7500
巴斯夫	德国等	自配	BASF^[7]	6000
浙江医药	中国	外购	BASF	5600
安迪苏	法国等	外购	BASF	5000
金达威	中国	外购	Roche	2900

公司	产地	上游中间体	生产工艺	产能/(t/a)
新和成	中国	自配	Roche^[7]	10000
帝斯曼	荷兰等	外购	Roche	7500
巴斯夫	德国等	自配	BASF^[7]	6000
浙江医药	中国	外购	BASF	5600
安迪苏	法国等	外购	BASF	5000
金达威	中国	外购	Roche	2900

底盘细胞	工程化手段	产量	文献
大肠杆菌	表达家鼠来源的β-胡萝卜素15，15′-氧化酶；优化最佳反应条件：pH、温度、底物浓度和表面活性剂	视黄醛产量：72 mg/L	[32]
大肠杆菌	选择密码子优化后的β-胡萝卜素15，15′-氧化酶；上调dxs，优化内源MEP途径；引入外源性MVA途径，提供IPP和DMAPP；筛选大肠菌株：MG1655、DH5a、X11-BLUE、S17-1和BL21；发酵条件优化：氧浓度、碳源和温度	视黄醛产量：136 mg/L	[33]
大肠杆菌	引入两个异源基因blhSR和RALDH2；删除内源编码醛还原酶的ybbO基因；采用mRNA稳定区工程方法；优化维生素A产生菌的培养条件:温度、pH和氧浓度	5 L生物反应器分批发酵；维甲酸产量： (8.20±0.05) mg/L	[34]
大肠杆菌	筛选了编码β-胡萝卜素裂解酶的基因：blh、brp和bcox；采用mRNA稳定区工程提高blh催化效率；共同表达合成视黄基棕榈酸酯路径基因LRAT和CRBP	分批补料发酵培养；视黄基棕榈酸酯产量为(69.96±2.64) mg/L	[35]
大肠杆菌	优化发酵条件：温度、pH、搅拌速度；筛选最佳表面活性剂	视黄醛产量：600 mg/L	[36]
大肠杆菌	过表达内源基因ybbO，增强醛→醇转化；敲除编码O-乙酰转移酶的基因cat	视黄醛、视黄醇、视黄醇醋酸酯产量的比例为6%、88%、6%	[37]
酿酒酵母	采用以木糖为碳源的酵母菌株；采用十二烷和橄榄油两相原位萃取	3 L生物反应器分批补料；视黄醇、视黄醛产量为1256、2094 mg/L	[38]
酿酒酵母	引入人来源RDH12和乳球菌noxE；优化发酵条件，以木糖为碳源，十二烷为萃取剂；筛选最适培养和储存条件，包括温度、光照和抗氧化剂	250 ml摇瓶发酵；视黄醇产量：123.1 mg/L	[39]
酿酒酵母	过表达酵母内源ENV9和截短的Hmg1；发酵添加1.44 mmol/L Fe²⁺以维持blh酶活性	视黄醇产量：443.43 mg/L	[40]

底盘细胞	工程化手段	产量	文献
大肠杆菌	表达家鼠来源的β-胡萝卜素15，15′-氧化酶；优化最佳反应条件：pH、温度、底物浓度和表面活性剂	视黄醛产量：72 mg/L	[32]
大肠杆菌	选择密码子优化后的β-胡萝卜素15，15′-氧化酶；上调dxs，优化内源MEP途径；引入外源性MVA途径，提供IPP和DMAPP；筛选大肠菌株：MG1655、DH5a、X11-BLUE、S17-1和BL21；发酵条件优化：氧浓度、碳源和温度	视黄醛产量：136 mg/L	[33]
大肠杆菌	引入两个异源基因blhSR和RALDH2；删除内源编码醛还原酶的ybbO基因；采用mRNA稳定区工程方法；优化维生素A产生菌的培养条件:温度、pH和氧浓度	5 L生物反应器分批发酵；维甲酸产量： (8.20±0.05) mg/L	[34]
大肠杆菌	筛选了编码β-胡萝卜素裂解酶的基因：blh、brp和bcox；采用mRNA稳定区工程提高blh催化效率；共同表达合成视黄基棕榈酸酯路径基因LRAT和CRBP	分批补料发酵培养；视黄基棕榈酸酯产量为(69.96±2.64) mg/L	[35]
大肠杆菌	优化发酵条件：温度、pH、搅拌速度；筛选最佳表面活性剂	视黄醛产量：600 mg/L	[36]
大肠杆菌	过表达内源基因ybbO，增强醛→醇转化；敲除编码O-乙酰转移酶的基因cat	视黄醛、视黄醇、视黄醇醋酸酯产量的比例为6%、88%、6%	[37]
酿酒酵母	采用以木糖为碳源的酵母菌株；采用十二烷和橄榄油两相原位萃取	3 L生物反应器分批补料；视黄醇、视黄醛产量为1256、2094 mg/L	[38]
酿酒酵母	引入人来源RDH12和乳球菌noxE；优化发酵条件，以木糖为碳源，十二烷为萃取剂；筛选最适培养和储存条件，包括温度、光照和抗氧化剂	250 ml摇瓶发酵；视黄醇产量：123.1 mg/L	[39]
酿酒酵母	过表达酵母内源ENV9和截短的Hmg1；发酵添加1.44 mmol/L Fe²⁺以维持blh酶活性	视黄醇产量：443.43 mg/L	[40]

改造策略	具体改造信息	结果	文献
碳源	葡萄糖、木糖	20.74 mg/L维生素A（44 g/L木糖）	[38]
温度、pH	温度：24~33℃ pH：5.5~8.0	视黄醛产量最大（pH 7.0，30℃），细胞量达到最大（27℃）	[36]
表面活性剂	Span20、Span40、Span60、Span80、 Tween20、Tween40、Tween80、Brij58、Triton X-100	细胞量最大，细胞积累量为15.2 g/L（10 g/L Span80）	[36]
搅拌速率	400、500、600、700、800 r/min	视黄醛产量最大为600 mg/L（600 r/min）	[36]
萃取剂	十二烷、大豆油和橄榄油	十二烷效果最佳	[39]
光	光照条件、黑暗条件	视黄醇浓度最高为124.6 mg/L（黑暗）	[39]