天然类胡萝卜素的生物合成研究进展

doi:10.13560/j.cnki.biotech.bull.1985.2021-1184

摘要/Abstract

摘要：

类胡萝卜素是广泛存在于高等植物、微生物和藻类中的一大类天然色素物质的总称,是重要的天然食用色素群之一。细胞工厂被广泛用于各种化学品、食品和药品等的生产,是绿色生物制造的中心环节。利用细胞工厂合成来源稀缺、获取难度大的高附加值天然类胡萝卜素,具有经济可行和环境友好等优势。类胡萝卜素在抗氧化、视力保护和抗癌等方面具有非常重要的作用。本文对生物合成类胡萝卜素细胞工厂构建过程中的类胡萝卜素基本合成路径解析、底盘细胞的选择和改造、途径的模块组装与适配及合成生物学策略等展开详细介绍。同时,对未来类胡萝卜素生物合成所面临的机遇和挑战进行了展望。

关键词: 类胡萝卜素, 虾青素, β-胡萝卜素, 叶黄素, 合成生物学

Abstract:

Carotenoids are the general term for a large class of natural pigment substances broadly in higher plants,microorganisms and algae,one of important natural pigments groups for food. Cell factories are widely used in the production of various chemicals,food and drugs,and are the central link of green biological manufacturing. It is economically feasible and environmentally friendly to synthesize source-scarce,difficult-to-be-obtained and high value-added natural carotenoids in cell factories. Carotenoids play important roles in anti-oxidation,eye-vision protection and anti-cancer. In the present review,the analysis of carotenoid biosynthesis pathway,the selection and modification of chassis cells,the module assembly and adaptation of pathway,and the strategy of synthetic biology during the construction of cell factory used for carotenoids biosynthesis are introduced in details. Future opportunities and challenges in the industrial production of carotenoids are discussed.

Key words: carotenoids, astaxanthin, β-carotene, lutein, synthetic biology

周琳, 梁轩铭, 赵磊. 天然类胡萝卜素的生物合成研究进展[J]. 生物技术通报, 2022, 38(7): 119-127.

ZHOU Lin, LIANG Xuan-ming, ZHAO Lei. Biosynthesis of Natural Carotenoids：Progress and Perspective[J]. Biotechnology Bulletin, 2022, 38(7): 119-127.

图/表 6

参考文献 59

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类胡萝卜素 Carotenoids	生物学功能 Biological function
虾青素 Astaxanthin	抗氧化、抗炎、抗癌、预防心血管疾病
β-胡萝卜素 β-carotene	抗氧化、预防夜盲症、抗肝硬化
叶黄素 Lutein	预防白内障和老年性黄斑变性、抗氧化、抗肿瘤
番茄红素 Lycopene	抗癌、预防心血管疾病、防辐射、抗氧化

类胡萝卜素 Carotenoids	生物学功能 Biological function
虾青素 Astaxanthin	抗氧化、抗炎、抗癌、预防心血管疾病
β-胡萝卜素 β-carotene	抗氧化、预防夜盲症、抗肝硬化
叶黄素 Lutein	预防白内障和老年性黄斑变性、抗氧化、抗肿瘤
番茄红素 Lycopene	抗癌、预防心血管疾病、防辐射、抗氧化

名称 Name	编号 No.	简写 Abbreviation	编码基因 Encoding gene
八氢番茄红素合成酶 Phytoene synthase	2.5.1.32	PSY	AtPSY
八氢番茄红素脱氢酶 Phytoene desaturase	1.3.5.5	PDS	AtPDS3
ξ-胡萝卜素脱氢酶 ζ-carotene desaturase	1.3.5.6	ZDS	AtZDS
番茄红素ε-环化酶 Lycopene ε-cyclase	5.5.1.18	ε-LCY（LCYe）	AtLUT2
番茄红素β-环化酶 Lycopene-β-cyclase	5.5.1.19	β-LCY（LCYb）	AtLYC
类胡萝卜素ε-羟化酶 Carotenoid ε-hydroxylase	1.14.14.158	LUT1	AtLUT1
β-胡萝卜素3-羟化酶 β-carotene 3-hydroxylase	1.14.15.24	CrtZ	Atβ-OHASE_1 Atβ-OHASE_2
β-环羟化酶 β-cyclohydroxylase	1.14	LUT5	AtCYP97A3
玉米黄素环氧化酶 Zeathanxin epoxidase	1.14.15.21	ZEP	AtABA1
堇菜黄质脱环氧化酶 Violaxanthin deepoxidase	1.23.5.1	VDE	NPQ1

名称 Name	编号 No.	简写 Abbreviation	编码基因 Encoding gene
八氢番茄红素合成酶 Phytoene synthase	2.5.1.32	PSY	AtPSY
八氢番茄红素脱氢酶 Phytoene desaturase	1.3.5.5	PDS	AtPDS3
ξ-胡萝卜素脱氢酶 ζ-carotene desaturase	1.3.5.6	ZDS	AtZDS
番茄红素ε-环化酶 Lycopene ε-cyclase	5.5.1.18	ε-LCY（LCYe）	AtLUT2
番茄红素β-环化酶 Lycopene-β-cyclase	5.5.1.19	β-LCY（LCYb）	AtLYC
类胡萝卜素ε-羟化酶 Carotenoid ε-hydroxylase	1.14.14.158	LUT1	AtLUT1
β-胡萝卜素3-羟化酶 β-carotene 3-hydroxylase	1.14.15.24	CrtZ	Atβ-OHASE_1 Atβ-OHASE_2
β-环羟化酶 β-cyclohydroxylase	1.14	LUT5	AtCYP97A3
玉米黄素环氧化酶 Zeathanxin epoxidase	1.14.15.21	ZEP	AtABA1
堇菜黄质脱环氧化酶 Violaxanthin deepoxidase	1.23.5.1	VDE	NPQ1

底盘细胞Chassis cells	类胡萝卜素Carotenoid	工程手段 Engineering measures
大肠杆菌 E. coli	番茄红素Lycopene	过表达戊二酸脱氢酶SucAB、琥珀酸脱氢酶SdhABCD 和转醛酶B TalB
	β-胡萝卜素β-carotene	对合成路径模块进行工程化改造;过表达α-酮戊二酸脱氢酶SucAB 和琥珀酸脱氢酶SdhABCD;过表达转醛酶TalB,增强磷酸戊糖途径
	玉米黄质Zeaxanthin	应用可调控基因间序列平衡异MVA 路径蛋白表达;使用IPP/FPP 响应启动子动态调控异源MVA 路径基因,减少有毒中间代谢物积累
	虾青素Astaxanthin	组合筛选路径基因启动子,促进番茄红素向β-胡萝卜素转化;OmpF 和TrxA 标签分别与截短蛋白N 端或C 端融合,实现CrtW 的稳定表达和质膜定位
酿酒酵母 S. cerevisiae	番茄红素Lycopene	CrtB 序列的筛选;CrtYB 与CrtE 的定向进化;调节Crt 编码基因拷贝数;构建非营养缺陷二倍体细胞
	β-胡萝卜β-carotene	以GAL 系统和HXT1 启动子时序调控胡萝卜素和竞争性鲨烯的合成
	虾青素Astaxanthin	CrtZ 和CrtW 协同定向进化;温敏调控系统控制关键基因表达
耶氏解脂酵母 Y. lipolytica	番茄红素Lycopene	引入异戊烯醇同化路径增强前体IPP/DMAPP 供给;增强脂质积累;增加IDI 拷贝
耶氏解脂酵母 Y. lipolytica	β-胡萝卜β-carotene	选择高度积累脂质的底盘;路径基因启动子的组合适配;发酵条件优化
	虾青素Astaxanthin	下调鲨烯合酶的表达;同时表达不同来源的CrtE;筛选CrtZ/CrtW 来源,调节表达量