Non-natural redox cofactors empowered biomanufacturing

doi:10.11949/0438-1157.20240589

Abstract

Abstract:

Biomanufacturing uses the functions of organisms to process and synthesize chemicals on a large scale. Compared to traditional approaches, biomanufacturing method is considered as a green and sustainable one because it normally consumes less energy, generates less wastes yet outputs higher added-value products. It is routine to design and construct microbial cell factories for biomanufacturing applications. However, microbial cell factories often fail to exhibit expected functions due to intrinsic crosstalk mediated by cofactors. This article briefly introduces conventional methods for regulating intracellular redox coenzyme levels and their limitations, highlights the necessity of creating non-natural coenzyme-mediated systems. Furthermore, with nicotinamide cytosine dinucleotide (NCD) and nicotinamide mononucleotide (NMN) as examples, we will discuss how to explore new biological components and redox modules to design new microbial cell factories. Finally, we predict the future directions and analysis the challenges of biomanufacturing based on non-natural cofactors.

Key words: biocatalysis, enzyme, synthetic biology, coenzyme engineering, non-natural redox cofactor

摘要：

生物制造以生物体机能进行大规模化学品加工与合成。这种生产方式相较于传统的化学制造，具有更低的能耗、更少的废物排放和更高的产品附加值，是一种绿色、可持续的生产方式。构建微生物细胞工厂是生物制造的具体实现形式之一。然而，微生物细胞工厂常因胞内辅酶层次的串扰而未能呈现出预期功能。简要介绍调控胞内氧化还原辅酶水平的常规方法及其局限，说明创制非天然辅酶介导系统的必要性。此外，重点以烟酰胺胞嘧啶二核苷酸和烟酰胺单核苷酸为例，介绍如何研制新型生物学元件、氧化还原模块进而设计新的微生物细胞工厂。最后，预测了基于非天然辅酶的生物制造未来发展方向，并分析了所面临的挑战。

关键词: 生物催化, 酶, 合成生物学, 辅酶工程, 非天然氧化还原辅酶

CLC Number:

Q 819

Xueying WANG, Yongjin ZHOU, Zongbao ZHAO. Non-natural redox cofactors empowered biomanufacturing[J]. CIESC Journal, 2024, 75(11): 4037-4047.

王雪颖, 周雍进, 赵宗保. 非天然氧化还原辅酶赋能生物制造[J]. 化工学报, 2024, 75(11): 4037-4047.

Figures/Tables 3

References 81

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酶	来源	突变体	天然辅酶	非天然辅酶	催化效率提高倍数^①	突变体辅酶偏好性^②
苹果酸酶^[37]	Escherichia coli	L310R/Q401C	NAD⁺	NCD⁺	55.2	429.4
D-乳酸脱氢酶^[50]	L. helveticus	V152R/I177K/N213I	NAD⁺	NCD⁺	6.5	41.9
P450-BM3^[51]	B. megaterium	F87A/R966D/W1046S/Q976E/Q1004E	NADP⁺	NCD⁺	1.1×10³	30.8
亚磷酸脱氢酶^[52]	Ralstonia sp.	I151R	NAD⁺	NCD⁺	0.57	3.1
亚磷酸脱氢酶^[53]	Ralstonia sp.	I151R/P176R/M207A	NAD⁺	NCD⁺	0.16	45.3
甲酸脱氢酶^[55]	Pseudomonas sp. 101	V198I/C256I/P260S/E261P/S381N/S383F	NAD⁺	NCD⁺	85.6	424.2
甲醛脱氢酶^[56]	P. putida	A192R/L223V/L236V	NAD⁺	NCD⁺	1.3	153.7
甲醇脱氢酶^[57]	B. stearothermophilus	Y171R/I196V/V237T/N240E/K241A	NAD⁺	NCD⁺	11.3	429
葡萄糖脱氢酶^[64]	B. subtilis	I195R/A93K/Y39Q/S17E	NADP⁺	NMN⁺	446.8	19.1
甘油醛-3-磷酸脱氢酶^[66]	S. mutans	P179K/F153S/S330R/I234E/G214E	NADP⁺	NMN⁺	7.3	5.5
NADH氧化酶^[67]	L. pentosus	I158S/D177W/G178E/V240L/P362H/V395L	NAD⁺	NMN⁺	10	0.5
谷胱甘肽还原酶^[47]	E. coli	I178T/R198M/R204L	NADP⁺	NMN⁺	4	1.1×10^-1
亚磷酸脱氢酶^[47]	P. stutzeri	A155N/E175A/A176F	NAD⁺	NMN⁺	110	3.7×10^-2
NADH氧化酶^[68]	L. lactits	I159T/D178N/A179F/I243E	NAD⁺	NMN⁺	250	2.8
m-丁二醇脱氢酶^[68]	K. pneumoniae	M189T/Y34Q/A87K	NAD⁺	NMN⁺	19	136.4
3α-羟基类固醇脱氢酶^[69]	Comamonas testosteroni	A70K	NAD⁺	NMN⁺	8.7	4.4×10⁴
P450-BM3^[70]	B. megaterium	S848R	NADP⁺	NMN⁺	2.1	—
葡萄糖-6-磷酸脱氢酶^[31]	Z. mobilis	S25A/S115A/P118S/Y417H/M421L	NADP⁺	NMN⁺	112	4.3×10^-3

酶	来源	突变体	天然辅酶	非天然辅酶	催化效率提高倍数^①	突变体辅酶偏好性^②
苹果酸酶^[37]	Escherichia coli	L310R/Q401C	NAD⁺	NCD⁺	55.2	429.4
D-乳酸脱氢酶^[50]	L. helveticus	V152R/I177K/N213I	NAD⁺	NCD⁺	6.5	41.9
P450-BM3^[51]	B. megaterium	F87A/R966D/W1046S/Q976E/Q1004E	NADP⁺	NCD⁺	1.1×10³	30.8
亚磷酸脱氢酶^[52]	Ralstonia sp.	I151R	NAD⁺	NCD⁺	0.57	3.1
亚磷酸脱氢酶^[53]	Ralstonia sp.	I151R/P176R/M207A	NAD⁺	NCD⁺	0.16	45.3
甲酸脱氢酶^[55]	Pseudomonas sp. 101	V198I/C256I/P260S/E261P/S381N/S383F	NAD⁺	NCD⁺	85.6	424.2
甲醛脱氢酶^[56]	P. putida	A192R/L223V/L236V	NAD⁺	NCD⁺	1.3	153.7
甲醇脱氢酶^[57]	B. stearothermophilus	Y171R/I196V/V237T/N240E/K241A	NAD⁺	NCD⁺	11.3	429
葡萄糖脱氢酶^[64]	B. subtilis	I195R/A93K/Y39Q/S17E	NADP⁺	NMN⁺	446.8	19.1
甘油醛-3-磷酸脱氢酶^[66]	S. mutans	P179K/F153S/S330R/I234E/G214E	NADP⁺	NMN⁺	7.3	5.5
NADH氧化酶^[67]	L. pentosus	I158S/D177W/G178E/V240L/P362H/V395L	NAD⁺	NMN⁺	10	0.5
谷胱甘肽还原酶^[47]	E. coli	I178T/R198M/R204L	NADP⁺	NMN⁺	4	1.1×10^-1
亚磷酸脱氢酶^[47]	P. stutzeri	A155N/E175A/A176F	NAD⁺	NMN⁺	110	3.7×10^-2
NADH氧化酶^[68]	L. lactits	I159T/D178N/A179F/I243E	NAD⁺	NMN⁺	250	2.8
m-丁二醇脱氢酶^[68]	K. pneumoniae	M189T/Y34Q/A87K	NAD⁺	NMN⁺	19	136.4
3α-羟基类固醇脱氢酶^[69]	Comamonas testosteroni	A70K	NAD⁺	NMN⁺	8.7	4.4×10⁴
P450-BM3^[70]	B. megaterium	S848R	NADP⁺	NMN⁺	2.1	—
葡萄糖-6-磷酸脱氢酶^[31]	Z. mobilis	S25A/S115A/P118S/Y417H/M421L	NADP⁺	NMN⁺	112	4.3×10^-3

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