Rapid construction and directed evolution of cell factories for carotenoid biosynthesis

doi:10.16085/j.issn.1000-6613.2020-1944

Abstract

Abstract:

Carotenoids have brilliant colors and strong antioxidant activity, which not only endow these compounds with high commercial value, but also accelerate the construction process for building efficient biosynthetic pathways in heterogeneous chassis. Carotenoids have become an ideal object to develop irrational design strategy. Here we critically review recent advances about carotenoid biosynthesis via irrational design strategies on pathway construction by parts optimization, modular compatibility, multidimensional regulation etc., as well as chassis evolution by DNA mutation and adaptive evolution. These strategies aims to break the bottleneck of carotenoid biosynthesis through improving the supply and utilization of precursor GGPP as well as relieving the stress from carotenoid accumulation on microbiological chassis. Based on the demand of employing unconventional chassis and expanding carotenoid kingdom, we also propose the development of the construction strategies for carotenoid microbiological cell factories towards the automation and intellectualization.

Key words: synthetic biology, cell factories, carotenoids, irrational design, pathway construction, chassis evolution

摘要：

类胡萝卜素具有丰富的颜色和强抗氧化活性，这赋予了该类产品较高的商业价值，加速了该类产品合成路径在异源底盘中的构建进程，使之成为发展非理性设计策略的有效研究对象。本文主要针对微生物合成类胡萝卜素细胞工厂构建中的瓶颈问题，从增强合成类胡萝卜素前体GGPP的供给利用以及缓解各类胡萝卜素产品对微生物底盘细胞的胁迫压力两个角度，系统综述非理性设计策略对元件优化、模块适配和多维调控等路径构建优化方法的丰富，以及对不同尺度的DNA变异和适应性进化等底盘进化手段的推动作用。并从扩展类胡萝卜素合成底盘物种的多样性和丰富非天然类胡萝卜素产品结构的可获得性的需求出发，对构建合成该类产品微生物细胞工厂自动化和智能化的发展趋势进行展望。

关键词: 合成生物学, 细胞工厂, 类胡萝卜素, 非理性设计, 路径构建, 底盘进化

CLC Number:

Q819

WANG Ying, QU Junze, LIANG Nan, HAO He, YUAN Yingjin. Rapid construction and directed evolution of cell factories for carotenoid biosynthesis[J]. Chemical Industry and Engineering Progress, 2021, 40(3): 1187-1201.

王颖, 曲俊泽, 梁楠, 郝鹤, 元英进. 合成类胡萝卜素细胞工厂的快速构建和定向进化[J]. 化工进展, 2021, 40(3): 1187-1201.

Figures/Tables 7

References 130

1	TAKAICHI S. Carotenoids in algae: distributions, biosyntheses and functions[J]. Marine Drugs, 2011, 9(6): 1101-1118.
2	CARAIL M, CARIS-VEYRAT C. Carotenoid oxidation products: from villain to saviour?[J]. Pure and Applied Chemistry, 2006, 78(8): 1493-1503.
3	AULDRIDGE M E, MCCARTY D R, KLEE H J. Plant carotenoid cleavage oxygenases and their apocarotenoid products[J]. Current Opinion in Plant Biology, 2006, 9: 315-321.
4	AHRAZEM O, GÓMEZ-GÓMEZ L, RODRIGO M, et al. Carotenoid cleavage oxygenases from microbes and photosynthetic organisms: features and functions[J]. International Journal of Molecular Sciences, 2016, 17(11): 1781.
5	MUSSAGY C U, WINTERBURN J, SANTOS-EBINUMA V C, et al. Production and extraction of carotenoids produced by microorganisms[J]. Applied Microbiology and Biotechnology, 2019, 103(3): 1095-1114.
6	GHAZI H, USHIO S, GOTO H, et al. Astaxanthin, a carotenoid with potential in human health and nutrition[J]. Journal of Natural Products, 2006, 3(69): 443-449.
7	BONGERS M, CHRYSANTHOPOULOS P K, BEHRENDORFF J B, et al. Systems analysis of methylerythritol-phosphate pathway flux in E. coli: insights into the role of oxidative stress and the validity of lycopene as an isoprenoid reporter metabolite[J]. Microbial Cell Factories, 2015, 14: 193.
8	WANG J, MENG H, XIONG Z, et al. Identification of novel knockout and up-regulated targets for improving isoprenoid production in E. coli[J]. Biotechnology Letters, 2014, 36(5): 1021-1027.
9	OSTROV N, JIMENEZ M, BILLERBECK S, et al. A modular yeast biosensor for low-cost point-of-care pathogen detection[J]. Science Advances, 2017, 3(6): e1603221.
10	WATSTEIN D M, MCNERNEY M P, STYCZYNSKI M P. Precise metabolic engineering of carotenoid biosynthesis in Escherichia coli towards a low-cost biosensor[J]. Metabolic Engineering, 2015, 31: 171-180.
11	YANG J, GUO L. Biosynthesis of β-carotene in engineered E. coli using the MEP and MVA pathways[J]. Microbial Cell Factories, 2014, 13(1): 160.
12	LI Y, LIN Z, HUANG C, et al. Metabolic engineering of Escherichia coli using CRISPR-Cas9 meditated genome editing[J]. Metabolic Engineering, 2015, 31: 13-21.
13	GAO S, TONG Y, ZHU L, et al. Iterative integration of multiple-copy pathway genes in Yarrowia lipolytica for heterologous β-carotene production[J]. Metabolic Engineering, 2017, 41: 192-201.
14	MA T, SHI B, YE Z, et al. Lipid engineering combined with systematic metabolic engineering of Saccharomyces cerevisiae for high-yield production of lycopene[J]. Metabolic Engineering, 2019, 52: 134-142.
15	LÓPEZ J, CATALDO V F, PEÑA M, et al. Build your bioprocess on a solid strain-β-carotene production in recombinant Saccharomyces cerevisiae[J]. Frontiers in Bioengineering and Biotechnology, 2019, 7: 1-9.
16	SUN T, MIAO L, LI Q, et al. Production of lycopene by metabolically-engineered Escherichia coli[J]. Biotechnology Letters, 2014, 36(7): 1515-1522.
17	SHI B, MA T, YE Z, et al. Systematic metabolic engineering of Saccharomyces cerevisiae for lycopene overproduction[J]. Journal of Agricultural and Food Chemistry, 2019, 67(40): 11148-11157.
18	WU Y, YAN P, LI Y, et al. Enhancing β-carotene production in Escherichia coli by perturbing central carbon metabolism and improving the NADPH supply[J]. Frontiers in Bioengineering and Biotechnology, 2020, 8: 585.
19	陶俊, 张上隆, 徐昌杰, 等. 类胡萝卜素合成的相关基因及其基因工程[J]. 生物工程学报, 2002, 18(3): 276-281.
	TAO Jun, ZHANG Shanglong, XU Changjie, et al. Gene and gene engineering of carotenoid biosynthesis[J]. Chinese Journal of Biotechnology, 2002, 18(3): 276-281.
20	LEE W, LEE D G. Lycopene-induced hydroxyl radical causes oxidative DNA damage in Escherichia coli[J]. Journal of Microbiology and Biotechnology, 2014, 24(9): 1232-1237.
21	LU Q, LIU J. Enhanced astaxanthin production in Escherichia colivia morphology and oxidative stress engineering[J]. Journal of Agricultural and Food Chemistry, 2019, 67(42): 11703-11709.
22	武陶, 张柏林, 毕昌昊. 细胞膜合成途径模块化调控与形态改造提高大肠杆菌β-胡萝卜素的积累与产量[J]. 生物工程学报, 2018, 34(5): 703-711.
	WU Tao, ZHANG Bolin, BI Changhao. Improving β-carotene production in Escherichia coli by modularized regulation of the membrane synthetic pathway and morphology engineering[J]. Chinese Journal of Biotechnology, 2018, 34(5): 703-711.
23	ZHAO J, LI Q, SUN T, et al. Engineering central metabolic modules of Escherichia coli for improving β-carotene production[J]. Metabolic Engineering, 2013, 17: 42-50.
24	SHEN H, CHENG B, ZHANG Y, et al. Dynamic control of the mevalonate pathway expression for improved zeaxanthin production in Escherichia coli and comparative proteome analysis[J]. Metabolic Engineering, 2016, 38: 180-190.
25	PARK S Y, BINKLEY R M, KIM W J, et al. Metabolic engineering of Escherichia coli for high-level astaxanthin production with high productivity[J]. Metabolic Engineering, 2018(49): 105-115.
26	WANG W, HE P, ZHAO D, et al. Construction of Escherichia coli cell factories for crocin biosynthesis[J]. Microbial Cell Factories, 2019, 18(1):120.
27	XIE W, LV X, YE L, et al. Construction of lycopene-overproducing Saccharomyces cerevisiae by combining directed evolution and metabolic engineering[J]. Metabolic Engineering, 2015, 30: 69-78.
28	WANG Z, LI X, YU C, et al. Continuous self-cycling fermentation leads to economical lycopene production by Saccharomyces cerevisiae[J]. Frontiers in Bioengineering and Biotechnology, 2020(8): 420.
29	XIE W, YE L, LV X, et al. Sequential control of biosynthetic pathways for balanced utilization of metabolic intermediates in Saccharomyces cerevisiae[J]. Metabolic Engineering, 2015, 28: 8-18.
30	ZHOU P, LI M, SHEN B, et al. Directed coevolution of β-carotene ketolase and hydroxylase and its application in temperature-regulated biosynthesis of astaxanthin[J]. Journal of Agricultural and Food Chemistry, 2019, 67(4): 1072-1080.
31	JIANG G, YANG Z, WANG Y, et al. Enhanced astaxanthin production in yeast via combined mutagenesis and evolution[J]. Biochemical Engineering Journal, 2020, 156: 107519.
32	CHAI F, WANG Y, MEI X, et al. Heterologous biosynthesis and manipulation of crocetin in Saccharomyces cerevisiae[J]. Microbial Cell Factories, 2017, 16(1): 54.
33	LIU T, DONG C, QI M, et al. Construction of a stable and temperature-responsive yeast cell factory for crocetin biosynthesis using CRISPR-Cas9[J]. Frontiers in Bioengineering and Biotechnology, 2020(8): 653.
34	SONG T, WU N, WANG C, et al. Crocetin overproduction in engineered Saccharomyces cerevisiaevia tuning key enzymes coupled with precursor engineering[J]. Frontiers in Bioengineering and Biotechnology, 2020(8): 578005.
35	LUO Z, LIU N, LAZAR Z, et al. Enhancing isoprenoid synthesis in Yarrowia lipolytica by expressing the isopentenol utilization pathway and modulating intracellular hydrophobicity[J]. Metabolic Engineering, 2020, 61: 344-351.
36	ZHANG X, WANG D, CHEN J, et al. Metabolic engineering of β-carotene biosynthesis in Yarrowia lipolytica[J]. Biotechnology Letters, 2020, 42(6): 945-956.
37	LARROUDE M, CELINSKA E, BACK A, et al. A synthetic biology approach to transform Yarrowia lipolytica into a competitive biotechnological producer of β-carotene[J]. Biotechnology and Bioengineering, 2018, 115(2): 464-472.
38	TRAMONTIN L R R, KILDEGAARD K R, SUDARSAN S, et al. Enhancement of astaxanthin biosynthesis in oleaginous yeast Yarrowia lipolyticavia microalgal pathway[J]. Microorganisms, 2019, 7(10): 472.
39	ZELCBUCH L, ANTONOVSKY N, BAR-EVEN A, et al. Spanning high-dimensional expression space using ribosome-binding site combinatorics[J]. Nucleic Acids Research, 2013, 41(9): e98.
40	ÖZAYDıN B, BURD H, LEE T S, et al. Carotenoid-based phenotypic screen of the yeast deletion collection reveals new genes with roles in isoprenoid production[J]. Metabolic Engineering, 2013, 15: 174-183.
41	OLSON M L, JOHNSON J, CARSWELL W F, et al. Characterization of an evolved carotenoids hyper-producer of Saccharomyces cerevisiae through bioreactor parameter optimization and Raman spectroscopy[J]. Journal of Industrial Microbiology & Biotechnology, 2016, 43(10): 1355-1363.
42	REYES L H, GOMEZ J M, KAO K C. Improving carotenoids production in yeast via adaptive laboratory evolution[J]. Metabolic Engineering, 2014, 21: 26-33.
43	YANG C, GAO X, JIANG Y, et al. Synergy between methylerythritol phosphate pathway and mevalonate pathway for isoprene production in Escherichia coli[J]. Metabolic Engineering, 2016, 37: 79-91.
44	YOSHIDA R, YOSHIMURA T, HEMMI H. Reconstruction of the “archaeal” mevalonate pathway from the methanogenic archaeon Methanosarcina mazei in Escherichia coli cells[J]. Applied and Environmental Microbiology, 2020, 86(6): e02889.
45	CHATZIVASILEIOU A O, WARD V, EDGAR S M, et al. Two-step pathway for isoprenoid synthesis[J]. Proc. Natl. Acad. Sci. USA, 2019, 116(2): 506-511.
46	CLOMBURG J M, QIAN S, TAN Z, et al. The isoprenoid alcohol pathway, a synthetic route for isoprenoid biosynthesis[J]. Proceedings of the National Academy of Sciences of the United States of America, 2019, 26(166): 12810-12815.
47	CHEN Y, XIAO W, WANG Y, et al. Lycopene overproduction in Saccharomyces cerevisiae through combining pathway engineering with host engineering[J]. Microbial Cell Factories, 2016, 15(1): 113.
48	KILDEGAARD K R, ADIEGO-PÉREZ B, DOMÉNECH BELDA D, et al. Engineering of Yarrowia lipolytica for production of astaxanthin[J]. Synthetic and Systems Biotechnology, 2017, 2(4): 287-294.
49	LIANG N, CHEN C, WANG Y, et al. Exploring catalysis specificity of phytoene dehydrogenase CrtI in carotenoid synthesis[J]. ACS Synthetic Biology, 2020, 9(7): 1753-1762.
50	LU Q, BU Y, LIU J. Metabolic engineering of Escherichia coli for producing astaxanthin as the predominant carotenoid[J]. Marine Drugs, 2017, 15(10): 296.
51	HENKE N A, HEIDER S A, PETERS-WENDISCH P, et al. Production of the marine carotenoid astaxanthin by metabolically engineered Corynebacterium glutamicum[J]. Marine Drugs, 2016, 14(7): 124.
52	WANG R, GU X, YAO M, et al. Engineering of β-carotene hydroxylase and ketolase for astaxanthin overproduction in Saccharomyces cerevisiae[J]. Frontiers of Chemical Science and Engineering, 2017, 11(1): 89-99.
53	ZHOU P, XIE W, LI A, et al. Alleviation of metabolic bottleneck by combinatorial engineering enhanced astaxanthin synthesis in Saccharomyces cerevisiae[J]. Enzyme and Microbial Technology, 2017, 100: 28-36.
54	SWEETLOVE L J, FERNIE A R. The role of dynamic enzyme assemblies and substrate channelling in metabolic regulation[J]. Nature Communications, 2018, 9(1): 2136.
55	KANG W, MA T, LIU M, et al. Modular enzyme assembly for enhanced cascade biocatalysis and metabolic flux[J]. Nature Communications, 2019, 10(1): 4248.
56	XU X, TIAN L, TANG S, et al. Design and tailoring of an artificial DNA scaffolding system for efficient lycopene synthesis using zinc-finger-guided assembly[J]. Journal of Industrial Microbiology & Biotechnology, 2020, 47(2): 209-222.
57	YE L, ZHU X, WU T, et al. Optimizing the localization of astaxanthin enzymes for improved productivity[J]. Biotechnology for Biofuels, 2018, 11(1): 278.
58	QU J, CAO S, WEI Q, et al. Synthetic multienzyme complexes, catalytic nanomachineries for cascade biosynthesis in vivo[J]. ACS Nano, 2019, 13(9): 9895-9906.
59	NOGUEIRA M, ENFISSI E M A, WELSCH R, et al. Construction of a fusion enzyme for astaxanthin formation and its characterisation in microbial and plant hosts: a new tool for engineering ketocarotenoids[J]. Metabolic Engineering, 2019(52): 243-252.
60	ZHANG X, NIE M, CHEN J, et al. Multicopy integrants of crt genes and co-expression of AMP deaminase improve lycopene production in Yarrowia lipolytica[J]. Journal of Biotechnology, 2019, 289: 46-54.
61	WU X, LI B, ZHANG W, et al. Genome-wide landscape of position effects on heterogeneous gene expression in Saccharomyces cerevisiae[J]. Biotechnology for Biofuels, 2017, 10(1): 189.
62	LARROUDE M, CELINSKA E, BACK A, et al. A synthetic biology approach to transform Yarrowia lipolytica into a competitive biotechnological producer of β-carotene[J]. Biotechnology and Bioengineering, 2018, 115(2): 464-472.
63	SU B, SONG D, YANG F, et al. Engineering a growth-phase-dependent biosynthetic pathway for carotenoid production in Saccharomyces cerevisiae[J]. Journal of Industrial Microbiology & Biotechnology, 2020, 47(4/5): 383-393.
64	XU J, XU X, XU Q, et al. Efficient production of lycopene by engineered E. Coli strains harboring different types of plasmids[J]. Bioprocess and Biosystems Engineering, 2018, 41(4): 489-499.
65	YE L, HE P, LI Q, et al. Type Ⅱs restriction based combinatory modulation technique for metabolic pathway optimization[J]. Microbial Cell Factories, 2017, 16(1): 47.
66	COUSSEMENT P, BAUWENS D, MAERTENS J, et al. Direct combinatorial pathway optimization[J]. ACS Synthetic Biology, 2016, 6(2): 224-232.
67	LIAN J, JIN R, ZHAO H. Construction of plasmids with tunable copy numbers in Saccharomyces cerevisiae and their applications in pathway optimization and multiplex genome integration[J]. Biotechnology and Bioengineering, 2016, 113(11): 2462-2473.
68	KANG C W, LIM H G, YANG J, et al. Synthetic auxotrophs for stable and tunable maintenance of plasmid copy number[J]. Metabolic Engineering, 2018, 48: 121-128.
69	YAMADA R, YAMAUCHI A, ANDO Y, et al. Modulation of gene expression by cocktail δ-integration to improve carotenoid production in Saccharomyces cerevisiae[J]. Bioresource Technology, 2018, 268: 616-621.
70	RONDA C, MAURY J, JAKOČIUNAS T, et al. CrEdit: CRISPR mediated multi-loci gene integration in Saccharomyces cerevisiae[J]. Microbial Cell Factories, 2015, 14(1): 97.
71	HOU S, QIN Q, DAI J. Wicket: a versatile tool for the integration and optimization of exogenous pathways in Saccharomyces cerevisiae[J]. ACS Synthetic Biology, 2018, 7(3): 782-788.
72	CUI Z, JIANG X, ZHENG H, et al. Homology-independent genome integration enables rapid library construction for enzyme expression and pathway optimization in Yarrowia lipolytica[J]. Biotechnology Bioengineering, 2019, 2(116): 354-363.
73	VOGL T, KICKENWEIZ T, PITZER J, et al. Engineered bidirectional promoters enable rapid multi-gene co-expression optimization[J]. Nature Communications, 2018, 9(1): 3589.
74	WU Z, ZHAO D, LI S, et al. Combinatorial modulation of initial codons for improved zeaxanthin synthetic pathway efficiency in Escherichia coli[J]. Microbiology Open, 2019, 8(12): e930.
75	JIN W, XU X, JIANG L, et al. Putative carotenoid genes expressed under the regulation of Shine-Dalgarno regions in Escherichia coli for efficient lycopene production[J]. Biotechnology Letters, 2015, 37(11): 2303-2310.
76	PFLEGER B F, PITERA D J, SMOLKE C D, et al. Combinatorial engineering of intergenic regions in operons tunes expression of multiple genes[J]. Nature Biotechnology, 2006, 24(8): 1027-1032.
77	LI X, TIAN G, SHEN H, et al. Metabolic engineering of Escherichia coli to produce zeaxanthin[J]. Journal of Industrial Microbiology & Biotechnology, 2015, 42(4): 627-636.
78	BLAZECK J, LIU L, REDDEN H, et al. Tuning gene expression in Yarrowia lipolytica by a hybrid promoter approach[J]. Applied and Environmental Microbiology, 2011, 77(22): 7905-7914.
79	BLAZECK J, GARG R, REED B, et al. Controlling promoter strength and regulation in Saccharomyces cerevisiae using synthetic hybrid promoters[J]. Biotechnology and Bioengineering, 2012, 109(11): 2884-2895.
80	NASERI G, BEHREND J, RIEPER L, et al. COMPASS for rapid combinatorial optimization of biochemical pathways based on artificial transcription factors[J]. Nature Communications, 2019, 10(1): 2615.
81	CUPERUS J T, LO R S, SHUMAKER L, et al. A tetO toolkit to alter expression of genes in Saccharomyces cerevisiae[J]. ACS Synthetic Biology, 2015, 4(7): 842-852.
82	GUO Y, DONG J, ZHOU T, et al. YeastFab: the design and construction of standard biological parts for metabolic engineering in Saccharomyces cerevisiae[J]. Nucleic Acids Research, 2015, 43(13): e88.
83	WU Y, ZHU R, MITCHELL L A, et al. In vitro DNA SCRaMbLE[J]. Nature Communications, 2018, 9(1): 1935.
84	LIU W, LUO Z, WANG Y, et al. Rapid pathway prototyping and engineering using in vitro and in vivo synthetic genome SCRaMbLE-in methods[J]. Nature Communications, 2018, 9(1): 1936.
85	QI D, JIN J, LIU D, et al. In vitro and in vivo recombination of heterologous modules for improving biosynthesis of astaxanthin in yeast[J]. Microbial Cell Factories, 2020, 19(1): 103.
86	AJIKUMAR P K, XIAO W H, TYO K E J, et al. Isoprenoid pathway optimization for taxol precursor overproduction in Escherichia coli[J]. Science, 2010, 330(6000): 70-74.
87	ZHANG C, SEOW V Y, CHEN X, et al. Multidimensional heuristic process for high-yield production of astaxanthin and fragrance molecules in Escherichia coli[J]. Nature Communications, 2018, 9(1): 1858.
88	LI N, HAN Z, O'DONNELL T J, et al. Production and excretion of astaxanthin by engineered Yarrowia lipolytica using plant oil as both the carbon source and the biocompatible extractant[J]. Appl. Microbiol. Biotechnol., 2020, 104(16): 6977-6989.
89	GOTO S, KOGURE K, ABE K, et al. Efficient radical trapping at the surface and inside the phospholipid membrane is responsible for highly potent antiperoxidative activity of the carotenoid astaxanthin[J]. Biochim. Biophys. Acta, 2001, 1512(2): 251-258.
90	VERWAAL R, JIANG Y, WANG J, et al. Heterologous carotenoid production in Saccharomyces cerevisiae induces the pleiotropic drug resistance stress response[J]. Yeast, 2010, 27(12): 983-998.
91	ZHOU P, XIE W, YAO Z, et al. Development of a temperature-responsive yeast cell factory using engineered Gal4 as a protein switch[J]. Biotechnology and Bioengineering, 2018, 115(5): 1321-1330.
92	DOSHI R, NGUYEN T, CHANG G. Transporter-mediated biofuel secretion[J]. Proceedings of the National Academy of Sciences, 2013, 110(19): 7642-7647.
93	WU T, LI S, YE L, et al. Engineering an artificial membrane vesicle trafficking system (AMVTS) for the excretion of β-carotene in Escherichia coli[J]. ACS Synthetic Biology, 2019, 8(5): 1037-1046.
94	WU T, YE L, ZHAO D, et al. Membrane engineering: a novel strategy to enhance the production and accumulation of β-carotene in Escherichia coli[J]. Metabolic Engineering, 2017, 43(A): 85-91.
95	WU T, YE L, ZHAO D, et al. Engineering membrane morphology and manipulating synthesis for increased lycopene accumulation in Escherichia coli cell factories[J]. Biotech, 2018, 8(6): 269.
96	LIU Y, LOW Z J, MA X, et al. Using biopolymer bodies for encapsulation of hydrophobic products in bacterium[J]. Metabolic Engineering, 2020, 61: 206-214.
97	CHEN Y, WANG Y, LIU M, et al. Primary and secondary metabolic effects of a key gene deletion (ΔYPL062W) in metabolically engineered terpenoid-producing Saccharomyces cerevisiae[J]. Appl. Environ. Microbiol., 2019, 85(7): e01990.
98	WANG M, LIU G, LIU H, et al. Engineering global transcription to tune lipophilic properties in Yarrowia lipolytica[J]. Biotechnology for Biofuels, 2018, 11(1): 115.
99	HUANG L, PU Y, YANG X, et al. Engineering of global regulator cAMP receptor protein (CRP) in Escherichia coli for improved lycopene production[J]. Journal of Biotechnology, 2015, 199: 55-61.
100	张雪, 张晓菲, 王立言, 等. 常压室温等离子体生物诱变育种及其应用研究进展[J]. 化工学报, 2014, 65(7): 2676-2684.
	ZHANG Xue, ZHANG Xiaofei, WANG Liyan, et al. Recent progress on atmospheric and room temperature plasma mutation breeding technology and its applications[J]. CIESC Journal, 2014, 65(7): 2676-2684.
101	JIN J, WANG Y, YAO M, et al. Astaxanthin overproduction in yeast by strain engineering and new gene target uncovering[J]. Biotechnology for Biofuels, 2018, 11(1): 215-230.
102	WANG H H, ISAACS F J, CARR P A, et al. Programming cells by multiplex genome engineering and accelerated evolution[J]. Nature, 2009, 460(7257): 894-898.
103	BARBIERI E M, MUIR P, AKHUETIEONI B O, et al. Precise editing at DNA replication forks enables multiplex genome engineering in eukaryotes[J]. Cell, 2017, 171(6): 1453-1467.
104	LI J, SHEN J, SUN Z, et al. Discovery of several novel targets that enhance β-carotene production in Saccharomyces cerevisiae[J]. Frontiers in Microbiology, 2017, 8: 1116.
105	BEUTTLER H, HOFFMANN J, JESKE M, et al. Biosynthesis of zeaxanthin in recombinant Pseudomonas putida[J]. Applied Microbiology and Biotechnology, 2011, 89(4): 1137-1147.
106	LIAN J, HAMEDIRAD M, HU S, et al. Combinatorial metabolic engineering using an orthogonal tri-functional CRISPR system[J]. Nature Communications, 2017, 8(1): 1688.
107	LIU D, LIU H, QI H, et al. Constructing yeast chimeric pathways to boost lipophilic terpene synthesis[J]. ACS Synthetic Biology, 2019, 8(4): 724-733.
108	JIA B, WU Y, LI B Z, et al. Precise control of SCRaMbLE in synthetic haploid and diploid yeast[J]. Nature Communications, 2018, 9(1): 1933.
109	WANG J, XIE Z X, MA Y, et al. Ring synthetic chromosome V SCRaMbLE[J]. Nature Communications, 2018, 9(1): 3783.
110	SUN X, REN L, BI Z, et al. Development of a cooperative two-factor adaptive-evolution method to enhance lipid production and prevent lipid peroxidation in Schizochytrium sp.[J]. Biotechnology for Biofuels, 2018, 11(1): 65.
111	SUN X, REN L, JI X, et al. Adaptive evolution of Schizochytrium sp. by continuous high oxygen stimulations to enhance docosahexaenoic acid synthesis[J]. Bioresource Technology, 2016, 211: 374-381.
112	TILLICH U M, WOLTER N, FRANKE P, et al. Screening and genetic characterization of thermo-tolerant Synechocystis sp.PCC6803 strains created by adaptive evolution[J]. BMC Biotechnology, 2014, 14(1): 66.
113	PERRINEAU M, ZELZION E, GROSS J, et al. Evolution of salt tolerance in a laboratory reared population of Chlamydomonas reinhardtii[J]. Environmental Microbiology, 2014, 16(6): 1755-1766.
114	LIN Y, CHANG J, LIN H, et al. Metabolic engineering a yeast to produce astaxanthin[J]. Bioresource Technology, 2017, 245: 899-905.
115	HEIDER S A E, WOLF N, HOFEMEIER A, et al. Optimization of the IPP precursor supply for the production of lycopene, decaprenoxanthin and astaxanthin by Corynebacterium glutamicum[J]. Frontiers in Bioengineering and Biotechnology, 2014, 2: 28.
116	ZHANG X, WANG D, DUAN Y, et al. Production of lycopene by metabolically engineered Pichia pastoris[J]. Biosci. Biotechnol. Biochem., 2020, 84(3): 463-470.
117	HASUNUMA T, TAKAKI A, MATSUDA M, et al. Single-stage astaxanthin production enhances the nonmevalonate pathway and photosynthetic central metabolism in Synechococcus sp. PCC 7002[J]. ACS Synthetic Biology, 2019, 8(12): 2701-2709.
118	YE X, ALBABILI S, KLOTI A, et al. Engineering the provitamin a (β-carotene) biosynthetic pathway into (carotenoid-free) rice endosperm[J]. Science, 2000, 287(5451): 303-305.
119	PAINE J A, SHIPTON C A, CHAGGAR S, et al. Improving the nutritional value of Golden Rice through increased pro-vitamin a content[J]. Nature Biotechnology, 2005, 23(4): 482-487.
120	ZHU Q, ZENG D, YU S, et al. From golden rice to aSTARice: bioengineering astaxanthin biosynthesis in rice endosperm[J]. Molecular Plant, 2018, 11(12): 1440-1448.
121	DIAO J, SONG X, ZHANG L, et al. Tailoring cyanobacteria as a new platform for highly efficient synthesis of astaxanthin[J]. Metabolic Engineering, 2020, 61: 275-287.
122	UMENO D, TOBIAS A V, ARNOLD F H. Diversifying carotenoid biosynthetic pathways by directed evolution[J]. Microbiol. Mol. Biol. Rev., 2005, 69(1): 51-78.
123	FURUBAYASHI M, IKEZUMI M, TAKAICHI S, et al. A highly selective biosynthetic pathway to non-natural C₅₀ carotenoids assembled from moderately selective enzymes[J]. Nature Communications, 2015, 6(1): 7534.
124	LI L, FURUBAYASHI M, WANG S, et al. Genetically engineered biosynthetic pathways for nonnatural C₆₀ carotenoids using C₅-elongases and C₅₀-cyclases in Escherichia coli[J]. Scientific Reports, 2019, 9: 2982.
125	DRUMMOND L, KSCHOWAK M J, BREITENBACH J, et al. Expanding the isoprenoid building block repertoire with an IPP methyltransferase from Streptomyces monomycini[J]. ACS Synthetic Biology, 2019, 8(6): 1303-1313.
126	LI L, FURUBAYASHI M, OTANI Y, et al. Nonnatural biosynthetic pathway for 2-hydroxylated xanthophylls with C₅₀-carotenoid backbone[J]. Journal of Bioscience and Bioengineering, 2019, 128(4): 438-444.
127	ZENG W, GUO L, XU S, et al. High-throughput screening technology in industrial biotechnology[J]. Trends in Biotechnology, 2020, 38(8): 888-906.
128	EXLEY K, REYNOLDS C R, SUCKLING L, et al. Utilising datasheets for the informed automated design and build of a synthetic metabolic pathway[J]. Journal of Biological Engineering, 2019, 8(13): 1-10.
129	ZHOU Y, LI G, DONG J, et al. MiYA, an efficient machine-learning workflow in conjunction with the YeastFab assembly strategy for combinatorial optimization of heterologous metabolic pathways in Saccharomyces cerevisiae[J]. Metabolic Engineering, 2018, 47: 294-302.
130	HAMEDIRAD M, CHAO R, WEISBERG S, et al. Towards a fully automated algorithm driven platform for biosystems design[J]. Nature Communications, 2019, 10(1): 5150.

底盘物种	产品	工程化手段	产量/mg·L^-1	产率/mg·g DCW^-1	参考文献
大肠杆菌	番茄红素	过表达磷酸戊糖途径和TCA循环关键蛋白戊二酸脱氢酶SucAB、琥珀酸脱氢酶SdhABCD和转醛酶B TalB，提高底盘NADPH和ATP的供给基于RBS序列调节dxs、idi和crt基因簇的转录水平	3520	50.6	[16]
	β-胡萝卜素	将合成路径划分为MEP路径、β-胡萝卜素合成、ATP合成、磷酸戊糖途径、TCA循环5个模块，分别进行工程化优化改造调节MEP路径模块和β-胡萝卜素合成模块表达强度，过表达dxs和idi 过表达α-酮戊二酸脱氢酶SucAB和琥珀酸脱氢酶SdhABCD，增强TCA循环，提高能量和还原力NADPH供给过表达转醛酶TalB，增强磷酸戊糖途径，提高NADPH供给	2100	59.9	[23]
	β-胡萝卜素	敲除葡萄糖-6-磷酸脱氢酶编码基因zwf，减弱碳代谢流进入磷酸戊糖途径；以半乳糖渗透酶系统（galP）替代葡萄糖磷酸转移酶系统（ptsHIcrr）负责葡萄糖转运，减少磷酸烯醇式丙酮酸的消耗，提高MEP路径前体供给敲除醛还原酶编码基因yjgB，降低NADPH的消耗；过表达nadK，将NADH转化为NADPH，提高NADPH供给	2579	NP	[18]
	β-胡萝卜素	引入异源MVA路径上调dxs、fni、GPPS2，优化内源MEP路径发酵条件优化，以甘油为碳源	3200	NP	[11]
	玉米黄质	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，避免有毒中间代谢物积累	722.5	23.2	[24]
	虾青素	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优化培养基碳源和发酵温度，降低IPTG诱导浓度基于代谢流分析选择过表达靶点ispD和ispF 引入毒素-抗毒素系统hok/sok，提高表达质粒稳定性	432.8	7.12	[25]
	藏红花酸	组合筛选CCD、ALD、UGT 体外酶活实验确定UGT功能	4.42	NP	[26]
酿酒酵母	番茄红素	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	1610	24.41	[27]
	番茄红素	过表达乙醇脱氢酶ADH2、乙醛脱氢酶ALD6和来自沙门氏菌的乙酰-CoA合成酶ACS，提高前体乙酰-CoA的供给过表达tHMGR，增强MVA路径过表达NADH激酶POS5，促进NADH向NADPH的转化增加CrtE和CrtI的拷贝数敲除YPL062W和EXG1 引入乙酰-CoA羧化酶ACC1突变体S659A/S1157A，过表达磷脂酸磷酸酶PAH1和二酰基甘油酰基转移酶DGA1，增强甘油三酯的合成过表达脂肪酸脱氢酶OLE1，增加不饱和脂肪酸的含量敲除Seipin编码基因FLD1，增大脂质体体积	2370	73.3	[14]
	番茄红素	启动子筛选过表达tHMGR，增强MVA路径筛选CrtE、CrtB、CrtI来源，增加CrtE和CrtI的拷贝数过表达NADH激酶POS5，促进NADH向NADPH的转化过表达乙醇脱氢酶ADH2、乙醛脱氢酶ALD6和来自沙门氏菌的乙酰-CoA合成酶ACS，提高前体乙酰-CoA的供给 ΔGAL80与ΔGAL1,7,10诱导系统的比较	3280	NP	[17]
	番茄红素	敲除YPL062W提高乙酰-CoA供给组合筛选CrtE、CrtB、CrtI来源调节CrtI表达水平选择底盘交配型工程化非路径相关靶点，即敲除ROX1和过表达INO2 引入异源血红蛋白VHb 连续自循环发酵	5880	NP	[28]
	β-胡萝卜素	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	1156	20.8	[29]
	虾青素	CrtZ和CrtW协同定向进化温敏调控系统控制关键基因表达	235.0	NP	[30]
	虾青素	等离子体诱变与适应性进化交替，促进菌株进化	404.8	NP	[31]
	藏红花酸	组合筛选CrtZ、CCD、ALD 发酵温度优化 CCD和ALD的过表达	6.278	NP	[32]
	藏红花酸	引入温敏调控系统控制藏红花酸合成，隔离细胞生长和生产调节CCD2和ALDH的拷贝数培养基碳源葡萄糖和半乳糖比例优化	NP	0.14	[33]
	藏红花酸	强化前体供应：敲除柠檬酸合酶CIT2、苹果酸合酶MLS1 CrtZ和CCD2的融合表达	12.43	NP	[34]
耶氏解脂酵母	番茄红素	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	4200	约170	[35]
	β-胡萝卜素	敲除KU70，增强同源重组效率敲除过氧化氢酶编码基因YALI0F30987、脂代谢相关基因LIP1、POX3-6，并作为后续表达盒整合位点过表达3个拷贝tHMGR、4个拷贝CrtYB和1个拷贝的CrtE、CrtI 过表达2个拷贝的MVA路径蛋白ERG10、ERG13、ERG12、ERG8、ERG19、IDI、ERG20 用受葡萄糖和甘油抑制的启动子ALK1p替换ERG9本源启动子，以实现ERG9下调氮源抑制发酵	3968	49.0	[13]
	β-胡萝卜素	敲除脂代谢基因POX2、MFE、POX3、LIP1，增强脂质积累过表达HMGR、ERG13，增强MVA路径增加3个拷贝的CrtYB、2个拷贝的CrtI和1个拷贝的CrtE 下调鲨烯合酶的表达	4500	57.5	[36]
	β-胡萝卜素	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	6500	89.6	[37]
	虾青素	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	285	6.0	[38]

底盘物种	产品	工程化手段	产量/mg·L^-1	产率/mg·g DCW^-1	参考文献
大肠杆菌	番茄红素	过表达磷酸戊糖途径和TCA循环关键蛋白戊二酸脱氢酶SucAB、琥珀酸脱氢酶SdhABCD和转醛酶B TalB，提高底盘NADPH和ATP的供给基于RBS序列调节dxs、idi和crt基因簇的转录水平	3520	50.6	[16]
	β-胡萝卜素	将合成路径划分为MEP路径、β-胡萝卜素合成、ATP合成、磷酸戊糖途径、TCA循环5个模块，分别进行工程化优化改造调节MEP路径模块和β-胡萝卜素合成模块表达强度，过表达dxs和idi 过表达α-酮戊二酸脱氢酶SucAB和琥珀酸脱氢酶SdhABCD，增强TCA循环，提高能量和还原力NADPH供给过表达转醛酶TalB，增强磷酸戊糖途径，提高NADPH供给	2100	59.9	[23]
	β-胡萝卜素	敲除葡萄糖-6-磷酸脱氢酶编码基因zwf，减弱碳代谢流进入磷酸戊糖途径；以半乳糖渗透酶系统（galP）替代葡萄糖磷酸转移酶系统（ptsHIcrr）负责葡萄糖转运，减少磷酸烯醇式丙酮酸的消耗，提高MEP路径前体供给敲除醛还原酶编码基因yjgB，降低NADPH的消耗；过表达nadK，将NADH转化为NADPH，提高NADPH供给	2579	NP	[18]
	β-胡萝卜素	引入异源MVA路径上调dxs、fni、GPPS2，优化内源MEP路径发酵条件优化，以甘油为碳源	3200	NP	[11]
	玉米黄质	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，避免有毒中间代谢物积累	722.5	23.2	[24]
	虾青素	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优化培养基碳源和发酵温度，降低IPTG诱导浓度基于代谢流分析选择过表达靶点ispD和ispF 引入毒素-抗毒素系统hok/sok，提高表达质粒稳定性	432.8	7.12	[25]
	藏红花酸	组合筛选CCD、ALD、UGT 体外酶活实验确定UGT功能	4.42	NP	[26]
酿酒酵母	番茄红素	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	1610	24.41	[27]
	番茄红素	过表达乙醇脱氢酶ADH2、乙醛脱氢酶ALD6和来自沙门氏菌的乙酰-CoA合成酶ACS，提高前体乙酰-CoA的供给过表达tHMGR，增强MVA路径过表达NADH激酶POS5，促进NADH向NADPH的转化增加CrtE和CrtI的拷贝数敲除YPL062W和EXG1 引入乙酰-CoA羧化酶ACC1突变体S659A/S1157A，过表达磷脂酸磷酸酶PAH1和二酰基甘油酰基转移酶DGA1，增强甘油三酯的合成过表达脂肪酸脱氢酶OLE1，增加不饱和脂肪酸的含量敲除Seipin编码基因FLD1，增大脂质体体积	2370	73.3	[14]
	番茄红素	启动子筛选过表达tHMGR，增强MVA路径筛选CrtE、CrtB、CrtI来源，增加CrtE和CrtI的拷贝数过表达NADH激酶POS5，促进NADH向NADPH的转化过表达乙醇脱氢酶ADH2、乙醛脱氢酶ALD6和来自沙门氏菌的乙酰-CoA合成酶ACS，提高前体乙酰-CoA的供给 ΔGAL80与ΔGAL1,7,10诱导系统的比较	3280	NP	[17]
	番茄红素	敲除YPL062W提高乙酰-CoA供给组合筛选CrtE、CrtB、CrtI来源调节CrtI表达水平选择底盘交配型工程化非路径相关靶点，即敲除ROX1和过表达INO2 引入异源血红蛋白VHb 连续自循环发酵	5880	NP	[28]
	β-胡萝卜素	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	1156	20.8	[29]
	虾青素	CrtZ和CrtW协同定向进化温敏调控系统控制关键基因表达	235.0	NP	[30]
	虾青素	等离子体诱变与适应性进化交替，促进菌株进化	404.8	NP	[31]
	藏红花酸	组合筛选CrtZ、CCD、ALD 发酵温度优化 CCD和ALD的过表达	6.278	NP	[32]
	藏红花酸	引入温敏调控系统控制藏红花酸合成，隔离细胞生长和生产调节CCD2和ALDH的拷贝数培养基碳源葡萄糖和半乳糖比例优化	NP	0.14	[33]
	藏红花酸	强化前体供应：敲除柠檬酸合酶CIT2、苹果酸合酶MLS1 CrtZ和CCD2的融合表达	12.43	NP	[34]
耶氏解脂酵母	番茄红素	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	4200	约170	[35]
	β-胡萝卜素	敲除KU70，增强同源重组效率敲除过氧化氢酶编码基因YALI0F30987、脂代谢相关基因LIP1、POX3-6，并作为后续表达盒整合位点过表达3个拷贝tHMGR、4个拷贝CrtYB和1个拷贝的CrtE、CrtI 过表达2个拷贝的MVA路径蛋白ERG10、ERG13、ERG12、ERG8、ERG19、IDI、ERG20 用受葡萄糖和甘油抑制的启动子ALK1p替换ERG9本源启动子，以实现ERG9下调氮源抑制发酵	3968	49.0	[13]
	β-胡萝卜素	敲除脂代谢基因POX2、MFE、POX3、LIP1，增强脂质积累过表达HMGR、ERG13，增强MVA路径增加3个拷贝的CrtYB、2个拷贝的CrtI和1个拷贝的CrtE 下调鲨烯合酶的表达	4500	57.5	[36]
	β-胡萝卜素	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	6500	89.6	[37]
	虾青素	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	285	6.0	[38]

底盘物种	优势	产品	产量 /mg·L^-1	产率 /mg·g DCW^-1	参考文献
恶臭假单胞菌（Pseudomonas putida）	化学品、有机溶剂耐受性高 NADPH再生快	玉米黄质	239^①	NP	[105]
谷氨酸棒杆菌（Corynebacterium glutamicum）	可利用多种碳源天然合成C₅₀类胡萝卜素	虾青素	NP	1.6	[51]
毕赤酵母（Pichia pastoris）	蛋白表达的优势宿主，翻译后修饰更接近天然蛋白	番茄红素	714	9.319	[116]
克鲁维酵母（Kluyveromyces marxianus）	耐热性好，可在25~52℃的温度下发酵单位碳源生物量转化率高	虾青素	NP	9.972	[114]
集胞藻（Synechocystis sp.）	光自养型微生物；单位碳源生物量转化率高	虾青素	NP	29.6^①	[121]

底盘物种	优势	产品	产量 /mg·L^-1	产率 /mg·g DCW^-1	参考文献
恶臭假单胞菌（Pseudomonas putida）	化学品、有机溶剂耐受性高 NADPH再生快	玉米黄质	239^①	NP	[105]
谷氨酸棒杆菌（Corynebacterium glutamicum）	可利用多种碳源天然合成C₅₀类胡萝卜素	虾青素	NP	1.6	[51]
毕赤酵母（Pichia pastoris）	蛋白表达的优势宿主，翻译后修饰更接近天然蛋白	番茄红素	714	9.319	[116]
克鲁维酵母（Kluyveromyces marxianus）	耐热性好，可在25~52℃的温度下发酵单位碳源生物量转化率高	虾青素	NP	9.972	[114]
集胞藻（Synechocystis sp.）	光自养型微生物；单位碳源生物量转化率高	虾青素	NP	29.6^①	[121]

[1]	GAO Cong, CHEN Chenghu, CHEN Xiulai, LIU Liming. Progress and challenges of engineering microorganisms to produce biobased monomers [J]. Chemical Industry and Engineering Progress, 2023, 42(8): 4123-4135.
[2]	GUO Feng, ZHANG Shangjie, JIANG Yujia, JIANG Wankui, XIN Fengxue, ZHANG Wenming, JIANG Min. Biotransformation of one-carbon resources by yeast [J]. Chemical Industry and Engineering Progress, 2023, 42(1): 30-39.
[3]	TAO Yuxuan, ZHANG Shangjie, JING Yiwen, XIN Fengxue, DONG Weiliang, ZHOU Jie, JIANG Yujia, ZHANG Wenming, JIANG Min. Recent advances in the construction strategy of methylotrophic Escherichia coli [J]. Chemical Industry and Engineering Progress, 2021, 40(7): 3932-3941.
[4]	SUN Wentao, LI Chun. Design and construction of microbial cell factory for biosynthesis of plant natural products [J]. Chemical Industry and Engineering Progress, 2021, 40(3): 1202-1214.
[5]	GUO Liang, GAO Cong, ZHANG Li, CHEN Xiulai, LIU Liming. Advances in the suitability of artificial metabolic pathways [J]. Chemical Industry and Engineering Progress, 2021, 40(3): 1252-1261.
[6]	LIU Weibing, YE Bangce. Progress of synthetic biology research and biological manufacturing of actinomycetes polyketides [J]. Chemical Industry and Engineering Progress, 2021, 40(3): 1226-1237.
[7]	MA Yueyuan, CHEN Jinchun, CHEN Guoqiang. Halophilic microorganisms as microbial chassis: applications and prospects [J]. Chemical Industry and Engineering Progress, 2021, 40(3): 1178-1186.
[8]	GAO Cong, GUO Liang, HU Guipeng, CHEN Xiulai, LIU Liming. Advances of metabolic flux regulation in microbial cell factories [J]. Chemical Industry and Engineering Progress, 2021, 40(12): 6807-6817.
[9]	Chen WANG, Meng ZHAO, Mingzhu DING, Ying WANG, Mingdong YAO, Wenhai XIAO. Application of biological scaffold system on synthetic biology [J]. Chemical Industry and Engineering Progress, 2020, 39(11): 4557-4567.
[10]	Pengcheng CHANG, Yang YU, Ying WANG, Chun LI. Combinatorial regulation strategies for efficient synthesis of terpenoids in Saccharomyces cerevisiae [J]. Chemical Industry and Engineering Progress, 2019, 38(01): 598-605.
[11]	YANG Kun, WANG Ying, LI Chun. Cell transporter protein and engineered applications [J]. Chemical Industry and Engineering Progress, 2017, 36(04): 1410-1417.
[12]	LIU Dingyu, MENG Jiao, WANG Zhiwen, CHEN Tao, ZHAO Xueming. Progress and application on multivariate modular metabolic engineering in metabolic engineering [J]. Chemical Industry and Engineering Progress, 2016, 35(11): 3619-3626.
[13]	XIAO Wenhai, ZHOU Sijie, WANG Ying, YUAN Yingjin. How to make biology more “engineering” [J]. Chemical Industry and Engineering Progree, 2016, 35(06): 1827-1836.
[14]	WAN Tao1，QI Haishan1，CHEN Yunlin2，WEN Jianping1. Research progress of biosynthesis of daptomycin and its derivative [J]. Chemical Industry and Engineering Progree, 2012, 31(07): 1581-1586.