高能燃料前体α-红没药烯的生物制造过程优化

doi:10.11949/0438-1157.20240276

化工学报 ›› 2024, Vol. 75 ›› Issue (12): 4702-4711.DOI: 10.11949/0438-1157.20240276

高能燃料前体α-红没药烯的生物制造过程优化

马治东¹(), 张亚鹏², 高慧鹏³, 李文强⁴, 吕波², 秦磊⁴, 张全³(), 李春¹^,²^,⁴()

^1.石河子大学化学化工学院/化工绿色过程省部共建国家重点实验室培育基地，新疆石河子 832003
^2.北京理工大学化学与化工学院生物化工研究所/医药分子科学与制剂工信部重点实验室，北京 100081
^3.中石化大连石油化工研究院有限公司，辽宁大连 116023
^4.清华大学化学工程系工业生物催化教育部重点实验室，北京 100084

收稿日期:2024-03-07 修回日期:2024-08-08 出版日期:2024-12-25 发布日期:2025-01-03
通讯作者: 张全,李春
作者简介:马治东（2000—），男，硕士研究生，zhidong_mashz@163.com
基金资助:
国家自然科学基金项目(22138006);中国石化课题(421127)

Optimization of biomanufacturing process of high-energy fuel precursor α-bisabolene

Zhidong MA¹(), Yapeng ZHANG², Huipeng GAO³, Wenqiang LI⁴, Bo LYU², Lei QIN⁴, Quan ZHANG³(), Chun LI¹^,²^,⁴()

^1.School of Chemistry and Chemical Engineering/State Key Laboratory Incubation Base for Green Processing of Chemical Engineering, Shihezi University, Shihezi 832003, Xinjiang, China
^2.Key Laboratory of Medical Molecule Science and Pharmaceutics Engineering, Ministry of Industry and Information Technology, Institute of Biochemical Engineering, School of Chemistry and Chemical Engineering, Beijing Institute of Technology, Beijing 100081, China
^3.Sinopec Dalian Research Institute of Petroleum and Petrochemicals, Dalian 116023, Liaoninng, China
^4.Key Laboratory for Industrial Biocatalysis, Ministry of Education, Department of Chemical Engineering, Tsinghua University, Beijing 100084, China

Received:2024-03-07 Revised:2024-08-08 Online:2024-12-25 Published:2025-01-03
Contact: Quan ZHANG, Chun LI

摘要/Abstract

摘要：

α-红没药烯是一种主要存在于红没药、柠檬等植物精油中的倍半萜化合物，因具备多支链、环状结构，其烷化产物红没药烷可作为航空燃料替代品，受到了国内外的广泛关注。以酿酒酵母为底盘细胞来合成α-红没药烯是一种绿色、可持续的备选生产方式。本研究在强化甲羟戊酸（mevalonate，MVA）合成途径的酿酒酵母中表达来自北美冷杉的α-红没药烯合酶AgBIS，利用葡萄糖诱导型启动子P _HXT1 对支路鲨烯合酶及脂肪酸合成途径进行动态下调，进一步系统增强MVA途径多个基因表达，并对辅因子供应与乙醇利用途径进行强化，使α-红没药烯在摇瓶中的产量达到510 mg/L。在3 L发酵罐中，通过乙醇补料发酵使α-红没药烯产量达到16.5 g/L，为目前报道的最高产量。本研究为微生物细胞工厂高效合成α-红没药烯提供了有效策略。

关键词: α-红没药烯, 酿酒酵母, 代谢工程, 微生物细胞工厂, 合成生物学

Abstract:

α-bisabolene is a sesquiterpene compound mainly found in plant essential oils such as myrrh and lemon. Due to its multi-branched and cyclic structure, its alkylation product bisabolane can be used as a substitute for aviation fuel and has attracted widespread attention at home and abroad. The biosynthesis of α-bisabolene using Saccharomyces cerevisiae as a chassis is a green and sustainable alternative production mode. In this study, a α-bisabolene synthase AgBIS from Abies grandis was expressed in S. cerevisiae with the enhanced key genes of the mevalonate (MVA) pathway. The glucose inducible promoter P _HXT1 was used to dynamically downregulate the endogenous squalene synthase and fatty acid synthesis pathway. Further systematic enhancement of the expression of multiple genes in the MVA pathway, cofactor supplementary and ethanol utilization pathway systematically, the α-bisabolene production reached 510 mg/L in the flask. In a 3 L fermenter, the highest reported production of α-bisabolene was achieved at 120 h with ethanol supplementation, reaching 16.5 g/L. This study provides an effective strategy and research foundation for the efficient biosynthesis of α-bisabolene by microbial cell factories.

Key words: α-bisabolene, Saccharomyces cerevisiae, metabolic engineering, microbial cell factory, synthetic biology

中图分类号:

Q 819

马治东, 张亚鹏, 高慧鹏, 李文强, 吕波, 秦磊, 张全, 李春. 高能燃料前体α-红没药烯的生物制造过程优化[J]. 化工学报, 2024, 75(12): 4702-4711.

Zhidong MA, Yapeng ZHANG, Huipeng GAO, Wenqiang LI, Bo LYU, Lei QIN, Quan ZHANG, Chun LI. Optimization of biomanufacturing process of high-energy fuel precursor α-bisabolene[J]. CIESC Journal, 2024, 75(12): 4702-4711.

图/表 9

参考文献 34

1	Zhao Y K, Zhu K, Li J, et al. High-efficiency production of bisabolene from waste cooking oil by metabolically engineered Yarrowia lipolytica [J]. Microbial Biotechnology, 2021, 14(6): 2497-2513.
2	Yeo S K, Ali A Y, Hayward O A, et al. β-bisabolene, a sesquiterpene from the essential oil extract of opoponax (Commiphora guidottii), exhibits cytotoxicity in breast cancer cell lines[J]. Phytotherapy Research, 2016, 30(3): 418-425.
3	Jou Y J, Hua C H, Lin C S, et al. Anticancer activity of γ-bisabolene in human neuroblastoma cells via induction of p53-mediated mitochondrial apoptosis[J]. Molecules, 2016, 21(5): 601.
4	Peralta-Yahya P P, Ouellet M, Chan R, et al. Identification and microbial production of a terpene-based advanced biofuel[J]. Nature Communications, 2011, 2: 483.
5	Baral N R, Kavvada O, Mendez-Perez D, et al. Techno-economic analysis and life-cycle greenhouse gas mitigation cost of five routes to bio-jet fuel blendstocks[J]. Energy & Environmental Science, 2019, 12(3): 807-824.
6	Wang P P, Wei W, Ye W, et al. Synthesizing ginsenoside Rh2 in Saccharomyces cerevisiae cell factory at high-efficiency[J]. Cell Discovery, 2019, 5: 5.
7	Liu C L, Xue K, Yang Y K, et al. Metabolic engineering strategies for sesquiterpene production in microorganism[J]. Critical Reviews in Biotechnology, 2022, 42(1): 73-92.
8	Meadows A L, Hawkins K M, Tsegaye Y, et al. Rewriting yeast central carbon metabolism for industrial isoprenoid production[J]. Nature, 2016, 537(7622): 694-697.
9	Westfall P J, Pitera D J, Lenihan J R, et al. Production of amorphadiene in yeast, and its conversion to dihydroartemisinic acid, precursor to the antimalarial agent artemisinin[J]. Proceedings of the National Academy of Sciences of the United States of America, 2012, 109(3): E111-E118.
10	Zhang Y, Song X H, Lai Y M, et al. High-yielding terpene-based biofuel production in Rhodobacter capsulatus [J]. ACS Synthetic Biology, 2021, 10(6): 1545-1552.
11	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.
12	Zhao B X, Zhang Y H, Wang Y P, et al. Biosynthesis of α-bisabolene from low-cost renewable feedstocks by peroxisome engineering and systems metabolic engineering of the yeast Yarrowia lipolytica [J]. Green Chemistry, 2023, 25(20): 8145-8159.
13	Zhang Y P, Wang J, Wang Z B, et al. A gRNA-tRNA array for CRISPR-Cas9 based rapid multiplexed genome editing in Saccharomyces cerevisiae [J]. Nature Communications, 2019, 10(1): 1053.
14	Gietz R D, Schiestl R H. High-efficiency yeast transformation using the LiAc/SS carrier DNA/PEG method[J]. Nature Protocols, 2007, 2(1): 31-34.
15	Chen H L, Li M J, Liu C Q, et al. Enhancement of the catalytic activity of isopentenyl diphosphate isomerase (IDI) from Saccharomyces cerevisiae through random and site-directed mutagenesis[J]. Microbial Cell Factories, 2018, 17(1): 65.
16	Donald K A, Hampton R Y, Fritz I B. Effects of overproduction of the catalytic domain of 3-hydroxy-3-methylglutaryl coenzyme A reductase on squalene synthesis in Saccharomyces cerevisiae [J]. Applied and Environmental Microbiology, 1997, 63(9): 3341-3344.
17	Bohlmann J, Crock J, Jetter R, et al. Terpenoid-based defenses in conifers: cDNA cloning, characterization, and functional expression of wound-inducible (E)-alpha-bisabolene synthase from grand fir (Abies grandis)[J]. Proceedings of the National Academy of Sciences of the United States of America, 1998, 95(12): 6756-6761.
18	Parveen I, Wang M, Zhao J P, et al. Investigating sesquiterpene biosynthesis in Ginkgo biloba: molecular cloning and functional characterization of (E, E)-farnesol and α-bisabolene synthases[J]. Plant Molecular Biology, 2015, 89(4/5): 451-462.
19	Mafu S, Karunanithi P S, Palazzo T A, et al. Biosynthesis of the microtubule-destabilizing diterpene pseudolaric acid B from golden larch involves an unusual diterpene synthase[J]. Proceedings of the National Academy of Sciences of the United States of America, 2017, 114(5): 974-979.
20	Lancaster J, Lehner B, Khrimian A, et al. An IDS-type sesquiterpene synthase produces the pheromone precursor (Z)- α-bisabolene in Nezara viridula [J]. Journal of Chemical Ecology, 2019, 45(2): 187-197.
21	Chan A P, Crabtree J, Zhao Q, et al. Draft genome sequence of the oilseed species Ricinus communis [J]. Nature Biotechnology, 2010, 28(9): 951-956.
22	Yuan J F, Ching C B. Dynamic control of ERG9 expression for improved amorpha-4,11-diene production in Saccharomyces cerevisiae [J]. Microbial Cell Factories, 2015, 14: 38.
23	Xie W P, Ye L D, Lv X M, et al. Sequential control of biosynthetic pathways for balanced utilization of metabolic intermediates in Saccharomyces cerevisiae [J]. Metabolic Engineering, 2015, 28: 8-18.
24	Dusséaux S, Wajn W T, Liu Y X, et al. Transforming yeast peroxisomes into microfactories for the efficient production of high-value isoprenoids[J]. Proceedings of the National Academy of Sciences of the United States of America, 2020, 117(50): 31789-31799.
25	Yu T, Zhou Y J, Huang M T, et al. Reprogramming yeast metabolism from alcoholic fermentation to lipogenesis[J]. Cell, 2018, 174(6): 1549-1558.e14.
26	Davies B S J, Wang H S, Rine J. Dual activators of the sterol biosynthetic pathway of Saccharomyces cerevisiae: similar activation/regulatory domains but different response mechanisms[J]. Molecular and Cellular Biology, 2005, 25(16): 7375-7385.
27	Ro D K, Paradise E M, Ouellet M, et al. Production of the antimalarial drug precursor artemisinic acid in engineered yeast[J]. Nature, 2006, 440(7086): 940-943.
28	van Rossum H M, Kozak B U, Pronk J T, et al. Engineering cytosolic acetyl-coenzyme A supply in Saccharomyces cerevisiae: pathway stoichiometry, free-energy conservation and redox-cofactor balancing[J]. Metabolic Engineering, 2016, 36: 99-115.
29	张鸿伟, 王鹏超. 微生物辅因子工程研究进展[J]. 中国生物工程杂志, 2023, 43(4): 112-122.
	Zhang H W, Wang P C. Research progress of microbial cofactor engineering[J]. China Biotechnology, 2023, 43(4): 112-122.
30	Asadollahi M A, Maury J, Patil K R, et al. Enhancing sesquiterpene production in Saccharomyces cerevisiae through in silico driven metabolic engineering[J]. Metabolic Engineering, 2009, 11(6): 328-334.
31	Hou J, Vemuri G N, Bao X M, et al. Impact of overexpressing NADH kinase on glucose and xylose metabolism in recombinant xylose-utilizing Saccharomyces cerevisiae [J]. Applied Microbiology and Biotechnology, 2009, 82(5): 909-919.
32	Paramasivan K, Mutturi S. Regeneration of NADPH coupled with HMG-CoA reductase activity increases squalene synthesis in Saccharomyces cerevisiae [J]. Journal of Agricultural and Food Chemistry, 2017, 65(37): 8162-8170.
33	陈孚江, 周景文, 史仲平, 等. 乙酰辅酶A合成代谢对酿酒酵母生理功能的影响[J]. 微生物学报, 2010, 50(9): 1172-1179.
	Chen F J, Zhou J W, Shi Z P, et al. Effect of acetyl-CoA synthase gene overexpression on physiological function of Saccharomyces cerevisiae [J]. Acta Microbiologica Sinica, 2010, 50(9): 1172-1179.
34	Huang Y L, Ye Z L, Wan X K, et al. Systematic mining and evaluation of the sesquiterpene skeletons as high energy aviation fuel molecules[J]. Advanced Science, 2023, 10(23): e2300889.

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质粒/菌株		基因型	来源
酿酒酵母	S59	CEN.PK2-1D, ΔGal80:: P _Gal10 -IDI1 (L141H,Y195F,W356C )-T _PGI1, P _Gal1-tHMG1-T _CYC3	课题组构建
	BS-GbTPS	S59, ΔYPRCtau3:: P _Gal1 -GbTPS-T _PGK1 -LEU2	本研究构建
	BS-PaTPS5	S59, ΔYPRCtau3:: P _Gal1 -PaTPS5-T _PGK1 -LEU2	本研究构建
	BS-NvTPS	S59, ΔYPRCtau3:: P _Gal1 -NvTPS-T _PGK1 -LEU2	本研究构建
	BS-RcTPS	S59, ΔYPRCtau3:: P _Gal1 -RcTPS-T _PGK1 -LEU2	本研究构建
	BS1	S59, ΔYPRCtau3:: P _Gal1 -AgBIS-T _PGK1 -LEU2	本研究构建
	BS2	BS1, ΔERG9:: P _HXT1 -ERG9-T _ERG9 -HIS3	本研究构建
	BS3	BS2, ΔACC1::P _HXT1 -ACC1-T _ACC1	本研究构建
	BS9	BS3, ΔLPP1, ΔDPP1:: P _Gal1 -UPC2-1-T _GPD1	本研究构建
	BS11	BS3, Δ308a:: P _Gal1 -ERG8-T _PRC1 -P _Gal10 -ERG19-T _EFM1	本研究构建
	BS11-174	BS11, ΔGRE3:: P_T_EF1 -t17POS5-T _CPS1 -TRP1	本研究构建
	BS11-A22	BS11, ΔGDH1:: P _TEF1-GDH2-T _PGK1	本研究构建
	BS13	BS11, Δ1622b::P _Gal1 -ERG20-T _RPL41 -P _Gal10 -ERG13-T _RPL3	本研究构建
	BS14	BS13, ΔURA3::P _Gal1 -ERG12-T _EBS1 -P _Gal10 -ERG10-T _NAT1	本研究构建
	BS17-A1	BS14, ΔGRE3::P _TEF1 -t17POS5-T _CPS1 -TRP1, Δ911b::P _Gal2 -ADH2-T _MRP49	本研究构建
	BS17-A2	BS17-A1, Δyprcdelta15::P _Gal1 -ACS1-T _ADH1 -P _Gal10-ERG10-T _VMA2	本研究构建
质粒	A1	pUC19, ΔYPRCtau3:: P _Gal1 -GbTPS-T _PGK1 -LEU2	本研究构建
	A2	pUC19, ΔYPRCtau3:: P _Gal1 -PaTPS5-T _PGK1 -LEU2	本研究构建
	A3	pUC19, ΔYPRCtau3:: P _Gal1 -NvTPS-T _PGK1 -LEU2	本研究构建
	A4	pUC19, ΔYPRCtau3:: P _Gal1 -RcTPS-T _PGK1 -LEU2	本研究构建
	A5	pUC19, ΔYPRCtau3:: P _Gal1 -AgBIS-T _PGK1 -LEU2	本研究构建
	A8	pUC19, ΔERG9:: P _HXT1 -ERG9-T _ERG9 -HIS3	课题组构建
	A9	pUC19, ΔACC1:: P _HXT1 -ACC1-T _ACC1	本研究构建
	A11	pUC19, Δ308a::P _Gal1 -ERG8-T _PRC1 -P _Gal10 -ERG19-T _EFM1	课题组构建
	A12	pUC19, Δ1622b::P _Gal1 -ERG20-T _RPL41 -P _Gal10 -ERG13-T _RPL3	课题组构建
	A13	pUC19, ΔURA3::P _Gal1 -ERG12-T _EBS1 -P _Gal10 -ERG10-T _NAT1	课题组构建
	A14	pUC19, ΔGDH1::P _TEF1-GDH2-T _PGK1	本研究构建
	A15	pUC19, ΔGRE3::P _TEF1 -t17POS5-T _CPS1 -TRP1	课题组构建
	A16	pUC19, Δyprcdelta15::P _Gal1 -ACS1-T _ADH1 -P _Gal10 -ERG10-T _VMA2	课题组构建
	A17	pUC19,Δ911b::P _Gal2 -ADH2-T _MRP49	本研究构建
	A18	pUC19,ΔDPP1::P _Gal1 -UPC2-1-T _GPD1	本研究构建

质粒/菌株		基因型	来源
酿酒酵母	S59	CEN.PK2-1D, ΔGal80:: P _Gal10 -IDI1 (L141H,Y195F,W356C )-T _PGI1, P _Gal1-tHMG1-T _CYC3	课题组构建
	BS-GbTPS	S59, ΔYPRCtau3:: P _Gal1 -GbTPS-T _PGK1 -LEU2	本研究构建
	BS-PaTPS5	S59, ΔYPRCtau3:: P _Gal1 -PaTPS5-T _PGK1 -LEU2	本研究构建
	BS-NvTPS	S59, ΔYPRCtau3:: P _Gal1 -NvTPS-T _PGK1 -LEU2	本研究构建
	BS-RcTPS	S59, ΔYPRCtau3:: P _Gal1 -RcTPS-T _PGK1 -LEU2	本研究构建
	BS1	S59, ΔYPRCtau3:: P _Gal1 -AgBIS-T _PGK1 -LEU2	本研究构建
	BS2	BS1, ΔERG9:: P _HXT1 -ERG9-T _ERG9 -HIS3	本研究构建
	BS3	BS2, ΔACC1::P _HXT1 -ACC1-T _ACC1	本研究构建
	BS9	BS3, ΔLPP1, ΔDPP1:: P _Gal1 -UPC2-1-T _GPD1	本研究构建
	BS11	BS3, Δ308a:: P _Gal1 -ERG8-T _PRC1 -P _Gal10 -ERG19-T _EFM1	本研究构建
	BS11-174	BS11, ΔGRE3:: P_T_EF1 -t17POS5-T _CPS1 -TRP1	本研究构建
	BS11-A22	BS11, ΔGDH1:: P _TEF1-GDH2-T _PGK1	本研究构建
	BS13	BS11, Δ1622b::P _Gal1 -ERG20-T _RPL41 -P _Gal10 -ERG13-T _RPL3	本研究构建
	BS14	BS13, ΔURA3::P _Gal1 -ERG12-T _EBS1 -P _Gal10 -ERG10-T _NAT1	本研究构建
	BS17-A1	BS14, ΔGRE3::P _TEF1 -t17POS5-T _CPS1 -TRP1, Δ911b::P _Gal2 -ADH2-T _MRP49	本研究构建
	BS17-A2	BS17-A1, Δyprcdelta15::P _Gal1 -ACS1-T _ADH1 -P _Gal10-ERG10-T _VMA2	本研究构建
质粒	A1	pUC19, ΔYPRCtau3:: P _Gal1 -GbTPS-T _PGK1 -LEU2	本研究构建
	A2	pUC19, ΔYPRCtau3:: P _Gal1 -PaTPS5-T _PGK1 -LEU2	本研究构建
	A3	pUC19, ΔYPRCtau3:: P _Gal1 -NvTPS-T _PGK1 -LEU2	本研究构建
	A4	pUC19, ΔYPRCtau3:: P _Gal1 -RcTPS-T _PGK1 -LEU2	本研究构建
	A5	pUC19, ΔYPRCtau3:: P _Gal1 -AgBIS-T _PGK1 -LEU2	本研究构建
	A8	pUC19, ΔERG9:: P _HXT1 -ERG9-T _ERG9 -HIS3	课题组构建
	A9	pUC19, ΔACC1:: P _HXT1 -ACC1-T _ACC1	本研究构建
	A11	pUC19, Δ308a::P _Gal1 -ERG8-T _PRC1 -P _Gal10 -ERG19-T _EFM1	课题组构建
	A12	pUC19, Δ1622b::P _Gal1 -ERG20-T _RPL41 -P _Gal10 -ERG13-T _RPL3	课题组构建
	A13	pUC19, ΔURA3::P _Gal1 -ERG12-T _EBS1 -P _Gal10 -ERG10-T _NAT1	课题组构建
	A14	pUC19, ΔGDH1::P _TEF1-GDH2-T _PGK1	本研究构建
	A15	pUC19, ΔGRE3::P _TEF1 -t17POS5-T _CPS1 -TRP1	课题组构建
	A16	pUC19, Δyprcdelta15::P _Gal1 -ACS1-T _ADH1 -P _Gal10 -ERG10-T _VMA2	课题组构建
	A17	pUC19,Δ911b::P _Gal2 -ADH2-T _MRP49	本研究构建
	A18	pUC19,ΔDPP1::P _Gal1 -UPC2-1-T _GPD1	本研究构建

引物	序列(5′-3′)
P _Gal1 -AgBIS-F1	GAAAAAACTATAATGGCTGGTGTTTCTGCTG
T _PGK1 - AgBIS-R1	GGAAAGCTTTTACAATGGCAATGGTTCGATCAAAC
AgBIS-T _PGK1 -F1	CCATTGTAAAAGCTTTCCCATGTCTCTACTGG
T _PGK1 -R1	AACGAACGCAGAATTTTCGAG
P _HXT1 -F1	GGTTCAAGCAGAAGAGACAACAATTG
P _HXT1 -ACC1-R1	AAGCTTTCTTCGCTCATGATTTTACGTATATCAACTAGTTGACGATTATG
ACC1-F1	ATGAGCGAAGAAAGCTTATTCGAG
ACC1-R1	GATGACTTTCCTCTTAGACTGGGAC
P _HXT1 -FAS1-R1	GTGGAGTAAGCGTCCATGATTTTACGTATATCAACTAGTTGACG
FAS1-F1	ATGGACGCTTACTCCACAAG
FAS1-R1	GATATAGATCACGCAATTCTTCAAAGTAG
GDH1L-P _TEF1 -F1	GAGACCAAAAAGAAAAAGAAGACATGGAGGCCCAGAATAC
GDH2-P _TEF1 -R1	GATTTTTGTTATCAAAAAGCATGGTTGTTTATGTTCGGATGTGATG
GDH2-F1	ATGCTTTTTGATAACAAAAATCGCGGTG
GDH2-R1	TCAAGCACTTGCCTCCGCTTC
GDH2-T _PGK1 -F1	GGAGGCAAGTGCTTGAAAGCTTTCCCATGTCTCTACTG
T _PGK1 -R1	AACGAACGCAGAATTTTCGAG
ADH2-F1	ATGTCTATTCCAGAAACTCAAAAAGC
ADH2-R1	CAAACTTATCGAGAGAAAGCTTATTTAGAAGTGTCAACAACGTATCTACC

引物	序列(5′-3′)
P _Gal1 -AgBIS-F1	GAAAAAACTATAATGGCTGGTGTTTCTGCTG
T _PGK1 - AgBIS-R1	GGAAAGCTTTTACAATGGCAATGGTTCGATCAAAC
AgBIS-T _PGK1 -F1	CCATTGTAAAAGCTTTCCCATGTCTCTACTGG
T _PGK1 -R1	AACGAACGCAGAATTTTCGAG
P _HXT1 -F1	GGTTCAAGCAGAAGAGACAACAATTG
P _HXT1 -ACC1-R1	AAGCTTTCTTCGCTCATGATTTTACGTATATCAACTAGTTGACGATTATG
ACC1-F1	ATGAGCGAAGAAAGCTTATTCGAG
ACC1-R1	GATGACTTTCCTCTTAGACTGGGAC
P _HXT1 -FAS1-R1	GTGGAGTAAGCGTCCATGATTTTACGTATATCAACTAGTTGACG
FAS1-F1	ATGGACGCTTACTCCACAAG
FAS1-R1	GATATAGATCACGCAATTCTTCAAAGTAG
GDH1L-P _TEF1 -F1	GAGACCAAAAAGAAAAAGAAGACATGGAGGCCCAGAATAC
GDH2-P _TEF1 -R1	GATTTTTGTTATCAAAAAGCATGGTTGTTTATGTTCGGATGTGATG
GDH2-F1	ATGCTTTTTGATAACAAAAATCGCGGTG
GDH2-R1	TCAAGCACTTGCCTCCGCTTC
GDH2-T _PGK1 -F1	GGAGGCAAGTGCTTGAAAGCTTTCCCATGTCTCTACTG
T _PGK1 -R1	AACGAACGCAGAATTTTCGAG
ADH2-F1	ATGTCTATTCCAGAAACTCAAAAAGC
ADH2-R1	CAAACTTATCGAGAGAAAGCTTATTTAGAAGTGTCAACAACGTATCTACC

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高能燃料前体α-红没药烯的生物制造过程优化

Optimization of biomanufacturing process of high-energy fuel precursor α-bisabolene

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