A homomeric geranyl diphosphate synthase-encoding gene from Camptotheca acuminata and its combinatorial optimization for production of geraniol in Escherichia coli
文献类型：期刊论文
| 作者 | Yang, LX; Jiang, LZ; Li, W; Yang, Y; Zhang, GL; Luo, YG; Luo, YG (reprint author), Chinese Acad Sci, Chengdu Inst Biol, Ctr Nat Prod Res, Chengdu 610041, Peoples R China.; Luo, YG (reprint author), Chinese Acad Sci, Shanghai Inst Organ Chem, State Key Lab Bioorgan & Nat Prod Chem, Shanghai 200032, Peoples R China. |
| 刊名 | JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY
 |
| 出版日期 | 2017 |
| 卷号 | 44期号:10页码:1431-1441 |
| 关键词 | Geranyl Diphosphate Synthase Camptotheca Acuminata Geraniol Metabolic Engineering Fermentation |
| DOI | 10.1007/s10295-017-1967-3 |
| 产权排序 | 1 |
| 文献子类 | Article |
| 英文摘要 | Geranyl diphosphate (GPP), the unique precursor for all monoterpenoids, is biosynthesized from isopentenyl diphosphate and dimethylallyl diphosphate via the head-to-tail condensation reaction catalyzed by GPP synthase (GPPS). Herein a homomeric GPPS from Camptotheca acuminata, a camptothecin-producing plant, was obtained from 5'- and 3'-rapid amplification of cDNA ends and subsequent overlap extension and convenient PCR amplifications. The truncate CaGPPS was introduced to replace ispA of pBbA5c-MevT(CO)-MBIS(CO, ispA), a de novo biosynthetic construct for farnesyl diphosphate generation, and overexpressed in Escherichia coli, together with the truncate geraniol synthase-encoding gene from C. acuminata (tCaGES), to confirm CaGPPS-catalyzed reaction in vivo. A 24.0 +/- 1.3 mg L-1 of geraniol was produced in the recombinant E. coli. The production of GPP was also validated by the direct UPLC-HRMSE analyses. The tCaGPPS and tCaGES genes with different copy numbers were introduced into E. coli to balance their catalytic potential for high-yield geraniol production. A 1.6-fold increase of geraniol production was obtained when four copies of tCaGPPS and one copy of tCaGES were introduced into E. coli. The following fermentation conditions optimization, including removal of organic layers and addition of new n-decane, led to a 74.6 +/- 6.5 mg L-1 of geraniol production. The present study suggested that the gene copy number optimization, i.e., the ratio of tCaGPPS and tCaGES, plays an important role in geraniol production in the recombinant E. coli. The removal and addition of organic solvent are very useful for sustainable high-yield production of geraniol in the recombinant E. coli in view of that the solubility of geraniol is limited in the fermentation broth and/or n-decane. |
| 学科主题 | Biotechnology & Applied Microbiology |
| 语种 | 英语 |
| 资助机构 | The authors wish to acknowledge the financial support in part by the KSCX2-EW-Q-6 and ZHTS-003 projects from the Chinese Academy of Sciences, the Applied and Basic Research Program of Sichuan Province (2015JY0058), and the 21172216 project from the National Natural Science Foundation of China. All authors have agreed to submit this manuscript to the Journal of Industrial Microbiology & Biotechnology. ; Chinese Academy of Sciences [KSCX2-EW-Q-6, ZHTS-003] ; The authors wish to acknowledge the financial support in part by the KSCX2-EW-Q-6 and ZHTS-003 projects from the Chinese Academy of Sciences, the Applied and Basic Research Program of Sichuan Province (2015JY0058), and the 21172216 project from the National Natural Science Foundation of China. All authors have agreed to submit this manuscript to the Journal of Industrial Microbiology & Biotechnology. ; Chinese Academy of Sciences [KSCX2-EW-Q-6, ZHTS-003] ; Applied and Basic Research Program of Sichuan Province [2015JY0058] ; Applied and Basic Research Program of Sichuan Province [2015JY0058] ; National Natural Science Foundation of China [21172216] ; National Natural Science Foundation of China [21172216] ; The authors wish to acknowledge the financial support in part by the KSCX2-EW-Q-6 and ZHTS-003 projects from the Chinese Academy of Sciences, the Applied and Basic Research Program of Sichuan Province (2015JY0058), and the 21172216 project from the National Natural Science Foundation of China. All authors have agreed to submit this manuscript to the Journal of Industrial Microbiology & Biotechnology. ; Chinese Academy of Sciences [KSCX2-EW-Q-6, ZHTS-003] ; The authors wish to acknowledge the financial support in part by the KSCX2-EW-Q-6 and ZHTS-003 projects from the Chinese Academy of Sciences, the Applied and Basic Research Program of Sichuan Province (2015JY0058), and the 21172216 project from the National Natural Science Foundation of China. All authors have agreed to submit this manuscript to the Journal of Industrial Microbiology & Biotechnology. ; Chinese Academy of Sciences [KSCX2-EW-Q-6, ZHTS-003] ; Applied and Basic Research Program of Sichuan Province [2015JY0058] ; Applied and Basic Research Program of Sichuan Province [2015JY0058] ; National Natural Science Foundation of China [21172216] ; National Natural Science Foundation of China [21172216] |
| 源URL | [http://210.75.237.14/handle/351003/29212]  |
| 专题 | 成都生物研究所_天然产物研究 |
| 通讯作者 | Luo, YG (reprint author), Chinese Acad Sci, Chengdu Inst Biol, Ctr Nat Prod Res, Chengdu 610041, Peoples R China.; Luo, YG (reprint author), Chinese Acad Sci, Shanghai Inst Organ Chem, State Key Lab Bioorgan & Nat Prod Chem, Shanghai 200032, Peoples R China. |
| 推荐引用方式 GB/T 7714 | Yang, LX,Jiang, LZ,Li, W,et al. A homomeric geranyl diphosphate synthase-encoding gene from Camptotheca acuminata and its combinatorial optimization for production of geraniol in Escherichia coli[J]. JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY,2017,44(10):1431-1441. |
| APA | Yang, LX.,Jiang, LZ.,Li, W.,Yang, Y.,Zhang, GL.,...&Luo, YG .(2017).A homomeric geranyl diphosphate synthase-encoding gene from Camptotheca acuminata and its combinatorial optimization for production of geraniol in Escherichia coli.JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY,44(10),1431-1441. |
| MLA | Yang, LX,et al."A homomeric geranyl diphosphate synthase-encoding gene from Camptotheca acuminata and its combinatorial optimization for production of geraniol in Escherichia coli".JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY 44.10(2017):1431-1441. |
入库方式： OAI收割
来源：成都生物研究所
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