Functional characterization of a geraniol synthase-encoding gene from Camptotheca acuminata and its application in production of geraniol in Escherichia coli

被引:40
作者
Chen, Fei [1 ,3 ]
Li, Wei [1 ,3 ]
Jiang, Liangzhen [1 ]
Pu, Xiang [1 ,3 ]
Yang, Yun [1 ,3 ]
Zhang, Guolin [1 ]
Luo, Yinggang [1 ,2 ]
机构
[1] Chinese Acad Sci, Chengdu Inst Biol, Ctr Nat Prod Res, Chengdu 610041, Peoples R China
[2] Chinese Acad Sci, Shanghai Inst Organ Chem, State Key Lab Bioorgan & Nat Prod Chem, Shanghai 200032, Peoples R China
[3] Univ Chinese Acad Sci, Beijing 100049, Peoples R China
基金
中国国家自然科学基金;
关键词
Geraniol synthase; Camptotheca acuminata; Geraniol; Camptothecin; Metabolic engineering; Fermentation; ALKALOID BIOSYNTHESIS; ADVANCED BIOFUEL; MICROBIAL-PRODUCTION; TERPENE SYNTHASES; HAIRY ROOTS; PATHWAY; PRECURSOR; LIMONENE; EXPRESSION; CATALYSIS;
D O I
10.1007/s10295-016-1802-2
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Geraniol synthase (GES) catalyzes the conversion of geranyl diphosphate (GPP) into geraniol, an acyclic monoterpene alcohol that has been widely used in many industries. Here we report the functional characterization of CaGES from Camptotheca acuminata, a camptothecin-producing plant, and its application in production of geraniol in Escherichia coli. The full-length cDNA of CaGES was obtained from overlap extension PCR amplification. The intact and N-terminus-truncated CaGESs were overexpressed in E. coli and purified to homogeneity. Recombinant CaGES showed the conversion activity from GPP to geraniol. To produce geraniol in E. coli using tCaGES, the biosynthetic precursor GPP should be supplied and transferred to the catalytic pocket of tCaGES. Thus, ispA(S80F), a mutant of farnesyl diphosphate (FPP) synthase, was prepared to produce GPP via the head-to-tail condensation of isoprenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP). A slight increase of geraniol production was observed in the fermentation broth of the recombinant E. coli harboring tCaGES and ispA(S80F). To enhance the supply of IPP and DMAPP, the encoding genes involved in the whole mevalonic acid biosynthetic pathway were introduced to the E. coli harboring tCaGES and the ispA(S80F) and a significant increase of geraniol yield was observed. The geraniol production was enhanced to 5.85 +/- 0.46 mg L-1 when another copy of ispA(S80F) was introduced to the above recombinant strain. The following optimization of medium composition, fermentation time, and addition of metal ions led to the geraniol production of 48.5 +/- 0.9 mg L-1. The present study will be helpful to uncover the biosynthetic enigma of camptothecin and tCaGES will be an alternative to selectively produce geraniol in E. coli with other metabolic engineering approaches.
引用
收藏
页码:1281 / 1292
页数:12
相关论文
共 42 条
[1]   Metabolic engineering of Escherichia coli for limonene and perillyl alcohol production [J].
Alonso-Gutierrez, Jorge ;
Chan, Rossana ;
Batth, Tanveer S. ;
Adams, Paul D. ;
Keasling, Jay D. ;
Petzold, Christopher J. ;
Lee, Taek Soon .
METABOLIC ENGINEERING, 2013, 19 :33-41
[2]   Plant terpenoid synthases: Molecular biology and phylogenetic analysis [J].
Bohlmann, J ;
Meyer-Gauen, G ;
Croteau, R .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95 (08) :4126-4133
[3]   Geraniol - A review of a commercially important fragrance material [J].
Chen, W. ;
Viljoen, A. M. .
SOUTH AFRICAN JOURNAL OF BOTANY, 2010, 76 (04) :643-651
[4]   BIOSYNTHESIS AND CATABOLISM OF MONOTERPENOIDS [J].
CROTEAU, R .
CHEMICAL REVIEWS, 1987, 87 (05) :929-954
[5]   Natural products for cancer chemotherapy [J].
Demain, Arnold L. ;
Vaishnav, Preeti .
MICROBIAL BIOTECHNOLOGY, 2011, 4 (06) :687-699
[6]   Characterization of two geraniol synthases from Valeriana officinalis and Lippia dulcis: Similar activity but difference in subcellular localization [J].
Dong, Lemeng ;
Miettinen, Karel ;
Goedbloed, Miriam ;
Verstappen, France W. A. ;
Voster, Alessandra ;
Jongsma, Maarten A. ;
Memelink, Johan ;
van der Krol, Sander ;
Bouwmeester, Harro J. .
METABOLIC ENGINEERING, 2013, 20 :198-211
[7]   Deoxyxylulose phosphate pathway to terpenoids [J].
Eisenreich, W ;
Rohdich, F ;
Bacher, A .
TRENDS IN PLANT SCIENCE, 2001, 6 (02) :78-84
[8]   Specificity of Ocimum basilicum geraniol synthase modified by its expression in different heterologous systems [J].
Fischer, Marc J. C. ;
Meyer, Sophie ;
Claudel, Patricia ;
Perrin, Mireille ;
Ginglinger, Jean Francois ;
Gertz, Claude ;
Masson, Jean E. ;
Werck-Reinhardt, Daniele ;
Hugueney, Philippe ;
Karst, Francis .
JOURNAL OF BIOTECHNOLOGY, 2013, 163 (01) :24-29
[9]   Metabolic Engineering of Monoterpene Synthesis in Yeast [J].
Fischer, Marc J. C. ;
Meyer, Sophie ;
Claudel, Patricia ;
Bergdoll, Marc ;
Karst, Francis .
BIOTECHNOLOGY AND BIOENGINEERING, 2011, 108 (08) :1883-1892
[10]   Terpenoid synthase structures: a so far incomplete view of complex catalysis [J].
Gao, Yang ;
Honzatko, Richard B. ;
Peters, Reuben J. .
NATURAL PRODUCT REPORTS, 2012, 29 (10) :1153-1175