Biosynthesis of Valerena-4,7（11）-diene in an Engineered Escherichia coli Strain

doi:10.13560/j.cnki.biotech.bull.1985.2017.01.014

Abstract

Abstract: Valerena-4,7（11）-diene is a bicyclic sesquiterpene and shows particularly profound sedative,stress reducing and anxiolytic effects,and it could be used to treat attention deficit hyperactivity disorder（ADHD）and other mental illness. To achieve the de novo biosynthesis of valerena-4,7（11）-diene in Escherichia coli,eight genes of Saccharomyces cerevisiae involved in converting acetyl-CoA into farnesyl diphosphate（FPP）from mevalonate pathway and the valerena-4,7（11）-diene synthase（VoTPS）gene from Valeriana officinalis were co-overexpressed in BL21（DE3）. Adopting the bipolar fermentation method of water phase and organic phase,the valerian-4,7（11）-diene was detected in organic phase by GC-MS,indicating that the synthesis of valerian-4,7（11）-diene was achieved initially in theE. coli. After primarily optimizing the IPTG concentration,the temperature of protein induction and choice of varied organic phases,the fermentation conditions were determined as follows：protein induced temperature was 20℃,the concentration of IPTG was 0.1 mmol/L,and organic phase was dodecane.

Key words: Escherichia coli, valerian-4,7（11）-diene, biosynthesis, valerian-4,7（11）-diene synthase（VoTPS）

LIU Chang, YIN Hua, ZHUANG Yi-bin, LIU Tao. Biosynthesis of Valerena-4,7（11）-diene in an Engineered Escherichia coli Strain[J]. Biotechnology Bulletin, 2017, 33(1): 129-134.

References

[1] Bryan WP, Hue TT, Benjamin P, et al. Enzymatic synthesis of valerena-4, 7（11）-diene by a unique sesquiterpene synthase from the valerian plant（Valeriana officinalis）[J]. FEBS Journal, 2012, 279：3136-3146.
[2] Takemoto H, Omameuda Y, Ito M, et al. Inhalation administration of Valerena-4, 7（11）-diene from Nardostachys chinensis roots ameliorates restraint stress-induced changes in murine behavior and stress-related factors[J]. Biol Pharm Bull, 2014, 37：1050-1055.
[3] Houghton PJ. The scientific basis for the reputed activity of Valerian[J]. Pharm Pharmacol, 1999, 51：505-512.
[4] Letchamo W, Ward W, Heard B, et al. Essential oil of Valeriana officinalis L. cultivars and their antimicrobial activity as influenced by harvesting time under commercial organic cultivation[J]. AgricFood Chem, 2004, 52：3915-3919.
[5] Wang Y, Jin L, Yu S, et al. Chemical constituents of plants from the genus Valeriana[J]. Mini Rev Org Chem, 2010, 7：161-172.
[6] Bos R, Woerdenbag HJ, et al. Composition of the essential oils from underground parts of Valeriana officinalis L. and several closely related taxa[J]. Flavour Fragr, 1997, 12：359-370.
[7] Khom S, Strommer B, Ramharter J, et al. Valerenic acid derivatives as novel subunit-selective GABAA receptor ligands- in vitro and in vivo characterization[J]. Br J Pharmacol, 2010, 161：65-78.
[8] Post RM. Transduction of psychosocial stress into the neurobiology of recurrent affective disorder[J]. Am J Psychiatry, 1992, 149：999-1010.
[9] Takemoto H, Yagura T, Ito M. Evaluation of volatile components from spikenard：valerena-4, 7（11）-diene is a highly active sedative compound[J]. Nat Med, 2009, 63：380-385.
[10] Lader M. Anxiolytic drugs：dependence, addiction and abuse[J]. Eur Neuropsychopharmacol, 1994, 4：85-91.
[11] Vukcević NP, Ercegović GV, Segrt Z, et al. Benzodiazepine poisoning in elderly[J]. Vojnosanit Pregl, 2016, 73：234-238.
[12] Kitayama T, Kawabata G, Ito M. Concise synthesis of valerena-4, 7（11）-diene, a highly active sedative, from valerenic acid[J]. Biosci Biotechnol Biochem, 2010, 74（9）：1963-1964.
[13] Chotani G, Dodge T, Hsu A, et al. The commercial production of chemicals using pathway engineering[J]. Biochim Biophys Acta, 2000, 1543（2）：434-455.
[14] Edgar S, Li FS, Qiao K, et al. Engineering of Taxadiene synthase for improved selectivity and yield of a key taxol biosynthetic intermediate[J]. ACS Synth Biol, 2016, doi:10. 1021/acssynbio. 6b00206.
[15] 田宁. 利用工程大肠杆菌生物法合成香叶醇的发酵工艺研究[D]. 青岛：中国海洋大学, 2015.
[16] 岳雪, 聂少振, 王素珍, 等. 合成生物学在天然药物和微生物药物开发中的应用[J]. 中国抗生素杂志, 2016, 41（8）：568-577.
[17] Yin H, Zhuang YB, Li EE, et al. Heterologous biosynthesis of costunolide in Escherichia coli and yield improvement[J]. Biotechnol Lett, 2015, 37：1249-1255.
[18] Peralta-Yahya PP, Ouellet M, Chan R, et al. Identification and microbial production of a terpene-based advanced biofuel[J]. Nat Commun, 2011, 2：483.
[19] 陈妙芬, 冯静仪, 刘秋琼, 等. GC法测定香茅中香叶醇的含量[J]. 中国药房, 2011（31）：2950-2951.
[20] 马媛媛, 何健民, 康永杰. 外源性蛋白在大肠杆菌中可溶性表达的策略综述[J]. 世界科技研究与发展, 2015, 37（5）：627-630.
[21] 田宁, 咸漠, 胡仰栋, 等. 产香叶醇重组大肠杆菌发酵培养基的优化[J]. 林产化学与工业, 2015, 35（4）：131-137.
[22] Becker A, Felgentreff F, Schröder H, et al. The anxiolytic effects of a Valerian extract is based on Valerenic acid[J]. BMC Complementary and Alternative Medicine, 2014, 14（1）：1-5.
[23] Vidal-Cantú GC, Jiménez-Hernández M, Rocha-González HI, et al. Role of 5-HT5A and 5-HT1B/1D receptors in the antinociception produced by ergotamine and valerenic acid in the rat formalin test[J]. Eur J Pharmacol, 2016, 781：109-116.
[24] Khom S, Hintersteiner J, Luger D, et al. Analysis of β-subunit-dependent GABAA receptor modulation and behavioral effects of Valerenic acid derivatives[J]. Journal of Pharmacology & Experimental Therapeutics, 2016, 357（3）：580-590.
[25] Liu Q, Majdi M, Cankar K, et al. Reconstitution of the costunolide biosynthetic pathway in yeast and Nicotiana benthamiana[J]. PLoS One, 2011, 6（8）：764-778.
[26] Ikezawa N, Göpfert JC, Nguyen DT, et al. Lettuce costunolide synthase（CYP71BL2）and its homolog（CYP71BL1）from sunflower catalyze distinct regio- and stereoselective hydroxylations in sesquiterpene lactone metabolism[J]. Journal of Biological Chemistry, 2011, 286（24）：21601-21611.