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| Study on soluble expression conditions of sesquiterpene synthase GsSTPS2 from Ganoderma sinense |
| Hits 550 Download times 409 Received:September 01, 2022 |
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| DOI
10.11656/j.issn.1673-9043.2022.06.19 |
| Key Words
Ganoderma sinense;sesquiterpene synthase;characterization;HS-SPME-GC-MS |
| Author Name | Affiliation | E-mail | | WEI Qianhe | Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China | | | WANG Qi | Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China | | | CAO Rui | Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China | | | WANG Lizhi | Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China | wanglz2013@tjutcm.edu.cn | | WANG Haiying | Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China | haiyingwang-99@163.com |
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| Abstract
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| [Objective] To explore the expression methods of sesquiterpene synthase GsSTPS2 from Ganoderma sinense. [Methods] Different expression vectors and strains were replaced,and the induced expression conditions of GsSTPS2 were optimized by screening at different OD600,temperatures,time and IPTG concentration. On this basis, the volatile components in the cell culture medium of recombinant Escherichia coli were analyzed by headspace solid phase microextraction(HS-SPME) and gas chromatography-mass spectrometry(GC-MS). [Results] The soluble expression of GsSTPS2 was the highest,reaching 28.07%,under the conditions of initial bacterial solution OD600 of 0.8,induction temperature of 18℃,induction time of 12 h,and final IPTG concentration of 1 mmol/L. GsSTPS2 catalyzed the production of sesquiterpene hydrocarbons Cadina-3,5-diene,β-Cadinene,cis-Muurola-4(15),5-diene and δ-Cadinene and sesquiterpene oxygenated derivatives gleenol,epicubenol and τ-muurolol,among which gleenol,as one of the main products of GsSTPS2,has the effects of killing termites,anti-helminth and plant growth regulation,and has good application value and prospects. [Conclusion] This study provides a reference for the expression of proteins with poor solubility or easy to form inclusion bodies,and lays a foundation for the production of gleenol by synthetic biological methods. |
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