Computer-Aid Directed Evolution of GPPS and PS Enzymes.
Pinene, a natural active monoterpene, is widely used as a flavoring agent, perfume, medicine, and biofuel. Although genetically engineered microorganisms have successfully produced pinene, to date, the biological yield of pinene is much lower than that of semiterpenes (isoprene) and sesquiterpenes (...
| Publicado en: | BioMed Research International pp. 1 - 8 |
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| Autores principales: | , , , , , , , |
| Formato: | pictorial research tables/charts Journal Article |
| Publicado: |
Wiley-Blackwell
4/21/2021
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| Acceso en línea: | Ver este registro en EBSCOhost |
| fields | @attributes: recordID: 1 pdfLink: plink: https://search.ebscohost.com/login.aspx?direct=true&db=ccm&AN=149920125&site=ehost-live header: @attributes: shortDbName: ccm uiTerm: 149920125 longDbName: CINAHL Complete uiTag: AN controlInfo: bkinfo: dissinfo: jinfo: jid: 23146133 FT2T jtl: BioMed Research International issn: 23146133 maglogo: N pubinfo: dt: 4/21/2021 pid: 480 pub: Wiley-Blackwell place: Malden, Massachusetts artinfo: ui: 149920125 149920125 149920125 10.1155/2021/6653500 149920125 ppf: 1 ppct: 7 formats: fmt: @attributes: type: P tig: atl: Computer-Aid Directed Evolution of GPPS and PS Enzymes. aug: au: Chen, Fei Cheng, Hong Zhu, Jiaqi Wang, Shiyu Zhang, Liancheng Dong, Haolong Liu, Gang Chen, Huipeng affil: Institutes of Physical Science and Information Technology, Anhui University, Hefei 230000, China sug: subj: Terpenes Metabolism Transferases Metabolism Bioinformatics In Vitro Studies Molecular Docking Simulation Methods Gene Expression Enzymes Mutation Cytotoxicity Tests, Immunologic Molecular Structure ab: Pinene, a natural active monoterpene, is widely used as a flavoring agent, perfume, medicine, and biofuel. Although genetically engineered microorganisms have successfully produced pinene, to date, the biological yield of pinene is much lower than that of semiterpenes (isoprene) and sesquiterpenes (farnesene). In addition to the low heterologous expression of geranyl pyrophosphate synthase (GPPS) and pinene synthase (PS), cytotoxicity due to accumulation of the monoterpene also limits the production of pinene in microorganisms. In this study, we attempted to use two strategies to increase the biological yield of pinene. By deleting the random coils of GPPS and PS alone or in combination, a strain with a 335% yield increase was obtained. Additionally, upon computer-guided molecular modeling and docking of GPPS with isopentenyl pyrophosphate (IPP), its substrate, the key sites located within the catalytic pocket for substrate binding, was predicted. After screening, a strain harboring the T273R mutation of GPPS was selected among a batch of mutations of the key sites with a 154% increase in pinene yield. pubtype: Academic Journal doctype: pictorial research tables/charts Journal Article ougenre: Article language: English refInfo: holdings: @attributes: islocal: N |
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