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promoter engineering enables precise metabolic regulation towards efficientβ-elemene production in Ogataea polymorpha

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Synthetic and Systems Biotechnology 2024年第2期9卷 234-241页

作者： Min Ye Jiaoqi Gao Jingjing Li Wei Yu Fan Bai Yongjin J.Zhou Division of Biotechnology Dalian Institute of Chemical PhysicsChinese Academy of Sciences457 Zhongshan RoadDalian116023PR China CAS Key Laboratory of Separation Science for Analytical Chemistry Dalian Institute of Chemical PhysicsChinese Academy of SciencesDalian116023PR China Dalian Key Laboratory of Energy Biotechnology Dalian Institute of Chemical PhysicsChinese Academy of SciencesDalian116023PR China University of Chinese Academy of Sciences Beijing100049PR China

Precisely controlling gene expression is beneficial for optimizing biosynthetic pathways for improving the ***,promoters in nonconventional yeasts such as Ogataea polymorpha are always limited,which results in incompatible gene ***,we expanded the promoter library in *** based on transcriptional data,among which 13 constitutive promoters had the strengths ranging from 0–55%of PGAP,the commonly used strong constitutive promoter,and 2 were growth phase-dependent ***,2 hybrid growth phase-dependent promoters were constructed and characterized,which had 2-fold higher ***,promoter engineering was applied to precisely regulate cellular metabolism for efficient production ofβ-*** glyceraldehyde-3-phosphate dehydrogenase gene GAP was downregulated to drive more flux into pentose phosphate pathway(PPP)and then to enhance the supply of acetyl-CoA by using phosphoketolase-phosphotransacetylase(PK-PTA)*** with the phase-dependent expression of synthase module(ERG20∼LsLTC2 fusion),the highest titer of 5.24 g/L with a yield of 0.037 g/(g glucose)was achieved in strain YY150U under fed-batch fermentation in shake *** work characterized and engineered a series of promoters,that can be used to fine-tune genes for constructing efficient yeast cell factories.

关键词： promoter library Growth phase-dependent promoters promoter engineering Precise metabolic regulation Ogataea polymorpha

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Screening and engineering of high-activity promoter elements through transcriptomics and red fluorescent protein visualization in Rhodobacter sphaeroides

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Synthetic and Systems Biotechnology 2021年第4期6卷 335-342页

作者： Tong Shi Lu Zhang Mindong Liang Weishan Wang Kefeng Wang Yue Jiang Jing Liu Xinwei He Zhiheng Yang Haihong Chen Chuan Li Dongyuan Lv Liming Zhou Biqin Chen Dan Li Li-Xin Zhang Gao-Yi Tan State Key Laboratory of Bioreactor Engineering(SKLBE) And School of BiotechnologyEast China University of Science and Technology(ECUST)Shanghai200237China State Key Laboratory of Microbial Resources and CAS Key Laboratory of Pathogenic Microbiology and Immunology Institute of MicrobiologyChinese Academy of Sciences(CAS)Beijing100101China Inner Mongolia Kingdomway Pharmaceutical Co. LtdHohhot010206China

The versatile photosyntheticα-proteobacterium Rhodobacter sphaeroides,has recently been extensively engineered as a novel microbial cell factory(MCF)to produce pharmaceuticals,nutraceuticals,commodity chemicals and even ***,there are no well-characterized high-activity promoters to modulate gene transcription during the engineering of *** this study,several native promoters from *** JDW-710(JDW-710),an industrial strain producing high levels of co-enzyme Q10(Q10)were selected on the basis of transcriptomic *** candidate promoters were then characterized by using gusA as a reporter *** native promoters,Prsp_7571 and Prsp_6124,showed 620%and 800%higher activity,respectively,than the tac promoter,which has previously been used for gene overexpression in *** addition,a Prsp_7571-derived synthetic promoter library with strengths ranging from 54%to 3200%of that of the tac promoter,was created on the basis of visualization of red fluorescent protein(RFP)expression in ***,as a demonstration,the synthetic pathway of Q10 was modulated by the selected promoter T334*in JDW-710;the Q10 yield in shake-flasks increased 28%and the production reached 226 mg/*** well-characterized promoters should be highly useful in current synthetic biology platforms for refactoring the biosynthetic pathway in ***-derived MCFs.

关键词： Rhodobacter sphaeroides promoter library Transcriptomics Co-enzyme Q_(10) Red fluorescent protein

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