Engineering Komagataella phaffii to biosynthesize cordycepin from methanol which drives global metabolic alterations at the transcription level  

作　　者：Huiping Tan Liang Wang Huiguo Wang Yanghao Cheng Xiang Li Huihui Wan Chenguang Liu Tian Liu Qian Li 

机构地区：[1]Department of Life and Health,Dalian University,No.10 Xuefu Street,Dalian Economic and Technological Development Zone,Dalian,116622,China [2]School of Biological Engineering,Dalian Polytechnic University,No.1 Qinggongyuan Road,Ganjingzi District,Dalian,116034,China [3]Analytical Instrumentation Centre,Dalian University of Technology,No.2 Linggong Road,Dalian,116024,China [4]State Key Laboratory of Microbial Metabolism,Joint International Research Laboratory of Metabolic&Developmental Science,School of Life Sciences and Biotechnology,Shanghai Jiao Tong University,800 Dongchuan Road,Minhang District,Shanghai,200240,China [5]School of Bioengineering,Dalian University of Technology,No.2 Linggong Road,Dalian,116024,China

出　　处：《Synthetic and Systems Biotechnology》2023年第2期242-252,共11页合成和系统生物技术（英文）

基　　金：support from Open Funding Project of State Key Laboratory of Microbial Metabolism(No.MMLKF20-09);Research Project of Applied Basic Research Program of Department of Science&Technology of Liaoning Province(2022JH2/101300137);the Project of Natural Science Foundation of Liaoning Province(2022-KF-15-02);Science Research Foundation of Educational Department of Liaoning Province(No.J2020099).

摘　　要：Cordycepin has the potential to be an alternative to the disputed herbicide glyphosate.However,current laborious and time-consuming production strategies at low yields based on Cordyceps militaris lead to extremely high cost and restrict its application in the field of agriculture.In this study,Komagataella phaffii(syn.Pichia pastoris)was engineered to biosynthesize cordycepin from methanol,which could be converted from CO_(2).Combined with fermentation optimization,cordycepin content in broth reached as high as 2.68±0.04 g/L within 168 h,around 15.95 mg/(L⋅h)in productivity.Additionally,a deaminated product of cordycepin was identified at neutral or weakly alkaline starting pH during fermentation.Transcriptome analysis found the yeast producing cordycepin was experiencing severe inhibition in methanol assimilation and peroxisome biogenesis,responsible for delayed growth and decreased carbon flux to pentose phosphate pathway(PPP)which led to lack of precursor supply.Amino acid interconversion and disruption in RNA metabolism were also due to accumu-lation of cordycepin.The study provided a unique platform for the manufacture of cordycepin based on the emerging non-conventional yeast and gave practical strategies for further optimization of the microbial cell factory.

关 键 词：CORDYCEPIN Komagataella phaffii Fermentation optimization TRANSCRIPTOME 

分 类 号：O65[理学—分析化学]