Komagataella phaffii is a methylotrophic yeast harboring a tightly regulated alcohol oxidase promoter(PAOX1),which is now widely used for recombinant protein production.During PAOX1 expression phase by methanol induct...Komagataella phaffii is a methylotrophic yeast harboring a tightly regulated alcohol oxidase promoter(PAOX1),which is now widely used for recombinant protein production.During PAOX1 expression phase by methanol induction,a methanol metabolism organelle peroxisome enlarged and occupied 80%of K.phaffii cell through peroxins functions of matrix protein import and organelle division.Using a K.phaffii expressing xylanase in this study,each of all 23 PEX genes of K.phaffii,encoding peroxin,was knockout to influence the peroxisome size,leading to changes of K.phaffii physiological status and recombinant xylanase expression.It was observed that PEX3 knockout reduced peroxisome size by 54.3%,increased xylanase expression by 29%,decreased apoptosis ratio by 70.6%.Transcriptome analysis revealed that PEX3 gene knockout decreased 18 other PEX genes of all three steps of peroxisome propagation,biogenesis,matrix protein import,and peroxisome fission.PEX3 gene knockout influenced expression of ribosomal subunit-related and protein transportation significantly based on gene function annotation and enrichment analysis.Additionally,Therefore,PEX3 gene knockout promoted xylanase folding correctly via Sec63 complex,and PDI1 significantly.In a summary,PEX3 gene knockout pro-vided a novel strategy to enhance recombinant xylanase by K.phaffii.展开更多
Protein ubiquitination regulates diverse cellular processes in eukaryotic organisms,from growth and development to stress response.Proteins subjected to ubiquitination can be found in virtually all subcellular locatio...Protein ubiquitination regulates diverse cellular processes in eukaryotic organisms,from growth and development to stress response.Proteins subjected to ubiquitination can be found in virtually all subcellular locations and organelles,including peroxisomes,singlemembrane and highly dynamic organelles ubiquitous in eukaryotes.Peroxisomes contain metabolic functions essential to plants and animals such as lipid catabolism,detoxification of reactive oxygen species(ROS),biosynthesis of vital hormones and cofactors,and photorespiration.Plant peroxisomes possess a complex proteome with functions varying among different tissue types and developmental stages,and during plant response to distinct environmental cues.However,how these diverse functions are regulated at the post-translational level is poorly understood,especially in plants.In this review,we summarized current knowledge of the involvement of protein ubiquitination in peroxisome protein import,remodeling,pexophagy,and metabolism,focusing on plants,and referencing discoveries from other eukaryotic systems when relevant.Based on previous ubiquitinomics studies,we compiled a list of 56 ubiquitinated Arabidopsis peroxisomal proteins whose functions are associated with all the major plant peroxisomal metabolic pathways.This discovery suggests a broad impact of protein ubiquitination on plant peroxisome functions,therefore substantiating the need to investigate this significant regulatory mechanism in peroxisomes at more depths.展开更多
基金supported by the Science and Technology Commission of Shanghai Municipality(No.22dz1205800)National Natural Science Foundation of China(No.31870045)Open Funding Project of the State Key Laboratory of Bioreactor Engineering.
文摘Komagataella phaffii is a methylotrophic yeast harboring a tightly regulated alcohol oxidase promoter(PAOX1),which is now widely used for recombinant protein production.During PAOX1 expression phase by methanol induction,a methanol metabolism organelle peroxisome enlarged and occupied 80%of K.phaffii cell through peroxins functions of matrix protein import and organelle division.Using a K.phaffii expressing xylanase in this study,each of all 23 PEX genes of K.phaffii,encoding peroxin,was knockout to influence the peroxisome size,leading to changes of K.phaffii physiological status and recombinant xylanase expression.It was observed that PEX3 knockout reduced peroxisome size by 54.3%,increased xylanase expression by 29%,decreased apoptosis ratio by 70.6%.Transcriptome analysis revealed that PEX3 gene knockout decreased 18 other PEX genes of all three steps of peroxisome propagation,biogenesis,matrix protein import,and peroxisome fission.PEX3 gene knockout influenced expression of ribosomal subunit-related and protein transportation significantly based on gene function annotation and enrichment analysis.Additionally,Therefore,PEX3 gene knockout promoted xylanase folding correctly via Sec63 complex,and PDI1 significantly.In a summary,PEX3 gene knockout pro-vided a novel strategy to enhance recombinant xylanase by K.phaffii.
基金supported by the Leading Innovative and Entrepreneur Team Introduction Program of Zhejiang(No.2019R01002)Key Research Project of Zhejiang Laboratory(No.2021PE0AC04)+3 种基金Scientific Research Fund of Zhejiang Provincial Education Department(No.Y202148338)Zhejiang University Student Research Practice Program(No.P2021041)to R.P.by the National Science Foundation(No.MCB 2148206)US Department of Energy(No.DEFG02-91ER20021)to J.H。
文摘Protein ubiquitination regulates diverse cellular processes in eukaryotic organisms,from growth and development to stress response.Proteins subjected to ubiquitination can be found in virtually all subcellular locations and organelles,including peroxisomes,singlemembrane and highly dynamic organelles ubiquitous in eukaryotes.Peroxisomes contain metabolic functions essential to plants and animals such as lipid catabolism,detoxification of reactive oxygen species(ROS),biosynthesis of vital hormones and cofactors,and photorespiration.Plant peroxisomes possess a complex proteome with functions varying among different tissue types and developmental stages,and during plant response to distinct environmental cues.However,how these diverse functions are regulated at the post-translational level is poorly understood,especially in plants.In this review,we summarized current knowledge of the involvement of protein ubiquitination in peroxisome protein import,remodeling,pexophagy,and metabolism,focusing on plants,and referencing discoveries from other eukaryotic systems when relevant.Based on previous ubiquitinomics studies,we compiled a list of 56 ubiquitinated Arabidopsis peroxisomal proteins whose functions are associated with all the major plant peroxisomal metabolic pathways.This discovery suggests a broad impact of protein ubiquitination on plant peroxisome functions,therefore substantiating the need to investigate this significant regulatory mechanism in peroxisomes at more depths.
