Advancing Yarrowia lipolytica sub-organelle engineering with endogenous mitochondrial targeting sequence.

IF 2 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biotechnology Letters Pub Date : 2025-05-08 DOI:10.1007/s10529-025-03590-8

Krutika Bhoir, K J Hemavathi, Gunjan Prakash

{"title":"Advancing Yarrowia lipolytica sub-organelle engineering with endogenous mitochondrial targeting sequence.","authors":"Krutika Bhoir, K J Hemavathi, Gunjan Prakash","doi":"10.1007/s10529-025-03590-8","DOIUrl":null,"url":null,"abstract":"Objective: The aim of the study was identification and validation of an endogenous mitochondrial targeting signal (MTS) sequence of Yarrowia lipolytica, for efficient compartmentalization of a target protein to mitochondria.Results: MTS from citrate synthase of Y. lipolytica (YlCISY-MTS) was identified, isolated and fused with green fluorescent protein (GFP) to direct it to the mitochondrial matrix. The efficiency of localization of GFP to mitochondrial matrix with YlCISY-MTS was compared with currently used MTS from Saccharomyces cerevisiae's cytochrome oxidase subunit IV. Confocal microscopy confirmed the targeted and greater GFP localization, underlining the potential of endogenous YlCISY-MTS for mitochondrial engineering in Y. lipolytica. The availability of endogenous MTS will evade the need of codon optimization of S. cerevisiae MTS for mitochondrial engineering in Y. lipolytica. This is the first report of an endogenous MTS of Y. lipolytica.Conclusion: An endogenous MTS of Y. lipolytica has been identified to facilitate the targeted delivery of a protein in the mitochondria enabling future advancements through leveraging the unique subcellular environment for metabolic engineering applications.","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 3","pages":"53"},"PeriodicalIF":2.0000,"publicationDate":"2025-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biotechnology Letters","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.1007/s10529-025-03590-8","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}

引用次数: 0

Abstract

Objective: The aim of the study was identification and validation of an endogenous mitochondrial targeting signal (MTS) sequence of Yarrowia lipolytica, for efficient compartmentalization of a target protein to mitochondria.

Results: MTS from citrate synthase of Y. lipolytica (YlCISY-MTS) was identified, isolated and fused with green fluorescent protein (GFP) to direct it to the mitochondrial matrix. The efficiency of localization of GFP to mitochondrial matrix with YlCISY-MTS was compared with currently used MTS from Saccharomyces cerevisiae's cytochrome oxidase subunit IV. Confocal microscopy confirmed the targeted and greater GFP localization, underlining the potential of endogenous YlCISY-MTS for mitochondrial engineering in Y. lipolytica. The availability of endogenous MTS will evade the need of codon optimization of S. cerevisiae MTS for mitochondrial engineering in Y. lipolytica. This is the first report of an endogenous MTS of Y. lipolytica.

Conclusion: An endogenous MTS of Y. lipolytica has been identified to facilitate the targeted delivery of a protein in the mitochondria enabling future advancements through leveraging the unique subcellular environment for metabolic engineering applications.

查看原文本刊更多论文

利用内源性线粒体靶向序列推进多脂耶氏菌亚细胞器工程。

目的：本研究的目的是鉴定和验证一种内源性线粒体靶向信号（MTS）序列，用于有效地将靶蛋白区隔到线粒体。结果：从Y. lipolytica的柠檬酸合成酶（ylcsis -MTS）中鉴定出MTS，分离并与绿色荧光蛋白（GFP）融合，引导其指向线粒体基质。利用ylczy -MTS将GFP定位到线粒体基质的效率与目前使用的来自酵母细胞色素氧化酶亚基IV的MTS进行了比较。共聚焦显微镜证实了GFP的靶向性和更强的定位，强调了内源性ylczy -MTS在脂肪酶酵母线粒体工程中的潜力。内源性MTS的可用性将避免酵母MTS密码子优化在脂质体线粒体工程中的需要。这是首次报道的内源性脂肪肝Y. MTS。结论：已经鉴定出一种内源性的脂质体Y. lipolytica MTS，可以促进线粒体中蛋白质的靶向递送，从而通过利用独特的亚细胞环境实现代谢工程应用的未来进展。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Biotechnology Letters 工程技术-生物工程与应用微生物

CiteScore

5.90

自引率

3.70%

发文量

108

审稿时长

1.2 months

期刊介绍： Biotechnology Letters is the world’s leading rapid-publication primary journal dedicated to biotechnology as a whole – that is to topics relating to actual or potential applications of biological reactions affected by microbial, plant or animal cells and biocatalysts derived from them. All relevant aspects of molecular biology, genetics and cell biochemistry, of process and reactor design, of pre- and post-treatment steps, and of manufacturing or service operations are therefore included. Contributions from industrial and academic laboratories are equally welcome. We also welcome contributions covering biotechnological aspects of regenerative medicine and biomaterials and also cancer biotechnology. Criteria for the acceptance of papers relate to our aim of publishing useful and informative results that will be of value to other workers in related fields. The emphasis is very much on novelty and immediacy in order to justify rapid publication of authors’ results. It should be noted, however, that we do not normally publish papers (but this is not absolute) that deal with unidentified consortia of microorganisms (e.g. as in activated sludge) as these results may not be easily reproducible in other laboratories. Papers describing the isolation and identification of microorganisms are not regarded as appropriate but such information can be appended as supporting information to a paper. Papers dealing with simple process development are usually considered to lack sufficient novelty or interest to warrant publication.