Effective expression of taste-modifying protein miraculin fusing FMDV 2A gene in Chlamydomonas reinhardtii

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal Pub Date : 2025-03-06 DOI:10.1016/j.bej.2025.109706

Chuan Wang, Xiaomei Wang, Ge Rang Lamu, Wei Liang, Yaomei Tian

{"title":"Effective expression of taste-modifying protein miraculin fusing FMDV 2A gene in Chlamydomonas reinhardtii","authors":"Chuan Wang, Xiaomei Wang, Ge Rang Lamu, Wei Liang, Yaomei Tian","doi":"10.1016/j.bej.2025.109706","DOIUrl":null,"url":null,"abstract":"<div><div>Miraculin is a taste modification protein derived from plants, which can change sour taste into sweet taste, and has great application potential in food industry. Currently, the extraction of miraculin presented some problems such as cumbersome process and low purity. <em>Chlamydomonas reinhardtii</em> is a potential expression system of exogenous proteins. However, the exogenous genes are integrated randomly in genome of <em>C. reinhardtii</em> during transformation, which leads to significant expression variability among transformants. The 2A peptide from FMDV virus has been proven to effectively improve the expression efficiency of exogenous proteins in multi-gene expression systems through self-cleavage mechanism. In presented study, the expression vector pSP108–2A-miraculin containing 2A-miraculin fusion gene was constructed and transformed into <em>C. reinhardtii</em> by glass bead method. Two positive strains were obtained through bleomycin resistance selecting. The detection results showed that miraculin gene was integrated into the genome of <em>C. reinhardtii</em> cells at the molecular level, and was successfully expressed at the protein level. This study was the first time to express the miraculin in <em>C. reinhardtii</em>, which provided an optional way to obtain miraculin.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"219 ","pages":"Article 109706"},"PeriodicalIF":3.7000,"publicationDate":"2025-03-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biochemical Engineering Journal","FirstCategoryId":"5","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1369703X25000804","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}

引用次数: 0

Abstract

Miraculin is a taste modification protein derived from plants, which can change sour taste into sweet taste, and has great application potential in food industry. Currently, the extraction of miraculin presented some problems such as cumbersome process and low purity. Chlamydomonas reinhardtii is a potential expression system of exogenous proteins. However, the exogenous genes are integrated randomly in genome of C. reinhardtii during transformation, which leads to significant expression variability among transformants. The 2A peptide from FMDV virus has been proven to effectively improve the expression efficiency of exogenous proteins in multi-gene expression systems through self-cleavage mechanism. In presented study, the expression vector pSP108–2A-miraculin containing 2A-miraculin fusion gene was constructed and transformed into C. reinhardtii by glass bead method. Two positive strains were obtained through bleomycin resistance selecting. The detection results showed that miraculin gene was integrated into the genome of C. reinhardtii cells at the molecular level, and was successfully expressed at the protein level. This study was the first time to express the miraculin in C. reinhardtii, which provided an optional way to obtain miraculin.

查看原文本刊更多论文

求助全文

约1分钟内获得全文求助全文

来源期刊

Biochemical Engineering Journal 工程技术-工程：化工

CiteScore

7.10

自引率

5.10%

发文量

380

审稿时长

34 days

期刊介绍： The Biochemical Engineering Journal aims to promote progress in the crucial chemical engineering aspects of the development of biological processes associated with everything from raw materials preparation to product recovery relevant to industries as diverse as medical/healthcare, industrial biotechnology, and environmental biotechnology. The Journal welcomes full length original research papers, short communications, and review papers* in the following research fields: Biocatalysis (enzyme or microbial) and biotransformations, including immobilized biocatalyst preparation and kinetics Biosensors and Biodevices including biofabrication and novel fuel cell development Bioseparations including scale-up and protein refolding/renaturation Environmental Bioengineering including bioconversion, bioremediation, and microbial fuel cells Bioreactor Systems including characterization, optimization and scale-up Bioresources and Biorefinery Engineering including biomass conversion, biofuels, bioenergy, and optimization Industrial Biotechnology including specialty chemicals, platform chemicals and neutraceuticals Biomaterials and Tissue Engineering including bioartificial organs, cell encapsulation, and controlled release Cell Culture Engineering (plant, animal or insect cells) including viral vectors, monoclonal antibodies, recombinant proteins, vaccines, and secondary metabolites Cell Therapies and Stem Cells including pluripotent, mesenchymal and hematopoietic stem cells; immunotherapies; tissue-specific differentiation; and cryopreservation Metabolic Engineering, Systems and Synthetic Biology including OMICS, bioinformatics, in silico biology, and metabolic flux analysis Protein Engineering including enzyme engineering and directed evolution.