Heterologous expression of algal fucosyltransferases and sulfotransferases in Escherichia coli

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal Pub Date : 2025-02-18 DOI:10.1016/j.bej.2025.109690

Ahmed Zayed , Benjamin Ledermann , Thomas Fischöder , Lothar Elling , Nicole Frankenberg-Dinkel , Roland Ulber

{"title":"Heterologous expression of algal fucosyltransferases and sulfotransferases in Escherichia coli","authors":"Ahmed Zayed , Benjamin Ledermann , Thomas Fischöder , Lothar Elling , Nicole Frankenberg-Dinkel , Roland Ulber","doi":"10.1016/j.bej.2025.109690","DOIUrl":null,"url":null,"abstract":"<div><div>Alternative to classical production of fucoidans, enzymatic synthesis has been proposed especially following genomic and bioinformatic investigations in <em>Ectocarpus siliculosus</em> and <em>Saccharina</em> sp. The present research aimed to assess the activity of four putative algal enzymes, including fucosyltransferases (FucTs_21 and FucTs_50) and carbohydrate sulfotransferases (STs_32 and STs_283) expressed in <em>E. coli</em> BL21 (DE3) for future application in lab-synthesized fucoidans. A specific activity of 0.47 pmol.min<sup>−1</sup>.µg<sup>−1</sup> was observed for FucTs_50 in the Glycosyltransferase activity assay, while multiplexed capillary electrophoresis (MP-CE) revealed that almost 32 % of GDP-L-fucose was affected. Additionally, the Universal Sulfotransferase Activity Kit was used to evaluate STs, and the results indicated that STs_283 and STs_32 exhibited specific activities at 2.29 and 2.38 pmol.min<sup>−1</sup>.µg<sup>−1</sup>, respectively. The results indicated correctly folded active enzymes with binding sites specific to their corresponding substrates.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"218 ","pages":"Article 109690"},"PeriodicalIF":3.7000,"publicationDate":"2025-02-18","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/S1369703X25000634","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

Alternative to classical production of fucoidans, enzymatic synthesis has been proposed especially following genomic and bioinformatic investigations in Ectocarpus siliculosus and Saccharina sp. The present research aimed to assess the activity of four putative algal enzymes, including fucosyltransferases (FucTs_21 and FucTs_50) and carbohydrate sulfotransferases (STs_32 and STs_283) expressed in E. coli BL21 (DE3) for future application in lab-synthesized fucoidans. A specific activity of 0.47 pmol.min⁻¹.µg⁻¹ was observed for FucTs_50 in the Glycosyltransferase activity assay, while multiplexed capillary electrophoresis (MP-CE) revealed that almost 32 % of GDP-L-fucose was affected. Additionally, the Universal Sulfotransferase Activity Kit was used to evaluate STs, and the results indicated that STs_283 and STs_32 exhibited specific activities at 2.29 and 2.38 pmol.min⁻¹.µg⁻¹, respectively. The results indicated correctly folded active enzymes with binding sites specific to their corresponding substrates.

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.