{"title":"Identification, heterologous expression, and characterisation of β-1,3-xylanase BcXyn26B from human gut bacterium Bacteroides cellulosilyticus WH2.","authors":"Sanae Hori, Fumiyoshi Okazaki","doi":"10.1007/s10529-024-03547-3","DOIUrl":null,"url":null,"abstract":"<p><p>The cell walls of red and green algae contain β-1,3-xylan, which is hydrolysed by the endo-type enzyme β-1,3-xylanase. Notably, only marine-bacteria-derived β-1,3-xylanases have been functionally characterised to date. In this study, we characterised the enzymatic properties of a potential β-1,3-xylanase (BcXyn26B) derived from the human gut bacterium, Bacteroides cellulosilyticus WH2. The codon optimized BcXyn26B gene was synthesised and expressed in Escherichia coli BL21(DE3). The recombinant protein was purified by a two-step purification process using Ni-affinity chromatography followed by anion exchange chromatography, and its enzymatic properties were characterised. The recombinant BcXyn26B exhibited specific hydrolytic activity against β-1,3-xylan and released various β-1,3-xylooligosaccharides, with β-1,3-xylobiose as the primary product. The optimum reaction temperature was 50 °C, higher than that for other enzymes derived from marine bacteria. This study represents the first report on the identification, heterologous expression, and characterisation of β-1,3-xylanase from human gut microbes. Notably, the substrate specificity of BcXyn26B indicates that human gut Bacteroides species possess an unknown β-1,3-xylan utilisation system.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 1","pages":"10"},"PeriodicalIF":2.0000,"publicationDate":"2024-12-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11611983/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biotechnology Letters","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.1007/s10529-024-03547-3","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
The cell walls of red and green algae contain β-1,3-xylan, which is hydrolysed by the endo-type enzyme β-1,3-xylanase. Notably, only marine-bacteria-derived β-1,3-xylanases have been functionally characterised to date. In this study, we characterised the enzymatic properties of a potential β-1,3-xylanase (BcXyn26B) derived from the human gut bacterium, Bacteroides cellulosilyticus WH2. The codon optimized BcXyn26B gene was synthesised and expressed in Escherichia coli BL21(DE3). The recombinant protein was purified by a two-step purification process using Ni-affinity chromatography followed by anion exchange chromatography, and its enzymatic properties were characterised. The recombinant BcXyn26B exhibited specific hydrolytic activity against β-1,3-xylan and released various β-1,3-xylooligosaccharides, with β-1,3-xylobiose as the primary product. The optimum reaction temperature was 50 °C, higher than that for other enzymes derived from marine bacteria. This study represents the first report on the identification, heterologous expression, and characterisation of β-1,3-xylanase from human gut microbes. Notably, the substrate specificity of BcXyn26B indicates that human gut Bacteroides species possess an unknown β-1,3-xylan utilisation system.
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.