Beroigui Oumaima, El Ghadraoui Lahsen, Errachidi Faouzi
Department of Biology, Functional Ecology and Environmental Engineering Laboratory, University Sidi Mohammed Ben Abdellah, Fez, Morocco.
J Basic Microbiol. 2023 Mar;63(3-4):427-438. doi: 10.1002/jobm.202200491. Epub 2023 Jan 27.
Inulinase is an enzyme that catalyzes inulin to d-fructose. This enzyme can be extracted from plants, but it is difficult to obtain it in large quantities, so its production cost is high. Therefore, microbial inulinase has great potential for industrial needs. In the last decade, there have been very few reports on actinobacterial inulinases, especially on purification and characterization of inulinase process extraction. This study aims to select actinomycetes that possess high inulinase activity from the soil. To screen inulinase-producing bacteria, modified Czapex-Dox agar supplemented with 1% inulin powder was used. The most effective isolate was Streptomyces sp. EFBO8, morphological and genotypic identification methods, confirmed that the strain is Streptomyces anulatus and that its nucleotide sequence has been deposited in GenBank under accession number OQ073700. To optimize inulinase production, kinetics were performed by using S. anulatus strain, which proved to be most productive with a value of 24,024 EU/mL. The enzyme was purified from the culture filtrate by precipitation with ammonium sulfate (NH ) SO , followed by column chromatography Sephadex (G-50) separation. Purified protein has a molecular mass of 3331.83 Da.
菊粉酶是一种将菊粉催化转化为d-果糖的酶。这种酶可以从植物中提取，但难以大量获取，因此其生产成本很高。所以，微生物菊粉酶在工业需求方面具有巨大潜力。在过去十年中，关于放线菌菊粉酶的报道极少，尤其是关于菊粉酶提取过程的纯化和特性研究。本研究旨在从土壤中筛选出具有高菊粉酶活性的放线菌。为筛选产菊粉酶的细菌，使用了添加1%菊粉粉末的改良察氏琼脂培养基。最有效的分离菌株是链霉菌属EFBO8，通过形态学和基因型鉴定方法，确认该菌株为环形链霉菌，其核苷酸序列已保存在GenBank中，登录号为OQ073700。为优化菊粉酶的生产，使用环形链霉菌菌株进行动力学研究，结果表明该菌株产酶量最高，为24,024 酶活力单位/毫升。通过硫酸铵((NH₄)₂SO₄)沉淀，随后用Sephadex (G - 50)柱色谱分离，从培养滤液中纯化该酶。纯化后的蛋白质分子量为3331.83 Da。
