{"title":"海洋几丁质酶 AfChi:针对球孢子菌和炭疽病的绿色防御管理。","authors":"Rajesh K M, Keyur Raval, Ritu Raval","doi":"10.1186/s13568-024-01786-1","DOIUrl":null,"url":null,"abstract":"<p><p>Anthracnose disease, caused by the Colletotrichum gloeosporioides species, affects vegetables, fruits, pulses, and cereals, leading to significant economic losses worldwide. Although many synthetic fungicides are used to control this pathogen, eco-friendly biological alternatives are gaining popularity. This study focuses on isolating and purifying chitinase ( Af Chi)from a marine bacterium and testing its antifungal efficacy against C. gloeosporioides spore germination by targeting the chitin in the fungal cell wall. The chitinase was purified from a marine bacterium A. faecalis from the Arabian Sea and had a molecular mass of 45 kDa and a specific activity of 84.6 U/mg. Af Chi worked best at 50 °C and pH 7.0 in Tris HCl buffer. Na<sup>+</sup> ion was the highest cofactor, highlighting the halophilic nature of this chitinase. K<sup>+</sup>, Ca<sup>2+</sup>, Cu<sup>2+</sup>, Mg<sup>2+</sup>, Mn<sup>2+</sup>, and EDTA also increased activity, while Fe<sup>3+</sup>, Zn<sup>2+</sup>, Co<sup>2+</sup>, and Pb<sup>2+</sup> decreased it. The K<sub>m</sub> and V<sub>max</sub> values were 1.87 µg/mL and 17.45 U/mL, respectively. Purified Af Chi at 10 mg/mL completely inhibited spore germination within 8 h and reduced the size of the spores.</p>","PeriodicalId":7537,"journal":{"name":"AMB Express","volume":"14 1","pages":"128"},"PeriodicalIF":3.5000,"publicationDate":"2024-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11579262/pdf/","citationCount":"0","resultStr":"{\"title\":\"Marine chitinase AfChi: green defense management against Colletotrichum gloeosporioides and anthracnose.\",\"authors\":\"Rajesh K M, Keyur Raval, Ritu Raval\",\"doi\":\"10.1186/s13568-024-01786-1\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Anthracnose disease, caused by the Colletotrichum gloeosporioides species, affects vegetables, fruits, pulses, and cereals, leading to significant economic losses worldwide. Although many synthetic fungicides are used to control this pathogen, eco-friendly biological alternatives are gaining popularity. This study focuses on isolating and purifying chitinase ( Af Chi)from a marine bacterium and testing its antifungal efficacy against C. gloeosporioides spore germination by targeting the chitin in the fungal cell wall. The chitinase was purified from a marine bacterium A. faecalis from the Arabian Sea and had a molecular mass of 45 kDa and a specific activity of 84.6 U/mg. Af Chi worked best at 50 °C and pH 7.0 in Tris HCl buffer. Na<sup>+</sup> ion was the highest cofactor, highlighting the halophilic nature of this chitinase. K<sup>+</sup>, Ca<sup>2+</sup>, Cu<sup>2+</sup>, Mg<sup>2+</sup>, Mn<sup>2+</sup>, and EDTA also increased activity, while Fe<sup>3+</sup>, Zn<sup>2+</sup>, Co<sup>2+</sup>, and Pb<sup>2+</sup> decreased it. The K<sub>m</sub> and V<sub>max</sub> values were 1.87 µg/mL and 17.45 U/mL, respectively. Purified Af Chi at 10 mg/mL completely inhibited spore germination within 8 h and reduced the size of the spores.</p>\",\"PeriodicalId\":7537,\"journal\":{\"name\":\"AMB Express\",\"volume\":\"14 1\",\"pages\":\"128\"},\"PeriodicalIF\":3.5000,\"publicationDate\":\"2024-11-20\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11579262/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"AMB Express\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://doi.org/10.1186/s13568-024-01786-1\",\"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}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"AMB Express","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.1186/s13568-024-01786-1","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
摘要
炭疽病是由球孢霉菌(Colletotrichum gloeosporioides)引起的,主要危害蔬菜、水果、豆类和谷物,给全世界造成重大经济损失。尽管许多合成杀菌剂被用来控制这种病原体,但环保型生物替代品正日益受到欢迎。本研究的重点是从一种海洋细菌中分离和纯化几丁质酶(Af Chi),并测试其通过靶向真菌细胞壁中的几丁质来抑制球孢子菌孢子萌发的抗真菌功效。几丁质酶是从阿拉伯海的海洋细菌 A. faecalis 中纯化出来的,其分子质量为 45 kDa,比活度为 84.6 U/mg。Af Chi 在温度为 50 °C、pH 值为 7.0 的 Tris HCl 缓冲液中效果最佳。Na+ 离子是最高的辅助因子,突出了这种几丁质酶的嗜盐性。K+、Ca2+、Cu2+、Mg2+、Mn2+ 和 EDTA 也能提高活性,而 Fe3+、Zn2+、Co2+ 和 Pb2+ 则会降低活性。Km 和 Vmax 值分别为 1.87 µg/mL 和 17.45 U/mL。纯化的 Af Chi 在 10 mg/mL 的浓度下可在 8 小时内完全抑制孢子萌发,并缩小孢子的体积。
Marine chitinase AfChi: green defense management against Colletotrichum gloeosporioides and anthracnose.
Anthracnose disease, caused by the Colletotrichum gloeosporioides species, affects vegetables, fruits, pulses, and cereals, leading to significant economic losses worldwide. Although many synthetic fungicides are used to control this pathogen, eco-friendly biological alternatives are gaining popularity. This study focuses on isolating and purifying chitinase ( Af Chi)from a marine bacterium and testing its antifungal efficacy against C. gloeosporioides spore germination by targeting the chitin in the fungal cell wall. The chitinase was purified from a marine bacterium A. faecalis from the Arabian Sea and had a molecular mass of 45 kDa and a specific activity of 84.6 U/mg. Af Chi worked best at 50 °C and pH 7.0 in Tris HCl buffer. Na+ ion was the highest cofactor, highlighting the halophilic nature of this chitinase. K+, Ca2+, Cu2+, Mg2+, Mn2+, and EDTA also increased activity, while Fe3+, Zn2+, Co2+, and Pb2+ decreased it. The Km and Vmax values were 1.87 µg/mL and 17.45 U/mL, respectively. Purified Af Chi at 10 mg/mL completely inhibited spore germination within 8 h and reduced the size of the spores.
期刊介绍:
AMB Express is a high quality journal that brings together research in the area of Applied and Industrial Microbiology with a particular interest in ''White Biotechnology'' and ''Red Biotechnology''. The emphasis is on processes employing microorganisms, eukaryotic cell cultures or enzymes for the biosynthesis, transformation and degradation of compounds. This includes fine and bulk chemicals, polymeric compounds and enzymes or other proteins. Downstream processes are also considered. Integrated processes combining biochemical and chemical processes are also published.