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筛选几丁质分解真菌并通过固态发酵技术优化几丁质酶高产的参数。

Screening of Chitinolytic Microfungi and Optimization of Parameters for Hyperproduction of Chitinase Through Solid-State Fermentation Technique.

机构信息

Department of Biochemistry and Biotechnology, University of Gujrat, Hafiz Hayat Campus, Gujrat, Punjab, Pakistan.

出版信息

Appl Biochem Biotechnol. 2024 Apr;196(4):1840-1862. doi: 10.1007/s12010-023-04663-y. Epub 2023 Jul 13.

Abstract

This study is intended for the production of chitinase enzyme from locally isolated fungal strains. Out of 10 isolated fungal strains from district Gujrat, Punjab, Pakistan, Aspergillus terreus SB3 (accession number ON738571) was found with maximum chitinolytic potential (80.8 U/mL/min). By applying central composite design (CCD) through response surface methodology (RSM) under solid-state fermentation (SSF), eight nutritional and physical parameters were optimized. Among these, temperature, substrate concentration, and pH were found as significant factors toward chitinase production in the first phase. Moisture and nitrogen source were found as significant factors during second phase of chitinase production. The effect of incubation period, inoculum size, and magnesium source was observed as non-significant. The chitinase activity was successfully enhanced more than 2 folds up to 198.5 U/mL/min at optimized conditions of 35 °C temperature, 4.5 pH, 20 g substrate concentration, 4-day incubation period, 55% moisture content, 4.5 mL inoculum size, 0.25 g ammonium sulfate, and 0.30 g magnesium sulfate using RSM design. It was also found that Ganoderma lucidum (bracket fungus) has more potential to be used for the production of chitinase compared to fish scales. The present study exhibited Aspergillus terreus SB3 (ON738571) as a potential indigenous strain capable for hyperproduction of chitinase through cheap fermentation technology that might be employed for the eradication of chitin-based sea waste to remove the marine pollution.

摘要

本研究旨在从当地分离的真菌菌株中生产几丁质酶。在从巴基斯坦旁遮普省古吉拉特地区分离的 10 株真菌菌株中,发现 Aspergillus terreus SB3(登录号 ON738571)具有最大的几丁质酶活力(80.8 U/mL/min)。通过响应面法(RSM)中的中心组合设计(CCD)在固态发酵(SSF)下,优化了 8 种营养和物理参数。在这些因素中,温度、底物浓度和 pH 被发现是几丁质酶生产的第一阶段的重要因素。在几丁质酶生产的第二阶段,发现水分和氮源是重要因素。培养时间、接种量和镁源的影响被观察为非显著因素。在优化条件下,温度为 35°C、pH 为 4.5、底物浓度为 20 g、培养时间为 4 天、水分含量为 55%、接种量为 4.5 mL、硫酸铵为 0.25 g 和硫酸镁为 0.30 g,几丁质酶活性成功提高了 2 倍以上,达到 198.5 U/mL/min。通过 RSM 设计还发现,与鱼鳞相比,灵芝(担子菌)更有潜力用于生产几丁质酶。本研究表明,Aspergillus terreus SB3(ON738571)是一种具有生产几丁质酶潜力的潜在本土菌株,通过廉价的发酵技术可以实现几丁质酶的高产,这可能用于消除基于几丁质的海洋废物,以消除海洋污染。

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