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从喜马拉雅土壤中分离出的嗜热栖热放线菌(PANG L)来源的耐热纤维素酶的特性分析

Characterization of Thermostable Cellulase from PANG L Isolated from the Himalayan Soil.

作者信息

Shyaula Manita, Regmi Sunil, Khadka Deegendra, Poudel Ram Chandra, Dhakal Agni, Koirala Devesh, Sijapati Jaishree, Singh Anjana, Maharjan Jyoti

机构信息

Central Department of Microbiology, Tribhuvan University, Kirtipur, Kathmandu, Nepal.

Nepal Academy of Science and Technology, Khumaltar, Lalitpur, Nepal.

出版信息

Int J Microbiol. 2023 Aug 31;2023:3615757. doi: 10.1155/2023/3615757. eCollection 2023.

Abstract

This study aimed to isolate, purify, and characterize a potential thermophilic cellulase-producing bacterium from the Himalayan soil. Eleven thermophilic bacteria were isolated, and the strain PANG L was found to be the most potent cellulolytic producer. Morphological, physiological, biochemical, and molecular characterization identified PANG L as . This is the first study on the isolation of thermostable cellulase-producing from the Himalayan soil. This bacterium was processed for the production of cellulase enzyme. The optimum conditions for cellulase production were achieved at 45°C after 48 h of incubation at pH 6.5 in media-containing carboxymethyl cellulose (CMC) and yeast extract as carbon and nitrogen sources, respectively, in a thermo-shaker at 100 rpm. The enzyme was partially purified by 80% ammonium sulphate precipitation followed by dialysis, resulting in a 1.52-fold purification. The optimal activity of partially purified cellulase was observed at a temperature of 60°C and pH 5. The cellulase enzyme was stable within the pH ranges of 3-5 and retained 67% of activity even at 55°C. Cellulase activity was found to be enhanced in the presence of metal ions such as Cd, Pb, and Ba. The enzyme showed the highest activity when CMC was used as a substrate, followed by cellobiose. The and values of the enzyme were 1.8 mg/ml and 10.92 g/ml/min, respectively. The cellulase enzyme obtained from PANG L had suitable catalytic properties for use in industrial applications.

摘要

本研究旨在从喜马拉雅土壤中分离、纯化并鉴定一种潜在的嗜热纤维素酶产生菌。分离出了11株嗜热细菌,发现菌株PANG L是最有效的纤维素分解产物。通过形态学、生理学、生物化学和分子特征鉴定,PANG L为……。这是首次关于从喜马拉雅土壤中分离嗜热纤维素酶产生菌的研究。对该细菌进行了纤维素酶的生产处理。在含有羧甲基纤维素(CMC)和酵母提取物分别作为碳源和氮源的培养基中,于45°C、pH 6.5条件下在恒温振荡器中以100 rpm培养48小时后,达到了纤维素酶生产的最佳条件。通过80%硫酸铵沉淀,然后透析对酶进行部分纯化,纯化倍数为1.52倍。部分纯化的纤维素酶在温度60°C和pH 5时观察到最佳活性。纤维素酶在pH 3 - 5范围内稳定,即使在55°C时仍保留67%的活性。发现金属离子如Cd、Pb和Ba的存在会增强纤维素酶的活性。当以CMC为底物时,该酶显示出最高活性,其次是纤维二糖。该酶的Km和Vmax值分别为1.8 mg/ml和10.92 g/ml/min。从PANG L获得的纤维素酶具有适用于工业应用的催化特性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e5c1/10484656/ab8780cd2549/IJMICRO2023-3615757.001.jpg

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