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源自HBB 7328的漆酶的生产、纯化及固定化及其在纺织染料脱色中的作用

Production, Purification and Immobilization of Laccase from HBB 7328 for its Role in Decolorization of Textile Dyes.

作者信息

Goyat Nikita, Kapoor Rajeev Kumar, Saharan Baljeet Singh, Kapoor Prexha, Kumari Kajal, Singh Namita

机构信息

Department of Microbiology, CCS Haryana Agricultural University, Hisar, 125004 India.

Department of Microbiology, Maharshi Dayanand University, Rohtak, Haryana 124001 India.

出版信息

Indian J Microbiol. 2024 Dec;64(4):1870-1883. doi: 10.1007/s12088-024-01260-3. Epub 2024 Apr 1.

Abstract

Laccase is an extracellular enzyme that is widely used in the decolonization of textile dyes in waste water. The aim of our study was to isolate, purify, characterize and immobilize the laccase enzyme produced by HBB 7328. Purified laccase enzyme was immobilized in polyacrylamide gel to explore its ability in decolonization of textile dyes. Laccase purification process was carried out by fractionation using ammonium sulphate (80%) followed by DEAE Sepharose column (30 × 3 cm) chromatography method. Recovery and fold purification in this step were 27.35 and 16.23%. Purified laccase (named as LAC1) revealed its optimum activity at pH 5.0 and 35 °C temperature, and displayed remarkable stability in the range of 30-40 °C and in the pH range (pH 3.0-7.0). The single bands on SDS-PAGE represent the purity of LAC1 with molecular weight of 60 kDa. Both free and immobilized laccase assessed for their ability to decolorize textile dyes. Free laccase decolorized Methyl red to 72.705%, Reactive orange to 57.851%, Reactive blue to 37.231%, Bromophenol blue to 24.412% however Immobilized laccase decolorized Methyl red to 89.823%, Reactive orange to 63.151%, Reactive blue to 59.548%, Bromophenol blue to 49.421% respectively. This study proposes the role of laccase from HBB 7328 in decolonization of textile dyes.

摘要

漆酶是一种胞外酶,广泛应用于废水中纺织染料的脱色处理。我们研究的目的是分离、纯化、表征并固定化由HBB 7328产生的漆酶。将纯化后的漆酶固定在聚丙烯酰胺凝胶中,以探究其对纺织染料的脱色能力。漆酶的纯化过程通过使用硫酸铵(80%)分级沉淀,随后采用DEAE琼脂糖柱(30×3 cm)层析法进行。此步骤中的回收率和纯化倍数分别为27.35%和16.23%。纯化后的漆酶(命名为LAC1)在pH 5.0和35℃温度下表现出最佳活性,并且在30 - 40℃范围和pH范围(pH 3.0 - 7.0)内具有显著的稳定性。SDS - PAGE上的单条带代表分子量为60 kDa的LAC1的纯度。对游离和固定化漆酶的纺织染料脱色能力进行了评估。游离漆酶使甲基红脱色72.705%,活性橙脱色57.851%,活性蓝脱色37.231%,溴酚蓝脱色24.412%;然而,固定化漆酶分别使甲基红脱色89.823%,活性橙脱色63.151%,活性蓝脱色59.548%,溴酚蓝脱色49.421%。本研究提出了来自HBB 7328的漆酶在纺织染料脱色中的作用。

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