Characterization of β -Glucosidase Produced by Aspergillus niger under Solid-State Fermentation and Partially Purified Using MANAE-Agarose.

Biotechnology Research International Pub Date : 2014-01-01 Epub Date: 2014-04-01 DOI:10.1155/2014/317092
Anderson Baraldo Junior, Diogo G Borges, Paulo W Tardioli, Cristiane S Farinas
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引用次数: 35

Abstract

β -Glucosidase (BGL) is a hydrolytic enzyme with specificity for a wide variety of glycoside substrates, being an enzyme with a large range of biotechnological applications. However, enzyme properties can be different depending both on the microorganism and the cultivation procedure employed. Therefore, in order to explore potential biocatalytical applications of novel enzymes, their characterization is essential. In this work, a BGL synthesized by a selected strain of Aspergillus niger cultivated under solid-state fermentation (SSF) was partially purified and fully characterized in terms of optimum pH, temperature, and thermostability. The single-step purification using MANAE-agarose in a chromatographic column yielded an enzyme solution with specific activity (17.1 IU/mg protein) adequate for the characterization procedures. Electrophoresis SDS-PAGE and size-exclusion chromatography analysis resulted in an estimated molecular mass of 60 kDa. Higher enzyme activities were found in the range between 40 and 65°C and between pH 4 and 5.5, indicating an interesting characteristic for application in the hydrolysis of lignocellulosic biomass for biofuels production. Thermostability studies of purified BGL resulted in half-lives at 37°C of 56.3 h and at 50°C of 5.4 h. These results provide support for further studies of this enzyme towards revealing its potential biotechnological applications.

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黑曲霉固态发酵产β -葡萄糖苷酶的特性及部分纯化
β -葡萄糖苷酶(BGL)是一种对多种糖苷底物具有特异性的水解酶,是一种具有广泛生物技术应用的酶。然而,酶的性质可以根据微生物和所采用的培养程序而有所不同。因此,为了探索新型酶的潜在生物催化应用,它们的表征是必不可少的。在这项工作中,由选定的黑曲霉在固态发酵(SSF)下培养的菌株合成的BGL进行了部分纯化,并在最佳pH,温度和热稳定性方面进行了充分的表征。在色谱柱上使用manae -琼脂糖进行一步纯化,得到比活性(17.1 IU/mg蛋白质)的酶溶液,足以用于表征程序。电泳SDS-PAGE和大小排除色谱分析结果估计分子质量为60 kDa。在40至65°C和pH 4至5.5之间的范围内发现较高的酶活性,这表明了用于生产生物燃料的木质纤维素生物质水解的有趣特性。纯化BGL的热稳定性研究结果表明,在37℃和50℃的半衰期分别为56.3 h和5.4 h。这些结果为进一步研究该酶以揭示其潜在的生物技术应用提供了支持。
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