Purification and Biochemical and Kinetic Properties of an Endo-Polygalacturonase from the Industrial Fungus Aspergillus sojae.

IF 1.2 Q2 Biochemistry, Genetics and Molecular Biology

Journal of Molecular Microbiology and Biotechnology Pub Date : 2017-01-01 Epub Date: 2017-04-28 DOI:10.1159/000460296

Dante Fratebianchi, Ivana Alejandra Cavello, Sebastián Fernando Cavalitto

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引用次数: 11

Abstract

An endo-polygalacturonase secreted by Aspergillus sojae was characterized after being purified to homogeneity from submerged cultures with orange peel as the sole carbon source by gel filtration and ion-exchange chromatographies. According to SDS-PAGE and analytical isoelectric focusing analyses, the enzyme presents a molecular weight of 47 kDa and pI value of 4.2. This enzyme exhibits considerable stability under highly acidic to neutral conditions (pH 1.5-6.5) and presents a half-life of 2 h at 50°C. Besides its activity towards pectin and polygalacturonic acid, the enzyme displays pectin-releasing activity, acting best in a pH range of 3.3-5.0. Thin-layer chromatographic analysis revealed that tri-galacturonate is the main enzymatic end product of polygalacturonic acid hydrolysis, indicating that it is an endo-polygalacturonase. The enzyme exhibits Michaelis-Menten kinetics, with KM and VMAX values of 0.134 mg/mL and 9.6 µmol/mg/min, respectively, and remained stable and active in the presence of SO2, ethanol, and various cations assayed except Hg2+.

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工业真菌大豆曲霉内切聚半乳糖醛酸酶的纯化及生化动力学性质研究。

以陈皮为唯一碳源，采用凝胶过滤和离子交换色谱法对大豆曲霉分泌的一种内聚半乳糖醛酸酶进行了纯化。根据SDS-PAGE和等电聚焦分析，该酶分子量为47 kDa, pI值为4.2。该酶在高酸性至中性条件下(pH 1.5-6.5)表现出相当大的稳定性，在50°C时半衰期为2小时。该酶除对果胶和聚半乳糖醛酸具有释放果胶的活性外，还具有释放果胶的活性，在3.3 ~ 5.0的pH范围内效果最佳。薄层色谱分析表明，三半乳糖醛酸是聚半乳糖醛酸水解的主要酶促终产物，表明其为内切型聚半乳糖醛酸酶。该酶具有Michaelis-Menten动力学，KM和VMAX值分别为0.134 mg/mL和9.6µmol/mg/min，在SO2、乙醇和除Hg2+外的各种阳离子存在下均保持稳定和活性。

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来源期刊

Journal of Molecular Microbiology and Biotechnology 生物-生物工程与应用微生物

CiteScore

3.90

自引率

0.00%

发文量

审稿时长

>12 weeks

期刊介绍： We are entering a new and exciting era of microbiological study and application. Recent advances in the now established disciplines of genomics, proteomics and bioinformatics, together with extensive cooperation between academic and industrial concerns have brought about an integration of basic and applied microbiology as never before.