从地衣芽孢杆菌 So-B3 中提取的耐热 α-淀粉酶的纯化和特性及其在水解生淀粉中的应用。

Purification and characterization of thermostable α-amylase produced from Bacillus licheniformis So-B3 and its potential in hydrolyzing raw starch.

机构信息

Molecular Biology and Genetics Department, Science Faculty, Dicle University, 21280 Diyarbakir, Turkey.

Food Processing Programme, Technical Science Vocational School, Mersin University, TR-33343 Yenişehir, Mersin, Turkey.

出版信息

Life Sci. 2021 Jan 1;264:118639. doi: 10.1016/j.lfs.2020.118639. Epub 2020 Oct 24.

Abstract

AIMS

This work was achieved to obtain the optimum culture conditions of the thermostable alpha-amylase produced by thermophilic Bacillus licheniformis SO-B3. Furthermore, the α-amylase was purified and then characterized, and also its kinetic parameters were determined.

MATERIALS AND METHODS

A new thermotolerant bacteria called Bacillus licheniformis SO-B3 employed in this work was isolated from a sample of thermal spring mud in Şırnak (Meyremderesi). Several parameters such as the impact of temperature, time, and pH on enzyme production were examined. Thin-Layer Chromatography (TLC) was employed to analyze the end-products of soluble starch hydrolysis, and the utilization of purified α-amylase in the clarification of unripe apple juices was studied.

KEY FINDINGS

The highest enzyme production conditions were determined as 35 °C, 36th hour, and pH 7.0. Thermostable α-amylase was purified by 70% ammonium sulfate precipitation, DEAE-cellulose ion-exchange chromatography, and dialysis, with a 51-purification fold and 30% yield recovery. The K and V values for this enzyme were 0.004 mM and 3.07 μmol min at 70 °C, respectively. The α-amylase's molecular weight was found as 74 kDa. In addition, α-amylase showed a good degradation rate for raw starch.

SIGNIFICANCE

It was hypothesized that Bacillus licheniformis SO-B3 could be used as an α-amylase source. These findings displayed that purified enzyme could be utilized in fruit juice industries for clarification of apple juice and raw starch hydrolyzing.

摘要

目的

本工作旨在获得嗜热地衣芽孢杆菌 SO-B3 产生的耐热α-淀粉酶的最佳培养条件。此外,对α-淀粉酶进行了纯化和特性分析,并测定了其动力学参数。

材料与方法

本工作中使用的新耐热细菌,即地衣芽孢杆菌 SO-B3,是从舍尔纳克(Meyremderesi)一处温泉泥样中分离得到的。考察了温度、时间和 pH 等参数对酶产生的影响。采用薄层层析(TLC)分析可溶性淀粉水解的终产物,并研究了纯化的α-淀粉酶在未成熟苹果汁澄清中的应用。

主要发现

确定了最佳产酶条件为 35°C、36 小时和 pH 7.0。采用 70%硫酸铵沉淀、DEAE-纤维素离子交换层析和透析对耐热α-淀粉酶进行纯化,酶的纯化倍数为 51,回收率为 30%。该酶在 70°C 时的 K 和 V 值分别为 0.004 mM 和 3.07 μmol·min。α-淀粉酶的分子量为 74 kDa。此外,α-淀粉酶对原淀粉具有良好的降解率。

意义

推测地衣芽孢杆菌 SO-B3 可用作α-淀粉酶的来源。这些发现表明,纯化后的酶可用于果汁工业,用于澄清苹果汁和水解原淀粉。

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索