Purification of bacterial inulinase in aqueous two-phase systems

被引：10

作者：

Temkov, Mishela ^{[1
,2
]}

Strinska, Hristina ^{[2
]}

Dobrev, Georgi ^{[2
]}

Velickova, Elena ^{[1
]}

Muresan, Vlad ^{[3
]}

Krastanov, Albert ^{[2
]}

机构：

[1] Ss Cyril & Methodius Univ Skopje, Dept Food Technol & Biotechnol, Fac Technol & Met, Rudjer Boskovic 16, Skopje 1000, Macedonia

[2] Univ Food Technol, Dept Biotechnol & Mol Biol, Fac Technol, Plovdiv, Bulgaria

[3] Univ Agr & Vet Med Cluj Napoca, Dept Food Engn, Fac Food Technol, Cluj Napoca, Romania

来源：

ENGINEERING IN LIFE SCIENCES | 2018年 / 18卷 / 11期

关键词：

Aqueous-two phase systems; Binodal curve; Inulinase; Polyethylene glycol; Purification; EXTRACELLULAR INULINASE; PHASE-DIAGRAMS; PROTEIN; HYDROLYSIS; EXTRACTION; SEPARATION; LIPASE; ENZYME; INVERTASE; PARTITION;

D O I：

10.1002/elsc.201700174

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

In this study, extracellular inulinase from Bacillus sp. 11/3 was partially purified and concentrated using aqueous two-phase system (ATPS). Two different phase forming salts and four types of polyethylene glycol (PEG) were used. Binodal curves and tie-length lines (TLLs) for eight ATPS were developed. For inulinase purification, concentrations of PEG and salt according to binodal curves (between 17 and 26%) were chosen. All ATPSs for inulinase purification were characterized. An ATPS consisted of 26% PEG1000 and 26% MgSO4 was found to be the most suitable for inulinase purification. This ATPS has 28.47% TLL, 1.03 of volume ratio, purification factor of 4.65 fold and recovery yield of 66.17%. On the SDS-PAGE electrophoresis two protein bands with molecular weight of around 24 and 56 kDa were observed. The partially purified enzymes had optimal activity at pH 8.0 and 6.5, optimal temperature at 30 and 70 degrees C and kinetic parameters K-m = 26.32 mmol and V-max = 526 mmol/min.

引用

页码：840 / 850

页数：11

共 48 条

[1] A Novel Aqueous Two Phase System Composed of a Thermo-Separating Polymer and an Organic Solvent for Purification of Thermo-Acidic Amylase Enzyme from Red Pitaya (Hylocereus polyrhizus) Peel [J].