Purification of bacterial inulinase in aqueous two-phase systems
| Autor: | Hristina Strinska, Mishela Temkov, Elena Velickova, Vlad Muresan, Georgi Dobrev, Albert Krastanov |
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| Rok vydání: | 2018 |
| Předmět: |
0106 biological sciences
Binodal chemistry.chemical_classification Environmental Engineering Aqueous solution Chromatography 010405 organic chemistry Chemistry Bioengineering Polyethylene glycol Bacillus sp 01 natural sciences 0104 chemical sciences Electrophoresis chemistry.chemical_compound Enzyme 010608 biotechnology PEG ratio Inulinase Research Articles Biotechnology |
| Zdroj: | Eng Life Sci |
| ISSN: | 1618-0240 |
| Popis: | 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% MgSO(4) 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°C and kinetic parameters K(m) = 26.32 mmol and V(max) = 526 mmol/min. |
| Databáze: | OpenAIRE |
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