Partitioning and purification of extracellular β-1,3-1,4-glucanase in aqueous two-phase systems |
| |
| Authors: | HE Guo-qing ZHANG Xiu-yan TANG Xing-jun CHEN Qi-he RUAN Hui |
| |
| Institution: | (1) Department of Biochemical and Pharmaceutical Technology, University of S?o Paulo, 05508-000 S?o Paulo, SP, Brazil;(2) Department of Chemical Engineering, University of S?o Paulo, 05508-000 S?o Paulo, SP, Brazil;(3) Department of Fundamental Chemistry, UFPE, 50740-540 Recife, PE, Brazil;(4) Laboratory of Immunopathology Keizo Asami (LIKA), UFPE, 50670-901 Recife, PE, Brazil;(5) Department of Chemical and Process Engineering “G. B. Bonino”, via Opera Pia 15, 16145 Genoa, Italy |
| |
| Abstract: | The partition behaviors of β-1,3-1,4-glucanase, α-amylase and neutral proteases from clarified and whole fermentation broths
ofBacillus subtilis ZJF-1A5 were investigated. An aqueous two-phase system (polyethylene glycol (PEG)/MgSO4) was examined with regard to the effects of PEG molecular weight (MW) and concentration, MgSO4 concentration, pH and NaCl concentration on enzyme partition and extraction. The MW and concentration of PEG were found to
have significant effects on enzyme partition and extraction with low MW PEG showing the greatest benefit in the partition
and extraction of β-glucanase with the PEG/MgSO4 system. MgSO4 concentration influenced the partition and extraction of β-glucanase significantly. pH had little effect on β-glucanase or
proteases partition but affected α-amylase partition when pH was over 7.0. The addition of NaCl had little effect on the partition
behavior of β-glucanase but had very significant effects on the partitioning of α-amylase and on the neutral proteases. The
partition behaviors of β-glucanase, α-amylase and proteases in whole broth were also investigated and results were similar
to those obtained with clarified fermentation broth. A two-step process for purifying β-glucanase was developed, which achieved
β-glucanase recovery of 65.3% and specific activity of 14027 U/mg, 6.6 times improvement over the whole broth. |
| |
| Keywords: | |
| 本文献已被 万方数据 SpringerLink 等数据库收录！ |
|
