Full Text:
<2954>
CLC number: Q814.1
On-line Access:
Received: 2005-03-17
Revision Accepted: 2005-04-28
Crosschecked: 0000-00-00
Cited: 3
Clicked: 6201
Partitioning and purification of extracellular β-1,3-1,4-glucanase in aqueous two-phase systems
| |||||||||||||
HE Guo-qing, ZHANG Xiu-yan, TANG Xing-jun, CHEN Qi-he, RUAN Hui. Partitioning and purification of extracellular β-1,3-1,4-glucanase in aqueous two-phase systems[J]. Journal of Zhejiang University Science B, 2005, 6(8): 825-831.
@article{title="Partitioning and purification of extracellular β-1,3-1,4-glucanase in aqueous two-phase systems",
author="HE Guo-qing, ZHANG Xiu-yan, TANG Xing-jun, CHEN Qi-he, RUAN Hui",
journal="Journal of Zhejiang University Science B",
volume="6",
number="8",
pages="825-831",
year="2005",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.2005.B0825"
}
%0 Journal Article
%T Partitioning and purification of extracellular β-1,3-1,4-glucanase in aqueous two-phase systems
%A HE Guo-qing
%A ZHANG Xiu-yan
%A TANG Xing-jun
%A CHEN Qi-he
%A RUAN Hui
%J Journal of Zhejiang University SCIENCE B
%V 6
%N 8
%P 825-831
%@ 1673-1581
%D 2005
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.2005.B0825
TY - JOUR
T1 - Partitioning and purification of extracellular β-1,3-1,4-glucanase in aqueous two-phase systems
A1 - HE Guo-qing
A1 - ZHANG Xiu-yan
A1 - TANG Xing-jun
A1 - CHEN Qi-he
A1 - RUAN Hui
J0 - Journal of Zhejiang University Science B
VL - 6
IS - 8
SP - 825
EP - 831
%@ 1673-1581
Y1 - 2005
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.2005.B0825
Abstract: The partition behaviors of β-1,3-1,4-glucanase, α-amylase and neutral proteases from clarified and whole fermentation broths of Bacillus 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.
[1] Edney, M.J., Marchylo, B.A., Macgregor, A.W., 1991. Structure of total barley-glucan. J. Inst. Brew., 97:39-44.
[2] Godfrey, T., Reichely, J., 1983. Industrial Enzymology. McMillan, London, p.466.
[3] Han, J.H., Lee, C.H., 1997. Effects of salts and poly(ethylene glycol)-palmitate on the partitioning of proteins and Bacillus subtilis neutral protease in aqueous two-phase systems. Colloids and Surfaces B: Biointerfaces, 9:109-116.
[4] Huddleston, J.G., Veide, A.K., 1991. The molecular basis of partitioning in aqueous two-phase systems. TIBTECH, 9:381-388.
[5] Jain, A., Johri, B.N., 1999. Partitioning of an extracellular xylanase produced by a thermophilic fungus Melanocarpus albomyces IIS-68 in an aqueous two-phase system. Bioresource Technology, 67:205-207.
[6] Jiao, Q.C., Liu, X., Chen, Y.Z., 1998. Studies on the separation and extraction of (-amylase and proteinase from fermented liquor of (-amylase by aqueous two-phase partitioning. Chem. J. Chinese Universities, 3:391 (in Chinese).
[7] Johansson, G., Reczey, K., 1998. Concentration and purification of β-glucosidase from Aspergillus niger by using aqueous two-phase partitioning. Journal of Chromatography B, 711:161-172.
[8] Miller, G.L., 1959. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal. Chem., 31:426-427.
[9] Pan, I.H., Li, Y.K., 2001. Rapid process for purification of an extracellular β-xylosidase by aqueous two-phase extraction. Journal of Chromatography B, 754:179-184.
[10] Planas, A., 2000. Bacterial 1,3-1,4-β-glucanases: Structure, function and protein engineering. Biochimica et Biophysica Acta, 1543:361-382.
[11] Schmidt, A.S., Ventom, A.M., Asenjo, J.A., 1994. Partitioning and purification of α-amylase in aqueous two-phase systems. Enzyme Microb. Technol., 16:131-142.
[12] Stone, B.A., Clarke, A.E., 1992. Chemistry and Biology of 1,3-β-Glucans. La Trobe University Press, Bundoora, Australia.
[13] Syu, M.J., Chen, Y.H., 1997. A study on the α-amylase fermentation performed by Bacillus amyloliquefaciens. Chemical Engineering Journal, 65:237-247.
[14] Xiao, J.X., Bao, Y.X., Fang, L., 2001. Phase behavior and protein partitioning in aqueous two-phase systems formed by mixtures of polymer and cationic-anionic surfactants. Chem. J. Chinese Universities, 22(1):112-114.
[15] White, W.B., Bird, H.R., Sunde, M.L., Marlett, J.A., 1983. Poult. Sci., 62:853-862.
[16] Woodward, J.R., Phillips, D.R., Fincher, G.B., 1983. Water soluble (1-3), (1-4)-β-D-glucans from barley (Hordeum vulgare) endosperm. I. Physicochemical properties. Carbohydr. Polym., 3:143-156.
[17] Zhang, L.X., Zhang, T.F., Li, L.Y., 1997. Method and Technology of Biochemistry Experiments. Higher Education Press, Beijing (in Chinese).
Open peer comments: Debate/Discuss/Question/Opinion
<1>
ramyadevi<ramyachem@rediffmail.com>
2010-11-12 15:02:29
it very nice and useful for my research work