Full Text:   <3500>

CLC number: TQ925

On-line Access: 

Received: 2005-06-07

Revision Accepted: 2005-09-03

Crosschecked: 0000-00-00

Cited: 9

Clicked: 6784

Citations:  Bibtex RefMan EndNote GB/T7714

-   Go to

Article info.
1. Reference List
Open peer comments

Journal of Zhejiang University SCIENCE B 2005 Vol.6 No.11 P.1087-1094


Effective extraction of elastase from Bacillus sp. fermentation broth using aqueous two-phase system

Author(s):  XU Ying, HE Guo-qing, LI Jing-jun

Affiliation(s):  Department of Food Science and Nutrition, Zhejiang University, Hangzhou 310029, China; more

Corresponding email(s):   gqhe@zju.edu.cn

Key Words:  Elastase, Aqueous two-phase system (ATPS), Bioseparation, Purification, Optimization, Response surface methodology (RSM)

XU Ying, HE Guo-qing, LI Jing-jun. Effective extraction of elastase from Bacillus sp. fermentation broth using aqueous two-phase system[J]. Journal of Zhejiang University Science B, 2005, 6(11): 1087-1094.

@article{title="Effective extraction of elastase from Bacillus sp. fermentation broth using aqueous two-phase system",
author="XU Ying, HE Guo-qing, LI Jing-jun",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T Effective extraction of elastase from Bacillus sp. fermentation broth using aqueous two-phase system
%A XU Ying
%A HE Guo-qing
%A LI Jing-jun
%J Journal of Zhejiang University SCIENCE B
%V 6
%N 11
%P 1087-1094
%@ 1673-1581
%D 2005
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.2005.B1087

T1 - Effective extraction of elastase from Bacillus sp. fermentation broth using aqueous two-phase system
A1 - XU Ying
A1 - HE Guo-qing
A1 - LI Jing-jun
J0 - Journal of Zhejiang University Science B
VL - 6
IS - 11
SP - 1087
EP - 1094
%@ 1673-1581
Y1 - 2005
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.2005.B1087

This paper presents the evaluation of an aqueous two-phase system (ATPS) for extracting elastase produced by Bacillus sp. EL31410. The elastase and cell partition behavior in polyethylene glycol (PEG)/salt systems was investigated. The suitable system for elastase extraction was PEG/KH2PO4-K2HPO4, in which elastase is mainly partitioned into the PEG-rich phase, while the cells remained in the other phase. The influence of defined system parameters (e.g. PEG molecular mass, pH, NaCl addition) on the partitioning behavior of elastase is described. The concentration of phase forming components, PEG and KH2PO4-K2HPO4, was optimized for elastase recovery by means of response surface methodology, and it was found that they greatly influenced extraction recovery. The optimal ATPS was 23.1% (w/w) PEG 2 000 and 11.7% (w/w) KH2PO4-K2HPO4. The predicted recovery was about 89.5%, so this process is suggested to be a rapid and convenient method for elastase extraction.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


[1] Balasubramaniam, D., Wilkinson, C., Cott, K.V., Zhang, C.M., 2003. Tobacco protein separation by aqueous two-phase extraction. J. Chromatogr. A, 989:119-129.

[2] Baskir, N.J., Hatton, T.A., Suter, U.W., 1989. Protein partitioning in two-phase aqueous polymer systems. Biotechnol. Bioeng., 34:541-558.

[3] Bim, M.A., Franco, T.T., 2000. Extraction in aqueous two-phase systems of alkaline xylanase produced by Bacillus pumilus and its application in kraft pulp bleaching. J. Chromatogr. B, 743:349-356.

[4] Bradford, M.M., 1976. A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem., 72:248.

[5] Box, G.E.P., Wilson, K.B., 1951. On the experimental attainment of optimum conditions. J. Roy. Stat. Soc. B, 13:145.

[6] Box, G.E.P., Hunter, W.G., 1978. Statistics for Experimenters. John Wiley and Sons, NY.

[7] Cascone, O., Andrews, B.A., Asenjo, J.A., 1991. Partitioning and purification of thaumatin in aqueous two-phase systems. Enzyme Microb. Technol., 13:629-635.

[8] Chen, Q.H., He, G.Q., Mokhtar, A.M.A., 2002. Optimization of medium composition for the production of elastase by Bacillus sp. EL31410 with response surface methodology. Enzyme Microb. Technol., 30:667-672.

[9] Diamond, A.D., Hsu, J.T., 1992. Aqueous two-phase systems for biomolecule separation. Adv. Biochem. Eng. Biotechnol., 47:89-135.

[10] Guan, Y.X., Mei, L.H., Zhu, Z.Q., 1994. Recovery of antibiotics by aqueous two-phase partition-partitioning behavior of pure acetylspiramycin solution in polyethylene glycol/potassium phosphate aqueous two-phase systems. Biotechnol. Techniques, 8:491-496.

[11] He, G.Q., Xu, Y., Chen, Q.H., Ruan, H., Li, J.J., 2004. Effect of temperature on batch elastase production by Bacillus sp. EL31410. J. Zhejiang Univ. Sci., 5:1583-1589.

[12] Héctor, M., Fernóndez, L., María, V.M., Elda, R., Fraile, M.J., Biscoglio, J.B., Osvaldo, C., 1995. Partition behavior and purification of a Mucor Bacilliformis acid protease in aqueous two-phase systems. Process Biochem., 30:615-621.

[13] Isabel del-Val, M., Cristina, O., 2003. Biphasic aqueous media containing polyethylene glycol for the enzymatic synthesis of oligosaccharides from lactose. Enzyme Microb. Technol., 33:118-126.

[14] Kimura, K., 2000. Simultaneous accumulation of low-molecular-mass RNA at the interface along with accumulation of high-molecular-mass RNA on aqueous two-phase system partitioning. J. Chromatogr. B, 743:421-429.

[15] Li, Q.M., He, B.L., 1992. Study on the partition of aminoacylase from Aspergillus oryzae in aqueous polyethylene glycol-salt two-phase system. Ion Exchange Absorp., 8:417-423.

[16] Li, C., Bai, J.H., Li, W., Cai, Z.L., Ouyang, F., 2001. Optimization of conditions for bacteriocin extraction in PEG/salt aqueous two-phase systems using statistical experimental designs. Biotechnol. Progr., 17:366-368.

[17] Li, M., Zhu, Z.Q., Rodrigues, A.E., 2002. Process integration of separation of amino acids by a temperature-induced aqueous two-phase system. Industrial. Eng. Chem. Res., 41:251-256.

[18] Marco, R.P., Alejandro, N., Enrique, G., Leobardo, S.C., 2000. Aroma compounds recovery from mycelial cultures in aqueous two-phase processes. J. Chromatogr. B, 743:403-408.

[19] Marcos, J.C., Fonseca, L.P., Ramalho, M.T., Cabral, J.M.S., 2002. Application of surface response analysis to the optimization of penicillin acylase purification in aqueous two-phase systems. Enzyme Microb. Technol., 31:1006-1013.

[20] Michelle, M.Z., Chad, A.Z., Shana, R.P., Lynn, R., 2001. Expression and partial characterization of an elastase from Chromobacterium violaceum. Vet. Microb., 80:63-74.

[21] Morihara, K., 1967. Elastolytic properties of various proteases from microbial origin. Arch. Biochem. Biophys., 120:68-78.

[22] Ohlsson, R., Hentschel, C.C., Williams, J.G., 1978. A rapid method for the isolation of circular DNA using an aqueous two-phase partition system. Nucleic. Acids. Res., 5:583-590.

[23] Pan, I.H., Yao, H.J., Li, Y.K., 2001. Effective extraction and purification of β-xylosidase from Trichoderma koningii fermentation culture by aqueous two-phase partitioning. Enzyme Microb. Technol., 28:196-201.

[24] Planas, J., Varelas, V., Tjerneld, F., Hahn-Hägerdal, B., 1998. Amine-based aqueous polymers for the simultaneous titration and extraction of lactic acid in aqueous two-phase systems. J. Chromatogr. B, 711:265-275.

[25] Sachar, L.A., 1955. Photometry method for estimation of elastase activity. Proc. Soc. Expeti. Biol. Med., 90:323-325.

[26] Shibata, Y., Fujimura, S., Nakamura, T., 1993. Purification and partial characterization of an elastolytic serine protease of Prevotella intermedia. Appl. Environm. Microbial., 59:2107-2111.

[27] Silgia, A.C., Adslberto, P.J., Inês, C.R., 2000. Partitioning of xylanolitic complex from Penicillium janthinellum by an aqueous two-phase systems. J. Chromatogr. B, 743:339-348.

[28] Tanuja, S., Srinivas, N.D., Raghava, K.R., Gowthaman, M.K., 1997. Aqueous two-phase extraction for downstream processing of amyloglucosidase. Process Biochem., 32:635-641.

[29] Tsai, Y.C., Jung, R.Y., Lin, S.F., 1988. Production and further characterization of an alkaline elastase production by alkalophilic Bacillus strain YaB. Appl. Environm. Microbial., 54:3156-3161.

[30] Yan, X.K., 2001. Bioseperation Engineering. Chemistry Industry Press, Beijing, p.169-187 (in Chinese).

[31] Yang, W.Y., Lin, C.D., Chu, I.M., Lee, C.J., 1994. Extraction of Cephalosporin C from whole broth and separation of desacetyl Cephalosporin C by aqueous two-phase partition. Biotechnol. Bioeng., 35:439-445.

Open peer comments: Debate/Discuss/Question/Opinion


Please provide your name, email address and a comment

Journal of Zhejiang University-SCIENCE, 38 Zheda Road, Hangzhou 310027, China
Tel: +86-571-87952783; E-mail: cjzhang@zju.edu.cn
Copyright © 2000 - 2024 Journal of Zhejiang University-SCIENCE