AQUEOUS TWO-PHASE EXTRACTION FOR THE PURIFICATION OF ALKALINE AGARASES FROM CULTURE EXTRACTS OF Pseudomonas aeruginosa AG LSL-11

被引:4
|
作者
Koti, Basawaraj A. [1 ]
Lakshmikanth, M. [2 ]
Manohar, S. [3 ]
Lalitha, J. [1 ]
机构
[1] Gulbarga Univ, Dept Biochem, Gulbarga 585106, Karnataka, India
[2] Heinrich Pette Inst Expt Virol & Immunol, Dept Cell Biol & Virol, Hamburg, Germany
[3] Avestagen Creator, ITPB, Bangalore, Karnataka, India
来源
PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY | 2012年 / 42卷 / 04期
关键词
agarase; aqueous two-phase system; Pseudomonas aeruginosa AG LSL-11; zymogram; PARTITION; SYSTEMS;
D O I
10.1080/10826068.2011.623210
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The agarases were purified for the first time an using aqueous two-phase system (ATPS) consisting of polyethylene glycol (PEG) and phosphate salt. The three extracellular, alkaline agarases produced by Pseudomonas aeruginosa AG LSL-11 were efficiently extracted into the top PEG-rich layer. The influencing factors on the partition of agarases-molecular weight of the PEG, system pH, system temperature, and NaCl concentration-were investigated. All the factors were found to have a significant effect on the partition of agarases except NaCl. The optimal ATPS parameters for the partitioning and purification of agarases were found to be 12% PEG 600 and 11.9% (w/w) phosphate salt at pH 8.0 and 4 degrees C. All three agarases were concentrated in the top PEG phase with 6.19-fold purity and 71.21% recovery. The ATPS was found to be more convenient and economical than the conventional ion-exchange chromatography (IEC) method for extraction of three agarases and could be significantly employed for the purification of agarases from fermentation broth.
引用
收藏
页码:364 / 377
页数:14
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