EFFECT OF SALTS AND POLYETHYLENE GLYCOLS ON THE PARTITIONING AND RECOVERY OF TRYPSIN FROM HYBRID CATFISH VISCERA IN AQUEOUS TWO-PHASE SYSTEMS

被引：14

作者：

Klomklao, Sappasith ^{[1
]}

Benjakul, Soottawat ^{[2
]}

Kishimura, Hideki ^{[3
]}

Osako, Kazufumi ^{[4
]}

Tanaka, Munehiko ^{[4
]}

机构：

[1] Thaksin Univ, Dept Food Sci & Technol, Fac Technol & Community Dev, Phattalung 93110, Thailand

[2] Prince Songkla Univ, Dept Food Technol, Fac Agroind, Hat Yai 90112, Songkhla, Thailand

[3] Hokkaido Univ, Lab Marine Prod & Food Sci, Res Fac Fisheries Sci, Hakodate, Hokkaido 0418611, Japan

[4] Tokyo Univ Marine Sci & Technol, Dept Food Sci & Technol, Minato Ku, Tokyo 1088477, Japan

来源：

JOURNAL OF FOOD BIOCHEMISTRY | 2010年 / 34卷 / 04期

关键词：

PYLORIC CECA; PURIFICATION; EXTRACTION; SPLEEN; PEROXIDASE; PROTEINASE; INHIBITORS; PHOSPHATE;

D O I：

10.1111/j.1745-4514.2009.00311.x

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The partitioning behavior of trypsin from hybrid catfish viscera in aqueous two-phase systems (ATPS) was studied. Factors such as polyethylene glycol (PEG) molecular mass and concentration, as well as types and concentration of salts, affected protein separation. Trypsin partitioned mainly in the top PEG-rich phase. ATPS formed by PEG of molecular weight 4,000 (20%, w/w) and NaH2PO4 (20%, w/w) showed the best capability for trypsin purification from hybrid catfish viscera. Under such conditions, the highest specific activity (30.05 units/mg protein) and purification (27.3-fold), were obtained. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis revealed that the enzyme after ATPS separation was near homogeneity and based on the activity staining, the band intensity of enzyme in ATPS fraction increased, indicating the greater specific activity of the viscera extract. The partitioned enzyme displayed optimal activity at pH 9.0 and 50C, respectively. The enzyme was stable up to 40C and within the pH range of 8-12. The enzyme exhibited a progressive decrease in activity with increasing NaCl concentration.

引用

页码：730 / 747

页数：18

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