Comparative studies on proteolytic activity of splenic extract from three tuna species commonly used in thailand

被引:105
作者
Klomklao, S
Benjakul, S [1 ]
Visessanguan, W
机构
[1] Prince Songkla Univ, Dept Food Technol, Fac Agroind, Hat Yai 90112, Thailand
[2] Natl Sci & Technol Dev Agcy, Natl Ctr Genet Engn & Biotechnol, Pathum Thani 12120, Thailand
关键词
D O I
10.1111/j.1745-4514.2004.05203.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
Proteolytic activities of splenic extract from three tuna species including skipjack tuna (Katsuwonus pelamis), yellowfin tuna (Thunnus albacores) and tongol tuna (Thunnus tonggol) were studied. Optimal activity of splenic extract from all tuna species was at pH 9.0 and 55C when casein was used as a substrate. Among all species tested, yellowfin tuna showed the highest activity, followed by skipjack tuna and tongol tuna. The proteolytic activity was strongly inhibited by soybean trypsin inhibitor, TLCK and partially inhibited by ethylenediaminetetraacetic acid. E-64, N-ethylmaleimide, iodoacetic acid, TPCK and pepstatin A showed no inhibition. The effect of NaCl and CaCl2 on proteolytic activity was also investigated. Activities continuously decreased as NaCl concentration increased, and no activity remained in the presence of 30% NaCl. On the other hand, activities increased as CaCl2 concentration increased. The highest activity was obtained in the presence of 1 mM CaCl2. SDS-substrate gel electrophoresis revealed that major proteinases in splenic extract from different tuna species were different in apparent molecular weights and sensitivity to TLCK. Although them major activity bands of all species were strongly inhibited by soybean trypsin inhibitor varying sensitivity to TLCK probably implied the differences in binding characteristic of enzyme to substrate and/or inhibitors. The results suggest that major proteinases in spleen of all tuna species were trypsin-like serine proteinases.
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页码:355 / 372
页数:18
相关论文
共 46 条
[1]  
An H, 2000, SEAFOOD ENZYMES, P641
[2]   ASSAY SYSTEMS AND CHARACTERIZATION OF PACIFIC WHITING (MERLUCCIUS-PRODUCTUS) PROTEASE [J].
AN, HJ ;
SEYMOUR, TA ;
WU, JW ;
MORRISSEY, MT .
JOURNAL OF FOOD SCIENCE, 1994, 59 (02) :277-281
[3]   ISOLATION AND CHARACTERIZATION OF PEPSIN FROM POLAR COD (BOREOGADUS-SAIDA) [J].
ARUNCHALAM, K ;
HAARD, NF .
COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY, 1985, 80 (03) :467-473
[4]   Partial purification and characterization of a thermostable trypsin from pyloric caeca of tambaqui (Colossoma macropomum) [J].
Bezerra, RS ;
Santos, JF ;
Paiva, PMG ;
Correia, MTS ;
Coelho, LCBB ;
Vieira, VLA ;
Carvalho, LB .
JOURNAL OF FOOD BIOCHEMISTRY, 2001, 25 (03) :199-210
[5]   REFINED CRYSTAL-STRUCTURE OF BOVINE BETA-TRYPSIN AT 1.8 A RESOLUTION .2. CRYSTALLOGRAPHIC REFINEMENT, CALCIUM-BINDING SITE, BENZAMIDINE BINDING-SITE AND ACTIVE-SITE AT PH 7.0 [J].
BODE, W ;
SCHWAGER, P .
JOURNAL OF MOLECULAR BIOLOGY, 1975, 98 (04) :693-717
[6]   Purification and characterization of a serine proteinase from the tuna pyloric caeca [J].
Byun, HG ;
Park, PJ ;
Sung, NJ ;
Kim, SK .
JOURNAL OF FOOD BIOCHEMISTRY, 2002, 26 (06) :479-494
[7]   Purification and characterization of two anionic trypsins from the hepatopancreas of carp [J].
Cao, MJ ;
Osatomi, K ;
Suzuki, M ;
Hara, K ;
Tachibana, K ;
Ishihara, T .
FISHERIES SCIENCE, 2000, 66 (06) :1172-1179
[8]   Partial characterization and activities of proteases from the digestive tract of discus fish (Symphysodon aequifasciata) [J].
Chong, ASC ;
Hashim, R ;
Chow-Yang, L ;
Ali, AB .
AQUACULTURE, 2002, 203 (3-4) :321-333
[9]   PHYSICO-CHEMICAL PROPERTIES OF BOVINE CHYMOTRYPSINOGEN B - A COMPARATIVE STUDY WITH TRYPSINOGEN AND CHYMOTRYPSINOGEN A [J].
DELAAGE, M ;
ABITA, JP ;
LAZDUNSKI, M .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1968, 5 (02) :285-+
[10]  
DeVecchi S, 1996, J FOOD BIOCHEM, V20, P193