Purification and characterization of a novel thermoacid-stable fibrinolytic enzyme from Staphylococcus sp. strain AJ isolated from Korean salt-fermented Anchovy-joet

Abstract

A novel Wbrinolytic enzyme (AJ) was puriWed from Staphylococcus sp. strain AJ screened from Korean salt-fermented Anchovy-jeot. Relative molecular weight of AJ was determined as 26 kDa by using SDS-PAGE and Wbrin zymography. Based on a 2D gel, AJ was found to consist of three active isoforms (pI 5.5-6.0) with the same N-terminal amino acid sequence. AJ exhibited optimum pH and temperature at 2.5-3.0 and 85°C, respectively. AJ kept 85% of the initial activity after heating at 100°C for 20 min on the zymogram gel. The Michaelis constant (K m ) and K cat values of AJ towards -casein were 0.38 mM and 19.73 s ¡1 , respectively. AJ cleaved the A -chain of Wbrinogen but did not aVect the B -and -chains, indicating that it is an -Wbrinogenase. The Wbrinolytic activity was inhibited by diisopropyl Xuorophosphate, indicating AJ is a serine protease. Interestingly, AJ was very stable at acidic condition, SDS, and heat (100°C), whereas it was easily degraded at neutral and alkaline conditions. In particular, AJ formed an active homo-dimer in the pH range from 7.0 to 8.0. To our knowledge, a similar combination of acid and heat stability has not yet been reported for other Wbrinolytic enzymes.

Characterization and Production of Thermostable and Acid-stable Extracellular Fibrinolytic Enzymes from Cordyceps militaris

“…Staphylococcus sp. strain AJ (Choi et al, 2009) showed unusual thermostablility at acid condition, whereas it was unstable at neutral and alkaline conditions. AJ kept 85% of the initial activity after heating at 100 o C for 20 min on the zymogram gel.…”

Purification and Characterization of a Novel Fibrinolytic Enzyme from Culture Supernatant of Pleurotus ostreatus

“…However, some fibrinolytic enzymes cannot hydrolyze all three chains of fibrinogen; AJ from Staphylococcus sp. and AMMP from Armillaria mellea can hydrolyze only the α chain [3,14]. The fibrinolytic enzyme from fermented shrimp paste cannot hydrolyze the α chain of fibrinogen [7].…”

Identification of fibrinogen-induced nattokinase WRL101 from Bacillus subtilis WRL101 isolated from Doenjang

“…E-mail: jjsong@kribb.re.kr or choi8850@naver.com. natto (Sumi et al, 1987(Sumi et al, , 1990Fujita et al, 1993;Chang et al, 2000;Kwon et al, 2011;Wang et al, 2011), shiokara (Sumi et al, 1995) in Japanese food, Chungkook-Jang (Kim et al, 1996), Doenjang (Kim and Choi, 2000) and Jeot-Gal (Kim et al, 1997;Choi et al, 2009b) in Korean food, douchi (Peng et al, 2003) in Chinese food and Tempeh (Kim et al, 2006) in Indonesian food. The fibrinolytic enzymes were successively obtained from different microorganisms, the most important among which is the genus Bacillus.…”