Purification and characterization of an extracellular protease from Penicillium chrysogenum Pg222 active against meat proteins

被引：41

作者：

Benito, MJ ^{[1
]}

Rodríguez, M ^{[1
]}

Núñez, F ^{[1
]}

Asensio, MA ^{[1
]}

Bermúdez, ME ^{[1
]}

Córdoba, JJ ^{[1
]}

机构：

[1] Univ Extremadura, Fac Vet, Caceres 10071, Spain

来源：

APPLIED AND ENVIRONMENTAL MICROBIOLOGY | 2002年 / 68卷 / 07期

关键词：

D O I：

10.1128/AEM.68.7.3532-3536.2002

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

An extracellular protease from Penicillium chrysogenum (Pg222) isolated from dry-cured ham has been purified. The purification procedure involved several steps: ammonium sulfate precipitation, ion-exchange chromatography, filtration, and separation by high-performance liquid chromatography. Based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis and gel filtration, the purified fraction showed a molecular mass of about 35 kDa. The hydrolytic properties of the purified enzyme (EPg222) on extracted pork myofibrillar proteins under several conditions were evaluated by SDS-PAGE. EPg222 showed activity in the range of 10 to 60degreesC in temperature, 0 to 3 M NaCl, and pH 5 to 7, with maximum activity at pH 6, 45degreesC, and 0.25 M NaCL Under these conditions the enzyme was most active against tropomyosin, actin, and myosin. EPg222 showed collagenolytic activity but did not hydrolyze myoglobin. EPg222 showed higher activity than other proteolytic enzymes like papain, trypsin, and Aspergillus oryzae protease. The N-terminal amino acid sequence was determined and was found to be Glu-Asn-Pro-Leu-Gln-Pro-Asn-Ala-Pro-Ser-Trp. This partial amino acid sequence revealed a 55% homology with serine proteases from Penicillium citrinum. The activity of this novel protease may be of interest in ripening and generating the flavor of dry-cured meat products.

引用

页码：3532 / 3536

页数：5

共 31 条

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