Alkaline Serine Protease Is an Exotoxin of Vibrio alginolyticus in Kuruma Prawn, Penaeus japonicus

1997 ◽  
Vol 34 (2) ◽  
pp. 110-117 ◽  
Author(s):  
Kuo-Kau Lee ◽  
Shu-Ru Yu ◽  
Ping-Chung Liu
Keyword(s):  
Serine Protease ◽  
Vibrio Alginolyticus ◽  
Alkaline Serine Protease ◽  
Penaeus Japonicus

Lethal Attribute of Serine Protease Secreted by Vibrio alginolyticus Strains in Kuruma Prawn Penaeus japonicus

2000 ◽  
Vol 55 (1-2) ◽  
pp. 94-99 ◽  
Author(s):  
Ferng-Ruey Chen ◽  
Ping-Chung Liu ◽  
Kuo-Kau Lee
Keyword(s):  
Serine Protease ◽  
Lethal Factor ◽  
Rabbit Antiserum ◽  
Vibrio Alginolyticus ◽  
Protein G ◽  
Haliotis Diversicolor Supertexta ◽  
Penaeus Japonicus ◽  
Pathogenic Vibrio ◽  
Precipitation Band ◽  
Diversicolor Supertexta

Abstract Toxicity of the extracellular products (ECP) and the lethal attribute of serine protease secreted by five pathogenic Vibrio alginolyticus strains from various sources in kuruma prawn Penaeus japonicus were studied. The ECPs of organisms originally isolated from diseased kuruma prawn or small abalone Haliotis diversicolor supertexta were more lethal (LD50 value of 0.48 or 0.41 μg protein/g prawn) than those from diseased tiger prawn P.monodon, yellowfin porgy Acanthopagrus latus or horse mackerel (LD50 value of 0.98 -1.17 μg protein/g prawn). All the ECPs manifested strong, weak and no activities against gelatin, sheep erythrocytes and chitin, respectively. In immunodiffusion tests using rabbit antiserum to a purified 33 kDa serine protease of strain Swy against ECP of each tested strain produced one single precipitation band in each treatment. Furtherm ore, the serine protease was suggested to be the dominant protease secreted by V. alginolyticus strains tested since the majority of enzymatic activity of the respective ECP was inhibited by phenylmethanesulfonyl fluoride (PMSF). A higher inhibition of serine protease activity by PMSF resulted in lower mortality rate of the ECPs injected into the prawns suggesting that the protease is one of the major lethal factor(s) secreted by Valginolyticus.

Molecular Cloning, Nucleotide Sequence, and Expression of the Structural Gene for Alkaline Serine Protease from AlkaliphilicBacillussp. 221

1992 ◽  
Vol 56 (9) ◽  
pp. 1455-1460 ◽  
Author(s):  
Hideto Takami ◽  
Tetsuo Kobayashi ◽  
Masato Kobayashi ◽  
Mami Yamamoto ◽  
Satoshi Nakamura ◽  
...  
Keyword(s):  
Nucleotide Sequence ◽  
Molecular Cloning ◽  
Serine Protease ◽  
Structural Gene ◽  
Alkaline Serine Protease

Production, purification and characterization of thermomycolase, the extracellular serine protease of the thermophilic fungus Malbranchea pulchella var. sulfurea

1976 ◽  
Vol 22 (2) ◽  
pp. 165-176 ◽  
Author(s):  
Poh Seng Ong ◽  
G. Maurice Gaucher
Keyword(s):  
Serine Protease ◽  
Thermophilic Fungus ◽  
Amino Acid Residues ◽  
Submerged Cultures ◽  
Ph Optimum ◽  
Alkaline Serine Protease ◽  
Associated Production ◽  
Specificity Study ◽  
Malbranchea Pulchella ◽  
The Stability

The thermophilic fungus Malbranchea pulchella produces a single extracellular, alkaline, serine protease when grown at 45 °C, on 2% casein as sole carbon source. The growth-associated production of protease in submerged cultures was inhibited by addition of glucose, amino acids, or yeast extract. A simple four-step purification which yields homogeneous protease in 78% yield is described. The protease has an isoelectric point of 6.0, a pH optimum of 8.5, and is completely inhibited by serine protease inhibitors. A specificity study with small synthetic ester substrates indicated that the protease preferentially hydrolyzed bonds situated on the carboxyl side of aromatic or apolar amino acid residues which are not β-branched, positively charged or of the D configuration. Peptidase substrates and others such as N-acetyl-L-tyrosine-ethyl ester were not hydrolyzed. The protease was stable over a broad range of pH (6.5–9.5 at 30 °C, 20 h), and was particularly thermostable (t1/2 = 110 min at 73 °C, pH 7.4) in the presence of Ca2+ (10 mM). Macromolecules and Ca2+ also provide protection against the significant autolysis which occurs at pure protease concentrations greater than 0.01 mg/ml, as well as against surface denaturation which is enhanced by the presence of a silicone antifoam agent. Hence the stability of protease in submerged cultures is rationalized.

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