Production of Extracellular Alkaline Serine Protease from Pediococcus acidilactici NCDC 252: Isolation, Purification, Physicochemical and Catalytic Characterization

2020 ◽  
Author(s):  
Poonam Bansal ◽  
Raman Kumar ◽  
Jasbir Singh ◽  
Suman Dhanda
Keyword(s):  
Serine Protease ◽  
Pediococcus Acidilactici ◽  
Alkaline Serine Protease ◽  
Catalytic Characterization

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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