Purification and characterization of an extracellular thiol-containing serine proteinase from Thermomyces lanuginosus

1992 ◽  
Vol 70 (2) ◽  
pp. 117-122 ◽  
Author(s):  
Sadiq Hasnain ◽  
Khosrow Adeli ◽  
Andrew C. Storer
Keyword(s):  
Molecular Mass ◽  
Serine Protease ◽  
Filamentous Fungus ◽  
Serine Proteinase ◽  
Thermomyces Lanuginosus ◽  
Kinetic Characterization ◽  
Purification And Characterization ◽  
Inhibitor Specificity ◽  
Protein Substrates

An extracellular protease produced by the filamentous fungus Thermomyces lanuginosus has been purified and characterized. The results indicate that the enzyme, which we have called humicolin, is a thiol-containing serine protease with a molecular mass of 38 000 kilodaltons. Secretion of humicolin, which is glycosylated, is tightly regulated by protein substrates. Kinetic characterization has revealed that humicolin activity is highly dependent upon the deprotonation of a group with a pKa of 6.6 and that the enzyme has a specificity for phenylalanine in the P1 position of the substrate.Key words: thiol-containing serine proteinase, characterization, kinetics, inhibitor, specificity.

scholarly journals Purification and characterization of an extracellular (1 → 6)-β-glucanase from the filamentous fungus Acremonium persicinum

1996 ◽  
Vol 316 (3) ◽  
pp. 841-846 ◽  
Author(s):  
Stuart M. PITSON ◽  
Robert J. SEVIOUR ◽  
Barbara M. McDOUGALL ◽  
Bruce A. STONE ◽  
Maruse SADEK
Keyword(s):  
Molecular Mass ◽  
Filamentous Fungus ◽  
Gel Filtration ◽  
Purification And Characterization ◽  
Eisenia Bicyclis ◽  
Ph Optimum ◽  
Hydrolysis Products ◽  
Sds Page ◽  
Hydrolysis Of

An endo-(1 → 6)-β-glucanase has been isolated from the culture filtrates of the filamentous fungus Acremonium persicinum and purified by (NH4)2SO4 precipitation followed by anion-exchange and gel-filtration chromatography. SDS/PAGE of the purified enzyme gave a single band with an apparent molecular mass of 42.7 kDa. The enzyme is a non-glycosylated, monomeric protein with a pI of 4.9 and pH optimum of 5.0. It hydrolysed (1 → 6)-β-glucans (pustulan and lutean), initially yielding a series of (1 → 6)-β-linked oligoglucosides, consistent with endo-hydrolytic action. Final hydrolysis products from these substrates were gentiobiose and gentiotriose, with all products released as β-anomers, indicating that the enzyme acts with retention of configuration. The purified enzyme also hydrolysed Eisenia bicyclis laminarin, liberating glucose, gentiobiose, and a range of larger oligoglucosides, through the apparent hydrolysis of (1 → 6)-β- and some (1 → 3)-β-linkages in this substrate. Km values for pustulan, lutean and laminarin were 1.28, 1.38, and 1.67 mg/ml respectively. The enzyme was inhibited by N-acetylimidazole, N-bromosuccinimide, dicyclohexylcarbodi-imide, Woodward's Regent K, 2-hydroxy-5-nitrobenzyl bromide, KMnO4 and some metal ions, whereas D-glucono-1,5-lactone and EDTA had no effect.

Purification and Characterization of Serine Proteinase from a Halophilic Bacterium,Filobacillussp. RF2-5

2005 ◽  
Vol 69 (1) ◽  
pp. 38-44 ◽  
Author(s):  
Kazumi HIRAGA ◽  
Yasushi NISHIKATA ◽  
Sirilak NAMWONG ◽  
Somboon TANASUPAWAT ◽  
Katsumi TAKADA ◽  
...  
Keyword(s):  
Serine Proteinase ◽  
Halophilic Bacterium ◽  
Purification And Characterization

Purification and characterization of a new serine protease (EF-SP2) with anti-plant viral activity from Eisenia foetida: Analysis of anti-plant viral activity of EF-SP2

2011 ◽  
Vol 46 (9) ◽  
pp. 1711-1716 ◽  
Author(s):  
Mitsuhiro Ueda ◽  
Takayuki Yamashita ◽  
Masami Nakazawa ◽  
Kazutaka Miyatake ◽  
Satoshi Ohki ◽  
...  
Keyword(s):  
Serine Protease ◽  
Eisenia Foetida ◽  
Purification And Characterization ◽  
Viral Activity
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