[Purification and physico-chemical properties of Streptomyces sp. 1349 collagenase and Streptomyces sp. 1382 keratinase].
The schemes of isolation and purification of collagenolytic enzymes of Streptomyces sp. 1349 and keratinolyte enzymes of Streptomyces sp. 1382, which include fractionation by ammonium sulphate separation on TSK-gels: ion-exchange chromatography on Toyopearl DEAE-650(M) and gel-filtration on Toyopearl HW-50, as well as highly efficient liquid chromatography. The purified enzyme preparations proved to be proteases of serine type (collagenase 2 and keratinases) as well as metalloproteases (collagenases 1 and 3). It has seen established that collagenases are enzymes of broad specificity, which are active in respect of proteins of both globular and fibrillar nature. And vice versa, keratinases are proteolytic enzymes of narrow specificity which hydrolyze native keratin. Molecular masses of purified enzyme preparations, from the data of SDS-PAAG are approximately 30-40 kDa (collagenases 1-3) and about 15-20 kDa (keratinases 1 and 2). It is shown that the charged aminoacid residues (about 85%) prevail in enzyme molecules. The enzymes are distinguished by pH- and thermooptima.
Ivanko
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Institute of Microbiology and Virology, National Academy of Sciences of Ukraine, 154 Zabolotny St., Kyiv, 03143, Ukraine.
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D000645:Ammonium Sulfate; D000837:Anion Exchange Resins; D002853:Chromatography, Liquid; D006863:Hydrogen-Ion Concentration; D003012:Microbial Collagenase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D008970:Molecular Weight; D010447:Peptide Hydrolases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D011108:Polymers; D012117:Resins, Synthetic; D013302:Streptomyces / Q000201:enzymology*; D013379:Substrate Specificity; D013696:Temperature
D000837:Anion Exchange Resins; D011108:Polymers; D012117:Resins, Synthetic; C075368:Toyopearl DEAE 650 M; C058606:Toyopearl 650; D010447:Peptide Hydrolases; C035855:keratinase; D003012:Microbial Collagenase; D000645:Ammonium Sulfate