A novel chitinase gene was cloned from Streptomyces albolongus ATCC 27414, and expressed successfully in Escherichia coli BL21. The recombinant enzyme (SaChiA4) belongs to glycoside hydrolases (GH) family 18 and consists of a catalytic domain and a chitin binding domain (CBD) in its C-terminus. SaChiA4 was purified homogeneously (specific activity of 66.2 U/mg with colloidal chitin as substrate), and showed a molecular mass of approximately 47 kDa. SaChiA4 showed its optimal activity at pH 5.0 and 55 °C and exhibited remarkable pH and temperature stability. SaChiA4 has been proved to have a higher specificity toward glycosides containing acetyl groups and hydrolyzes the substrates in a non-processive manner with higher ability to produce (GlcNAc)2 and GlcNAc. The results indicated that SaChiA4 is a novel endo-type chitinase, which has potential applications in the treatment of chitin wastes and the production of (GlcNAc)2
Cloning, characterization and substrate degradation mode of a novel chitinase from Streptomyces albolongus ATCC 27414
Secundo F;
2018
Abstract
A novel chitinase gene was cloned from Streptomyces albolongus ATCC 27414, and expressed successfully in Escherichia coli BL21. The recombinant enzyme (SaChiA4) belongs to glycoside hydrolases (GH) family 18 and consists of a catalytic domain and a chitin binding domain (CBD) in its C-terminus. SaChiA4 was purified homogeneously (specific activity of 66.2 U/mg with colloidal chitin as substrate), and showed a molecular mass of approximately 47 kDa. SaChiA4 showed its optimal activity at pH 5.0 and 55 °C and exhibited remarkable pH and temperature stability. SaChiA4 has been proved to have a higher specificity toward glycosides containing acetyl groups and hydrolyzes the substrates in a non-processive manner with higher ability to produce (GlcNAc)2 and GlcNAc. The results indicated that SaChiA4 is a novel endo-type chitinase, which has potential applications in the treatment of chitin wastes and the production of (GlcNAc)2File | Dimensione | Formato | |
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Descrizione: Cloning, characterization and substrate degradation mode of a novel chitinase from Streptomyces albolongus
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