Purification and characterization of a new thermophilic collagenase from Nocardiopsis dassonvillei NRC2aza and its application in wound healing.
Int J Biol Macromol
; 116: 801-810, 2018 Sep.
Article
en En
| MEDLINE
| ID: mdl-29746969
ABSTRACT
A thermostable metallo-collagenase enzyme (150â¯kDa), recently identified in a newly isolated actinomycestes strain (Nocardiopsis dassonvillei NRC2aza), has been purified from natural source, characterized to have application in wound healing. A simple 3 step purification procedure gave an increase of purity by 6.23 fold with a specific activity of 387.2â¯Uâ¯mg-1. The enzyme activity showed stability across a range of pH (7.0-8.5) and temperature (40-55⯰C) with optima at pHâ¯8.0 and 60⯰C, respectively. Activators include Mg+2, Ca+2, Zn+2, Na+, K+ and Ba+2, while Mn+2, Co+2, Ni+2and Ag+ ions gave partial inhibition. Full inhibition was given by other tested ions and metalloproteinase inhibitors. Broad substrate specificity was demonstrated including activity against a native collagen. The Km and Vmax of the enzyme using azocollagen were 5.5â¯mg/ml and 1280â¯U, respectively. The purified collagenase enhanced wound closure in vitro and in vivo and the repair process was dose dependent. Topical application of the purified collagenase (either of 25 or 50â¯U) to cutaneous wounds significantly accelerated the rate of wound healing and the formation of granulation tissue. Hence, the purified collagenase has a great potential as a therapeutic agent in wound care and collagen related diseases.
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Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Proteínas Bacterianas
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Cicatrización de Heridas
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Heridas y Lesiones
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Actinobacteria
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Colagenasas
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Fibroblastos
Límite:
Animals
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Humans
Idioma:
En
Revista:
Int J Biol Macromol
Año:
2018
Tipo del documento:
Article
País de afiliación:
Egipto