NISCAIR Online Periodicals Repository

NISCAIR ONLINE PERIODICALS REPOSITORY (NOPR)  >
NISCAIR PUBLICATIONS >
Research Journals >
Indian Journal of Biotechnology (IJBT) >
IJBT Vol.07 [2008] >
IJBT Vol.07(3) [July 2008] >


Title: Extracellular collagenase from Rhizoctonia solani: Production, purification and characterization
Authors: Hamdy, Hossam S
Keywords: Collagenase
Production
Purification
Characterization
Rhizoctonia solani
Issue Date: Jul-2008
Publisher: CSIR
Abstract: Potentiality of R. solani grown on Sabouraud-glucose-collagen medium to produce glycosylated metallo-proteinase with collagenolytic activity was optimized and maximum production (212.33 U/mL) was recorded after 108 h of submerged incubation (175 rpm) at pH 5.5 and 30ºC of temperature. Two-step column chromatography technique on DEAE-cellulose and Sephadex G₁₅₀ was adopted to purify the partially purified enzyme produced by ammonium sulfate (40%, w/v) precipitation. Yield of purification was 60.49% of the original activity with specific activity of 18064.7 × 10³ U/mg protein and 18.72-folds of purification. The purified enzyme showed maximum activity at 40ºC and pH 5 ,which was stimulated by ions of Ca, Co, Cu, K, Mg, Na or Zn and inhibited by ions of Fe and Hg. Metal composition of the purified enzyme revealed that it contains Ca²⁺ and Zn²⁺. Tm (midpoint of thermal inactivation) was recorded at 65ºC and 55ºC after 1 and 6 h of exposure, respectively and T₁/₂ was found to be 7 and 5 weeks at ‒15ºC and 4ºC, respectively. Molecular mass, Kmg/mL, Vmax of the enzyme were found to be 66 ± 4 kDa, 0.033 kDa and 0.28 U/mg min⁻¹, respectively. Activities toward gelatin and casein were also detected.
Page(s): 333-340
ISSN: 0972-5849
Source: IJBT Vol.07(3) [July 2008]

Files in This Item:

File Description SizeFormat
IJBT 7(3) 333-340.pdf515.29 kBAdobe PDFView/Open
 Current Page Visits: 714 
Recommend this item

 

National Knowledge Resources Consortium |  NISCAIR Website |  Contact us |  Feedback

Disclaimer: NISCAIR assumes no responsibility for the statements and opinions advanced by contributors. The editorial staff in its work of examining papers received for publication is helped, in an honorary capacity, by many distinguished engineers and scientists.

CC License Except where otherwise noted, the Articles on this site are licensed under Creative Commons License: CC Attribution-Noncommercial-No Derivative Works 2.5 India

Copyright © 2012 The Council of Scientific and Industrial Research, New Delhi. All rights reserved.

Powered by DSpace Copyright © 2002-2007 MIT and Hewlett-Packard | Compliant to OAI-PMH V 2.0

Home Page Total Visits: 511124 since 06-Feb-2009  Last updated on 11-Apr-2014Webmaster: nopr@niscair.res.in