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Xylanases from Cryptococcus flavus isolate I-11: Enzymatic profile, isolation and heterologous expression of CfXYN1 in Saccharomyces cerevisiae

Skorupa Parachin, Nadia LU ; Siqueira, Saulo; de Faria, Fabricia Paula; Goncalves Torres, Fernando Araripe and Pepe de Moraes, Lidia Maria (2009) In Journal of Molecular Catalysis B: Enzymatic 59(1-3). p.52-57
Abstract
The aim of this study was to characterize the xylanolytic activity of Cryptococcus flavus isolate I-11. This microorganism was isolated from the Brazilian Cerrado, and enzyme plate assays showed that it also produces amylase and CMCase activity. The xylanolytic production of C. flavus isolate I-11 was improved by using a suitable combination of the carbon and nitrogen sources, reaching 130 U/mL A zymogram assay was performed showing three xylanase activity bands. The cDNA of one xylanase gene, CfXYN1, was obtained and preliminary expression analysis was performed on RNA samples collected after yeast growth on different carbon sources. This indicated that the CfXYN1 gene is transcribed in the presence of xylose, sugar cane bagasse and... (More)
The aim of this study was to characterize the xylanolytic activity of Cryptococcus flavus isolate I-11. This microorganism was isolated from the Brazilian Cerrado, and enzyme plate assays showed that it also produces amylase and CMCase activity. The xylanolytic production of C. flavus isolate I-11 was improved by using a suitable combination of the carbon and nitrogen sources, reaching 130 U/mL A zymogram assay was performed showing three xylanase activity bands. The cDNA of one xylanase gene, CfXYN1, was obtained and preliminary expression analysis was performed on RNA samples collected after yeast growth on different carbon sources. This indicated that the CfXYN1 gene is transcribed in the presence of xylose, sugar cane bagasse and carboxymethyl cellulose, but not in the presence of glucose, as carbon source. The cDNA of CfXYN1 was cloned and expressed in Saccharomyces cerevisiae. The recombinant enzyme was partially characterized and showed an optimum at a pH of 3.0 and temperature of 50 degrees C. The recombinant enzyme retained 70% of its initial activity after pre-incubation for 30 min at the optimum pH and temperature. Computational analysis predicted a molecular weight of 21.2 kDa, and an isoelectric point of 7.02. The Cfxyn1p has 209 amino acids, including a signal peptide consisting of 16 amino acids. (C) 2009 Elsevier B.V. All rights reserved. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Enzymatic profile, Xylanase, Cryptococcus flavus
in
Journal of Molecular Catalysis B: Enzymatic
volume
59
issue
1-3
pages
52 - 57
publisher
Elsevier
external identifiers
  • wos:000266128100006
  • scopus:64649091369
ISSN
1873-3158
DOI
10.1016/j.molcatb.2008.12.018
language
English
LU publication?
yes
id
26964f45-6f29-4079-800b-1829e1e8864f (old id 1425934)
date added to LUP
2009-06-26 14:42:39
date last changed
2017-12-10 03:51:24
@article{26964f45-6f29-4079-800b-1829e1e8864f,
  abstract     = {The aim of this study was to characterize the xylanolytic activity of Cryptococcus flavus isolate I-11. This microorganism was isolated from the Brazilian Cerrado, and enzyme plate assays showed that it also produces amylase and CMCase activity. The xylanolytic production of C. flavus isolate I-11 was improved by using a suitable combination of the carbon and nitrogen sources, reaching 130 U/mL A zymogram assay was performed showing three xylanase activity bands. The cDNA of one xylanase gene, CfXYN1, was obtained and preliminary expression analysis was performed on RNA samples collected after yeast growth on different carbon sources. This indicated that the CfXYN1 gene is transcribed in the presence of xylose, sugar cane bagasse and carboxymethyl cellulose, but not in the presence of glucose, as carbon source. The cDNA of CfXYN1 was cloned and expressed in Saccharomyces cerevisiae. The recombinant enzyme was partially characterized and showed an optimum at a pH of 3.0 and temperature of 50 degrees C. The recombinant enzyme retained 70% of its initial activity after pre-incubation for 30 min at the optimum pH and temperature. Computational analysis predicted a molecular weight of 21.2 kDa, and an isoelectric point of 7.02. The Cfxyn1p has 209 amino acids, including a signal peptide consisting of 16 amino acids. (C) 2009 Elsevier B.V. All rights reserved.},
  author       = {Skorupa Parachin, Nadia and Siqueira, Saulo and de Faria, Fabricia Paula and Goncalves Torres, Fernando Araripe and Pepe de Moraes, Lidia Maria},
  issn         = {1873-3158},
  keyword      = {Enzymatic profile,Xylanase,Cryptococcus flavus},
  language     = {eng},
  number       = {1-3},
  pages        = {52--57},
  publisher    = {Elsevier},
  series       = {Journal of Molecular Catalysis B: Enzymatic},
  title        = {Xylanases from Cryptococcus flavus isolate I-11: Enzymatic profile, isolation and heterologous expression of CfXYN1 in Saccharomyces cerevisiae},
  url          = {http://dx.doi.org/10.1016/j.molcatb.2008.12.018},
  volume       = {59},
  year         = {2009},
}