Identifying and characterizing the most significant beta-glucosidase of the novel species Aspergillus saccharolyticus
(2012) In Canadian Journal of Microbiology 58(9). p.1035-1046- Abstract
- The newly discovered fungal species Aspergillus saccharolyticus was found to produce a culture broth rich in beta-glucosidase activity. In this present work, the main beta-glucosidase of A. saccharolyticus responsible for the efficient hydrolytic activity was identified, isolated, and characterized. Ion exchange chromatography was used to fractionate the culture broth, yielding fractions with high beta-glucosidase activity and only 1 visible band on an SDS-PAGE gel. Mass spectrometry analysis of this band gave peptide matches to beta-glucosidases from aspergilli. Through a polymerase chain reaction approach using degenerate primers and genome walking, a 2919 bp sequence encoding the 860 amino acid BGL1 polypeptide was determined. BGL1 of... (More)
- The newly discovered fungal species Aspergillus saccharolyticus was found to produce a culture broth rich in beta-glucosidase activity. In this present work, the main beta-glucosidase of A. saccharolyticus responsible for the efficient hydrolytic activity was identified, isolated, and characterized. Ion exchange chromatography was used to fractionate the culture broth, yielding fractions with high beta-glucosidase activity and only 1 visible band on an SDS-PAGE gel. Mass spectrometry analysis of this band gave peptide matches to beta-glucosidases from aspergilli. Through a polymerase chain reaction approach using degenerate primers and genome walking, a 2919 bp sequence encoding the 860 amino acid BGL1 polypeptide was determined. BGL1 of A. saccharolyticus has 91% and 82% identity with BGL1 from Aspergillus aculeatus and BGL1 from Aspergillus niger, respectively, both belonging to Glycoside Hydrolase family 3. Homology modeling studies suggested beta-glucosidase activity with preserved retaining mechanism and a wider catalytic pocket compared with other beta-glucosidases. The bgl1 gene was heterologously expressed in Trichoderma reesei QM6a, purified, and characterized by enzyme kinetics studies. The enzyme can hydrolyze cellobiose, p-nitrophenyl-beta-D-glucoside, and cellodextrins. The enzyme showed good thermostability, was stable at 50 degrees C, and at 60 degrees C it had a half-life of approximately 6 h. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/3146811
- author
- Sorensen, Annette ; Ahring, Birgitte K. ; Luebeck, Mette ; Ubhayasekera, Wimal LU ; Bruno, Kenneth S. ; Culley, David E. and Luebeck, Peter S.
- organization
- publishing date
- 2012
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- beta-glucosidase, Aspergillus saccharolyticus, enzyme characterization, thermostability, biomass hydrolysis
- in
- Canadian Journal of Microbiology
- volume
- 58
- issue
- 9
- pages
- 1035 - 1046
- publisher
- National Research Council Canada
- external identifiers
-
- wos:000308368400002
- scopus:84875806354
- ISSN
- 0008-4166
- DOI
- 10.1139/W2012-076
- language
- English
- LU publication?
- yes
- id
- 91e8e1b5-a2ac-4e60-ac0d-b873888baad0 (old id 3146811)
- date added to LUP
- 2016-04-01 13:18:15
- date last changed
- 2022-03-06 05:09:36
@article{91e8e1b5-a2ac-4e60-ac0d-b873888baad0,
abstract = {{The newly discovered fungal species Aspergillus saccharolyticus was found to produce a culture broth rich in beta-glucosidase activity. In this present work, the main beta-glucosidase of A. saccharolyticus responsible for the efficient hydrolytic activity was identified, isolated, and characterized. Ion exchange chromatography was used to fractionate the culture broth, yielding fractions with high beta-glucosidase activity and only 1 visible band on an SDS-PAGE gel. Mass spectrometry analysis of this band gave peptide matches to beta-glucosidases from aspergilli. Through a polymerase chain reaction approach using degenerate primers and genome walking, a 2919 bp sequence encoding the 860 amino acid BGL1 polypeptide was determined. BGL1 of A. saccharolyticus has 91% and 82% identity with BGL1 from Aspergillus aculeatus and BGL1 from Aspergillus niger, respectively, both belonging to Glycoside Hydrolase family 3. Homology modeling studies suggested beta-glucosidase activity with preserved retaining mechanism and a wider catalytic pocket compared with other beta-glucosidases. The bgl1 gene was heterologously expressed in Trichoderma reesei QM6a, purified, and characterized by enzyme kinetics studies. The enzyme can hydrolyze cellobiose, p-nitrophenyl-beta-D-glucoside, and cellodextrins. The enzyme showed good thermostability, was stable at 50 degrees C, and at 60 degrees C it had a half-life of approximately 6 h.}},
author = {{Sorensen, Annette and Ahring, Birgitte K. and Luebeck, Mette and Ubhayasekera, Wimal and Bruno, Kenneth S. and Culley, David E. and Luebeck, Peter S.}},
issn = {{0008-4166}},
keywords = {{beta-glucosidase; Aspergillus saccharolyticus; enzyme characterization; thermostability; biomass hydrolysis}},
language = {{eng}},
number = {{9}},
pages = {{1035--1046}},
publisher = {{National Research Council Canada}},
series = {{Canadian Journal of Microbiology}},
title = {{Identifying and characterizing the most significant beta-glucosidase of the novel species Aspergillus saccharolyticus}},
url = {{http://dx.doi.org/10.1139/W2012-076}},
doi = {{10.1139/W2012-076}},
volume = {{58}},
year = {{2012}},
}