Production and Characterization of Keratinolytic Protease from New Wool-Degrading Bacillus Species Isolated from Egyptian Ecosystem

Novel keratin-degrading bacteria were isolated from sand soil samples collected from Minia Governorate, Egypt. In this study, the isolates were identified as Bacillus amyloliquefaciens MA20 and Bacillus subtilis MA21 based on morphological and biochemical characteristics as well as 16S rRNA gene seq...

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Main Authors: Mohamed A. Hassan, Bakry M. Haroun, Amro A. Amara, Ehab A. Serour
Format: Article
Language:English
Published: Hindawi Limited 2013-01-01
Series:BioMed Research International
Online Access:http://dx.doi.org/10.1155/2013/175012
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spelling doaj-63fce8990975468abcf8c3969b7d32732020-11-24T23:41:35ZengHindawi LimitedBioMed Research International2314-61332314-61412013-01-01201310.1155/2013/175012175012Production and Characterization of Keratinolytic Protease from New Wool-Degrading Bacillus Species Isolated from Egyptian EcosystemMohamed A. Hassan0Bakry M. Haroun1Amro A. Amara2Ehab A. Serour3Protein Research Department, Genetic Engineering and Biotechnology Research Institute, City of Scientific Research and Technological Applications, New Borg Al-Arab, P.O. Box. 21934, Alexandria, EgyptBotany and Microbiology Department, Faculty of Science (Boys), Al-Azhar University, Cairo, EgyptProtein Research Department, Genetic Engineering and Biotechnology Research Institute, City of Scientific Research and Technological Applications, New Borg Al-Arab, P.O. Box. 21934, Alexandria, EgyptProtein Research Department, Genetic Engineering and Biotechnology Research Institute, City of Scientific Research and Technological Applications, New Borg Al-Arab, P.O. Box. 21934, Alexandria, EgyptNovel keratin-degrading bacteria were isolated from sand soil samples collected from Minia Governorate, Egypt. In this study, the isolates were identified as Bacillus amyloliquefaciens MA20 and Bacillus subtilis MA21 based on morphological and biochemical characteristics as well as 16S rRNA gene sequencing. B. amyloliquefaciens MA20 and B. subtilis MA21 produced alkaline keratinolytic serine protease when cultivated in mineral medium containing 1% of wool straight off sheep as sole carbon and nitrogen source. The two strains were observed to degrade wool completely to powder at pH 7 and 37°C within 5 days. Under these conditions the maximum activity of proteases produced by B. amyloliquefaciens MA20 and B. subtilis MA21 was 922 and 814 U/ml, respectively. The proteases exhibited optimum temperature and pH at 60°C and 9, respectively. However, the keratinolytic proteases were stable in broad range of temperature and pH values towards casein Hammerstein. Furthermore the protease inhibitor studies indicated that the produced proteases belong to serine protease because of their sensitivity to PMSF while they were inhibited partially in presence of EDTA. The two proteases are stable in most of the used organic solvents and enhanced by metals suggesting their potential use in biotechnological applications such as wool industry.http://dx.doi.org/10.1155/2013/175012
collection DOAJ
language English
format Article
sources DOAJ
author Mohamed A. Hassan
Bakry M. Haroun
Amro A. Amara
Ehab A. Serour
spellingShingle Mohamed A. Hassan
Bakry M. Haroun
Amro A. Amara
Ehab A. Serour
Production and Characterization of Keratinolytic Protease from New Wool-Degrading Bacillus Species Isolated from Egyptian Ecosystem
BioMed Research International
author_facet Mohamed A. Hassan
Bakry M. Haroun
Amro A. Amara
Ehab A. Serour
author_sort Mohamed A. Hassan
title Production and Characterization of Keratinolytic Protease from New Wool-Degrading Bacillus Species Isolated from Egyptian Ecosystem
title_short Production and Characterization of Keratinolytic Protease from New Wool-Degrading Bacillus Species Isolated from Egyptian Ecosystem
title_full Production and Characterization of Keratinolytic Protease from New Wool-Degrading Bacillus Species Isolated from Egyptian Ecosystem
title_fullStr Production and Characterization of Keratinolytic Protease from New Wool-Degrading Bacillus Species Isolated from Egyptian Ecosystem
title_full_unstemmed Production and Characterization of Keratinolytic Protease from New Wool-Degrading Bacillus Species Isolated from Egyptian Ecosystem
title_sort production and characterization of keratinolytic protease from new wool-degrading bacillus species isolated from egyptian ecosystem
publisher Hindawi Limited
series BioMed Research International
issn 2314-6133
2314-6141
publishDate 2013-01-01
description Novel keratin-degrading bacteria were isolated from sand soil samples collected from Minia Governorate, Egypt. In this study, the isolates were identified as Bacillus amyloliquefaciens MA20 and Bacillus subtilis MA21 based on morphological and biochemical characteristics as well as 16S rRNA gene sequencing. B. amyloliquefaciens MA20 and B. subtilis MA21 produced alkaline keratinolytic serine protease when cultivated in mineral medium containing 1% of wool straight off sheep as sole carbon and nitrogen source. The two strains were observed to degrade wool completely to powder at pH 7 and 37°C within 5 days. Under these conditions the maximum activity of proteases produced by B. amyloliquefaciens MA20 and B. subtilis MA21 was 922 and 814 U/ml, respectively. The proteases exhibited optimum temperature and pH at 60°C and 9, respectively. However, the keratinolytic proteases were stable in broad range of temperature and pH values towards casein Hammerstein. Furthermore the protease inhibitor studies indicated that the produced proteases belong to serine protease because of their sensitivity to PMSF while they were inhibited partially in presence of EDTA. The two proteases are stable in most of the used organic solvents and enhanced by metals suggesting their potential use in biotechnological applications such as wool industry.
url http://dx.doi.org/10.1155/2013/175012
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