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BioMed Research International
Volume 2013 (2013), Article ID 175012, 14 pages
Production and Characterization of Keratinolytic Protease from New Wool-Degrading Bacillus Species Isolated from Egyptian Ecosystem
1Protein Research Department, Genetic Engineering and Biotechnology Research Institute, City of Scientific Research and Technological Applications, New Borg Al-Arab, P.O. Box. 21934, Alexandria, Egypt
2Botany and Microbiology Department, Faculty of Science (Boys), Al-Azhar University, Cairo, Egypt
3King Abdulaziz City for Science and Technology, Riyadh, Saudi Arabia
Received 2 December 2012; Revised 7 March 2013; Accepted 28 March 2013
Academic Editor: Periasamy Anbu
Copyright © 2013 Mohamed A. Hassan et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Scanning electron microscope of Bacillus amyloliquefaciens MA20 and Bacillus subtilis MA21
The supplementary data include the characterization of Bacillus amyloliquefaciens MA20 and Bacillus subtilis MA21 using scanning electron microscope as in figure 1 which indicates to the bacterial size of the 2 Bacillus strains which measured by slime view program software.
DNA isolation of Bacillus amyloliquefaciens MA20 and Bacillus subtilis MA21
The genomic DNA of B. amyloliquefaciens MA20 and B. subtilis MA21 were isolated and purified. The DNA was analyzed by gel electrophoresis using 1% (w/v) agarose gel containing ethidium bromide soluble in TBE buffer were used. The DNA ladder was loaded in gel for detecting the DNA. The DNA was investigated under UV light using gel documentation system and photographed as in figure 2.
16S ribosomal RNA (rRNA)
The amplified 16S rRNA gene from the DNA of B. amyloliquefaciens MA20 and B. subtilis MA21 were determined using 2% agarose gel. The PCR products were about 380 bp in compare to DNA ladder (Gene ruler 50 bp – 1031 bp DNA ladder) (Figure 3).
Production of keratinolytic proteases
The keratinolytic proteases were produced from B. amyloliquefaciens MA20 and B. subtilis MA21 using medium which containing wool as sole carbon and nitrogen source. The wool was degraded to powder after incubation for 5 days (figure 4).