Biochemical and Molecular Characterization of a Thermostable Alkaline Metallo-Keratinase from <i>Bacillus</i> sp. Nnolim-K1
Keratinases are considerably gaining momentum in green technology because of their endowed robustness and multifaceted application potentials, such as keratinous agro-wastes valorization. Therefore, the production of novel keratinases from relatively nonpathogenic bacteria grown in agro-wastes formu...
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MDPI AG
2020-08-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/8/9/1304 |
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| author | Nonso E. Nnolim Lindelwa Mpaka Anthony I. Okoh Uchechukwu U. Nwodo |
| author_facet | Nonso E. Nnolim Lindelwa Mpaka Anthony I. Okoh Uchechukwu U. Nwodo |
| author_sort | Nonso E. Nnolim |
| collection | DOAJ |
| description | Keratinases are considerably gaining momentum in green technology because of their endowed robustness and multifaceted application potentials, such as keratinous agro-wastes valorization. Therefore, the production of novel keratinases from relatively nonpathogenic bacteria grown in agro-wastes formulated medium is cost-effective, and also imperative for the sustainability of thriving bioeconomy. In this study, we optimized keratinase production by <i>Bacillus</i> sp. Nnolim-K1 grown in chicken feather formulated medium. The produced keratinase (KerBNK1) was biochemically characterized and also, the keratinase-encoding gene (<i>kerBNK1</i>) was amplified and sequenced. The optimal physicochemical conditions for extracellular keratinase production determined were 0.8% (<i>w/v</i>) xylose, 1.0% (<i>w/v</i>) feather, and 3.0% (<i>v/v</i>) inoculum size, pH 5.0, temperature (25 °C) and agitation speed (150 rpm). The maximum keratinase activity of 1943.43 ± 0.0 U/mL was achieved after 120 h of fermentation. KerBNK1 was optimally active at pH and temperature of 8.0 and 60 °C, respectively; with remarkable pH and thermal stability. KerBNK1 activity was inhibited by ethylenediamine tetra-acetic acid and 1,10-phenanthroline, suggesting a metallo-keratinase. The amplified <i>kerBNK1</i> showed a band size of 1104 bp and the nucleotide sequence was submitted to the GenBank with accession number MT268133. <i>Bacillus</i> sp. Nnolim-K1 and the keratinase displayed potentials that demand industrial and biotechnological exploitations. |
| first_indexed | 2024-03-10T16:46:39Z |
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| language | English |
| last_indexed | 2024-03-10T16:46:39Z |
| publishDate | 2020-08-01 |
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| series | Microorganisms |
| spelling | doaj.art-292f482092ec4523a1f9a6667d65d0382023-11-20T11:30:57ZengMDPI AGMicroorganisms2076-26072020-08-0189130410.3390/microorganisms8091304Biochemical and Molecular Characterization of a Thermostable Alkaline Metallo-Keratinase from <i>Bacillus</i> sp. Nnolim-K1Nonso E. Nnolim0Lindelwa Mpaka1Anthony I. Okoh2Uchechukwu U. Nwodo3SAMRC Microbial Water Quality Monitoring Centre, University of Fort Hare, Alice 5700, South AfricaSAMRC Microbial Water Quality Monitoring Centre, University of Fort Hare, Alice 5700, South AfricaSAMRC Microbial Water Quality Monitoring Centre, University of Fort Hare, Alice 5700, South AfricaSAMRC Microbial Water Quality Monitoring Centre, University of Fort Hare, Alice 5700, South AfricaKeratinases are considerably gaining momentum in green technology because of their endowed robustness and multifaceted application potentials, such as keratinous agro-wastes valorization. Therefore, the production of novel keratinases from relatively nonpathogenic bacteria grown in agro-wastes formulated medium is cost-effective, and also imperative for the sustainability of thriving bioeconomy. In this study, we optimized keratinase production by <i>Bacillus</i> sp. Nnolim-K1 grown in chicken feather formulated medium. The produced keratinase (KerBNK1) was biochemically characterized and also, the keratinase-encoding gene (<i>kerBNK1</i>) was amplified and sequenced. The optimal physicochemical conditions for extracellular keratinase production determined were 0.8% (<i>w/v</i>) xylose, 1.0% (<i>w/v</i>) feather, and 3.0% (<i>v/v</i>) inoculum size, pH 5.0, temperature (25 °C) and agitation speed (150 rpm). The maximum keratinase activity of 1943.43 ± 0.0 U/mL was achieved after 120 h of fermentation. KerBNK1 was optimally active at pH and temperature of 8.0 and 60 °C, respectively; with remarkable pH and thermal stability. KerBNK1 activity was inhibited by ethylenediamine tetra-acetic acid and 1,10-phenanthroline, suggesting a metallo-keratinase. The amplified <i>kerBNK1</i> showed a band size of 1104 bp and the nucleotide sequence was submitted to the GenBank with accession number MT268133. <i>Bacillus</i> sp. Nnolim-K1 and the keratinase displayed potentials that demand industrial and biotechnological exploitations.https://www.mdpi.com/2076-2607/8/9/1304Keratinasechicken featherbiodegradationthiol groupvalorizationbioeconomy |
| spellingShingle | Nonso E. Nnolim Lindelwa Mpaka Anthony I. Okoh Uchechukwu U. Nwodo Biochemical and Molecular Characterization of a Thermostable Alkaline Metallo-Keratinase from <i>Bacillus</i> sp. Nnolim-K1 Microorganisms Keratinase chicken feather biodegradation thiol group valorization bioeconomy |
| title | Biochemical and Molecular Characterization of a Thermostable Alkaline Metallo-Keratinase from <i>Bacillus</i> sp. Nnolim-K1 |
| title_full | Biochemical and Molecular Characterization of a Thermostable Alkaline Metallo-Keratinase from <i>Bacillus</i> sp. Nnolim-K1 |
| title_fullStr | Biochemical and Molecular Characterization of a Thermostable Alkaline Metallo-Keratinase from <i>Bacillus</i> sp. Nnolim-K1 |
| title_full_unstemmed | Biochemical and Molecular Characterization of a Thermostable Alkaline Metallo-Keratinase from <i>Bacillus</i> sp. Nnolim-K1 |
| title_short | Biochemical and Molecular Characterization of a Thermostable Alkaline Metallo-Keratinase from <i>Bacillus</i> sp. Nnolim-K1 |
| title_sort | biochemical and molecular characterization of a thermostable alkaline metallo keratinase from i bacillus i sp nnolim k1 |
| topic | Keratinase chicken feather biodegradation thiol group valorization bioeconomy |
| url | https://www.mdpi.com/2076-2607/8/9/1304 |
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