Purification and properties of a novel fungal alkaline keratinase from Cunninghamella echinulata

More, Sunil S.; Divyalakshmi, S S; Prakash, Sahana N; Umashankar, Swathi U; Vishwakarma, Jyothi V

doi:10.5505/tjb.2013.37928

Cited by 15 publications

(7 citation statements)

References 35 publications

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“…However, few enzymes are active outside this range, and even maximum enzyme production of these enzymes is achieved under highly alkaline or acidic conditions [32,33]. In line with the present study, More et al, isolated the keratinase from Cunninghamella echinulata, which showed the most activity at the pH values of 4.5 and 10.5, citing the possibility of the formation of two isoforms of keratinase [34], while our puri ed enzyme has only one isoform that is active in both acidic and alkaline conditions. Our keratinase, compared to the KerA commercial enzyme, which is extracted from B. licheniformis PW-1, has a higher thermal and pH stability as well as activity over a wide range of pH.…”

Section: Discussionsupporting

confidence: 87%

Characterization of a Novel Keratinase With Surfactant Stability in a Wide Range of pH Activity From a Native Isolate, Bacillus Mojavensis FUM125

Jeddi

Sharifmoghadam

Asoodeh

et al. 2021

Preprint

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Keratinases are enzymes with the most diverse sources and applications. Different forms of keratinase have been applied in environment and variety of industries, highlighting the necessity for novel potential keratinases, which could be applicable in variety of industries. Accordingly, the present study aimed to identify and characterize a novel keratinase producing bacterium with high potential in variety of industries. In the present study, the native isolate of Bacillus sp. FUM125 was isolated, identified and optimized for the keratinolytic activity. The keratinase was purified and characterized using biochemical assays. The Bacillus sp. FUM125 isolate was identified as Bacillus mojavensis R-OH-1 with 99.8% similarity. The isolate showed the maximum keratinolytic activity at pH of 8.5 after 24-hour incubation at 37°C (2.1-fold enzyme production). According to the biochemical analysis, the keratinase belonged to a serine protease family, whit 33.5 kDa molecular weight and was stable in a wide range of pH and temperature with maximum keratinolytic activity at 60°C and pH 8. Among the metal ions, K+, Ca2+, Na2+ and Mg2+ increased the enzyme activity. The activity was increased by the reducing agents of DTT and beta-mercaptoethanol. Based on the substrate profile findings, the enzyme was active in various soluble and insoluble substrates. The enzyme showed a half-life of 98 min in the optimal temperature and the ratio of keratinolytic:caseinolytic to be 0.95. Our enzyme with higher temperature and pH stability compared to existing commercial enzymes can be considered as a potential candidate for use in various industries.

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Section: Discussionsupporting

confidence: 87%

Characterization of a Novel Keratinase With Surfactant Stability in a Wide Range of pH Activity From a Native Isolate, Bacillus Mojavensis FUM125

Jeddi

Sharifmoghadam

Asoodeh

et al. 2021

Preprint

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“…Few reports are available on purification and characterization of keratinase enzyme produced by microorganisms Lysobacter sp. (Allpress et al, 2002), Bacillus licheniformis (Suntornsuk et al, 2005), Bacillus thrungenesis (Sivakumar 2012), Streptomyces albus (Esawy 2007), Microsporum gypseum (Raju et al, 2007), Trichophyton simmi (Singh 1997), Aspergillus parasiticus (Anitha and Palanivelu, 2012), Cunnighamella echinulata (More et al, 2013). However most of the study belong to bacterial isolates or dermatophytes, therefore the purification and characterization of keratinase from nonpathogenic microorganisms is important because of their capacity to degrade the ker-atin and various application.…”

Section: Introductionmentioning

confidence: 99%

Isolation of Chrysosporium indicum from poultry soil for keratinase enzyme, its purification and partial characterization

Kumar

Mahal

2021

JANS

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Keratinases are produced by microorganisms as fungi, actinomycetes and bacteria and have the capacity to degrade tough insoluble keratin proteins, including feathers. Feathers are waste produced from the poultry industry worldwide and accumulated as solid waste. Therefore, keratinolytic fungal strains Chrysosporium indicum were isolated by hair baiting method from poultry farm soil of Punjab, India. Isolated C. indicum were screened for proteolytic activity on skimmed milk agar. Field Emission Scanning Electron Microscopy (FeSEM) analysis confirmed morphological characters as C. indicum. Fourier transform infrared spectroscopy analysis was studied for the structural and mechanism analysis of feather degraded during keratinase production. Keratinase enzyme was purified 48.03% recovery by ammonium sulphate precipitation, dialysis for desalting and chromatography. Diethylaminoethyl sepharose (DEAE sepharose) and Sephadex-G75 column were used to perform chromatography and partial characterization of the keratinase for temperature, pH, and substrate. The maximum keratinase activity was observed at 500C, at pH 10. The maximum enzyme activity of 289.1 U/ml was observed with keratin powder as substrate and minimum enzyme activity 67.1 U/ml with keratin azure. This is the first report on the purification and characterization of keratinase by C. indicum using DEAE sepharose as affinity chromatography for the purification of the keratinase enzyme.

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“…Baweja and Singh (2017) reported ~ 53.0 kDa alkaline protease from Bacillus pumilus MP27. Likewise, keratinase of B. pumilus KS12 of 45 kDa was reported by Rajput et al (2010), whereas, alkaline protease of 33 kDa was found in Cunninghamella echinulata (More et al 2013).…”

Section: Expression and Purification Of Recombinant Enzymementioning

confidence: 79%

Molecular and biochemical characterization of a thermostable keratinase from Bacillus altitudinis RBDV1

et al. 2018

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A thermostable keratinase designated as KBALT was purified from RBDV1 from a poultry farm in Gujarat, India. The molecular weight of the native KBALT (nKBALT) purified using ammonium sulfate and ion exchange and gel permeation chromatography with a 40% yield and 80-fold purification was estimated to be ~ 43 kDa. The gene for KBALT was successfully cloned, sequenced and expressed in. Recombinant KBALT (rKBALT) when purified using a single step Ni-NTA His affinity chromatography achieved a yield of 38.20% and a 76.4-fold purification. Comparison of the deduced amino acid sequence of rKBALT with known proteases of species and inhibitory effect of PMSF suggest that rKBALT was a subtilisin-like serine protease. Both native and rKBALT exhibited higher activity at 85 °C and pH 8.0 in the presence of Mg, Mn, Zn, Ba and Fe metal ions. Interestingly, 70% of their activity was retained at temperatures ranging from 35 to > 95 °C. The keratinolytic activity of both nKBALT and rKBALT was enhanced in the presence of reducing agents. They exhibited broad substrate specificity towards various protein substrates. KBALT was determined for its kinetic properties by calculating its (0.61 mg/ml) and (1673 U/mg/min) values. These results suggest KBALT as a robust and promising contender for enzymatic processing of keratinous wastes in waste processing plants.

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Purification and properties of a novel fungal alkaline keratinase from Cunninghamella echinulata

Cited by 15 publications

References 35 publications

Characterization of a Novel Keratinase With Surfactant Stability in a Wide Range of pH Activity From a Native Isolate, Bacillus Mojavensis FUM125

Characterization of a Novel Keratinase With Surfactant Stability in a Wide Range of pH Activity From a Native Isolate, Bacillus Mojavensis FUM125

Isolation of Chrysosporium indicum from poultry soil for keratinase enzyme, its purification and partial characterization

Molecular and biochemical characterization of a thermostable keratinase from Bacillus altitudinis RBDV1

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