Purification and characterization of keratinase from a new Bacillus subtilis strain

Cai, Chenggang; Chen, Jishuang; Qi, Jiong-jiong; Yin, Yanlong; Zheng, Xiaodong

doi:10.1631/jzus.b0820128

Cited by 87 publications

(48 citation statements)

References 36 publications

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“…Different organic solvents such as ethanol, methanol, isopropyl alcohol and detergents such as tween-20 and tween-80 inhibited keratinase activity to some degree. Similar results were found for keratinase from Bacillus subtillis [35]. But the enzyme from O. corvina was stable in the presence of glycerol; while enzyme from Bacillus subtilis was not.…”

Section: Characterization Of the O Corvina Feather Degrading Enzymessupporting

confidence: 82%

“…Keratinase activity assay: Keratinase activity was measured as described [35] with a few modifications. Keratin azure (Sigma-Aldrich) was used as the substrate.…”

Section: Determination Of Enzyme Activitymentioning

confidence: 99%

“…The keratinase activity of O. corvina was decreased by SDS and triton x-100 detergents, as was the enzyme from Streptomyces pactum [40]. Reducing agents like DTT and β-mercaptoethanol generally enhance keratinase activity because the addition of reducing agents can break disulfide bonds to help sulfitolysis [35]. But keratinase activity of O. corvina was partially inhibited by DTT and β-mercaptoethanol after pre-incubation for 15 min.…”

Section: Characterization Of the O Corvina Feather Degrading Enzymesmentioning

confidence: 99%

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Production and Characterization of Keratinolytic Proteases Produced by Onygena corvina

Huang

Busk²,

Lange

2015

Fungal Genom Biol

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Poultry farms produce huge quantities of feather waste that causes serious local disposal and accumulative problems and results in environmental pollution. Feathers are composed of β-keratin rich protein and are highly resistant to degradation. The feather degradation potential of the non-pathogenic fungus Onygena corvina was investigated by cultivating this fungus in a medium with duck feathers as the sole carbon and nitrogen source. O. corvina secreted a high level of alkaline protease and keratinase sufficient for degrading duck feathers completely. The optimal conditions for feather keratinolysis were 25ºC, initial pH 8 and feather concentration of 15 g/l. The maximum protease and keratinase activities were found to be 1435 and 72 U/ml, respectively. The protease was active over a broad pH (pH 6-11) and temperature (40-60ºC) range. The keratinase was sensitive to serine protease inhibitors and organic solvents and inhibited by most metal ions, but was stimulated by Ca 2+ and Fe 2+ . Zymogram analysis showed that O. corvina secreted mainly proteases with molecular weight of approximately 35 and 20 kDa. Compared to the fungus Trichoderma asperellum, O. corvina showed higher potential and could be of relevance for bioconverting feather waste into high value products.

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Section: Characterization Of the O Corvina Feather Degrading Enzymessupporting

confidence: 82%

“…Keratinase activity assay: Keratinase activity was measured as described [35] with a few modifications. Keratin azure (Sigma-Aldrich) was used as the substrate.…”

Section: Determination Of Enzyme Activitymentioning

confidence: 99%

Section: Characterization Of the O Corvina Feather Degrading Enzymesmentioning

confidence: 99%

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Production and Characterization of Keratinolytic Proteases Produced by Onygena corvina

Huang

Busk²,

Lange

2015

Fungal Genom Biol

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“…In order to obtain colonies, the bacteria were first cultured on nutrient agar by streaking and then incubated at 37 ℃ for 24h (9,10). After characterization with Gram staining and morphology, the colonies were transferred and incubated in skim milk agar to separate hemolytic bacteria (11). Keratinase-producing isolates were cultured in liquid FMB medium containing 100 g of powdered feathers for 72h.…”

Section: Resultsmentioning

confidence: 99%

Isolation and Molecular Identification of Keratinase-Producing Bacteria from the Sludge of Qeshm Island

2018

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Background and Objectives: Keratinase is an enzyme commonly used in the production of detergents, cosmetics, drugs, leather, and other industries. Considering the high cost of traditional methods for decomposition of feather, hair, hooves, nails, and wool that contain high levels of keratin, their biodegradation with keratinase-producing bacteria can be a valuable solution. The present study aimed for isolation and molecular identification of keratinase-producing bacteria in Qeshm Island and Peyposht village in Iran.Methods: Water and sludge samples from the Qeshm Island and Peyposht village were collected. The bacteria isolates were screened for keratinase production using the Lowry method. Effect of pH and temperature was assessed on the production of keratinase and on the growth of the isolates. Colony-polymerase chain reaction was used for molecular identification of the isolates.Results: Bacillus berevis and Enterobacter cloacae were isolated in this study. Keratinase production in B. berevis was highest at pH 7.5 and 35 °C. In addition, the highest level of enzyme production by E. cloacae was observed at pH 7 and 37 °C.Conclusion: It seems that the bacterial strains isolated from sludge in the study area have relatively favorable keratinase production capacity.

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“…Nevertheless, enzymatic keratinolysis in vitro, conducted in the absence of red-ox potential of living microbial cells usually requires additional reducing agents to support disulfide bonds cleavage. This could be attained either by introducing chemical reducers into reaction environment or through initial substrate pretreatment [22,23].…”

Section: Introductionmentioning

confidence: 99%

Enzymatic Degradation of Pretreated Pig Bristles with Crude Keratinase of Bacillus cereus PCM 2849

Łaba

Chorążyk

Pudło

et al. 2016

Waste Biomass Valor

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Purpose The aim of the study was to apply and optimize the process of bioconversion of pig bristle waste using keratinolytic enzymes of Bacillus cereus PCM 2849, and to evaluate the amino acid composition of the resultant hydrolysate. Methods Hydrolysis with concentrated culture fluid of B. cereus was applied for bioconversion of pig bristles, after thermo-chemical pretreatment with sulfite. The effect of substrate concentration, sulfite concentration during pretreatment and reaction temperature on the release of amino acids was determined using Box-Behnken design. Amino acid composition of the obtained hydrolysate was determined by HPLC. Structural condition and substructural changes of the residual substrate were evaluated with SEM microscopy and FTIR spectroscopy. Results The applied enzymatic preparation for bristle biodegradation was verified to contain multiple proteases of a wide molecular weight range. A regression model was developed, in which influential parameters were: linear effect of substrate concentration, followed by quadratic effects of reaction temperature, substrate concentration and pretreatment. Optimum reaction conditions were also determined. The resultant hydrolysate was rich in branched-chain amino acids. Residual substrate was detriorated and sulfitolytic cleavage of disulfides and alteration of protein secondary structures was confirmed. Conclusions Application of B. cereus crude keratinase allowed for partial hydrolysis of pig bristles, preceded by sulfitolytic pretreatment. A regression model was built to describe the process of hydrolysis to release free amino acids, at constant enzyme load. Hydrolysis in given conditions allowed to obtain hydrolysate rich in branched chain amino acids. The presented process poses an alternate way of management over pig bristles, a hard-to-degrade keratinous waste.

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Purification and characterization of keratinase from a new Bacillus subtilis strain

Cited by 87 publications

References 36 publications

Production and Characterization of Keratinolytic Proteases Produced by Onygena corvina

Production and Characterization of Keratinolytic Proteases Produced by Onygena corvina

Isolation and Molecular Identification of Keratinase-Producing Bacteria from the Sludge of Qeshm Island

Enzymatic Degradation of Pretreated Pig Bristles with Crude Keratinase of Bacillus cereus PCM 2849

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