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Journal ArticleDOI

Isolation and characterization of a new keratinolytic bacterium that exhibits significant feather-degrading capability

15 Sep 2009-African Journal of Biotechnology (Academic Journals)-Vol. 8, Iss: 18, pp 4590-4596
TL;DR: Potential biotechnological applications of this bacterium that involve hydrolysis of keratin, including the improvement of the nutritional properties of feathers (and other keratins) used as supplementary feedstuffs are suggested.
Abstract: A novel bacterium, Bacillus licheniformis K-19, which produces a large amount of akeratinase that is extremely thermostable and has a broad resistance to pH, was isolated and characterized. The maximum amount of keratinase activity (about 224 Uml-1) was produced at 37°C when the bacterium was cultured for 72 h in broth containing feather meal with initial pH of 7.5. The keratinase activity was observed over a wide range of temperatures (30 - 90°C) and pH values (pH 6 - 10). It was optimal at 60°C and pH 7.5 - 8 respectively. These results suggest potential biotechnological applications of this bacterium that involve hydrolysis of keratin, including the improvement of the nutritional properties of feathers (and other keratins) used as supplementary feedstuffs. Key words: Bacillus licheniformis, chicken feather, keratin, keratinolytic protease.

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Citations
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Journal ArticleDOI
TL;DR: Alkaline proteases for the textile and detergent industries, as well as for the degradation of keratin-rich wastes are given a special focus in this review.
Abstract: The types and sources of proteolytic enzymes, enzyme assays, strategies for fermentation yield improvement, and novel proteases and their applications in industrial sectors are widely covered in this review. We give a special focus on alkaline proteases for the textile and detergent industries, as well as for the degradation of keratin-rich wastes.

11 citations


Cites background from "Isolation and characterization of a..."

  • ...…Chen and Wang (2008); Abusham et al. (2009); Infante et al. (2010); Chen et al. (2011) Azokeratin 37 – 60 440 – 450 15 min – 30 min Riffel et al. (2003a); Xu et al. (2009); Zhang et al. (2009); Chen et al. (2011) Casein 30 – 60 280 – 660 10 min – 1 h Kamal et al. (1995); Garcia-Kirchner et…...

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  • ...…(1999); Kanekar et al. (2002); Nadeem et al. (2007); Saran et al. (2007b); Devi et al. (2008); Tang et al. (2008); Abusham et al. (2009); Fang et al. (2009); Mala and Srividya (2010) Liquid medium Intact feathers Tatineni et al. (2008); Xu et al. (2009); Jaouadi et al. (2010) Azocasein Reddy at al....

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01 Jan 2015
TL;DR: The study shows the potential of the feather degrading isolate to carry out environmentally safe disposal of poultry waste and also to produce amino acids from a cheap raw material.
Abstract: A large amount of feather waste is generated from the poultry industry. This waste containing a considerable amount of keratin can be utilized by microorganisms thus leading to its environmentally safe disposal. The objective of the study was to isolate and identify a feather degrading bacterium from poultry farm soil, optimize the cultural conditions and partial purification of keratinase enzyme using ammonium sulphate precipitation, followed by exploring the applications. The study was carried out by collecting soil samples, enrichment using an appropriate enrichment medium, followed by screening, identification and then optimizing cultural conditions. The isolated keratinolytic bacterium carried out complete degradation of the feather waste within 4 weeks and was found to be of Bacillus sp. by biochemical identification. The bacterium presented optimum growth at pH 6.8 and a temperature of 37 oC. The keratinase enzyme activity was found to be 8.80U/ml/min for isolate P and 13.60U/ml/min for isolate W. The study shows the potential of the feather degrading isolate to carry out environmentally safe disposal of poultry waste and also to produce amino acids from a cheap raw material.

11 citations

Journal ArticleDOI
F Iruolaje, J Ogbeba, M Tula, J Ijebor, B Dogo 
TL;DR: These isolates are therefore promising organisms for the management of chicken feather waste through biotechnological processes.
Abstract: Aims: To isolate and identify feather degrading bacteria from soils collected from the feather dumping site. Study Design: Isolation and preliminary identification of bacterial isolates with keratinolytic potentials. Place and Duration of Study: Mudalawal poultry processing site, Bauchi state, Nigeria, between January 2014 to October, 2014. Methods: Soil samples from feather dumping sites were screened for bacterial growth. The Original Research Article Iruolaje et al.; JABB, 5(2): 1-9, 2016; Article no.JABB.22082 2 isolated bacteria were identified based on their morphological and biochemical characteristics and were tested for their ability to degrade whole intact feather and powdered feather samples. Results: The results showed that three (3) of the isolates belonged to the genus Bacillus which include; Bacillus licheniformis, Bacillus subtilis and Bacillus cereus, while one isolate is a Gramnegative rod, Serratia marcencens. Although B. licheniformis demonstrated higher feather degrading ability but with no statistical difference from other isolates in degrading both powdered and intact whole feathers samples. Synergy among the isolates to degrade powdered feather showed no statistical difference (p=0.317). Also, synergy among all the isolates resulted in degradation of intact feather significantly higher than that of S. marcescens, but not significantly different from those of B. licheniformis (p=0.389), B. subtilis (p=0.096) and B. cereus (p=0.096). Moreover, the keratinolytic ability of the isolates on intact whole feather was time dependent. It was also observed that the rate of degradation of powder feather samples was significantly higher than the rate of degradation of intact whole feather by all the isolates (p=0.026). Conclusion: These isolates are therefore promising organisms for the management of chicken feather waste through biotechnological processes.

9 citations


Cites background from "Isolation and characterization of a..."

  • ...However, production of feather meal is an expensive process which destroys certain amino acids, yielding a product with poor digestibility and variable nutritional quality [1,2]....

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01 Jan 2014
TL;DR: The keratinases produced by the fungi could be useful in decomposition of keratin-wastes and could find applications in leather, pharmaceutical and cosmetics industries as well.
Abstract: A feather degrading fungi was newly isolated from marine sponges collected from Kulasekarapattinam, Tuticori district, Tamil Nadu using agar mineral medium containing soluble keratin as a substrate and it was identified by its morphological features as Scopulariopsis brevicaulis. The selected growth medium contained feather meal as a sole source of carbon and nitrogen at pH 7.5 and temperature at 28±2 0 C. Purification was carried through ammonium sulfate precipitation and chromatography via Sephadex G- 100 column using with soluble keratin as a substrate, 10.0-fold purification with a yield of 22.8% was obtained. The partially purified keratinase has a molecular mass of 28±0.5 kDa, optimum pH in the neutral region 7 to 7.5 and optimum temperature of 50 0 C.The keratinases produced by the fungi could be useful in decomposition of keratin-wastes and could find applications in leather, pharmaceutical and cosmetics industries as well.

8 citations

Journal Article
TL;DR: From this study, Bacillus megaterium isolated from feather dumped soil can be used as a potential candidate for degradation of feather and for can be use as additives in poultry field.
Abstract: Feather keratin is highly resistant to degradation, but some keratinase producing microorganisms can easily degrade these insoluble keratins. These keratinase producing species have an important application in removal of poultry waste and recycled into valuable byproduct. A feather-degrading bacterium with high keratinase activity was isolated and identified as Bacillus megaterium (A1). The selected organism showed keratinase activity of 72.875 IU/mg and the protein content of 4 mg/ml. Maximum enzyme production was observed on 96 hr. The intense feather degrading was achieved in 35°C and initial pH adjusted to 7.5. Among the extra nitrogen or carbon sources used, significant improvement in yield of keratinase was obtained when grown in medium containing 2% feather meal. The protein profile was analyzed in SDS-PAGE showed multiple bands. Zymography analysis showed a single band with molecular weight of 46 KDa which corresponds to keratinase activity. Immobilized cells of B. megaterium (A1) had highest durability to degrade feather keratin. Fourier Transform Infrared Spectroscopy (FTIR) showed that the change in the functional group was catalyzed by the unique enzymes of B. megaterium (A1). From this study, Bacillus megaterium isolated from feather dumped soil can be used as a potential candidate for degradation of feather and for can be used as additives in poultry field.

7 citations

References
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Journal ArticleDOI
TL;DR: The purified keratinase hydrolyzes a broad range of substrates and displays higher proteolytic activity than most proteases and is a useful enzyme for promoting the hydrolysis of feather keratin and improving the digestibility of feather meal.
Abstract: A keratinase was isolated from the culture medium of feather-degrading Bacillus licheniformis PWD-1 by use of an assay of the hydrolysis of azokeratin. Membrane ultrafiltration and carboxymethyl cellulose ion-exchange and Sephadex G-75 gel chromatographies were used to purify the enzyme. The specific activity of the purified keratinase relative to that in the original medium was approximately 70-fold. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis and Sephadex G-75 chromatography indicated that the purified keratinase is monomeric and has a molecular mass of 33 kDa. The optimum pH and the pI were determined to be 7.5 and 7.25, respectively. Under standard assay conditions, the apparent temperature optimum was 50°C. The enzyme is stable when stored at −20°C. The purified keratinase hydrolyzes a broad range of substrates and displays higher proteolytic activity than most proteases. In practical applications, keratinase is a useful enzyme for promoting the hydrolysis of feather keratin and improving the digestibility of feather meal. Images

363 citations


"Isolation and characterization of a..." refers background or methods or result in this paper

  • ...Keratinolytic activity was measured using insoluble azokeratin as a substrate (Lin et al., 1992)....

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  • ...…B. licheniformis K-19 can secrete a large amount of a keratinase that is more thermostable and has broader pH resistance than other keratinolytic proteases from Bacillus reported previously (Lin et al., 1992; Cheng et al., 1995; Lin et al., 1999; Suntornsuk et al., 2003; Suntornsuk et al., 2005)....

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  • ...licheniformis K-19 can secrete a large amount of a keratinase that is more thermostable and has broader pH resistance than other keratinolytic proteases from Bacillus reported previously (Lin et al., 1992; Cheng et al., 1995; Lin et al., 1999; Suntornsuk et al., 2003; Suntornsuk et al., 2005)....

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  • ...Many Bacillus species have been reported to produce keratinolytic proteases (Lin et al., 1992; Cheng et al., 1995; Lin et al., 1999; Manczinger et al., 2003; Suntornsuk and Suntornsuk, 2003; Zerdani et al., 2004; Suntornsuk et al., 2005)....

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  • ...The keratinase of B. licheniformis PWD-1, which was isolated from a poultry waste digester, and the gene (kerA) that encodes this keratinase have been isolated and characterized (Williams et al. 1990, Lin et al., 1992, Cheng et al. 1995, Lin et al., 1995)....

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Journal ArticleDOI
TL;DR: A feather-degrading culture was enriched with isolates from a poultry waste digestor and adapted to grow with feathers as its primary source of carbon, sulfur, and energy, indicating a potential biotechnique for degradation and utilization of feather keratin.
Abstract: A feather-degrading culture was enriched with isolates from a poultry waste digestor and adapted to grow with feathers as its primary source of carbon, sulfur, and energy. Subsequently, a feather-hydrolytic, endospore-forming, motile, rod-shaped bacterium was isolated from the feather-degrading culture. The organism was Gram stain variable and catalase positive and demonstrated facultative growth at thermophilic temperatures. The optimum rate of growth in nutrient broth occurred at 45 to 50°C and at pH 7.5. Electron microscopy of the isolate showed internal crystals. The microorganism was identified as Bacillus licheniformis PWD-1. Growth on hammer-milled-feather medium of various substrate concentrations was determined by plate colony count. Maximum growth (approximately 109 cells per ml) at 50°C occurred 5 days postinoculation on 1% feather substrate. Feather hydrolysis was evidenced as free amino acids produced in the medium. The most efficient conditions for feather fermentation occurred during the incubation of 1 part feathers to 2 parts B. licheniformis PWD-1 culture (107 cells per ml) for 6 days at 50°C. These data indicate a potential biotechnique for degradation and utilization of feather keratin.

335 citations

Journal Article

303 citations


"Isolation and characterization of a..." refers methods in this paper

  • ...Azokeratin was synthesized based on the methodology described for azoalbumin (Tomarelli et al., 1949)....

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Journal ArticleDOI
TL;DR: A novel feather-degrading microorganism was isolated from poultry waste, producing a high keratinolytic activity when cultured on broth containing native feather, and complete feather degradation was achieved during cultivation.
Abstract: A novel feather-degrading microorganism was isolated from poultry waste, producing a high keratinolytic activity when cultured on broth containing native feather. Complete feather degradation was achieved during cultivation. The bacterium presents potential use for biotechnological processes involving keratin hydrolysis. Chryseobacterium sp. strain kr6 was identified based on morphological and biochemical tests and 16S rRNA sequencing. The bacterium presented optimum growth at pH 8.0 and 30 degrees C; under these conditions, maximum feather-degrading activity was also achieved. Maximum keratinase production was reached at 25 degrees C, while concentration of soluble protein was similar at both 25 and 30 degrees C. Reduction of disulfide bridges was also observed, increasing with cultivation time. The keratinase of strain kr6 was active on azokeratin and azocasein as substrates, and presented optimum pH and temperature of 7.5 and 55 degrees C, respectively. The keratinase activity was inhibited by 1,10-phenanthroline, EDTA, Hg(2+), and Cu(2+) and stimulated by Ca(2+).

280 citations


"Isolation and characterization of a..." refers background in this paper

  • ..., 2005), Thermoanaerobacter (Riessen and Antranikian, 2001), Chryseobacterium (Riffel et al., 2003), Flavobacterium (Riffel and Brandelli, 2002; Nam et al....

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  • ...…et al., 2003; Suntornsuk and Suntornsuk, 2003; Zerdani et al., 2004; Suntornsuk et al., 2005), Thermoanaerobacter (Riessen and Antranikian, 2001), Chryseobacterium (Riffel et al., 2003), Flavobacterium (Riffel and Brandelli, 2002; Nam et al., 2002) and Vibrio (Sangali and Brandelli, 2000)....

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  • ...The microbial conversion of feather wastes is a potential technique for the degradation of feathers and their utilization as a feedstuff (Sangali and Brandelli, 2000; Riffel et al., 2003)....

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  • ...Keratinolytic activity has been reported for various bacterial genera, such as Bacillus (Williams et al., 1990; Lin et al., 1999; Manczinger et al., 2003; Suntornsuk and Suntornsuk, 2003; Zerdani et al., 2004; Suntornsuk et al., 2005), Thermoanaerobacter (Riessen and Antranikian, 2001), Chryseobacterium (Riffel et al., 2003), Flavobacterium (Riffel and Brandelli, 2002; Nam et al., 2002) and Vibrio (Sangali and Brandelli, 2000)....

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Journal ArticleDOI
TL;DR: The enzyme from F. islandicum AW-1 is a novel, thermostable keratinolytic serine protease that showed higher specific activity for the keratinous substrates than other proteases and catalyzed the cleavage of peptide bonds more rapidly following the reduction of disulfide bridges in feather keratin by 10 mM dithiothreitol.
Abstract: A native-feather-degrading thermophilic anaerobe was isolated from a geothermal hot stream in Indonesia. Isolate AW-1, identified as a member of the species Fervidobacterium islandicum, was shown to degrade native feathers (0.8%, w/v) completely at 70 °C and pH 7 with a maximum specific growth rate (0.14 h–1) in Thermotoga-Fervidobacterium (TF) medium. After 24 h of culture, feather degradation led to an increase in free amino acids such as histidine, cysteine and lysine. Moreover, nutritionally essential amino acids such as tryptophan and methionine, which are rare in feather keratin, were also produced as microbial metabolites. A homomultimeric membrane-bound keratinolytic protease (>200 kDa; 97 kDa subunits) was purified from a cell extract of F. islandicum AW-1. The enzyme exhibited activity toward casein and soluble keratin optimally at 100 °C and pH 9, and had a half-life of 90 min at 100 °C. The enzyme showed higher specific activity for the keratinous substrates than other proteases and catalyzed the cleavage of peptide bonds more rapidly following the reduction of disulfide bridges in feather keratin by 10 mM dithiothreitol. Therefore, the enzyme from F. islandicum AW-1 is a novel, thermostable keratinolytic serine protease.

263 citations


"Isolation and characterization of a..." refers background in this paper

  • ...…et al., 2003; Suntornsuk and Suntornsuk, 2003; Zerdani et al., 2004; Suntornsuk et al., 2005), Thermoanaerobacter (Riessen and Antranikian, 2001), Chryseobacterium (Riffel et al., 2003), Flavobacterium (Riffel and Brandelli, 2002; Nam et al., 2002) and Vibrio (Sangali and Brandelli, 2000)....

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  • ..., 2003), Flavobacterium (Riffel and Brandelli, 2002; Nam et al., 2002) and Vibrio (Sangali and Brandelli, 2000)....

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  • ...Keratinolytic activity has been reported for various bacterial genera, such as Bacillus (Williams et al., 1990; Lin et al., 1999; Manczinger et al., 2003; Suntornsuk and Suntornsuk, 2003; Zerdani et al., 2004; Suntornsuk et al., 2005), Thermoanaerobacter (Riessen and Antranikian, 2001), Chryseobacterium (Riffel et al., 2003), Flavobacterium (Riffel and Brandelli, 2002; Nam et al., 2002) and Vibrio (Sangali and Brandelli, 2000)....

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