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

doi:10.5897/AJB09.545

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

Journal Article•DOI•

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

Bo Xu, Qiaofang Zhong, Xianghua Tang, Yunjuan Yang, Zunxi Huang - Show less +1 more

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.

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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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Feather degrading actinomycetes: a biotechnological approach to environmental clean-up

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T. Ganesan

01 Jan 2011

3 citations

Dissertation•

Production, Characterization, and Potential Application of A Keratinolytic Alkaline Protease Produced by Alkaliphilic Vibrio Sp.

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Muhammed Seid

01 May 2011

3 citations

Journal Article•

Isolation and characterization of keratinolytic bacteria from soil samples of poultry waste dumping sites.

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Alvin T Reyes, I. D. Ambita, J. L. Batalon, B. L. Aba, A. Cortes, C. G. Macabecha, Andrew D. Montecillo - Show less +3 more

01 Jan 2018-International Journal of Agricultural Technology

TL;DR: The keratinolytic bacteria isolated from soil samples which containing the degrading feathers was investigated and two isolates exhibited conspicuous keratinase activity belonged to Bacillus cereus species.

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Abstract: The keratinolytic bacteria isolated from soil samples which containing the degrading feathers was investigated. Thirteen bacterial isolates were selected and were subjected to preliminary screening through protease assay using Milk Agar Medium. Six isolates were detected positive for protease activity and were further characterized via biochemical and microscopic assays. Isolates were grown in basal medium containing feathers as sole nutrient source and the degree of feather degradation was monitored. Two isolates exhibited conspicuous keratinase activity. DNA from these two candidate organisms were isolated and subjected to PCR using 16S rRNA specific primers. PCR products were sequenced and analysis revealed that both of them belong to Bacillus cereus species. Isolation of potential keratinolytic microorganisms could have potential biotechnological used especially in processes which involved keratin hyrdolysis.

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2 citations

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

...Dymatic hydrolysis by microorganisms that possess keratinolytic activity represents an attractive alternative to improve the nutritional value of feather wastes (Xu et al., 2009)....
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Partial Characterization of Keratinase of Thermophilic Bacteria from Three Hotsprings of North Sumatra

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Dwi Suryanto, Kabul Warsito, Erman Munir, Siti Khadijah Nasution

01 Jan 2015

TL;DR: Keratin is insoluble protein and resistant to degradation by most proteolytic enzymes, one major source of keratin is feather of poultry, where this material is usually dumped to the environment and microbial keratinolytic hydrolysis represents an attractive alternative to improve the nutritional value of feather wastes.

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Abstract: Keratin is insoluble protein and resistant to degradation by most proteolytic enzymes. One major source of keratin is feather of poultry, where this material is usually dumped to the environment. Microbial keratinolytic hydrolysis represents an attractive alternative to improve the nutritional value of feather wastes. In this study, keratinolytic bacteria of Penen, Semangat Gunung, and Sipaholon hotprings of North Sumatra, Indonesia were isolated. Sixteen keratinolityc bacteria were isolated from the hotsprings. Three isolates KW05, SQ04, and WR03 were choosen and partial characterized for their crude keratinase activity. KW05 showed to have high keratinolytic activity. Assay of the crude enzyme in different pH and temperature showed that KW05 was more active in pH 5.0, while SQ04 and WR03 were in pH 6.5. Crude keratinase of all three isolates showed more active in 60°C.

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1 citations

Journal Article•DOI•

Partial Characterization of Keratinolytic Activity of Local Novel Bacteria Isolated from Feather Waste

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Dwi Suryanto, Hilda Walida, Siti Khadijah Nasution, Erman Munir

31 Mar 2017-Journal of Pure and Applied Microbiology

TL;DR: Three keratinolytic bacterial isolates were characterized partially for their keratinase activity and showed that keratinae activity of isolate A4 was optimum at 20% of ammonium sulphate, while B4 and B6 were more active at 70%.

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Abstract: Three keratinolytic bacterial isolates were characterized partially for their keratinase activity. Bacterial isolates were grown in feather meal agar. Ammonium sulfate precipitation followed by dialysis was performed to know the bacterial isolate keratinase activity in differet pH and temperature. Identification of the bacteria was done by using their 16S rRNA gene sequences. The result showed that bacterial growth was coinciding with keratinase activity. Precipitation with ammonium sulfate showed that keratinae activity of isolate A4 was optimum at 20% of ammonium sulphate, while B4 and B6 were more active at 70%. Keratinase activity increased after dialysis. Keratinase of A4 showed to have optimum activity at temperature of 45oC and pH=8, B4 was optimum at temperature of 35oC and pH=7, while B6 was optimum at temperature of 40oC and pH=7, respectively. Identification of the bacterial isolates using 16S rRNA gen showed that A4, B4, and B6 were closed to Leclercia adecarboxylata strain M-X17B, Azotobacter chroococcum strain ABA-1, and Stenotrophomonas maltophilia strain BIW by 97%, 99%, and 98%, respectively. Two bacteria L. adecarboxylata and A. chroococcum were firstly reported to produce keratinase.

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1 citations

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Journal Article•DOI•

Purification and Characterization of a Keratinase from a Feather-Degrading Bacillus licheniformis Strain

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Xiang Lin¹, Chung-Ginn Lee¹, Ellen S. Casale¹, Jason C. H. Shih¹•Institutions (1)

North Carolina State University¹

01 Oct 1992-Applied and Environmental Microbiology

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.

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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

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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 Article•DOI•

Isolation, identification, and characterization of a feather-degrading bacterium.

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C. M. Williams¹, C. S. Richter¹, J. M. MacKenzie¹, Jason C. H. Shih¹•Institutions (1)

North Carolina State University¹

01 Jun 1990-Applied and Environmental Microbiology

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.

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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.

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335 citations

Journal Article•

The use of azoalbumin as a substrate in the colorimetric determination or peptic and tryptic activity.

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R M Tomarelli, J Charney, M L Harding

01 Mar 1949-Journal of Laboratory and Clinical Medicine

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 Article•DOI•

Characterization of a new keratinolytic bacterium that completely degrades native feather keratin.

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Alessandro Riffel¹, Françoise S. Lucas², Philipp Heeb², Adriano Brandelli¹•Institutions (2)

Universidade Federal do Rio Grande do Sul¹, University of Lausanne²

28 Feb 2003-Archives of Microbiology

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.

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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+).

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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 Article•DOI•

Native-feather degradation by Fervidobacterium islandicum AW-1, a newly isolated keratinase-producing thermophilic anaerobe

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Gae Won Nam¹, Dong Woo Lee¹, Han Seoung Lee¹, Nam Lee¹, Byoung-Chan Kim¹, Eun Ah Choe¹, Jae Kwan Hwang¹, Maggy Thenawidjaja Suhartono², Yu Ryang Pyun¹ - Show less +5 more•Institutions (2)

Yonsei University¹, Bogor Agricultural University²

16 Oct 2002-Archives of Microbiology

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.

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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.

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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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