• Journal of Microbiology and Biotechnology
• /
• v.28 no.2
• /
• pp.314-322
• /
• 2018

Bacillus subtilis 8 is highly efficient at degrading feather keratin. We observed integrated feather degradation over the course of 48 h in basic culture medium while studying the entire process with scanning electron microscopy. Large amounts of ammonia, sulfite, and $\text\tiny{L}$-cysteic acid were detected in the fermented liquid. In addition, four enzymes (gamma-glutamyltranspeptidase, peptidase T, serine protease, and cystathionine gamma-synthase) were identified that play an important role in this degradation pathway, all of which were verified with molecular cloning and prokaryotic expression. To the best of our knowledge, this report is the first to demonstrate that cystathionine gamma-synthase secreted by B. subtilis 8 is involved in the decomposition of feather keratin. This study provides new data characterizing the molecular mechanism of feather degradation by bacteria, as well as potential guidance for future industrial utilization of waste keratin.

• Journal of Environmental Science International
• /
• v.19 no.9
• /
• pp.1077-1082
• /
• 2010

This study was performed to investigate the nutritional conditions controlling keratinase activity in Bacillus megaterium F7-1. B. megaterium F7-1 produced keratinase using chicken feather as a sole source of carbon, nitrogen and sulfur. Addition of the feather medium with glucose enhanced keratinase production (68.9 U/ml), compared to control without glucose (63.2 U/ml). The synthesis of keratinase was repressed by addition of $NH_4Cl$ in B. megaterium F7-1. The highest keratinase production (70.9 U/ml) was obtained with the feather medium containing glucose and $MgSO_4{\cdot}7H_2O$. Keratinase was produced in the absence of feather (4.9 U/ml), indicating its constitutive synthesis. Feather degradation resulted in free SH group formation. B. megaterium F7-1 effectively degraded chicken feather meal (86%), whereas duck feather, human nail, human hair and sheep wool displayed relatively low degradation rates (8-34%).

• Journal of Environmental Science International
• /
• v.21 no.2
• /
• pp.253-261
• /
• 2012

We isolated and characterized novel duck feather-degrading bacteria producing keratinase. Twelve strains were isolated from soil and faces at poultry farm, and decayed feathers. They were identified as Bacillus methylotrophicus, Pseudomonas geniculata, Pseudomonas hibiscicola, Exiquobacterium profundum, Bacillus pumilus, Bacillus amyloliquefaciens, Chryseobacterium indologenes, Bacillus thuringiensis, Thermomonas koreensis, respectively, by phenotypic characters and 16S rRNA gene analysis. Generally, the level of keratinase production was not proportional to feather degradation rate. The highest keratinolytic activity was observed in the culture inoculated with Chryseobacterium indologenes D27. Although all strains did not degrade human hair, strains tested effectively degraded chicken feather(53.8-91.4%), wool(40.4-93.0%) and human nail (51.0-82.9%). These results suggest that strains isolated could be not only used to improve the nutritional value of recalcitrant feather waste but also is a potential candidate for biotechnological processes of keratin hydrolysis.

• Asian-Australasian Journal of Animal Sciences
• /
• v.10 no.2
• /
• pp.210-214
• /
• 1997

Ruminal degradation characteristics and intestinal availability of crude protein (CP) in four animal-origin feeds (fish meal, meat meal, viscera meal, feather meal) were estimated by mobile nylon bag technique. Three ruminally and duodenally cannulated Holstein dairy cows (average body wt. 550kg) fed a diet containing 40% concentrate and 60% orchard grass hay on a dry matter (DM) basis. Assuming that the outflow rate of diet in rumen is 5% per hour (k =0.05), contents of quickly degradable CP (QDP), slowly degradable CP (SDP), and undegradable CP (UDP) in the rumen were 27.6%, 9.4%, 63.0% for fish meal, 34.3% 28.1%, 37,6% for meat meal, 43.9%, 12.5%, 43.6% for viscera meal, and 14.4%, 15.8%, 69.8% for feather meal, respectively. Intestinal CP degradability was 51.0% for fish meal, 27.2% for meat meal, 37.9% for viscera meal and 56.2% for feather meal. Available UDP in the intestinal tract was contained 288 g, 217 g, 246 g and 423 g per kilogram DM of diet in fish meal, meat meal, viscera meal and feather meal, respectively.

• Microbiology and Biotechnology Letters
• /
• v.40 no.4
• /
• pp.303-309
• /
• 2012

Keratinases are of particular interest because of their action on insoluble keratins and generally on a broad range of protein substrates. Alkalophilic and neutrophilic actinomycete strains isolated from different soil samples, rich in keratinaceous substances were screened for keratinolytic activity. An alkalophilic isolate, TBG-S13A5, was found to possess good keratinolytic activity and was able to utilize feather as the sole carbon and nitrogen source. TBG-S13A5 exhibited an off-white aerial mass color with a rectus-flexibilis type of spore chain. The morphological, microscopical and biochemical characters were comparable with that of Streptomyces albidoflavus. Fatty acid methyl ester profiling (FAME) and 16S rDNA sequence analysis confirmed its identity as a strain of S. albidoflavus. Under submerged fermentation conditions, maximum protease production was recorded on the $5^{th}$ day of incubation at $30^{\circ}C$, using basal broth of pH 9.0 with 0.25% (w/v) white chicken feather. This strain could affect feather degradation when the initial pH was 8 and above and maximum protease production was recorded when the initial pH was around 10.5. The effectiveness of the crude enzyme in destaining and leather dehairing were also demonstrated.

• Journal of Environmental Science International
• /
• v.19 no.10
• /
• pp.1307-1314
• /
• 2010

This study was conducted to isolate and characterize a novel feather-degrading bacterium producing keratinase activity. A strain K9 was isolated from soil at poultry farm and identified as Xanthomonas sp. K9 by phenotypic characters and 16S rRNA gene analysis. The cultural conditions for the keratinase production were 0.3% fructose, 0.1% gelatin, 0.04% $K_2HPO_4$, 0.06% $KH_2PO_4$, 0.05% NaCl and 0.01% $FeSO_4$ with an initial pH 8.0 at $30^{\circ}C$ and 200 rpm. In an optimized medium containing 0.1% chicken feather, production yield of keratinase was approximately 8-fold higher than the yield in basal medium. The strain K9 effectively degraded chicken feather meal (67%) and duck feather (54%), whereas human nail and human hair showed relatively low degradation rates (13-22%). Total free amino acid concentration in the cell-free supernatant was about 25.799 mg/l. Feather hydrolysate produced by the strain K9 stimulated growth of red pepper, indicating Xanthomonas sp. K9 could be not only used to increase the nutritional value of chicken feather but also a potential candidate for the development of natural fertilizer applicable to crop plant soil.

• Korean Journal of Microbiology
• /
• v.46 no.1
• /
• pp.86-92
• /
• 2010

Thirty-one chicken feather-degrading bacteria were isolated from wasted feather, compost and wastewater in a chicken farm. These isolates were categorized as Firmicutes (21 strains), ${\gamma}$-proteobacteria (4 strains), Actinobacteria (4 strains), and Bacteroidetes (2 strains) by 16S rRNA gene sequence analysis. We examined the feather-degrading isolates for degradation in the 2% of chicken feather meal. The strain Chryseobacterium sp. FBF-7, Stenotrophomonas maltophilia FBS-4, and Lysinibacillus sp. FBW-3 were selected as a keratinolytic protein degrading bacteria which showed the highest feather degradation of 75-90%. The characteristics of amino acids extracted from chicken feather meal by using keratinolytic protein degrading isolates and chemical method with $Ca(OH)_2$ were analyzed. Total amino acid content of strain Chryseobacterium sp. FBF-7 was 1,661.6 ${\mu}mol$/ml, which was the highest and it was similar with chemical method. And essential amino acid content of total amino acid was thirty-seven percent (619.3 ${\mu}mol$/ml) and 596.9 ${\mu}mol$/ml for keratinolytic protein degrading isolates and chemical method, respectively. The major amino acids were valine, glutamic acid, aspartic acid, glycine, and proline by the strain Chryseobacterium sp. FBF-7 and especially, higher contents of aspartic acid, threonine, serine, cysteine, and tyrosine were detected compared with chemical method.

• Journal of Animal Science and Technology
• /
• v.44 no.1
• /
• pp.105-112
• /
• 2002

In order to investigate the effects of heat treatment of three animal by-products(feather meal, tallow meal, viscera meal) on in situ ruminal degradation characteristics and gastrointestinal availability of dietary crude protein(CP), three ruminally and duodenally cannulated dry Holstein cows were employed. Cows were fed a diet containing 60% concentrate and 40% orchard grass hay, and had free access to water and mineral block. Experimental feeds were processed for 4 hr at 149$^{\circ}C$ in a forced-air oven, and were passed through a 1-mm screen. Degradation kinetics of feed protein in the rumen were fitted to an exponential type model, and intestinal availability was estimated by the mobile nylon bag technique. Effective CP degradabilities in the rumen for feather meal, tallow meal and viscera meal were 30.2%, 75.0% and 56.4% at 5% passage rate per hour(k=0.05), respectively. In addition, heat treatment increased effective ruminal CP degradability on feather meal and viscera meal treatments, whereas decreased in tallow meal treatment(P$<$0.05). Gastrointestinal CP disappearances of feather meal, tallow meal and viscera meal were 56.2%, 18.6%, and 37.9%, respectively. In addition, heat treatment decreased the gastrointestinal CP disappearance on feather meal and viscera meal treatment, but increased in tallow meal treatment(P$<$0.05). Intestinal availability of rumen undegradable protein(A-UDP) was 80.4% for feather meal, 83.8% for tallow meal and 86.9% for viscera meal. In addition, heat treatment increased A-UDP on feather meal and tallow meal treatment, 94.0% and 91.3%, respectively, but decreased on viscera meal treatment, 76.5%(P$<$0.05).

• Mycobiology
• /
• v.33 no.2
• /
• pp.121-123
• /
• 2005

Extracellular keratinase isolated from Aspergillus flavus K-03 was immobilized on calcium alginate. The properties and reaction activities of free and immobilized keratinase with calcium alginate were characterized. The immobilized keratinase showed proteolytic activity against soluble azo-casein and azo-keratin, and insoluble feather keratin. Heat stability and pH tolerance of keratinase were greatly enhanced by immobilization. It also displayed a higher level of heat stability and an increased tolerance toward alkaline pHs compared with free keratinase. During the durability test at $40^{\circ}C$, 48% of the original enzyme activity of the immobilized keratinase was remained after 7 days of incubation. The immobilized keratinase exhibited better stability, thus increasing its potential for use in industrial application.

• Journal of Microbiology and Biotechnology
• /
• v.27 no.12
• /
• pp.2190-2198
• /
• 2017

Thermoactinomyces sp. strain YT06 was isolated from poultry compost and observed to degrade integral chicken feathers completely at $60^{\circ}C$, resulting in the formation of 3.24 mg/ml of free amino acids from 50 ml of culture containing 10 g/l chicken feathers. Strain YT06 could grow and secrete keratinase using feather as the only carbon and nitrogen sources without other supplement, but complementation of 10 g/l sucrose and 4 g/l $NaNO_3$ increased the production of the keratinolytic enzyme. The maximum protease activity obtained was 110 U/ml and for keratinase was 42 U/ml. The keratinase maintained active status over a broad pH (pH 8-11) and temperature ($60-75^{\circ}C$). It was inhibited by serine protease inhibitors and most metal ions; however, it could be stimulated by $Mn^{2+}$ and the surfactant Tween-20. A reductive agent (${\beta}$-mercaptoethanol) was observed to cleave the disulfide bond of keratin and improve the access of the enzyme to the keratinaceous substrate. Zymogram analysis showed that strain YT06 primarily secreted keratinase with a molecular mass of approximately 35 kDa. The active band was assessed by MALDI-TOF mass spectrometry and was observed to be completely identical to an alkaline serine protease from Thermoactinomyces sp. Gus2-1. Thermoactinomyces sp. strain YT06 shows great potential as a novel candidate in enzymatic processing of hard-to-degrade proteins into high-value products, such as keratinous wastes.