Chryseobacterium aquifrigidense keratinase liberated essential and nonessential amino acids from chicken feather degradation.

Keratinous biomass valorization for value-added products presents a high prospect in ecological management and the advancement of the bio-economy. Consequently, soil samples from the poultry dumpsite were collected. The bacteria isolated on the basal salt medium were screened for keratinolytic activity. The potent chicken feathers degrading bacteria were identified through 16S rRNA gene sequencing and phylogenetic analysis. Fermentation process conditions were optimized, and the amino acid compositions of the feather hydrolysate were likewise quantified. Ten (10) proteolytic bacteria evaluated on skimmed milk agar showed intact chicken feather degradation ranging from 33% (WDS-03) to 88% (FPS-09). The extracellular keratinase activity ranged from 224.52 ± 42.46 U/mL (WDS-03) to 834.55 ± 66.86 U/mL (FPS-07). Based on 16S rRNA gene sequencing and phylogenetic analysis, the most potent keratinolytic isolates coded as FPS-07, FPS-09, FPS-01, and WDS-06 were identified as Chryseobacterium aquifrigidense FANN1, Chryseobacterium aquifrigidense FANN2, Stenotrophomonas maltophilia ANNb, and Bacillus sp. ANNa, respectively. C aquifrigidense FANN2 maximally produced keratinase (1460.90 ± 26.99 U/mL) at 72 h of incubation under optimal process conditions of pH (6), inoculum side (5%; v/v), temperature (30°C), and chicken feather (25 g/L). The feather hydrolysate showed a protein value of 67.54%, with a relative abundance of arginine (2.84%), serine (3.14%), aspartic acid (3.33%), glutamic acid (3.73%), and glycine (2.81%). C. aquifrigidense FANN2 yielded high keratinase titre and dismembered chicken feathers into amino acids-rich hydrolysate, highlighting its significance in the beneficiation of recalcitrant keratinous wastes into dietary proteins as potential livestock feed supplements.

Purification and properties of a keratinolytic metalloprotease from Microbacterium sp.

AIMS: This study was developed to purify and to characterize a keratinolytic protease from the bacterium Microbacterium sp. strain kr10. METHODS AND RESULTS: Enzyme purification was carried out by sequential liquid chromatography on Sephadex G-100 and Q-Sepharose columns. The purification was about 255-fold, with a yield of 34%, as determined with azocasein as substrate. The molecular weight of the enzyme was estimated as 42,000 Da by SDS-PAGE. The enzyme had pH and temperature optima of 7.5 and 50 degrees C respectively. This keratinase was inhibited by EDTA and 1,10-phenanthroline, and analysis of metal content indicates that Zn(2+) and Mg(2+) are present. A 2(2) factorial design was developed to investigate the effect of keratinase and mercaptoacetate concentration on feather keratinolysis. Statistical analysis showed that both variables have a significant effect on hydrolysis of keratin. CONCLUSIONS: A new keratinase produced by Microbacterium sp. was purified and characterized. SIGNIFICANCE AND IMPACT OF THE STUDY: This keratinolytic enzyme offers an interesting potential for the hydrolysis of keratin wastes to be used as feed supplement or bioconversion to added-value products.

Degradation of chicken feathers by Chrysosporium georgiae.

Using a baiting technique, Chrysosporium georgiae was isolated from chicken feathers. Twenty-eight different fungal isolates were evaluated for their ability to produce keratinase enzymes using a keratin-salt agar medium containing either white chicken feathers or a prepared feather keratin suspension (KS). The Chrysosporium species were able to use keratin and grow at different rates. Chrysosporium georgiae completely degraded the added keratin after 9 days of incubation. Degradation of feathers by C. georgiae was affected by several cultural factors. Highest keratinolytic activity occurred after 3 weeks of incubation at 6 and 8 pH at 30 degrees C. Chrysosporium georgiae was able to degrade white chicken feathers, whereas bovine and human hair and sheep wool were not degraded and did not support fungal growth. Addition of 1% glucose to the medium containing keratin improved fungal growth and increased enzyme production. Higher keratin degradation resulted in high SH accumulation and the utilization of the carbohydrate carbon in the medium resulted in high keto-acid accumulation but decreased ammonia accumulation. Supplementation of the keratin-salt medium with minerals such as NH4Cl and MgSO4 slightly increased mycelial growth, but decreased production of extracellular keratinase. Keratinase enzymes were very poorly produced in the absence of keratin, indicating its inducible nature. Analysis of endocellular keratinases in the mycelial homogenate indicated higher activity of intracellular keratinase as compared to the extracellular enzyme in culture filtrates. Chrysosporium georgiae was the most superior for keratinase production among the Chrysosporium species tested in the presence or absence of glucose. It produced more of the intracellular enzymes than the exocellular ones.

The isolation of salmonellae from poultry environmental samples by several enrichment procedures using plating media with and without novobiocin.

A two-part study was conducted to examine the efficacy of several enrichment-broth techniques and of plating media for detecting salmonellae from poultry environmental samples. The data are reported on pooled samples collected from five poultry houses. The samples were cultured for salmonellae, using up to four different enrichment procedures and employing plating media with and without novobiocin. The primary enrichment-broth procedures were: 1) buffered peptone water preenrichment to Hajna's tetrathionate (TT) broth; and 2) direct inoculation in TT broth. The delayed secondary-enrichment procedure involved prolonged incubation at room temperature and transfer of the primary broths. The plating media consisted of: 1) xylose lysine desoxycholate agar (XLD); 2) xylose lysine desoxycholate agar containing 15 or 20 micrograms per mL of novobiocin (XLDN); 3) brilliant green sulfapyridine agar (BGSP); and 4) brillant green agar containing 20 micrograms per mL of novobiocin (BGN). Of the 94 Salmonella-positive recoveries from the enrichment broths in which complete comparisons could be made, an average of 75% were recovered from the primary enrichment broths and an average of 86% were recovered from the delayed secondary-enrichment broths. Of the 254 Salmonella-positive isolations in which complete comparisons could be made, an average of 65% were isolated on the plating media without novobiocin and an average of 97% were isolated on the plating media containing novobiocin. Overall, the delayed secondary enrichment and enteric plates supplemented with novobiocin significantly improved Salmonella detection from the farm environmental samples.