There is an increasing demand for cost-effective and ecologically-friendly methods for valorization of poultry feather waste, in which keratinolytic bacteria present a great potential. Feather-degrading bacteria were isolated from living poultry and a single strain, identified as Kocuria rhizophila p3-3, exhibited significant keratinolytic properties. The bacterial strain effectively degraded up to 52% of chicken feathers during 4 days of culture at 25 °C. Zymographic analysis revealed the presence of two dominating proteolytic enzymes in the culture fluid. Culture conditions were optimized in order to maximize the liberation of soluble proteins and free amino acids. A two-step procedure was used, comprising a Plackett–Burman screening design, followed by a Box–Behnken design. Concentration of feather substrate, MgSO4 and KH2PO4 were the most influential parameters for the accumulation of soluble proteins in culture K. rhizophila p3-3, while feathers and MgSO4 also affected the concentration of amino acids. The resultant raw hydrolysate supernatant, prior to and after additional treatments, was rich in phenylalanine, histidine, arginine and aspartic acid. Additionally the hydrolysate exhibited radical-scavenging activity and ferric reducing power.Electronic supplementary materialThe online version of this article (10.1186/s13568-018-0538-y) contains supplementary material, which is available to authorized users.
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.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.