Abstract
Incubation parameters used for the creation of a protein lysate from enzymatically degraded waste feathers using crude keratinase produced by the Laceyella sacchari strain YNDH were optimized using the Response Surface Methodology (RSM); amino acids quantification was also estimated. The optimization elevated the total protein to 2089.5 µg/ml through the application of the following optimal conditions: a time of 20.2 h, a feather concentration (conc.) of 3 g%, a keratinase activity of 24.5 U/100 ml, a pH of 10, and a cultivation temperature of 50 °C. The produced Feather Protein Lysate (FPL) was found to be enriched with essential and rare amino acids. Additionally, this YNDH enzyme group was partially purified, and some of its characteristics were studied. Crude enzymes were first concentrated with an Amicon Ultra 10-k centrifugal filter, and then concentrated proteins were applied to a "Q FF" strong anion column chromatography. The partially purified enzyme has an estimated molecular masses ranging from 6 to 10 kDa. The maximum enzyme activity was observed at 70 °C and for a pH of 10.4. Most characteristics of this protease/keratinase group were found to be nearly the same when the activity was measured with both casein and keratin-azure as substrates, suggesting that these three protein bands work together in order to degrade the keratin macromolecule. Interestingly, the keratinolytic activity of this group was not inhibited by ethylenediamine tetraacetic acid (EDTA), phenylmethanesulfonyl fluoride (PMSF), or iron-caused activation, indicating the presence of a mixed serine–metallo enzyme type.
Highlights
In the poultry industry, feathers are a significant byproduct since they account for 5–7% of the body weight of the chicken
All cultures were conducted in triplicate, and the obtained results were averaged
The present study clearly shows that feather degradation by keratinase provided by the Laceyella sacchari YNDH strain is cost-effective and a viable method to efficiently utilize disregarded feather waste
Summary
Feathers are a significant byproduct since they account for 5–7% of the body weight of the chicken. Several million tons of feathers are estimated to be created annually from the global poultry industry[2]. This protein source should be used in a large scale and be converted into value added product. A cost-effective alternative way consists of biodegradation of the feathers by microorganisms using keratinase. Feather biodegradation by microbes producing keratinase or by direct enzymes formed by numerous microorganisms is considered to be an environmentally friendly approach to keratin waste recycling[7,8]. This study explores a strategic cost-effective optimization protocol for the enzymatic production of FPL enriched with different amino acids from waste feathers using Laceyella sacchari YNDH keratinase. The intensive characterization of partially purified applied enzymes was explored
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