2017 ; Vol.27-12: 2190~2198
|Author||Lin Wang, Yuting Qian, Yun Cao, Ying Huang, Zhizhou Chang, Hongying Huang|
|Place of duty||Circular Agriculture Research Center, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, P.R. China,Nanjing Institute of Agricultural Sciences in Jiangsu Hilly Area, Nanjing 210046, P.R. China|
|Title||Production and Characterization of Keratinolytic Proteases by a Chicken Feather-Degrading Thermophilic Strain, Thermoactinomyces sp. YT06|
J. Microbiol. Biotechnol.2017 ;
|Abstract||Thermoactinomyces sp. strain YT06 was isolated from poultry compost and observed to degrade
integral chicken feathers completely at 60°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 NaNO3 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°C). It was inhibited by serine protease inhibitors and most
metal ions; however, it could be stimulated by Mn2+ and the surfactant Tween-20. A reductive
agent (β-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.|
|Key_word||Thermophile, keratinase, feather degradation, characterization|
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