2018 ; Vol.28-2: 314~322
|Author||Zhoufeng He, Rong Sun, Zizhong Tang, Tongliang Bu, Qi Wu, Chenlei Li, Hui Chen|
|Place of duty||College of Life Science, Sichuan Agricultural University, Ya’an 625014, P.R. China|
|Title||Biodegradation of Feather Waste Keratin by the Keratin-Degrading Strain Bacillus subtilis 8|
J. Microbiol. Biotechnol.2018 ;
|Abstract||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 L-cysteic
acid were detected in the fermented liquid. In addition, four enzymes (gammaglutamyltranspeptidase,
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.|
|Key_word||Bacillus subtilis, keratin, purification, degradation mechanism, prokaryotic expression|
Ramnani P, Singh R, Gupta R. 2005. Keratinolytic potential of Bacillus licheniformis RG1: structural and biochemical mechanism of feather degradation. Can. J. Microbiol. 51: 191196.
Feng J. 1995. Development and utilization of feather meal protein feed. Feed Ind. 1995: 42-43.
Gradisar H, Friedrich J, Križaj I, Jerala R. 2005. Similarities and specificities of fungal keratinolytic proteases: comparison of keratinases of Paecilomyces marquandii and Doratomyces microsporus to some known proteases. Appl. Environ. Microbiol. 71: 3420-3426.
Lopes FC, Silva LADE, Tichota DM, Daroit DJ, Velho RV, Pereira JQ, et al. 2011. Production of proteolytic enzymes by a keratin-degrading Aspergillus niger. Enzyme Res. 2011:487093.
Huang Y, Busk PK, Lange L. 2015. Production and characterization of keratinolytic proteases produced by Onygena corvina. Fungal Genom. Biol. 4: 119.
Ignatova Z, Gousterova A, Spassov G, Nedkov P. 1999. Isolation and partial characterisation of extracellular keratinase from a wool degrading thermophilic actinomycete strain Thermoactinomyces candidus. Can. J. Microbiol. 45: 217-222.
Jayalakshmi T, Krishnamoorthy P, Ramesh G, Sivamani P. 2010. Isolation and screening of a feather-degrading keratinolytic actinomycetes from Actinomyces sp. J. A m. S ci. 6: 45-48.
Matikevičienė V, Grigiškis S, Levišauskas D, Sirvydytė K, Dižavičienė O, Masiliūnienė D, et al. 2013. Optimization of keratinase production by Actinomyces fradiae 119 and its applicaion in degradation of keratin containing wastes. Proceedings of the 8th International Scientific and Practical Conference. Environment Technology Resources 1: 294-300.
Williams CM, Richter C, Mackenzie J, Shih JC. 1990. Isolation, identification, and characterization of a featherdegrading bacterium. Appl. Environ. Microbiol. 56: 1509-1515.
Joshi S G, T ejashwini M M, Rev ati N, Sridevi R, R oma D . 2007. Isolation, identification and characterization of a feather degrading bacterium. Int. J. Poultry Sci. 6: 689-693.
Sudhir MR. 2016. Isolation and identification of feather degrading bacteria from poultry farm soil and characterization of enzyme. PhD Thesis. Narsee Monjee Institute of Management Studies, India.
Mazotto AM, Coelho RRR, Cedrola SML, De Lima MF, Couri S, De Souza EP, et al. 2011. Keratinous production by three Bacillus spp. using feather meal and whole feather as substrate in a submerged fermentation. Enzyme Res. 2011: 1-7.
Prakash P, Jayalakshmi SK, Sreeramulu K. 2010. Purification and characterization of extreme alkaline, thermostable keratinase, and keratin disulfide reductase produced by Bacillus halodurans PPKS-2. Appl. Microbiol. Biotechnol. 87:625-633.
Qi Z. 2012. Research on keratin-degrading bacteria with high disulfide bond-reducing capacity. Master degree thesis. Chinese Academy of Agricultural Sciences, Beijing, China.
Neilson KA, Ali NA, Muralidharan S, Mirzaei M, Mariani M, Assadourian G, et al. 2011. Less label, more free: approaches in label-free quantitative mass spectrometry. Proteomics 11: 535-553.
Vermeij P, Kertesz MA. 1999. Pathways of assimilative sulfur metabolism in Pseudomonas putida. J. Bacteriol. 181:5833-5837.
Pillai P, Archana G. 2008. Hide depilation and feather disintegration studies with keratinolytic serine protease from a novel Bacillus subtilis isolate. Appl. Microbiol. Biotechnol. 78:643-650.
Tiwary E, Gupta R. 2010. Subtilisin-γ-glutamyl transpeptidase:a novel combination as ungual enhancer for prospective topical application. J. Pharm. Sci. 99: 4866-4873.
Liu Q, Zhang T, Song N, Li Q, Wang Z, Zhang X, et al. 2014. Purification and characterization of four key enzymes from a feather-degrading Bacillus subtilis from the gut of tarantula Chilobrachys guangxiensis. Int. Biodeterior. Biodegradation 96: 26-32.
Wang J, Zhu S, Xu C. 2002. Biological Chemistry, 3rd Ed. Higher Education Press, Beijing, China.
Kunert J, Stransky Z. 1988. Thiosulfate production from cystine by the keratinolytic prokaryote Streptomyces fradiae. Arch. Microbiol. 150: 600-601.
Suh HJ, Lee HK. 2001. Characterization of a keratinolytic serine protease from Bacillus subtilis KS-1. J. Protein Chem. 20: 165-169.
Strauch KL, Miller CG. 1983. Isolation and characterization Salmonella typhimurium mutants lacking a tripeptidase (peptidase T). J. Bacteriol. 154: 763-771.
Orlowski M, Meister A. 1970. The γ-glutamyl cycle: a possible transport system for amino acids. Proc. Natl. Acad. Sci. USA 67: 1248-1255.
Sharma R, Gupta R. 2012. Coupled action of γ-glutamyl transpeptidase-glutathione and keratinase effectively degrades feather keratin and surrogate prion protein, Sup 35NM. Bioresour. Technol. 120: 314-317.
Clausen T, Huber R, Prade L, Wahl MC, Messerschmidt A. 1998. Crystal structure of Escherichia coli cystathionine-γsynthase at 1.5 Å resolution. EMBO J. 17: 6827-6838.
Wallsgrove RM, Lea PJ, Miflin BJ. 1983. Intracellular localization of aspartate kinase and the enzymes of threonine and methionine biosynthesis in green leaves. Plant Physiol. 71: 780-784.
Tu G, Sun Y. 1998. Biochemical mechanism of the degradation of keratin in Streptomyces. Acta Agric. Univ. Jiangxiensis 20: 164-169.