Keratinase production and keratin degradation by a mutant strain of Bacillus subtilis

• Published in: Journal of Zhejiang University. B. Science 2008-01, Vol.9 (1), p.60-67
• Main Authors: Cai, Cheng-gang, Lou, Bing-gan, Zheng, Xiao-dong
• Format: Article
• Language: English
• Subjects: Amino Acids - biosynthesis; Bacillus subtilis - genetics; Bacillus subtilis - metabolism; Biomedical and Life Sciences; Biomedicine; Biotechnology; Culture Media; Hydrogen-Ion Concentration; Keratins - metabolism; Mutation; Peptide Hydrolases - biosynthesis; 杆状菌; 角蛋白酶; 角蛋白降解; 遗传变异
• ISSN: 1673-1581; 1862-1783
• DOI: 10.1631/jzus.b061620

A new feather-degrading bacterium was isolated from a local feather waste site and identified as Bacillus subtilis based on morphological, physiochemical, and phylogenetic characteristics. Screening for mutants with elevated keratinolytic activity using N-methyl-N＇-nitro-N-nitrosoguanidine mutagenesis resulted in a mutant strain KD-N2 producing keratinolytic activity about 2.5 times that of the wild-type strain. The mutant strain produced inducible keratinase in different substrates of feathers, hair, wool and silk under submerged cultivation. Scanning electron microscopy studies showed the degradation of feathers, hair and silk by the keratinase. The optimal conditions for keratinase production include initial pH of 7.5, inoculum size of 2% （v/v）, age of inoculum of 16 h, and cultivation at 23 ℃. The maximum keratinolytic activity of KD-N2 was achieved after 30 h. Essential amino acids like threonine, valine, methionine as well as ammonia were produced when feathers were used as substrates. Strain KD-N2, therefore, shows great promise of finding potential applications in keratin hydrolysis and keratinase production.