The invention discloses a higher-substrate-specificity keratinase mutant and a preparation method thereof, belonging to the field of enzyme engineering. Analysis is performed to obtain the optimal mutant site to perform site-directed mutagenesis, thereby obtaining the excellent mutant with higher keratinase enzyme activity and substrate specificity. The keratinase and mutant thereof can effectively hydrolyze feather, wool and other non-water-soluble keratin substrates, and can be used for leather textile industry and feed industry.

Higher-substrate-specificity keratinase mutant and preparation method thereof

The invention discloses keratinase with improved thermal stability and specific activity as well as a preparation method and application thereof, and belongs to the field of genetic engineering. The keratinase having wide industrial application prospect is obtained by mutating a leading peptide cleavage site of keratin and transforming the N-terminal of the keratin. The t1/2 of the keratinase at the temperature of 60 DEG C is 20 minutes and is increased by nearly two times in comparison with that of wild type keratinase. Meanwhile, the specific activity of a keratinase mutant is increased by nearly 50% in comparison with that of the wild type keratinase, so that a good foundation is laid for the application of the keratinase.

Keratinase with improved thermal stability and specific activity as well as preparation method and application thereof

The invention discloses a keratinase mutant with improved thermal stability and a preparation method thereof, and belongs to the field of enzyme engineering. Thermal stability and substrate specificity of the keratinase are improved by interchanging N ends or C ends of two different keratinases with higher homology from the same bacterial strain. The keratinase and the mutant thereof can effectively hydrolyze water-insoluble keratinase substrates such as feathers, wools, and the like, and can be used for leather spinning industry and feed industry.

Keratinase mutant with improved thermal stability and preparation method thereof

The invention relates to the technical field of microorganisms, and particularly discloses Bacillus subtilis 3-2 and application thereof The Bacillus subtilis 3-2 is collected in China Center for Typical Culture Collection (CCTCC) On November 8, 2013, with the collection number of CCTCC M2013555 The 16SrDNA nucleotide sequence of the Bacillus subtilis 3-2 is shown as SEQIDNO: 1 The Bacillus subtilis 3-2 can generate keratinase; determination shows that after the Bacillus subtilis 3-2 is cultured for 3 days, the activity of the keratinase reaches 4643 U/ml, so that Bacillus subtilis 3-2 R1 can be used for generating the keratinase Therefore, the Bacillus subtilis 3-2 has important values for researching and utilizing of the keratinase

Bacillus subtilis 3-2 and application thereof

The invention provides a keratinase. Amino acid sequence of the keratinase is an amino acid sequence as shown in SEQ ID NO.2 or is an amino acid sequence which is obtained through deletion, mutation and recombination based on SEQ ID NO.2 and which has a keratinase activity. The invention also provides a keratinase-coding gene. Nucleotide sequence of the gene is a nucleotide sequence as shown in SEQ ID NO.1 or is a nucleotide sequence which is artificially designed according to deletion, mutation and recombination of keratinase amino acid. The keratinase provided by the invention can effectively hydrolyze water-insoluble keratin substrates such as feather, wool and the like, and can be used in leather textile industry and feed industry.

Keratinase and its coding gene

The invention discloses keratinase with improved thermal stability and application thereof, and belongs to the field of genetic engineering. The keratinase (ker) gene of bacillus licheniformis BBE11-1 is subjected to site-directed mutation by using a site-directed mutation technology and is cloned and linked to a bacillus subtilis expression vector pMA 0911; and bacillus subtilis WB600 is transformed, and a bacillus subtilis WB600-pMA 0911-kerTB capable of producing the keratinase with better thermal stability can be obtained through purification tests. The keratinase expressed by a strain at T1/2 at 60 DEG C lasts for 78min, which is improved nearby 9 folds compared with wild type keratinase at T1/2. The keratinase with improved thermal stability lays an excellent foundation for the application of keratinase.

Protease with improved thermal stability as well as construction method and application thereof

The invention discloses stenotrophomonas maltophilia for generating keratinase and an application of stenotrophomonas maltophilia. A strain for generating keratinase can efficiently degrade feathers within 24 hours, the activity of the keratinase subjected to fermentation and primary optimization can reach 150U/mL. Multiple kinds of amino acids can be produced after the degraded feathers are subjected to amino acid analysis, and the degraded feathers can replace grain crops such as soybeans and the like to be used as a main nitrogen source of feeds. Meanwhile, a fermenting enzyme solution of the stenotrophomonas maltophilia has a better treatment effect on wool. The stenotrophomonas maltophilia for generating the keratinase, screened in the invention, has better application prospects in the industries of feeds and textiles.

Stenotrophomonas maltophilia for generating keratinase and application of stenotrophomonas maltophilia

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