Study On Extraction,Purification And Characterization Of Bacillus Subtilis Strain C3-41

The current research of laccase is mainly concentrated in the laccase from white rot fungus,but with the increase of bacterial laccase results showed that,compared with the laccase from white rot fungi,bacterial laccase has high temperature resistance,alkali resistance,rich in resources,so it is easy to realize the industrialized production of the advantages of expression in industrial host organisms,Spore laccase produced by Bacillus has high optimum temperature,high thermal stability,wide pH range and other characteristics,which has wide application prospect.(1)Bacillus C3-41 laccase properties preliminary research results show that in several laccase detection system,3mL buffer system composed of 1.8mL sodium hydrogen pHospHate citric acid buffer,0.2mL 0.6mM/LABTS and 1mL laccase samples atpH4 is the best detection systemfor spore laccase.the laccase activity and protein content in three different samples of the comparison shows that the main parts of Bacillus spHaericus C3-41 producing laccase is bacillus,and the spore laccase was endoenzyme.(2)Characterization of Bacillus C3-41 spore laccase showed that pH has great influence on spore laccase activity,the maximum activity of laccase was pH4;temperature stability of the spore laccase enzyme activity is good,at a high temperature is maintained at a high level,the optimum temperature of 50?;when the system of metal ions in the final concentration of 0.2mmol/L,Fe2+ and Fe3+ have inhibitory effect on the activity of laccase,Cu2+,Zn2+,Ba2+,Ca2+ have a great effect on the activity of laccase,which Ca2+ the most obvious promoting effect;add working DTT,?-Mercaptoethanol,EDTA,PMSF were added to the enzyme solution,DTT,and?-Mercaptoethanol were found to inhibit the enzyme activity,and the inhibitory effect of PMSF was the weakest.(3)Study on decolorization characteristics of Bacillus C3-41 produced by Bacillus laccase of reactive brilliant blue KN-R and reactive brilliant red-3B,malachite green,three kinds of dyes,When the dye concentration is not more than 2g/L,The decolorization rate of reactive brilliant blue was increased with the increase of dye concentration,the decolorization rate was 87.8% when the dye concentration was the highest;The decolorization rate of reactive red dye concentration decreases with theincrease of dye concentration,the decolorization rate was 72.8% when the dye concentration was the highest;the decolorization rate of malachite green dye concentration with little change,except when the concentration is 2g/L the decolorization rate was 82.2%,the decolorization rate was more than 86% other concentration.(4)Compared with purified laccase,spore laccase had higher temperature tolerance and pH toleranceThe results of this study show that the Bacillus subtilis C3-41 spore laccase has good value in the decolorization of dyes,the enzymatic properties of laccase and the cost reduction in engineering application.