Study On Starch And Protein Degradation Of Strips By Bacillus Ubtilis

Some strains could be isolated from Zimbabwe tobacco and we identified amylase fungusprotease highest strains. We identified the strains by physiological and biochemical experimentsand16SrRNA sequence analysis. the16SrRNA similarities of these strains（Protease-producing strainBDT-3and Amylase-producing strain BAWS-1） and bacillus subtilis all reached levels above99%,and we identified these strains as bacillus subtilis. Protease enzyme activity for52.10±0.13U/mL.Amylase enzyme activity for13.69±0.07U/mL. The two strains may play an important role indegradation of the protein and starch in Zimbabwe tobacco.In order to improve the fermentation enzyme production ability, optimal choose the twofermentation, BDT-3medium: corn flour4.544%, peptone1.43%, starch0.178%,（NH4）2SO40.4%, NaCl1%, NH4Cl0.13%, CaCl20.27%, Na2HPO4*12H2O0.4%; BAWS-1medium: beefextract5.5%, peptone2.0%, CaCl20.101%, KH2PO40.3%, MgSO40.5%, NaCl1.458%, K2HPO40.0596%.The author investigates the enzymology character of BDT-3strain and BAWS-1strain. BDT-3:the enzyme activity in pH7.5when reach maximum in3hours, can maintain the strains of theintact enzyme Fe²⁺live, metal ions of protease have larger activation function. BAWS-1:In pH for7.0with have the largest enzyme, in within3hours, this enzyme can keep more than80%of theenzyme, Ca²⁺and Mn²⁺of amylase have larger stimulative effect.Adopt L9（34） orthogonal design schemes are discussed for fermentation of coarse tobacco fluidadditives, enzyme degradation temperature, relative humidity, and duration of the tobacco leafprotein and starch content of influence. The influence of protein degradation significant order: time>humidity> fermented additives> temperature, optimal combination for: the processing time72h,relative humidity80%, enzyme bath additives for40%, temperature for45℃. According to the bestcombination experiment on tobacco protein degradation rate can reach9.16%. The influence ofstarch biodegradation significant order: temperature> fermented additives> humidity> time,optimal combination for: the processing time96h, enzyme liquid add than is40%, temperature for40℃, relative humidity for60%. According to the best combination experiment, the tobacco leaf inthe degradation of amylum rate can reach haemorrhage. Mixed add protease and amylase explainafter test, protein composition, protein content decreased7.93%，starch content decreased7.93%;And the reducing sugar content in tobacco increased2.14%, the amount of nitrogen contentincreased by0.9%.