Studying One Kind Of Bacillus J4-1Fermentation Production Of The Vanillin

Mainly existing in sweet pod of abram plants pod, the vanillin is a kind of high-grade spices whose production is the largest and application is the most wide in the world, at the same time, it can also be used in food, medicine, chemical industry and agriculture, etc. The methods producing of vanillin are mainly natural extract, chemical synthesis and biological transformation, and chemical synthesis method has been applied to industrial production. The biotransformation method includes cell engineering, enzyme catalysis and microbial fermentation method. The vanillin produced by microbial fermentation is a natural vanillin, with the smell of pure, safety and expensive. In this study, we used microbial flask fermentation method to produce vanillin and wanted to increase the vanillin yield by optimizing the production conditions, but also hope to provide a reference for the subsequent fermentation tank experiments.In this research, we have screened many strains from the soil of wu han jian min pharmaceutical factory which had a high tolerance for ferulic acid and also can convert ferulic acid into vanillin.By shaking flask fermentation, we also further screened one strain that the production of the vanillin was the highest, and named J4-1. Then, the fermentation medium compositions and fermentation conditions, the strain medium compositions and culture conditions, the substrate conditions were optimiz ed. Measuring the vanillin content by high performance liquid chromatography. according to the measured results, we tried to find out culture conditions which can make the vanillin production reached the highest level. At the same time, J4-1bacteria strain was identified by China Typical Culture Collection (CCTCC).The final screening results showed that there were103strains which can tolerate ferulic acid and40strains which can convert the ferulic acid into the vanillin. To determine the best seed of culture medium and the growth of the conditions:maltose peptone medium (maltose50g/L, peptone lOg/L, and sodium chloride5g/L), incubation temperature30℃, speed of100rpm, incubation time24h; the shaking flask production conditions for the optimum medium composition and fermentation conditions:the chaff20g/L, soybean meal3g/L, and inorganic salts (CaCl20.5g/L, K2HPO40.5g/L, MgSO4·7H2O0.5g/L, FeSO4·7H2O0.01g/L), medium initial pH=10.0,50mL of culture broth fluid volume, temperature40℃, speed of100rpm, light cultured for8days; substrate conditions:the concentration of12.5g/L trans-ferulic acid20mL. The concentration of the Vanillin in Fermentation Broth reached749.21mg/L, the substrate molar conversion rate was17.21%, while the concentration of the Vanillin kept22.84mg/L before optimization, so that the strain transformation ability after optimization was improved by32.80times, substrate molar ratio (1.94%) increased by8.87times. J4-1bacteria was identified as Bacillus fresh methanol Bacillus(Bacillus methylotrophicus).