Study On The Fermentation Production And Application Of Fish Attractant Acetoin

Acetoin is a special spice substance with a creamy aroma.As an important platform compound,has also been widely concerned and used in many fields.In recent years,the use of microbial fermentation to produce acetoin becomes more and more popular.Bacillus subtilis is probiotics,it can produce acetoin?2,3-butanediol,and the production processes are safe and non-polluting.In this study,the Bacillus subtilis168 is the metabolic study of the bacteria,the process is safety requirements.Optimizes its medium composition about fermentation of acetoin,separation extraction conditions and other optimization,and finally get the acetoin pure products.Sprayed the acetoin in homemade zebrafish feed,observe the effect of zebrafish seduction,and compared with the market common seduction.The results show that the effect of acetoin seduction is effective and it can be used as a seduction agent in production.The main results are as follows:1?Through the High Performance Liquid Chromatography?HPLC?detection to make the main fermentation products of acetoin and by-products 2,3-butanediol standard curve,the standard curve of the relevant R² are 0.9997 and 0.9998,respectively,indicating that feasibility and linear relationship is high.2?Screening of the formula for the birth of the acetoin medium for Bacillus subtilis 168fermented acetoin medium for single-factor optimization,obtained medium components for glucose 80 g,yeast extract powder10 g,K₂HPO₄ 10 g,MnSO₄ 0.08 g,corn pulp dry powder 10 g,pure water 1000 mL.By optimizing the medium,the yield of the acetoin is 28.73 g·L^-1.3?The response surface obtains the medium components:glucose 85.58 g,yeast extract powder10 g,K₂HPO₄ 10 g,urea 5.5 g,MgSO4 1 g,MnSO₄ 0.08 g,corn pulp dry powder5 g,pure water 1000 mL.High Performance Liquid Chromatography detection of the acetoin content increased to 34.50 g·L^-1,and the recovery rate is 90.7%.4?The separation of the metabolites of Bacillus subtilis 168 of the separation and extraction.Take fermentation liquid centrifugation,rotary evaporator swing speed of 60 rpm,water bath temperature 45?,pressure 45 hpa under the condition of rotation concentration 10 times,methanol 1:1 extraction three times,extraction on the rotary evaporator swing at the swing speed of 60 rpm,water bath temperature 45?,pressure 100 hpa,concentrated to 3 mL.Through the decompression of silicone column tomography,the use of different fluid elution,can be the product of the eye of the acetoin completely eluded,and contains a significant reduction in hetero-peaks.High Performance Liquid Chromatography preparation collects the acetoin,and makes pure products without hybrid peaks by decompression spin-steaming.5?Analysis of the sequencing of the transcription of Bacillus subtilis168 under low-carbon stress.The result shows that,compared with the high carbon group,the genes?-acetyl lactic acid decarboxylase,?-acetyl lactic acid synthetase,L-alanine dehydrogenase,pyruvate kinase related genes were down-regulation.The action of 2,3-butanediol dehydrogenase is up-regulation.6?Wither Bacillus subtilis168 metabolites acetoin as a seduction agent effect assessment test.The result shows that the advantage of acetoin as a seduction agent is obvious and the best addition of acetoin is 0.6%.Compared with the common feeding agents in the market,the process of fermentation is simple and easy to operate,safe and non-polluting,the fermentation cycle is short,the product is easy to obtain,and the effect is remarkable as the inducing agent.Which shows that the acetoin as a seduction agent production plan is feasible,the effect is significant.Above all,Bacillus subtilis 168 fermentation process of acetoin are safe and non-pollution,The fermentation materials are cheap and easy.The purity and recovery rate of High Performance Liquid Chromatography method to collect the acetoin product is high.Acetoin as attractant is remarkable and 0.6%for the best content,it can be used as attractant applied in aquaculture feed additives,provide the basis for developing new type of aquatic animals attractant.