M Kind Of Chinese Traditional Medicine Enzyme Glucose Injection Process Of Legal System Optimization And Microbial Growth Differences

Objective:This study use the orthogonal design to optimize the process of Broken rice, Rice, Corn enzymatic hydrolysis glucose injection, which focused on some important steps, including,liquefaction and saccharification, anion and cation exchange resin. And through the whole process of microbial detection including the total number of colonies, mold yeast count and metabolite HPLC detection to build microbial growth state and metabolites trend.Methods:In this study,the preferred optimum process using orthogonal design and analysis of variance, DE value, glucose purity as the standard to measure liquefaction-saccharification, anion and cation exchange.Pharmacopoeia2010edition Appendix microbial limit test as the main method.Reference the national security standard GB47892-2010and GB478915-2010to detect the total number of colonies and fungal yeast colony numbers. The use of high performance liquid chromatography to establish the metabolite fingerprintingof the optimal process sample,calculate the similarity of the atlas by2004A version of the traditional medicine fingerprint similarity evaluation system.Results:1.Broken Rice, Rice, Corn optimal process:thermostable alpha-amylase dosage60u.g-1、60u.g-1、30u.g-1; Liquefaction time:40min.40min.30min; Saccharifying enzyme:150u.g-1、120u.g-1、120u.g-1; Pullulanase:0.30apu.g-1、0.25apu.g-1、0.25apu.g-1; Saccharification time48h、48h、60h; Anionic column: three groups, two groups, three groups; The ion resin loaded for8g (+)/9g (-); pH is4.5(+)/10(-), the flow rate are lOmL/min. 2.The total number of colonies of raw materials and process:raw materials(broken rice、com、rice1.74×106、1.73×105、1.46×105)→rinsing (2.72×103、1.95×103、1.53×103)→liquefaction(3.01×102、0.79×102、0.82×102)→saccharification (4.27×102、1.38×102、0.92×102)→activated carbon bleaching (0.1×102、×0.1×102、0.1×102)→cation-anion column (...0...)→pin activated carbon (...0...)→crystallization (...0...);The mildew and yeast colony count results of raw materials and the whole process:Raw broken rice, com, rice, sequentially (mildew4.29×104yeast6.89×104)(6.26×103、8.90×103)(8.84×102、1.08×103)→rinsing(6.01×102、7.35×102)(1.01×102、2.36×102)→liquefaction (0、1.46×102)(0、0.45×102)(0、0.33×102)→saccharification(0.2.16×102)(0、0.56×102)(0、0.65×102)→activated carbon bleaching (0、0.03×102)(0、0.02×102)(0、0.03×102)→cation-anion column (...0...)→pin activated carbon (...0...)→crystallization (...0...);By preliminary isolation and identification Mucor, Aspergillus flavus and Aspergillus niger, Penicillium and yeast present in the feed.The metabolites’ fingerprint similarity analysis the results:the broken rice similarity range0.01-0.99, rice similarity range0.024-0.922, corn similarity range0-0.928, The overall large span illustrate process have a certain role to the removal of metabolites.Conclusion:The optimization method is simple and feasible. Though the microbial status tracking and detection of parallel process,to clarify the state of growth of the microorganisms in production.from a macro point to establish the system of microbial cells and metabolites in the whole process,so that the microbial growth status and changes in Metabolites are clarified,from the perspective of microbial affirmed each impurity links’role.