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The kanamycin A B fractionation and purification of new technologies

Author: LiZhongQin
Tutor: GuoYangHao
School: Fuzhou University
Course: Biochemistry and Molecular Biology
Keywords: Kanamycin Multivariate dissociation constant An ion exchange resin Competitive adsorption Crystal liquor Separation and purification Chromatographic separation
CLC: TQ465
Type: Master's thesis
Year: 2002
Downloads: 278
Quote: 0
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Abstract


The most domestic kanamycin production process 732 to # strongly acidic cation exchange resin static adsorption, dynamic elution process. Defects in the process, the work kanamycin A, B component separation and purification of new technology. Kanamycin A, B are the yuan weak base water-soluble compounds. This work is piecewise linear regression analysis based on the principle of buffer system, the algorithm of simultaneous equations and Rosenbrock algorithm based on the acid-base titration formula processing kanamycin A, B potential titration data were used. The estimated from kanamycin A diverse agreement from the value of the dissociation constant reported in the literature. This thesis is the first comprehensive reports the kanamycin B dissociation constants: kanamycin A, B ionization simulation under different pH the valence distribution coefficient curve (pH-distribution coefficient curve). Screened eight different types of strongly acidic and weakly acidic cation exchange resin as raw material, the fermentation liquid. Macroporous weak acid resin having a D-186 kanamycin high adsorption total less impurity adsorption and card B component enrichment rate characteristics. To optimize the use of orthogonal experimental design method on column adsorption preconcentration conditions (pH 7.0, flow rate 2.0V/V/h) and impurity elution conditions (concentration of ammonium sulfate solution 1.0% (v / v), pH7.5, flow rate 1.25V/V/h). D-186 resin and 732 the resin four-poster series dynamic adsorption elution process can effectively enriched the B component of the fermentation broth kanamycin. When the fermentation is not normal, it is possible to guarantee the quality of the products of the card alkali. D-186 resin is separated from the crystallization mother liquor kanamycin A, B component. Optimize the use of orthogonal experimental design method on the column adsorption preconcentration conditions (concentration of the mother liquor diluted 12,600 u / ml, pH7.0 velocity 1.5VN / h) and impurities elution conditions (concentration of ammonium sulfate solution 1.5% (V / V) , pH7.5, the flow rate 1.75V/V/h). The dynamic adsorption elution process route three columns in series, so that kanamycin A, B component of the total recovery of 80% and 93%, respectively, the impurity removal string of up to 99%. (Concentration 30 to 350 000 u / ml kanamycin B component 枓 enriched from the fermentation broth and crystal liquor was concentrated mixed in proportion to card B component content of about 35%), using anion exchange resin H401 column chromatography. Optimize the operating conditions of the chromatographic separation: ① particle diameter of 80 mesh (0.20mm), the degree of crosslinking of 2.5 H401 anion exchange resin: The amount of ② on the column 1.05ug/ml: ③ eluent NH4OH concentration of 0.6% (V / V ): ④ elution flow rate 0.5ml/min. The card A purity of the product reached 99.5%, the card B purity of the product is more than 90%.

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