茁霉多糖5L扩大发酵液提取及脱色工艺研究

发布时间：2018-06-01 17:56

  本文选题：茁霉多糖 + 扩大发酵　； 参考：《甘肃农业大学》2012年硕士论文

【摘要】：本实验以出芽短梗霉诱变菌种D1-11为出发菌，制备茁霉多糖发酵液，在小试（100ml）发酵工艺的基础上，设计了5L扩大发酵生产工艺方案，分别研究了接种量和转速对扩大发酵的影响，对比研究了扩大发酵和小试发酵液在生物量、多糖产量、蛋白含量及色素等各项指标上的差异；研究粉末活性炭法，硅藻土法，有机溶剂法，双氧水法等对茁霉多糖发酵液脱色效果，以脱色率和糖损耗率为指标，比较几种脱色方法的单因素实验结果，选择出较适宜的脱色方法，再通过响应面设计，研究确定该方法的最佳工艺参数；研究酶-NaCl-醇法，微波-碱水法以及超声波-盐水法等三种多糖提取方法，通过单因素及响应面数据分析得出最优提取方法及工艺。本实验主要研究结果如下： （1）扩大发酵生产茁霉多糖工艺条件为：装液量3.5L，接种量12%，初始pH6.5，温度30℃，发酵时间5d，在此工艺下制得的发酵液中多糖含量达到3.66g/L。 （2）硅藻土法最大脱色率87.43%，糖损耗率13.94%；有机溶剂法脱色最大脱色率69.22%，糖损耗率18.46%；H2O2法最大脱色率80.61%，糖损耗率18.24%；根据响应面结果，粉末活性炭法脱色最优工艺参数为pH6.3，脱色时间42min，脱色温度48℃，酶添加量1%，得出最大脱色率为88.58%，几种脱色方法中，粉末活性炭脱色率最高，糖损耗最小13.59%，简单快捷，成本低，适合工业化生产。 （3）酶-NaCl-醇法提取的最佳工艺条件是pH4，酶用量1.5%，提取温度55℃，NaCl用量1%，得出最大提取率为64.16%；微波辅助法提取的最佳工艺条件是碱水浓度0.5mol/L，，微波功率P40，提取温度70℃，微波时间90s，得出最大提取率为78.66%；超声波辅助法提取的最佳工艺条件是超声功率40%，超声时间10min，提取温度70℃，盐水浓度0.6mol/L，得出最大提取率为79.43%；提取率最高的是超声波辅助盐水法，其次是微波辅助碱水法，酶-NaCl-醇法最低，根据响应面实验结果分析，超声波-盐水法提取率最高，综合考虑，超声波辅助盐水提取法最优。
[Abstract]:In this experiment, the mutant strain D1-11 was used as the starting strain to prepare the fermentation broth of pullulan polysaccharide. On the basis of the experiment, the 5L expanded fermentation process was designed, and the effects of inoculation amount and rotational speed on the expanded fermentation were studied, respectively. The differences of biomass, polysaccharide yield, protein content and pigment between expanded fermentation and small scale fermentation broth were studied, including powder activated carbon method, diatomite method, organic solvent method, etc. The decolorization effect of hydrogen peroxide method on the fermentation broth of pullulan was studied. The decolorization rate and sugar loss rate were taken as indexes, the results of single factor experiments of several decolorization methods were compared, the more suitable decolorization method was selected, and then the response surface design was adopted. Three extraction methods of polysaccharides, enzyme NaCl-alcohol method, microwave-alkali water method and ultrasonic salt-water method, were studied. The optimum extraction method and process were obtained by single factor and response surface data analysis. The main results of this experiment are as follows: 1) the conditions of expanded fermentation were as follows: the amount of liquid was 3.5L, the inoculation amount was 12g, the initial pH was 6.5, the temperature was 30 鈩

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