酵母葡聚糖的研制
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摘要
本论文在现有酵母葡聚糖两种提取工艺的基础上,提出了高压—碱法提取工艺,对提取条件进行了优化、对提取物特性进行了分析。对甲酸水解制备酵母水溶性葡聚糖的工艺进行了优化,对其生物活性做了初步研究。
采用碱法、高压法和高压—碱法三种不同工艺提取酵母葡聚糖,通过对提取物的得率、蛋白质、多糖、脂类、灰分含量等进行分析,表明高压—碱法去除蛋白质效果好且多糖含量高;研究了丙酮干燥法、无水乙醇干燥法和真空冷冻干燥法对产品品质的影响,结果表明丙酮干燥法效果最好。
对酵母葡聚糖含量的测定方法进行研究。分别比较H_2SO_4和TFA在不同条件下水解酵母不溶性葡聚糖对测定结果的影响,实验表明,采用H_2SO_4进行水解优于TFA,最佳水解条件为:H_2SO_4浓度2.7mol/L、水解温度100℃、时间3h。
通过L_9(3~4)正交实验对高压—碱法提取工艺进行了优化。适宜的提取工艺条件为:碱处理浓度6%,压力0.1MPa,时间30min,酵母葡聚糖提取率为23.30%,提取物中多糖含量为91.85%。经红外光谱与纸层析分析结果表明:产品只含葡萄糖一种单糖,糖苷键为β型。
酵母葡聚糖的水不溶性制约了其在制药工业的应用。选择甲酸水解的方法制备酵母水溶性葡聚糖,采用L_9(3~4)正交设计对水解条件进行了优化,适宜的条件为:温度为80℃,时间为1.5h,甲酸用量为1:10(w/v),甲酸浓度90%。酵母水溶性葡聚糖得率为80.15%,经红外光谱分析结果表明:水解前后产品结构未发生改变。
研究了酵母水溶性葡聚糖对各种供试菌的抑菌作用。结果表明,酵母水溶性葡聚糖对细菌的抑制效果明显,对酵母菌、霉菌无抑菌效果;最小抑菌浓度分别为:大肠杆菌75mg/mL,鸡沙门氏菌37.5mg/mL,枯草芽孢杆菌37.5mg/mL,金黄色葡萄球菌37.5mg/mL。酵母水溶性葡聚糖最适抑菌pH范围为6.0～7.0,热稳定性良好。
In this paper,the high-pressure-alkali extraction technology was put forward on the basis of the two extraction technologies for yeast glucan,and the investigations were carried out on the optimum of the extraction conditions,the character of the extractive. Then the preparation technology of yeast soluble glucan was optimized,which hydrolyzed by methyl acid,and the pilot study was carried through on the biology activity of yeast soluble glucan.
The yeast glucan was extracted by three different technologies:alkali method, high-pressure method and the high-pressure-alkali method,and the yield,protein content, polysaccharide content,lipid content,ash content of the three extractions was determined. The results showed:the high-pressure-alkali method was the better method for the protein removing,and its extraction's polysaccharide content was high too.The effects of the dehydrated ethanol desiccation method,the acetone desiccation method and the vacuum freeze-drying method on the extraction were studied.The result was:the acetone desiccation method was the best way.
The method for determination of yeast glucan content was investigated.The influence of different acid hydrolysis on result of determination was compared when Yeast insoluble glucan was hydrolyzed by H_2SO_4 and TFA in different hydrolysis conditions.The results showed that H_2SO_4 was more suitable than TFA as hydrolysis acid and the best hydrolysis conditions for Yeast insoluble glucan were obtained:2.7mol/L H_2SO_4,100℃,3h.
The high-pressure-alkali extraction technology was optimized by orthogonal design. The result was that:alkali concentration 6%,pressure 0.1MPa and time 30min.The yield of yeast glucan was 23.30%and the polysaccharide content of the extraction was91.85%.The analysis results by IR and PC(Paper chromatography) showed the extraction only consisting glucose,and the extraction containingβ-glucoside.
The application of Yeast glucan in the pharmaceutical industry was restricted by its water insoluble nature,so the yeast soluble glucan was prepared by methyl acid hydrolyzing,and hydrolysis conditions was optimized by orthogonal design.The optimum conditions were:the hydrolysis temperature 80℃,time 1.5h,the addition of methyl acid 1:10 and the concentration of methyl acid 90%.The yield of yeast soluble glucan was 80.15%,the analysis results by IR showed that:the structure of the product was not changed after hydrolyzing by methyl acid.
The antimicrobial effect of the yeast soluble glucan against various microbe was studied.The results showed that the yeast soluble glucan could effectively inhibit the growth of bacteria,yet to mould and yeast,there was no effect.The minimum inhibitory concentration(MIC) of different bacterias were:Escherichia.Coli 75mg/mL,Salmonella gallinarum 37.5mg/mL,Bacillus subtilus 37.5mg/mL,Staphalococcus aureus 37.5mg/mL, and the pH range of inhibiting bacteria was 6.0～7.0.Moreover,the heat stability is good.
采用碱法、高压法和高压—碱法三种不同工艺提取酵母葡聚糖,通过对提取物的得率、蛋白质、多糖、脂类、灰分含量等进行分析,表明高压—碱法去除蛋白质效果好且多糖含量高;研究了丙酮干燥法、无水乙醇干燥法和真空冷冻干燥法对产品品质的影响,结果表明丙酮干燥法效果最好。
对酵母葡聚糖含量的测定方法进行研究。分别比较H_2SO_4和TFA在不同条件下水解酵母不溶性葡聚糖对测定结果的影响,实验表明,采用H_2SO_4进行水解优于TFA,最佳水解条件为:H_2SO_4浓度2.7mol/L、水解温度100℃、时间3h。
通过L_9(3~4)正交实验对高压—碱法提取工艺进行了优化。适宜的提取工艺条件为:碱处理浓度6%,压力0.1MPa,时间30min,酵母葡聚糖提取率为23.30%,提取物中多糖含量为91.85%。经红外光谱与纸层析分析结果表明:产品只含葡萄糖一种单糖,糖苷键为β型。
酵母葡聚糖的水不溶性制约了其在制药工业的应用。选择甲酸水解的方法制备酵母水溶性葡聚糖,采用L_9(3~4)正交设计对水解条件进行了优化,适宜的条件为:温度为80℃,时间为1.5h,甲酸用量为1:10(w/v),甲酸浓度90%。酵母水溶性葡聚糖得率为80.15%,经红外光谱分析结果表明:水解前后产品结构未发生改变。
研究了酵母水溶性葡聚糖对各种供试菌的抑菌作用。结果表明,酵母水溶性葡聚糖对细菌的抑制效果明显,对酵母菌、霉菌无抑菌效果;最小抑菌浓度分别为:大肠杆菌75mg/mL,鸡沙门氏菌37.5mg/mL,枯草芽孢杆菌37.5mg/mL,金黄色葡萄球菌37.5mg/mL。酵母水溶性葡聚糖最适抑菌pH范围为6.0～7.0,热稳定性良好。
In this paper,the high-pressure-alkali extraction technology was put forward on the basis of the two extraction technologies for yeast glucan,and the investigations were carried out on the optimum of the extraction conditions,the character of the extractive. Then the preparation technology of yeast soluble glucan was optimized,which hydrolyzed by methyl acid,and the pilot study was carried through on the biology activity of yeast soluble glucan.
The yeast glucan was extracted by three different technologies:alkali method, high-pressure method and the high-pressure-alkali method,and the yield,protein content, polysaccharide content,lipid content,ash content of the three extractions was determined. The results showed:the high-pressure-alkali method was the better method for the protein removing,and its extraction's polysaccharide content was high too.The effects of the dehydrated ethanol desiccation method,the acetone desiccation method and the vacuum freeze-drying method on the extraction were studied.The result was:the acetone desiccation method was the best way.
The method for determination of yeast glucan content was investigated.The influence of different acid hydrolysis on result of determination was compared when Yeast insoluble glucan was hydrolyzed by H_2SO_4 and TFA in different hydrolysis conditions.The results showed that H_2SO_4 was more suitable than TFA as hydrolysis acid and the best hydrolysis conditions for Yeast insoluble glucan were obtained:2.7mol/L H_2SO_4,100℃,3h.
The high-pressure-alkali extraction technology was optimized by orthogonal design. The result was that:alkali concentration 6%,pressure 0.1MPa and time 30min.The yield of yeast glucan was 23.30%and the polysaccharide content of the extraction was91.85%.The analysis results by IR and PC(Paper chromatography) showed the extraction only consisting glucose,and the extraction containingβ-glucoside.
The application of Yeast glucan in the pharmaceutical industry was restricted by its water insoluble nature,so the yeast soluble glucan was prepared by methyl acid hydrolyzing,and hydrolysis conditions was optimized by orthogonal design.The optimum conditions were:the hydrolysis temperature 80℃,time 1.5h,the addition of methyl acid 1:10 and the concentration of methyl acid 90%.The yield of yeast soluble glucan was 80.15%,the analysis results by IR showed that:the structure of the product was not changed after hydrolyzing by methyl acid.
The antimicrobial effect of the yeast soluble glucan against various microbe was studied.The results showed that the yeast soluble glucan could effectively inhibit the growth of bacteria,yet to mould and yeast,there was no effect.The minimum inhibitory concentration(MIC) of different bacterias were:Escherichia.Coli 75mg/mL,Salmonella gallinarum 37.5mg/mL,Bacillus subtilus 37.5mg/mL,Staphalococcus aureus 37.5mg/mL, and the pH range of inhibiting bacteria was 6.0～7.0.Moreover,the heat stability is good.
引文
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