摘要： 本课题以产阮假丝酵母为主要研究对象。首先，通过单因素实验来确定产阮假丝酵母的最佳自溶条件。然后通过单因素实验和正交实验确定产阮假丝酵母中的最佳破壁条件以及复合酶解的最佳反应条件，运用紫外分光光度法对提取出的β-葡聚糖粗品的含量进行测定。然后将提取出的β-葡聚糖粗品通过透析以及索氏抽提得到进一步纯化的β-葡聚糖，最后通过冷冻干燥得到β-葡聚糖初品。运用羟甲基化方法对纯化后的β-葡聚糖进行增溶研究，最后对初化后的β-葡聚糖进行成分测定。实验结果表明:最终自溶温度为60℃、溶液呈中性、pH7，自溶的时间为24h的反应条件最优、物料液比1:10，氯化钠添加量6%，为最佳反应条件;最终超声波破壁的破碎时间为60min，超声波破壁的破碎温度50℃，超声波破壁的破碎功率为60%，物料比1:15，为最佳破碎条件;最终确定选用碱性蛋白酶和果胶酶复合酶解以及添加量为1%为最佳反应条件。考虑到氯化钠对细胞自溶有一定影响，可能对复合酶解也会产生影响。结果发现氯化钠对葡聚糖的提取有一定促进作用。检测提取纯化的β-葡聚糖蛋白质含量：蛋白质含量为0。03%，总糖含量为88。0%，甘露糖含量为0。6%，还原糖含量为0。8%，脂质含量为4。5%，β-葡聚糖含量为83。2%。79591

毕业论文关键字：产朊假丝酵母；β-葡聚糖；紫外分光光度法；冷冻干燥。

Extraction and purification of β -glucan from candida utilis

Abstract: In this study， Candida vulgaris is the main research object。 First， the best autolysis conditions of Candida spp。 Were determined by single factor experiments。 The optimal reaction conditions and the optimum reaction conditions were determined by single factor experiment and orthogonal experiment。 The content of crude β-glucan was determined by ultraviolet spectrophotometry。 Determination。 And then the extracted β-glucan crude by dialysis and Soxhlet extraction to obtain further purification of β-glucan， and finally by freeze-drying to get β-glucan first product。 The purified β-glucan was solubilized by the method of hydroxymethylation。 Finally， the composition of β-glucan was determined。 The results showed that the final autolysis temperature was 60 ℃， the solution was neutral， pH7， the autolysis time was 24 h， the reaction condition was the best， the ratio of material to liquid was 1:10， and the amount of sodium chloride was 6% The final broken ultrasonic breaking time of 60min， ultrasonic broken wall breaking temperature of 50 ℃， ultrasonic broken broken power of 60%， material ratio of 1:15， for the best crushing conditions; the final selection of the use of alkaline protease and Pectinase complex enzymatic hydrolysis and the addition of 1% for the best reaction conditions。 Considering that sodium chloride has a certain effect on cell autolysis， it may also have an effect on the complex enzymatic hydrolysis。 The results showed that sodium chloride had a certain effect on the extraction of dextran。 The content of purified β-glucan was determined as follows: protein content was 0。03%， total sugar content was 88。0%， mannose content was 0。6%， reducing sugar content was 0。8%， lipid content was 4。5%， β-poly The sugar content was 83。2%。

Key Words: candida utilis Beta glucan; Ultraviolet spectrophotometry; freeze drying 。

目  录

1 引言 1

1。1产朊假丝酵母 1

1。2β-葡聚糖的提取 1

1。2。1酵母自溶的原理 3

1。2。2超声波破壁的影响因素