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1、葡聚糖的产品化,Company Logo,2,1.Introduction 【标志性成分及含量】每100克含：粗多糖48.1克、维生素C 7.9克 推荐服用人群：免疫力低下者、受辐射危害者、便秘患者、心血管病人。辅助放化疗，有一定的同步减毒增效的作用。,27g,成本价：13g，0.8元，得率按11%计算，27g的成本价为15.1元,Company Logo,3,很有研究价值,Company Logo,4,Biological activity of the polysaccharide glucan 1.anti - tumor 葡聚糖抗肿瘤效用是刺激免疫系统的结果，而不是直接对肿瘤的毒性细胞
2、产生作用。这种效用相对于传统治疗的优势在于，免疫细胞具有识别和摧毁“坏”细胞和“外来入侵者”的能力，所以葡聚糖对健康组织和细胞的危害很小。 2.improve function of immunological competence ;anti-virus 20世纪80年代，哈佛大学的研究陆续表明，-1，3-D-葡聚糖可帮助人们抵抗多种病原菌的侵害，如金色葡萄球菌 3.Reduce the radiation damage 美国空军辐射生物学研究所的研究表明，酵母葡聚糖 可以很好地帮助辐射后免疫系统的恢复和抵抗能力的快速提高。 在给予致死剂量的辐射后，对照组的小鼠在2周后全部死亡， 而服用酵母
3、葡聚糖的小鼠的死亡率只有40。,Company Logo,6,1.Introduction,（13）糖苷键连接主链，侧链是（16）糖苷键连接至少4个葡萄糖残基。 G Guizand等人用X射线法研究证明了酵母不溶性葡聚糖是以三股螺旋存在的，三条多糖链平行排列缠绕，通过链间氢键作用处于稳定状态。 约有1500个-（13）-葡萄糖残基，和190个-（16）葡萄糖单位,Company Logo,7,the cell wall of Saccharomyces cerevisiae represents up to 20% of the cell dry weight. Around 60% of t
4、his total corresponds to the -glucans.,Klis, Mol, Hellingwerf,& Brul, 2002,Company Logo,8,1.Introduction acid and alkaline washings lead to degradation of glucose chains extraction with hot water and enzyme an additional high-pressure homogenization step to aid in disrupting the yeast cell wall,sati
5、sfactory yields preservation of the glucose chains of the polymer.,Liu, Wang, Cui, & Liu, 2008). Freimund et al. (2003) Liu et al. (2008),Company Logo,9,insolubility in water,derivatization of the glucan molecule to carboxymethyl-glucan (CM-G).,Slamenov et al.,2003,Company Logo,10,2. Experimental Pr
6、ocedures,Company Logo,11,2. Experimental Procedures,2.1.Autolysis and hot water treatment,Company Logo,12,2. Experimental Procedures,2.2. Sonication tests were conducted at 20 kHz and 150W in an ice bath via a 32 full factorial design, with three replicates of the central point. The times studied, i
7、n minutes, were 2, 4 and 6 for cell dilutions of 10%, 15% and 20% in distilled water,respectively. After centrifugation at 4500g for 15 min at 10,the slides were prepared and submitted to Gram-staining to differentiate intact cells (violet) from those with ruptured walls (pink),Company Logo,13,2. Ex
8、perimental Procedures,2.3. Lipid extraction For extraction with petroleum ether, the sample was wrapped with filter paper and positioned so that the petroleum ether passed throughout the sample for 2 h under reflux. The recovered residue was washed two times with acetone1:1(w/v), centrifuged at 4500
9、g for 5 min, and then stored at 4 .,Company Logo,14,2.3.1. Extraction of (13)(16)-D-glucan,2.4. Proteolysis of the cell wall optimum conditions is 5 h at 55 and pH 7.5 tests with 0.05 U, 0.1 U, 0.2 U, 0.3 U, 0.4 U and 0.5 U of enzyme per gram of cell wall in a 20% aqueous suspension At the end of th
10、e reaction time, the enzyme was inactivated at 85 for 20 min. The material was cooled to 25 and washed with centrifugation at 4500g for 5 min The supernatants were collected for detection of soluble proteins, and the precipitate of each treatment was used for total nitrogen determination,Company Log
11、o,15,2.5. Dialysis and lyophilization Dialysis was performed for 48 h against distilled water, under mild agitation, with frequent water replacements. After dialysis, the material was frozen at -20 and dried under vacuum at 80 . The residual moisture after lyophilization was determined by drying at
12、102 until a constant weight was achieved.,Company Logo,16,2.6. Carboxymethylation of -D-glucan The derivatization of non-dialyzed water-insoluble (13)(16)-D-glucan was performed as described by Sandula, Kogan, Kacurakov, and Machov (1999), using monochloro acetic acid（氯乙酸）. The substitution degree (DS) of the carboxymethylated product was determined by potentiometric titration with potassium hydroxide solution（KOH）,CH2CLCOOH,GLUCAN,Thank You !,
