酵母细胞壁多糖提取纯化鉴定技术及其应用研究进展

Research Progress on Application,Extraction,Purification and Identification of Yeast Cell Wall Polysaccharide

酵母细胞壁多糖的主要活性成分是β-葡聚糖和甘露聚糖,在动物机体内与肠道受体竞争吸附病原菌以及在肠道降解提供能量,改善动物肠道环境;也会利用其结构与Fe 2+离子螯合抑制羟自由基的产生以及防止脂质过氧化连锁反应等,提高抗氧化能力;其特有的化学位点会通过多种分子间作用力吸附霉菌毒素;β-葡聚糖和甘露聚糖通过作用于巨噬细胞调节多种细胞因子,通过多个途径增强机体的特异性和非特异性免疫力。目前在养殖业中酵母细胞壁多糖可用于替代金霉素、硫酸黏杆菌素等抗生素控制鸡的坏死性肠炎、减少水产动物细菌感染、降低猪、牛等幼畜腹泻率、预防各种老化性疾病以及促进动物生长发育。对酵母细胞壁多糖现有的提取方法进行比较,提取酵母细胞壁β-葡聚糖,选用超声辅助酶解产率最高、效果最好;用低浓度的氢氧化钠(NaOH)溶液提取甘露聚糖以及酶碱法提取酵母细胞壁多糖效果最好。分离纯化技术包括有脱蛋白、脱色、单一多糖的分离,将其分离纯化效果分别比较,除去酵母细胞壁多糖中蛋白质的最佳方法是Sevage法;脱除色素的最佳方法是颗粒活性炭吸附法;单一多糖分离纯化采用DEAE-纤维素A52阴离子交换柱和Sephadex G-100柱联用的方法最佳。酵母细胞壁多糖结构鉴定一般采用高效液相色谱法(HPLC)测定多糖的单糖组成,高效凝胶渗透色谱测定多糖的均匀性和分子质量,傅立叶变换红外光谱(FT-IR)、气相色谱-质谱(GC-MS)和核磁共振(NMR)测定多糖的精细结构。文章通过对酵母细胞壁多糖的作用及应用、提取、分离纯化和鉴定技术进行阐述,为酵母细胞壁多糖作为抗生素替代品在畜牧业生产中的推广应用提供理论依据,为抗生素替代品的研发提供新的思路。

The main active components of yeast cell wall polysaccharide areβ-glucan and mannan,which compete with intestinal receptors to adsorb pathogenic bacteria and to provide energy in intestinal degradation to improve the intestinal environment of animals.They can also be used to chelate with Fe 2+ions to inhibit the production of hydroxyl radicals and prevent lipid peroxide chain reaction to improve antioxidant capacity.And its unique chemical sites will adsorb mycotoxins by a variety of intermolecular interactions.β-glucan and mannan enhance the specific and nonspecific immunity of the body by acting on macrophages and regulating a variety of cytokines.Currently,it can be used in aquaculture to replace chlortetracycline,mucin sulfate and other antibiotics to control the necrotizing enteritis of chickens,reduce bacterial infection in aquatic animals,and reduce the diarrhea rate of young animals such as pigs and cattle,prevent all kinds of aging diseases and promote the growth and development of animals.The results of the existing extraction methods of yeast cell wall polysaccharide were compared.The ultrasonic assisted enzyme hydrolysis rate was the highest and the effect was the best whenβ-glucan was extracted from yeast cell wall.The extraction of mannan by low concentration sodium hydroxide(NaOH)solution and the extraction of yeast cell wall polysaccharide by enzyme-alkali method were the best.The separation and purification techniques include deprotein,decolorization and separation of single polysaccharide.The effect of isolation and purification is compared.The best method to remove the protein from yeast cell wall polysaccharide is Sevage method.The best method of pigment removal is granular activated carbon adsorption,and the method of separation and purification of single polysaccharide by DEAE-cellulose A52 anion exchange column and Sephadex G-100 column is the best.Compared with the existing identification techniques of yeast cell wall polysaccharides,the structure identification of yeast cell wall polysaccharides includes that determination of monosaccharide composition of polysaccharides by high performance liquid chromatography,and uniformity and molecular weight of polysaccharides by high performance gel permeation chromatography.The fine structure of polysaccharide was determined by fourier transformation infrared spectrometer(FT-IR),chromatographic mass spectrometry(GC-MS)and nuclear magnetic resonance(NMR).This review commences with the effects and applications extraction,purification and identification techniques of yeast cell wall polysaccharides.It can provide a theoretical basis for the popularization and application of yeast cell wall polysaccharides as an antibiotic substitute in animal husbandry production and offer a new thinking direction for the research and development of antibiotic substitutes.