紫皮石斛寡糖的酶法制备及单糖组成分析  

Enzymatic Preparation and Oligosaccharide and Composition Analysis of Monosaccharides from Dendrobium devonianum

作　　者：蒋明星 易梦镯 李翠 赵云洁 李欢 蔡胤 殷根深 JIANG Ming-xing;YI Meng-zhuo;LI Cui;ZHAO Yun-jie;LI Huan;CAI Yin;YIN Gen-shen(Coll.of Agric.&Life Sci.,Kunming Uni.,Kunming 650214)

机构地区：[1]昆明学院农学与生命科学学院,云南昆明650214

出　　处：《微生物学杂志》2022年第3期54-60,共7页Journal of Microbiology

基　　金：昆明学院引进人才科研项目(YJL18006);云南省教育厅科学研究基金项目(2021J0709);云南省大学生创新训练项目(202011393002)。

摘　　要：从多糖提取后的紫皮石斛(Dendrobium devonianum Paxt.)渣上分离到一株能够水解石斛多糖的菌株,为丰富寡糖食品新原料种类,对该菌株的发酵产酶活性进行分析。利用酶法制备紫皮石斛寡糖,并进行小试和中试,采用硅胶柱色谱对寡糖进行初步分离,PMP衍生化后通过HPLC分析单糖组成。结果表明,水解石斛多糖的菌株为粗糙脉孢菌(Neurospora crassa)。该菌株液态发酵至第4天酶活最高,为0.1 U/μL,最佳酶解时间为24 h,粗酶液能够耐受30%的乙醇。通过粗酶液生物转化石斛多糖的小试和中试成功制备了大量的石斛寡糖,并对各组分进行了初步分离,单糖组成分析表明其主要由甘露糖组成,并含有少量的葡萄糖或果糖。通过粗糙脉孢菌酶法制备得到的紫皮石斛寡糖主要为甘露寡糖。A strain capable of hydrolyzing Dendrobium devonianum polysaccharides was isolated and obtained from residues extracted from D.devonianum polysaccharides.In order to enrich the types of new oligosaccharide food raw materials,the fermentation enzyme activity of this strain was analyzed.D.devonianum oligosaccharides were prepared by enzymatic method,and small-scale and pilot-scale tests were carried out.The oligosaccharides were preliminarily separated by silica gel column chromatography,and the composition of monosaccharide was analyzed by HPLC after PMP derivatization.The results showed that strain that hydrolyzed D.devonianum polysaccharides was Neurospora crassa.The enzyme activity was the highest at the 4th day of liquid fermentation,which was 0.1 U/μL.The best enzymatic hydrolysis time was 24 h,and the crude enzyme solution could tolerate 30% ethanol.A large number of D.devonianum oligosaccharides were successfully prepared in the small-scale and pilot-scale biotransformation of D.devonianum polysaccharides with crude enzyme solution,and preliminary separation of the components was carried out.Analysis of monosaccharide composition showed that it was mainly composed of mannose,with a small amount of glucose or fructose.D.devonianum oligosaccharides prepared by N.crassa enzymatic method are mainly mannose oligosaccharides.

关 键 词：紫皮石斛 寡糖 粗糙脉孢菌 酶法制备 单糖组成分析 

分 类 号：Q936[生物学—微生物学]