CAJ | 학술논문

将香菇多糖和0.02 mol/L三氟乙酸( TFA)溶液按照1 mg：200μL比例混合并于100℃水解,然后利用快速蛋白液相色谱(FPLC)技术对分别水解2,4,6 h的水解产物中不同聚合度寡糖的组成与丰度进行分析,从而确定最佳水解时间；用确定的最佳时间对水解残留多糖重复水解3次,直至多糖沉淀消失为止,然后分别对水解产物进行FPLC分离并依次收集每批次的七糖.对七糖进行ESI-CID-MS/MS分析,结合β-1,3-二糖、七糖及β-1,6-葡聚寡糖二糖的ESI-CID-MS/MS质谱特征,可获得每次水解产物中寡糖的结构信息,进而获得香菇多糖的水解机制；最后用P4凝胶柱对水解产物进行色谱分离,获得具有不同聚合度的全序列寡糖.结果表明,香菇多糖的最佳水解时间为4 h,在不同批次的水解过程中香菇寡糖的结构由β-1,3/1,6-寡糖过渡为纯β-1,3-寡糖.
장향고다당화0.02 mol/L삼불을산( TFA)용액안조1 mg：200μL비례혼합병우100℃수해,연후이용쾌속단백액상색보(FPLC)기술대분별수해2,4,6 h적수해산물중불동취합도과당적조성여봉도진행분석,종이학정최가수해시간；용학정적최가시간대수해잔류다당중복수해3차,직지다당침정소실위지,연후분별대수해산물진행FPLC분리병의차수집매비차적칠당.대칠당진행ESI-CID-MS/MS분석,결합β-1,3-이당、칠당급β-1,6-포취과당이당적ESI-CID-MS/MS질보특정,가획득매차수해산물중과당적결구신식,진이획득향고다당적수해궤제；최후용P4응효주대수해산물진행색보분리,획득구유불동취합도적전서렬과당.결과표명,향고다당적최가수해시간위4 h,재불동비차적수해과정중향고과당적결구유β-1,3/1,6-과당과도위순β-1,3-과당.
To acquire the mechanism of lentinan hydrolysis and full series of gluco-oligosaccharides from hy-drolyzed lentinan, the hydrolysis time of lentinan treated with 0. 02 mol/L trifluoroacetic acid( TFA) at 100℃was optimized through analysis the distribution of different degree of polymerization ( DP ) oligosaccharides at 2, 4, and 6 h. And then lentinan hydrolysis was repeated another three times until no precipitation can be ob-served under optimal hydrolysis time. The mass spectrometry character of 7-mer oligosaccharides from all the four batch hydrolysis products was analyzed using ESI-CID-MS/MS based on the mass spectrometry character of 7-mer β-1,3-andβ-1,6-gluco-oligosaccharides, and acquiring a hydrolysis mechanism. Finally, all the four batch hydrolysis products were combined and separated with P4 column and full series of gluco-oligosacchaides from 2-mer to 13-mer were acquired. The optimal time for lentinan hydrolysis was 4 h, andβ-1,3/1,6-gluco-oligosaccharides in hydrolysis lentinan was observed at the first batch, and then the amount ofβ-1,3/1,6-glu-co-oligosaccharides reduced with the increasing of hydrolysis times, and there only observedβ-1,3-gluco-oligo-saccharides at the fourth hydrolysis products. This results also suggested that the triple helix structure zone containing β-1,3/1,6-linkage of lentinan is loose, but the only containing β-1,3-linkage triple helix structure zone in lentinan is tight. The theory acquired in this work will supply useful information for preparation gluco-oligosaccharides in large-scale and it also expand the gluco-oligosaccharides library for exploring protein-carbo-hydrate interaction using carbohydrate microarray.