Purification Of Functional Oligosaccharide And Their Biological Activities

Effects of different separation conditions on isolation of cello-oligosaccharidesusing Sephadex LH-20was studied, and purified cellotriose, cellotetraose,cellopentose, cellohexose were analyzed by infrared spectrum and mass spectrum.Effects of cello-oligosaccharides and xylooligosaccharides on proliferation ofprobiotics such as Lactobacillus acidophilus and Bifidobacterium odolescentis wereinvestigated, then metabolic products of probiotics and metabolic mechanism wereanalyzed. The results showed as follow:Good separation performance could be obtained using sephadex LH-20,1.6cm×100cm column with injection concentration of33.77g/L(2mL),30℃andelution rate of0.3mL/min. To get more pure cellotriose～cellohexose, furtherisolation was needed. Then the degree of purity of cellotriose was97.5%,91.88%for cellotetraose and98.55%for cellopentose.Infrared spectrum of purified cellotriose～cellohexose showed cello-oligo-saccharides was a pyranose.3600cm-1～3200cm-1was the stretching vibration ofO-H;1600cm-1was the stretching vibration of C=O;1158cm-1,1068cm-1were thestretching vibration of C-O;1029cm-1was the stretching vibration of C-O-C;897cm-1was characteristic absorption band of C-H belonging to a β–pyranose.Purified cellotriose～cellohexose were analyzed by electron pray ionizationmass spectrometry(ESI-MS) with negative ion mode. Under the ESI negative ionmode, the fragments were formed in the three ways: breaking of unit of sugarring,containing reducing end group and middle unit of sugar ring; breaking ofglycosidic bond linking the reducing end group,losing a glucose residue; losing awater molecule.Glucose, cellobiose, cellotriose, cello-oligosaccharides, xylose and xylooligo-saccharides were applied to cultivate Lactobacillus acidophilus. The glucose was thebest carbon source for Lactobacillus acidophilus, followed by cellobiose, cello-oligosaccharides and cellotriose. The multiplication times of Lactobacillusacidophilus employed glucose, cellobiose, cellooligosaccharides and cellotriose ascarbon sources were5.84,5.04,3.48and3.2times, respectively. Lactobacillusacidophilus could not utilize xylose and xylooligosaccharides. The metabolicproducts were lactic acid, acetic acid and propionic acid.Glucose, xylose, xylooligosaccharides, cellobiose and cello-oligosaccharideswere applied to cultivate Bifidobacterium odolescentis. The glucose was the bestcarbon source for Bifidobacterium odolescentis, followed by xylooligosaccharidesand xylose. The multiplication times of Bifidobacterium odolescentis employed glucose, xylooligosaccharides and xylose as carbon sources were17,8.7and5.5times,respectively. Bifidobacterium odolescentis could not utilize cello-oligo-saccharides. The main metabolic products were acetic acid and lactic acid, andpropionic acid.Lactobacillus acidophilus and Bifidobacterium odolescentis were mainprobiotics of alimentary canal. Cello-oligosaccharides and xylooligosaccharidescould be mixed to cultivate Lactobacillus acidophilus and Bifidobacteriumodolescentis effectively to balance the microecology of alimentary canal.