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一种胞外多糖的分离与物理特性分析

Isolation and physical characterization of an exocellular polysaccharide.

作者信息

Tuinier R, Zoon P, Olieman C, Stuart M A, Fleer G J, de Kruif C G

机构信息

NIZO Food Research, The Netherlands.

出版信息

Biopolymers. 1999 Jan;49(1):1-9. doi: 10.1002/(SICI)1097-0282(199901)49:1<1::AID-BIP1>3.0.CO;2-B.

Abstract

The physical properties of a polysaccharide produced by the lactic acid bacterium Lactococcus lactis subsp. cremoris strain NIZO B40 were investigated. Separation of the polysaccharide from most low molar mass compounds in the culture broth was performed by filtration processes. Residual proteins and peptides were removed by washing with a mixture of formic acid, ethanol, and water. Gel permeation chromatography (GPC) was used to size fractionate the polysaccharide. Fractions were analyzed by multiangle static light scattering in aqueous 0.10 M NaNO3 solutions from which a number- (Mn) and weight-averaged (Mw) molar mass of (1.47 +/- 0.06).10(3) and (1.62 +/- 0.07).10(3) kg/mol, respectively, were calculated so that Mw/Mn approximately 1.13. The number-averaged radius of gyration was found to be 86 +/- 2 nm. From dynamic light scattering an apparent z-averaged diffusion coefficient was obtained. Upon correcting for the contributions from intramolecular modes by extrapolating to zero wave vector a hydrodynamic radius of 86 +/- 4 nm was calculated. Theoretical models for random coil polymers show that this z-averaged hydrodynamic radius is consistent with the z-averaged radius of gyration, 97 +/- 3 nm, as found with GPC.

摘要

对乳酸乳球菌乳脂亚种NIZO B40菌株产生的一种多糖的物理性质进行了研究。通过过滤过程从培养液中的大多数低摩尔质量化合物中分离出该多糖。用甲酸、乙醇和水的混合物洗涤以去除残留的蛋白质和肽。使用凝胶渗透色谱法(GPC)对多糖进行尺寸分级。在0.10 M NaNO3水溶液中通过多角度静态光散射对级分进行分析,据此计算出数均摩尔质量(Mn)为(1.47±0.06)×10³ kg/mol,重均摩尔质量(Mw)为(1.62±0.07)×10³ kg/mol,使得Mw/Mn约为1.13。发现数均回转半径为86±2 nm。通过动态光散射获得了表观z平均扩散系数。通过外推至零波矢校正分子内模式的贡献后,计算出流体力学半径为86±4 nm。无规卷曲聚合物的理论模型表明,该z平均流体力学半径与GPC测得的z平均回转半径97±3 nm一致。

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