HPLC分析植物乳杆菌胆盐水解酶降解特性

Degradation characteristics of BSH-1 from Lactobacillus plantarum by HPLC

本文建立了同时检测6种胆盐的HPLC方法,并对两株植物乳杆菌来源的胆盐水解酶BSH-1降解特性进行了分析。HPLC色谱条件如下:Sepax Bio-C18(3μm,4.6 mm×150 mm);流速:0.6 m L/min;检测波长:205 nm;流动相A:水/TFA=100/0.1,流动相B:乙腈/TFA=100/0.09;洗脱梯度如下:0 min^8 min,B 25%~35%;8 min^30 min,B 35%~48%;30 min^30.5 min,B48%~25%;30.5 min^40 min,B 25%。结果表明,本文建立的HPLC法可以快速准确的测定BSH-1对胆盐的降解活力及偏好,有利于筛选具有高效降解胆盐的乳酸菌。两种植物乳杆菌来源的BSH-1对GC、GCDC、GDC这三种胆盐可以进行降解,尤其是GDC的降解率高达99.9%,但是对TAC、TCDC、TDC几乎不能降解,也即是来源于植物乳杆菌的BSH-1对降解甘氨系列胆酸盐的能力是高于牛磺系列胆酸盐。

A HPLC method for simultaneous quantitative analysis of six salt biles was established. Then, the degradation characteristics of BSH-1 from Lactobacillus plantarum were investigated. The optimum chromatographic conditions were as follows： Sepax Bio-C18 column （3 p.m, 4.6 mm × 150 ram） with gradient elution using acetotrile and 0.1% aqueous TFA （0 min-8 min, B25% -35% ; 8 min-30min, B35% -48% ; 30 min -30.5 min, B48% -25% ; 30.5 min - 40 min, B 25 % ） , flow rate of 0.6 mL/min with detective wave 205 nm. The results showed that the degradation characteristics of BSH1 from Lactobacillus plantarum could be observed directly. Two BSH-1 from different Lactobacillus plantarum could degrade three kinds of salt biles GC, GCDC and GDC, especially the degradation ratio of GDC reached up to 99.9%. However, other salt biles TAC, TCDC and TDC could not be degraded by BSH-1. In other words, glycine salt biles were easier to be degraded compared with taurine salt biles.