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用于表征铜绿假单胞菌UG2在玉米油上产生的鼠李糖脂混合物的高效液相色谱法

High-performance liquid chromatography method for the characterization of rhamnolipid mixtures produced by pseudomonas aeruginosa UG2 on corn oil.

作者信息

Mata-Sandoval J C, Karns J, Torrents A

机构信息

Department of Civil and Environmental Engineering, University of Maryland at College Park, 20742, USA.

出版信息

J Chromatogr A. 1999 Dec 24;864(2):211-20. doi: 10.1016/s0021-9673(99)00979-6.

DOI:10.1016/s0021-9673(99)00979-6
PMID:10669288
Abstract

A HPLC method was developed to quantify rhamnolipid species in a bacterial biosurfactant mixture. The biosurfactant mixtures containing mainly 3-[3'-(L-rhamnopyranosyl-oxy)decanoyloxy]decanoic acid (RhC10C10), 3-[3'-(2'-O-alpha-L-rhamnopyranosyl-oxy)decanoyloxy]decanoic acid (Rh2C10C10), 3-[3'-(2'-O-alpha-L-rhamnopyranosyl-oxy)decanoyloxy]dodecanoic acid (Rh2C10C12), and a dehydrogenated variety of the latter, 3-[3'-(2'-O-alpha-L-rhamnopyranosyl-oxy)decanoyloxy]dodecenoic acid (Rh2C10C12-H2), were isolated from Pseudomonas aeruginosa UG2 cultures grown on corn oil as sole carbon. The rhamnolipid species were identified and quantified after their derivatization to the corresponding phenacyl esters. To confirm the reliability of the HPLC method, the biosurfactant mixtures and the HPLC isolated species were further analyzed. Mass spectroscopy (electrospray ionization and atmospheric pressure chemical ionization techniques) was used to confirm their molecular mass, gas chromatography to verify their fatty acid content, and a colorimetric assay to quantify the rhamnose content.

摘要

开发了一种高效液相色谱(HPLC)方法来定量分析细菌生物表面活性剂混合物中的鼠李糖脂种类。从以玉米油作为唯一碳源生长的铜绿假单胞菌UG2培养物中分离出主要含有3-[3'-(L-鼠李糖吡喃糖氧基)癸酰氧基]癸酸(RhC10C10)、3-[3'-(2'-O-α-L-鼠李糖吡喃糖氧基)癸酰氧基]癸酸(Rh2C10C10)、3-[3'-(2'-O-α-L-鼠李糖吡喃糖氧基)癸酰氧基]十二烷酸(Rh2C10C12)以及后者的一种脱氢变体3-[3'-(2'-O-α-L-鼠李糖吡喃糖氧基)癸酰氧基]十二碳烯酸(Rh2C10C12-H2)的生物表面活性剂混合物。将鼠李糖脂种类衍生化为相应的苯甲酰酯后进行鉴定和定量。为了确认HPLC方法的可靠性,对生物表面活性剂混合物和HPLC分离出的种类进行了进一步分析。使用质谱(电喷雾电离和大气压化学电离技术)确认其分子量,使用气相色谱验证其脂肪酸含量,并使用比色法对鼠李糖含量进行定量。

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