Multiplatform Mass Spectrometry-Based Approach Identifies Extracellular Glycolipids of the Yeast Rhodotorula babjevae UCDFST 04-877
Multiplatform Mass Spectrometry-Based Approach Identifies Extracellular Glycolipids of the Yeast Rhodotorula babjevae UCDFST 04-877
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DOI：
10.1021/acs.jnatprod.6b00497
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发表时间：
2016-10-01
影响因子：
5.1
通讯作者：
Fiehn, Oliver
中科院分区：
文献类型：
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作者：
Cajka, Tomas;Garay, Luis A.;Fiehn, Oliver
基于多平台质谱的方法用于阐明产油酵母红酵母 UCDFST 04-877 产生的具有生物表面活性剂特性的细胞外脂质。当在台式生物反应器中生长时，该菌株分泌 8.6 +/- 0.1 g/L 细胞外脂质，培养基中含有 100 g/L 葡萄糖，不添加疏水性底物（如油酸）。非靶向反相液相色谱-四极杆/飞行时间质谱 (QTOFMS) 检测到质量为 574-716 Da 的天然糖脂分子。水解成脂肪酸和糖成分并通过亲水相互作用色谱-QTOFMS 分析后，发现细胞外脂质由羟基脂肪酸和糖醇组成。衍生化和手性分离气相色谱-质谱（GC-MS）鉴定这些成分为D-阿拉伯糖醇、D-甘露醇、(R)-3-羟基肉豆蔻酸酯、(R)-3-羟基棕榈酸酯和(R)-3-羟基硬脂酸酯。为了将这些子结构重新组装成在初始筛选中检测到的完整糖脂，对潜在结构进行了计算机乙酰化，以匹配观察到的摩尔质量，随后使用 Mass Frontier 软件程序通过匹配预测和观察到的 MS/MS 碎片来进行表征。该酵母菌株鉴定了十一种羟基脂肪酸的乙酰化糖醇酯。
A multiplatform mass spectrometry-based approach was used for elucidating extracellular lipids with biosurfactant properties produced by the oleaginous yeast Rhodotorula babjevae UCDFST 04-877. This strain secreted 8.6 +/- 0.1 g/L extracellular lipids when grown in a benchtop bioreactor fed with 100 g/L glucose in medium without addition of hydrophobic substrate, such as oleic acid. Untargeted reversed-phase liquid chromatography-quadrupole/time-of-flight mass spectrometry (QTOFMS) detected native glycolipid molecules with masses of 574-716 Da. After hydrolysis into the fatty acid and sugar components and hydrophilic interaction chromatography-QTOFMS analysis, the extracellular lipids were found to consist of hydroxy fatty acids and sugar alcohols. Derivatization and chiral separation gas chromatography-mass spectrometry (GC-MS) identified these components as D-arabitol, D-mannitol, (R)-3-hydroxymyristate, (R)-3-hydroxypalmitate, and (R)-3-hydroxystearate. In order to assemble these substructures back into intact glycolipids that were detected in the initial screen, potential structures were in-silico acetylated to match the observed molar masses and subsequently characterized by matching predicted and observed MS/MS fragmentation using the Mass Frontier software program. Eleven species of acetylated sugar alcohol esters of hydroxy fatty acids were characterized for this yeast strain.