Extending our tools and resources in the non-conventional industrial yeast Xanthophyllomyces dendrorhous through the application of metabolite profiling methodologies.
Extending our tools and resources in the non-conventional industrial yeast Xanthophyllomyces dendrorhous through the application of metabolite profiling methodologies.
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DOI：
10.1007/s11306-017-1313-9
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Fraser PD
中科院分区：
文献类型：
--
作者：
Alcalde E;Fraser PD
Xanthophyllomyces dendrorhous是一种非传统的工业酵母。它在酵母菌中具有独特的能力来产生香叶基香叶基焦磷酸衍生的萜类化合物，如类胡萝卜素，特别是高价值的色素虾青素。为了利用现代工业生物技术方法充分开发黄酵母的工业潜力，需要在该生物体中进一步开发“组学”资源，以建立现在测序和注释的基因组。为了实现这一目标，本研究开发并实施了一种有效的代谢物分析系统，包括淬灭、提取和相关的GC-MS和UPLC分析。通过分析以实验室规模在液体振荡培养物中培养的黄孢酵母在滞后期、指数期以及稳定期早期和晚期的稳态代谢物变化，测试并验证了与GC-MS和UPLC分析兼容的四种淬灭方法和五种提取方法。为黄酵母创建了定制的自动质谱去卷积和鉴定系统（AMDIS）库，库中存在400多种化合物，其中78种在极性和非极性提取物中进行常规检测和定量。一个初步的生化网络已经建成。在一个标准化的实验室生长周期中，代谢物水平的变化已经被确定为未来菌株改良方法和初始生化网络构建的参考点。相关性分析表明，虾青素的形成与中间代谢的不同部分（例如TCA循环中间体和氨基酸形成）、“短”饱和脂肪酸和β-胡萝卜素正相关，而其他代谢产物响应于虾青素的产生而减少。中间代谢的这些部门提供了潜在的未来目标的操纵，导致产生的菌株与给定的萜类化合物的滴度提高。总之，一个强大的代谢产物分析系统的黄酵母是到位，以进一步我们的理解和潜在的利用这种未充分利用的工业酵母。本文的在线版本（10.1007/s11306-017-1313-9）包含补充材料，可供授权用户使用。
Xanthophyllomyces dendrorhous is a non-conventional industrial yeast. It has the unique ability among yeasts to produce geranylgeranyl pyrophosphate derived terpenoids such as carotenoids and in particular the high value pigment astaxanthin. In order to fully exploit the industrial potential of Xanthophyllomyces using modern industrial biotechnology approaches the further development of “omic” resources in this organism are required to build on the now sequenced and annotated genome. To contribute to this goal, the present study has developed and implemented an efficient metabolite profiling system comprised of, quenching, extraction and associated GC–MS and UPLC analysis. Four quenching methods and five extraction methods compatible with GC–MS and UPLC profiling were tested and validated by analysing steady state metabolite changes of Xanthophyllomyces cultivated at laboratory scale in liquid shake culture at lag, exponential and early and late stationary phases. A customised Automated Mass Spectral Deconvolution and Identification System (AMDIS) library has been created for Xanthophyllomyces, over 400 compounds are present in the library of which 78 are detected and quantified routinely in polar and non-polar derived extracts. A preliminary biochemical network has been constructed. Over a standardised laboratory growth cycle, changes in metabolite levels have been determined to create reference point for future strain improvement approaches and the initial biochemical network construction. Correlation analysis has illustrated that astaxanthin formation correlates positively with different sectors of intermediary metabolism (e.g. the TCA cycle intermediates and amino acid formation), “short” saturated fatty acids and β-carotene, while other metabolites are reduced in response to astaxanthin production. These sectors of intermediary metabolism offer potential future targets for the manipulation resulting in the generation of strains with improved titres of given terpenoids. In summary a robust metabolite profiling system for Xanthophyllomyces is in place to further our understanding and potential exploitation of this underutilised industrial yeast. The online version of this article (10.1007/s11306-017-1313-9) contains supplementary material, which is available to authorized users.
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