Quintilla Raquel, Kolecka Anna, Casaregola Serge, Daniel Heide M, Houbraken Jos, Kostrzewa Markus, Boekhout Teun, Groenewald Marizeth
Westerdijk Fungal Biodiversity Institute, Uppsalalaan 8, 3584 Utrecht, The Netherlands; Laboratory of Molecular Cell Biology, Institute of Botany and Microbiology, KU Leuven, B-3001 Leuven, Belgium; Department of Molecular Microbiology, VIB, Kasteelpark Arenberg 31, B-3001 Leuven-Heverlee, Flanders, Belgium.
Westerdijk Fungal Biodiversity Institute, Uppsalalaan 8, 3584 Utrecht, The Netherlands.
Int J Food Microbiol. 2018 Feb 2;266:109-118. doi: 10.1016/j.ijfoodmicro.2017.11.016. Epub 2017 Nov 24.
Since food spoilage by yeasts causes high economic losses, fast and accurate identifications of yeasts associated with food and food-related products are important for the food industry. In this study the efficiency of the matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) to identify food related yeasts was evaluated. A CBS in-house MALDI-TOF MS database was created and later challenged with a blinded test set of 146 yeast strains obtained from food and food related products. Ninety eight percent of the strains were correctly identified with log score values>1.7. One strain, Mrakia frigida, gained a correct identification with a score value<1.7. Two strains could not be identified at first as they represented a mix of two different species. These mixes were Rhodotorula babjevae with Meyerozyma caribbica and Clavispora lusitaniae with Debaryomyces hansenii. After separation, all four species could be correctly identified with scores>1.7. Ambiguous identifications were observed due to two incorrect reference mass spectra's found in the commercial database BDAL v.4.0, namely Candida sake DSM 70763 which was re-identified as Candida oleophila, and Candida inconspicua DSM 70631 which was re-identified as Pichia membranifaciens. MALDI-TOF MS can distinguish between most of the species, but for some species complexes, such as the Kazachstania telluris and Mrakia frigida complexes, MALDI-TOF MS showed limited resolution and identification of sibling species was sometimes problematic. Despite this, we showed that the MALDI-TOF MS is applicable for routine identification and validation of foodborne yeasts, but a further update of the commercial reference databases is needed.
由于酵母导致的食品腐败会造成巨大经济损失,因此快速、准确地鉴定与食品及食品相关产品相关的酵母对于食品工业来说至关重要。在本研究中,评估了基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)鉴定食品相关酵母的效率。创建了一个CBS内部MALDI-TOF MS数据库,随后用从食品和食品相关产品中获得的146株酵母菌株组成的盲测集对其进行验证。98%的菌株通过对数分数值>1.7被正确鉴定。一株嗜冷嗜冷酵母(Mrakia frigida)以分数值<1.7获得了正确鉴定。最初有两株菌株无法鉴定,因为它们代表了两种不同物种的混合。这些混合菌株是巴氏红酵母(Rhodotorula babjevae)与加勒比梅奇酵母(Meyerozyma caribbica)以及葡萄牙棒孢酵母(Clavispora lusitaniae)与汉逊德巴利酵母(Debaryomyces hansenii)。分离后,所有四个物种都能以分数>1.7被正确鉴定。由于在商业数据库BDAL v.4.0中发现了两个错误的参考质谱图,出现了模糊鉴定,即清酒假丝酵母(Candida sake)DSM 70763被重新鉴定为嗜油假丝酵母(Candida oleophila),以及隐匿假丝酵母(Candida inconspicua)DSM 70631被重新鉴定为膜醭毕赤酵母(Pichia membranifaciens)。MALDI-TOF MS可以区分大多数物种,但对于一些物种复合体,如特鲁氏卡扎奇酵母(Kazachstania telluris)和嗜冷嗜冷酵母复合体,MALDI-TOF MS显示出有限的分辨率,并且有时难以鉴定亲缘物种。尽管如此,我们表明MALDI-TOF MS适用于食源性酵母的常规鉴定和验证,但需要进一步更新商业参考数据库。