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多相分析,包括基质辅助激光解吸电离飞行时间质谱,对(某部分)菌株冻干长期保存的评估

Polyphasic, Including MALDI-TOF MS, Evaluation of Freeze-Drying Long-Term Preservation on (Section ) Strains.

作者信息

Rodriguez Rodrigo, Santos Carla, Simões Marta F, Soares Célia, Santos Cledir, Lima Nelson

机构信息

CEB-Centre of Biological Engineering, Micoteca da Universidade do Minho, University of Minho, 4710-057 Braga, Portugal.

State Key Laboratory of Lunar and Planetary Sciences, Space Science Institute. Macau University of Science and Technology, Avenida Wai Long, Taipa, Macau SAR 999078, China.

出版信息

Microorganisms. 2019 Aug 25;7(9):291. doi: 10.3390/microorganisms7090291.

Abstract

This study aims to evaluate the effect of freeze-drying and long-term storage on the biotechnological potential of section strains. Twelve selected strains were freeze-dried and aged by accelerated storage, at 37 °C in the dark, for 2 and 4 weeks. To assess possible changes as a consequence of the ageing in the freeze-drying ampoules, morphological characteristics, mycotoxins and enzymes production, matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALTI-TOF MS) spectra, and M13 phage probe fingerprinting were used as part of a polyphasic approach. Phenotypical changes were observed; nevertheless, they did not substantially affect the potential biotechnological use of these strains. The activity of hydrolytic enzymes (protease, carboxymethylcellulase, xylanase, pectinase and mannanase) was maintained or increased after freeze-drying. MALDI-TOF MS data originated spectra that grouped, for the majority of samples, according to strain independently of preservation time point. M13 profiles revealed the presence of some genetic polymorphisms after preservation. However, the three studied times still clustered for more than 50% of strains. Our results show that the studied strains maintain their biotechnological potential after preservation, with minimal phenotypic alterations. These findings provide evidence that freeze-drying preservation is a suitable option to preserve biotechnologically relevant aspergilli strains from section , and one should consider that the observed effects might be species/strain-dependent.

摘要

本研究旨在评估冷冻干燥和长期储存对部分曲霉菌株生物技术潜力的影响。选取了12株菌株进行冷冻干燥,并在37℃黑暗条件下通过加速储存进行老化处理,分别老化2周和4周。为了评估冷冻干燥安瓿瓶老化可能产生的变化,采用了多相方法,包括形态特征、霉菌毒素和酶的产生、基质辅助激光解吸/电离飞行时间质谱(MALTI-TOF MS)光谱以及M13噬菌体探针指纹图谱。观察到了表型变化;然而,这些变化并未对这些菌株的潜在生物技术用途产生实质性影响。冷冻干燥后,水解酶(蛋白酶、羧甲基纤维素酶、木聚糖酶、果胶酶和甘露聚糖酶)的活性得以维持或提高。MALDI-TOF MS数据生成了光谱,对于大多数样品,这些光谱根据菌株进行分组,与保存时间点无关。M13图谱显示保存后存在一些基因多态性。然而,在三个研究时间点,仍有超过50%的菌株聚类在一起。我们的结果表明,所研究的菌株在保存后仍保持其生物技术潜力,且表型改变最小。这些发现证明,冷冻干燥保存是保存部分具有生物技术相关性的曲霉菌株的合适选择,并且应该考虑到观察到的影响可能因物种/菌株而异。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9dcb/6780240/b208cd60b920/microorganisms-07-00291-g001.jpg

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