VTT Technical Research Centre of Finland Ltd, Espoo, Finland.
PLoS One. 2020 Aug 7;15(8):e0236822. doi: 10.1371/journal.pone.0236822. eCollection 2020.
Various marine fungi have been shown to produce interesting, bioactive compounds, but scaling up the production of these compounds can be challenging, particularly because little is generally known about how the producing organisms grow. Here we assessed the suitability of using 100-well BioScreen plates or 96-well plates incubated in a robot hotel to cultivate eight filamentous marine fungi, six sporulating and two non-sporulating, to obtain data on growth and substrate (glucose, xylose, galactose or glycerol) utilisation in a high throughput manner. All eight fungi grew in both cultivation systems, but growth was more variable and with more noise in the data in the Cytomat plate hotel than in the BioScreen. Specific growth rates between 0.01 (no added substrate) and 0.07 h-1 were measured for strains growing in the BioScreen and between 0.01 and 0.27 h-1 for strains in the plate hotel. Three strains, Dendryphiella salina LF304, Penicillium chrysogenum KF657 and Penicillium pinophilum LF458, consistently had higher specific growth rates on glucose and xylose in the plate hotel than in the BioScreen, but otherwise results were similar in the two systems. However, because of the noise in data from the plate hotel, the data obtained from it could only be used to distinguish between substrates which did or did not support growth, whereas data from BioScreen also provided information on substrate preference. Glucose was the preferred substrate for all strains, followed by xylose and galactose. Five strains also grew on glycerol. Therefore it was important to minimise the amount of glycerol introduced with the inoculum to avoid misinterpreting the results for growth on poor substrates. We concluded that both systems could provide physiological data with filamentous fungi, provided sufficient replicates are included in the measurements.
各种海洋真菌已被证明能产生有趣的、具有生物活性的化合物，但大规模生产这些化合物具有挑战性，特别是因为一般来说，人们对产生这些化合物的生物的生长方式知之甚少。在这里，我们评估了使用 100 孔生物筛选板或在机器人旅馆中孵育的 96 孔板来培养 8 种丝状海洋真菌的适宜性，这 8 种真菌中有 6 种是孢子形成的，2 种是非孢子形成的，以便以高通量的方式获得生长和基质（葡萄糖、木糖、半乳糖或甘油）利用的数据。这 8 种真菌在两种培养系统中都能生长，但在 Cytomat 板旅馆中的数据中，生长更为多变，噪声更大。在生物筛选中生长的菌株的比生长速率在 0.01（未添加基质）至 0.07 h-1 之间，在板旅馆中生长的菌株的比生长速率在 0.01 至 0.27 h-1 之间。三株菌株，Dendryphiella salina LF304、Penicillium chrysogenum KF657 和 Penicillium pinophilum LF458，在板旅馆中的葡萄糖和木糖上的比生长速率始终高于生物筛选中的比生长速率，但在两种系统中，结果相似。然而，由于板旅馆中数据的噪声，从中获得的数据只能用于区分是否支持生长的基质，而生物筛选的数据还提供了基质偏好的信息。所有菌株都优先使用葡萄糖，其次是木糖和半乳糖。五株菌株也能在甘油上生长。因此，重要的是要尽量减少接种物中引入的甘油量，以避免对较差基质上的生长结果产生误解。我们得出的结论是，两种系统都可以为丝状真菌提供生理数据，只要在测量中包含足够的重复。
