Faculty of Science and Technology, Norwegian University of Life Sciences, Postbox 5003, 1432, Ås, Norway.
Faculty of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, Postbox 5003, 1432, Ås, Norway.
Appl Microbiol Biotechnol. 2018 Jun;102(11):4915-4925. doi: 10.1007/s00253-018-8920-x. Epub 2018 Apr 11.
Recent developments in molecular biology and metabolic engineering have resulted in a large increase in the number of strains that need to be tested, positioning high-throughput screening of microorganisms as an important step in bioprocess development. Scalability is crucial for performing reliable screening of microorganisms. Most of the scalability studies from microplate screening systems to controlled stirred-tank bioreactors have been performed so far with unicellular microorganisms. We have compared cultivation of industrially relevant oleaginous filamentous fungi and microalga in a Duetz-microtiter plate system to benchtop and pre-pilot bioreactors. Maximal glucose consumption rate, biomass concentration, lipid content of the biomass, biomass, and lipid yield values showed good scalability for Mucor circinelloides (less than 20% differences) and Mortierella alpina (less than 30% differences) filamentous fungi. Maximal glucose consumption and biomass production rates were identical for Crypthecodinium cohnii in microtiter plate and benchtop bioreactor. Most likely due to shear stress sensitivity of this microalga in stirred bioreactor, biomass concentration and lipid content of biomass were significantly higher in the microtiter plate system than in the benchtop bioreactor. Still, fermentation results obtained in the Duetz-microtiter plate system for Crypthecodinium cohnii are encouraging compared to what has been reported in literature. Good reproducibility (coefficient of variation less than 15% for biomass growth, glucose consumption, lipid content, and pH) were achieved in the Duetz-microtiter plate system for Mucor circinelloides and Crypthecodinium cohnii. Mortierella alpina cultivation reproducibility might be improved with inoculation optimization. In conclusion, we have presented suitability of the Duetz-microtiter plate system for the reproducible, scalable, and cost-efficient high-throughput screening of oleaginous microorganisms.
近年来，分子生物学和代谢工程的发展使得需要测试的菌株数量大幅增加，这使得高通量筛选微生物成为生物工艺开发的重要步骤。可扩展性对于进行可靠的微生物筛选至关重要。迄今为止，大多数从微孔板筛选系统到控制搅拌罐生物反应器的可扩展性研究都是针对单细胞微生物进行的。我们比较了工业相关的油脂丝状真菌和微藻在 Duetz-微量滴定板系统中的培养，以及台式和预中试生物反应器。最大葡萄糖消耗率、生物量浓度、生物量的脂质含量、生物量和脂质产率值表明，Mucor circinelloides（差异小于 20%）和 Mortierella alpina（差异小于 30%）丝状真菌具有良好的可扩展性。Crypthecodinium cohnii 在微量滴定板和台式生物反应器中的最大葡萄糖消耗和生物量生产速率相同。由于该微藻在搅拌生物反应器中的剪切应力敏感性，生物量浓度和生物量的脂质含量在微量滴定板系统中明显高于台式生物反应器。尽管如此，与文献报道相比，在 Duetz-微量滴定板系统中获得的 Crypthecodinium cohnii 发酵结果令人鼓舞。Mucor circinelloides 和 Crypthecodinium cohnii 在 Duetz-微量滴定板系统中实现了良好的重现性（生物量生长、葡萄糖消耗、脂质含量和 pH 的变异系数小于 15%）。Mortierella alpina 的培养重现性可能通过接种优化得到改善。总之，我们已经证明了 Duetz-微量滴定板系统适用于重现性、可扩展性和成本效益高的油脂微生物高通量筛选。
