Development of a novel microplate for high-throughput screening and optimization of DHA producing strains based on CFD technology
Shao-Jie He1, Lin-Hui Yang1, Yuan-Hang Du1, Ling-Ling Tong1, Yue Wang1, Dong-Sheng Guo1*
1School of Food Science and Pharmaceutical Engineering, Nanjing Normal University, Nanjing, China
*Corresponding Author: Dong-Sheng Guo, School of Food Science and Pharmaceutical Engineering, Nanjing Normal University, No. 1 Wenyuan Road, Nanjing 210023, China. Email :guodongs@njnu.edu.cn
Abstract : Microtiter plates are suitable for screening and process development of most microorganisms. They are currently the container of choice for high-throughput and small-scale microbial culture, but require optimization for specific work. This research presents a novel type of microtiter plate was developed using computational fluid dynamics (CFD) technology. The new plate provides high oxygen supply and optimal mixing effects for the fermentation culture of docosahexaenoic acid (DHA) producing strains, surpassing the conventional method of strains screening with shake flasks, which is insufficient. the shape of the microtiter plate was modified, and baffles were introduced to improve mass transfer and oxygen supply effects in the vibrating bioreactor. CFD technology was used to model the new plate’s characteristics, establishing the superiority of hexagonal microtiter plates with six baffles. Parameters in the incubation process, such as vibration frequency and liquid load, were optimized, and the final result achieved a KLa of 0.61s-1 and a volume power input of 2364 w/m3, which was 4-5 times better than the original 96-well plate. The culture results optimized by the model were also verified. Therefore, this new microtiter plate provides a powerful tool for future high-throughput screening of strains.
Keywords : docosahexaenoic acid, computational fluid dynamics, high-throughput screening, microplate
Abbreviations : CFD, computational fluid dynamics; DHA, docosahexaenoic acid; KLa, Oxygen transfer coefficient; ε, average turbulent dissipation rate; k, turbulent flow kinetic energy; RSM, response surface methodology; MSG, monosodium glutamate; Yeast, yeast extract; Glu, glucose.