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Generic Protocol for Optimization of Heterologous Protein Production Using Automated Microbioreactor Technology
Published on: December 15, 2017
Optimization of Fermentation Conditions for Enhanced Single Cell Protein Production by Rossellomorea marisflavi NDS
Hui Zhang1,2, Wenwen Zhang1,2, Wen Zhang1,2
1Microbial Development and Metabolic Engineering Laboratory, School of Marine Science, Ningbo University, Ningbo 315211, China.
Abstract:
Microbial proteins offer a sustainable alternative for animal nutrition. Rossellomorea marisflavi NDS, a bacterium isolated from seawater, was previously identified as a promising candidate due to its high protein content. This study aimed to enhance its single cell protein production through systemic fermentation optimization. Single-factor optimization in shake flask determined the optimal conditions to be: a salinity of 20‱ NaCl, a temperature of 32 °C, and an initial pH of 7.3, and a medium composed of 1% (w/v) corn flour, 1% peptone, 0.3% beef extract, and 0.2% KCl. Scaling up to a 10 L bioreactor demonstrated that a two-stage agitation strategy (150 rpm for the first 20 h followed by 180 rpm for the remaining 12 h) enhanced single cell protein yield. Furthermore, allowing the pH to fluctuate freely was more beneficial for protein production than maintaining a constant pH of 7.3 ± 0.02. Under these optimized conditions, the biomass composition (wet weight) was determined to be 2.3767 ± 0.0205% crude ash, 15.6013 ± 0.0082% crude protein, 0.1023 ± 0.0026% crude lipid, and 2.6997 ± 0.0021% carbohydrates. Amino acid analysis revealed a rich profile, with lysine and glutamic acid being the predominant essential and non-essential amino acids, respectively. Fatty acids analysis indicated that C14:1n5 was the most dominant. These findings underscore the potential of R. marisflavi NDS as a high-quality dietary protein supplement and provide a solid foundation for its industrial-scale production.

