scholarly journals Optimization of Pre-Inoculum, Fermentation Process Parameters and Precursor Supplementation Conditions to Enhance Apigenin Production by a Recombinant Streptomyces albus Strain

Fermentation ◽  
2021 ◽  
Vol 7 (3) ◽  
pp. 161
Author(s):  
Simona Barbuto Ferraiuolo ◽  
Odile Francesca Restaino ◽  
Ignacio Gutiérrez-del-Río ◽  
Riccardo Ventriglia ◽  
Marcella Cammarota ◽  
...  

Streptomyces albus J1074-pAPI (Streptomyces albus-pAPI) is a recombinant strain constructed to biotechnologically produce apigenin, a flavonoid with interesting bioactive features that up to now has been manufactured by extraction from plants with long and not environmentally friendly procedures. So far, in literature, only a maximum apigenin concentration of 80.0 µg·L−1 has been obtained in shake flasks. In this paper, three integrated fermentation strategies were exploited to enhance the apigenin production by Streptomyces albus J1074-pAPI, combining specific approaches for pre-inoculum conditions, optimization of fermentation process parameters and supplementation of precursors. Using a pre-inoculum of mycelium, the apigenin concentration increased of 1.8-fold in shake flask physiological studies. In 2L batch fermentation, the aeration and stirring conditions were optimized and integrated with the new inoculum approach and the apigenin production reached 184.8 ± 4.0 µg·L−1, with a productivity of 2.6 ± 0.1 μg·L−1·h−1. The supplementation of 1.5 mM L-tyrosine in batch fermentations allowed to obtain an apigenin production of 343.3 ± 3.0 µg·L−1 in only 48 h, with an increased productivity of 7.1 ± 0.1 μg·L−1·h−1. This work demonstrates that the optimization of fermentation process conditions is a crucial requirement to increase the apigenin concentration and productivity by up to 4.3- and 10.7-fold.

2009 ◽  
Vol 76 (3) ◽  
pp. 744-750 ◽  
Author(s):  
Rintze M. Zelle ◽  
Erik de Hulster ◽  
Wendy Kloezen ◽  
Jack T. Pronk ◽  
Antonius J. A. van Maris

ABSTRACT A recent effort to improve malic acid production by Saccharomyces cerevisiae by means of metabolic engineering resulted in a strain that produced up to 59 g liter−1 of malate at a yield of 0.42 mol (mol glucose)−1 in calcium carbonate-buffered shake flask cultures. With shake flasks, process parameters that are important for scaling up this process cannot be controlled independently. In this study, growth and product formation by the engineered strain were studied in bioreactors in order to separately analyze the effects of pH, calcium, and carbon dioxide and oxygen availability. A near-neutral pH, which in shake flasks was achieved by adding CaCO3, was required for efficient C4 dicarboxylic acid production. Increased calcium concentrations, a side effect of CaCO3 dissolution, had a small positive effect on malate formation. Carbon dioxide enrichment of the sparging gas (up to 15% [vol/vol]) improved production of both malate and succinate. At higher concentrations, succinate titers further increased, reaching 0.29 mol (mol glucose)−1, whereas malate formation strongly decreased. Although fully aerobic conditions could be achieved, it was found that moderate oxygen limitation benefitted malate production. In conclusion, malic acid production with the engineered S. cerevisiae strain could be successfully transferred from shake flasks to 1-liter batch bioreactors by simultaneous optimization of four process parameters (pH and concentrations of CO2, calcium, and O2). Under optimized conditions, a malate yield of 0.48 ± 0.01 mol (mol glucose)−1 was obtained in bioreactors, a 19% increase over yields in shake flask experiments.


Author(s):  
I. W. Kularathne ◽  
C. A. Gunathilaka ◽  
A. C. Ratnaweera ◽  
C. S. Kalpage ◽  
S. Rajapakse ◽  
...  

2019 ◽  
Vol 66 (6) ◽  
pp. 930-938 ◽  
Author(s):  
Mengge Ning ◽  
Shujie Zhang ◽  
Yanli Xie ◽  
Wei Wang ◽  
Yajun Gao

2014 ◽  
Vol 2014 ◽  
pp. 1-11 ◽  
Author(s):  
Saprativ P. Das ◽  
Debasish Das ◽  
Arun Goyal

The statistical optimization of different fermentation process parameters in SSF of mixed MAA and organosolv pretreated 1% (w v−1) wild grass, namely, recombinant Clostridium thermocellum hydrolytic enzymes’ volume (GH5 cellulase, GH43 hemicellulase), fermentative microbes’ inoculum volume (Saccharomyces cerevisiae, Candida shehatae), pH, and temperature, was accomplished by Taguchi orthogonal array design. The optimized parameters in 100 mL of fermentation medium were (%, v v−1) as follows: 1.0, recombinant GH5 cellulase (5.7 mg−1, 0.45 mg mL−1); 2.0, recombinant GH43 hemicellulase (3.7 U mg−1, 0.32 mg mL−1); 1.5, S. cerevisiae (3.9 × 108 cells mL−1); 0.25, C. shehatae (2.7 × 107 cells mL−1); pH, 4.3; and temperature, 35∘C. pH with p-value 0.001 was found to be the most significant factor affecting SSF. The ethanol titre obtained in Taguchi optimized shake flask SSF was 2.0 g L−1 implying a 1.3-fold increase as compared to ethanol titre of 1.5 g L−1 in unoptimized shake flask SSF. A 1.5-fold gain in ethanol titre (3.1 g L−1) was obtained with the same substrate concentration in lab scale bioreactor on scaling up the shake flask SSF with Taguchi optimized process parameters.


2007 ◽  
Vol 36 (3) ◽  
pp. 366-370 ◽  
Author(s):  
Eun-Jung Kim ◽  
Yeong-Hoon Kim ◽  
Jong-Won Kim ◽  
Hyo-Hyung Lee ◽  
Yu-Jin Ko ◽  
...  

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