Influence of Substrate Feeding and Process Parameters on Production of Coenzyme Q<sub>10</sub> Using <i>Paracoccus denitrificans</i> ATCC 19367 Mutant Strain P-87

Abstract

Coenzyme Q 10 (CoQ 10 ), an important antioxidant molecule playing a major role in electron transport chain, has been commercially produced by fermentation process for the use in oral nutraceutical formulations. Constructing the high-yielding CoQ 10 producing strains is a pre-requisite for cost-effective production. A superior mutant strain P-87 generated from Paracoccus denitrificans ATCC 19367, which showed 1.25-fold improvement in specific CoQ 10 content higher than the wild type strain at shake flask level, was selected to carry out the studies on CoQ 10 yield improvement through fermenter process optimization. In the course of study, initially the cane-molasses-based medium and fed-batch fermentation strategy using pHBA in combination with sucrose were standardized in shake flask using wild type strain. This strategy was subsequently translated at 2 L laboratory fermenter while optimizing the fermentation process parameters using improved mutant strain P-87. Under optimized fermentation condition, mutant strain P-87 produced 49.85 mg/L of CoQ10 having specific content of 1.63 mg/g of DCW, which was 1.36 folds higher than the specific CoQ 10 content of wild-type strain under similar optimized condition. The temperature and DO were found to be critical parameters for CoQ 10 production by mutant strain P-87. The optimum temperature was found to be 32˚C and the optimum DO concentration to be maintained throughout the fermentation cycle was found to be 30% of air saturation. Overall, a new cost-effective process has been established for the production of CoQ 10 using the cheaper substrate "cane molasses" and higher CoQ 10 producing mutant strain P-87. *Corresponding author. P. Tokdar et al.967

Coenzyme Q10 and its Effective Sources

“…CoQ10, can be produced by chemical synthesis, extraction from biological tissues (plants and animal) and microbial fermentation (Laplante et al, 2009). In the wake of environmental awareness, the chemical options became least desirable due to inherent uses of solvents and chemicals in the process (Tokdar et al, 2014).…”

“…Microbial production offers an environmentally benign option based on the enzymatic catalysis at the cellular level for CoQ10 assembly. Moreover, this approach is attractive to the industry because the process is easy to control at a relatively low production cost (Tokdar et al, 2014). However, due to the limits of CoQ10 accumulation in cells, strain improvements have been made using genetic engineering (using recombinant nucleic acid technology), chemical mutagenesis and high hydrostatic pressure treatment (Kim et al, 2015).…”

“…Industrial production of CoQ10 have predominantly relied on bacterial and yeast mutants due to their higher CoQ10 content (Tokdar et al, 2014). The isolation of strains by mutagenesis and selection on inhibitors has shown to be the most successful strategy to enhance CoQ10 yields (Yen and Shih, 2009).…”

“…Recently CoQ10 found a wide range of therapeutic applications (Tokdar et al, 2014;Langsjoen, 1994). (Jankowski et al, 2016) Extensive research has been conducted to increase CoQ10 production to meet growing demands for this product.…”

Novel biotechnologies for nitrous oxide abatement

“…Likewise, a study assessing the effect of O 2 levels on the production of CoQ 10 in P. denitrificans CCM 982 showed that the highest yield (1.2 mg g −1 ) was obtained at an oxygen concentration of 2.5%, with a decrease in the coenzyme content to 0.43 mg g −1 when the O 2 levels increased to 21% (Kaplan et al, 1993). Recently, the CoQ 10 biosynthesis capacity of a mutant strain of P. denitrificans (P-87) has been evaluated using the precursor parahydroxy benzoic acid (Tokdar et al, 2014). This study revealed a maximum specific cell content of 1.63 mg g −1 , which was only 1.8-fold higher than that obtained in our study with no particular optimization strategy.…”