Bioconversion of vitamin D3 into calcitriol by Actinomyces hyovaginalis isolate CCASU- A11-2

Abstract

Vitamin D3 is a fat-soluble prohormone that is activated inside the liver to produce 25-hydroxyvitamin D3 (calcidiol), and in the kidney to produce the fully active 1α, 25-dihydroxy vitamin D3 (calcitriol). A previous work piloted in our laboratory, resulted in a successful recovery of a local soil-promising Actinomyces hyovaginalis isolate CCASU-A11-2 capable of converting vitamin D3 into calcitriol. Despite the rising amount of research on vitamin D3 bioconversion into calcitriol, further deliberate studies on this topic can significantly contribute to the improvement of such a bioconversion process. Therefore, this work aimed to improve the bioconversion process, using the study isolate, in a 14 L laboratory fermenter (4 L fermentation medium composed of fructose (15 g/L), defatted soybean (15 g/L), NaCl (5 g/L), CaCO3 2 g/L); K2HPO4, (1 g/L) NaF (0.5 g/L) and initial of pH 7.8) where different experiments were undertaken to investigate the effect of different culture conditions on the bioconversion process. Using the 14 L laboratory fermenter, the calcitriol production was increased by about 2.5-fold (32.8 µg/100 mL) to that obtained in the shake flask (12.4 µg/100 mL). The optimal bioconversion conditions were inoculum size of 2% v/v, agitation rate of 200 rpm, aeration rate of 1 vvm, initial pH of 7.8 (uncontrolled); addition of vitamin D3 (substrate) 48 h after the start of the main culture. In conclusion, the bioconversion of vitamin D3 into calcitriol in a laboratory fermenter showed a 2.5-fold increase as compared to the shake flask level where, the important factors influencing the bioconversion process were the aeration rate, inoculum size, the timing of substrate addition, and the fixed pH of the fermentation medium. So, those factors should be critically considered for the scaling-up of the biotransformation process.

Integrated Synthesis of Calcitriol and 26,27-Hexadeutero Calcitriol

“…25 In addition, bioconversion of vitamin D 3 into calcitriol ( 1 ) has been actively investigated for potential bioindustrial manufacture of calcitriol ( 1 ). 26 27…”

Characterization of vitamin D3 biotransformation by the cell lysate of Actinomyces hyovaginalis CCASU-A11-2

“…An aliquot (1 mL) of the obtained seed culture was used to inoculate the 50 mL medium used for the main culture of study isolate contained in a 250 mL Erlenmeyer flask (main culture), incubated at 30 °C and 200 rpm for 48 h. Then centrifugation was carried out at 5000 rpm for 10 min, the sediment was re-suspended in 10 mL phosphate buffered salts solution, and transferred to a 50 mL beaker. The cells were then lysed via sonication using a probe sonicator device adjusted at 70% amplitude as previously reported (Abbas et al 2011b ).…”

“…The extraction process was carried out as previously reported (Bligh and Dyer 1959 ), and (Abbas et al 2011b ). This experiment was done in triplicate and both the mean and standard deviations were calculated.…”