Optimization of culture conditions for the bioconversion of vitamin D3 to 1α,25-dihydroxyvitamin D3 usingPseudonocardia autotrophica ID 9302

Kang, Daehyun; Lee, Hong Sub; Park, Joon-Tae; Bang, Ji Sun; Hong, Soon-Kwang; Kim, Tae-Yong

doi:10.1007/bf02932307

Cited by 14 publications

(9 citation statements)

References 21 publications

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“…Our Findings revealed pH 7.8 was optimum for the biotransformation process. Such a finding was similar to that obtained by Kang et al 2006 and Fujii et al 2009 . Also, it was found that the optimum temperature for the enzyme activity was 28 °C.…”

Section: Discussionsupporting

confidence: 93%

“…Also, it was found that the optimum temperature for the enzyme activity was 28 °C. However, a considerable decline in calcitriol production occurred at 37 °C as previously documented (Kang et al 2006 ). It is worth noting that, using the enzymes extracted from cells produced in optimized fermentation, would properly have higher enzyme level and activity as compared to those obtained from in shake flask.…”

Section: Discussionsupporting

confidence: 77%

“…Bioconversion of vitamin D3 offers a more cost-effective alternative to chemical synthesis. Despite this, within the Actinomycetales order, specifically the genera Streptomyces and Amycolata , only a limited number of microorganisms have demonstrated the capability to conduct this conversion (Sasaki et al 1991 , 1992 ) (Sawada et al 2004 ; Kang et al 2006 ) (Takeda et al 2006 ). An isolated strain, A. hyovaginalis CCASU-A11-2, exhibited the ability to transform vitamin D3 into calcitriol (Abbas et al 2011a ).…”

Section: Introductionmentioning

confidence: 99%

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

Abbas,

Elkhatib,

Aboulwafa

et al. 2024

AMB Expr

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A former work conducted in our Lab, lead to in a effective scale up of vitamin D3 bioconversion into calcitriol by Actinomyces (A.) hyovaginalis isolate CCASU-A11-2 in Lab fermenter (14 L) resulting in 32.8 µg/100 mL of calcitriol. However, the time needed for such a bioconversion process was up to 5 days. Therefore, the objective of this study was to shorten the bioconversion time by using cell-free lysate and studying different factors influencing bioconversion. The crude cell lysate was prepared, freeze-dried, and primarily fractionated into nine fractions, of which, only three fractions, 50, 100, and 150 mM NaCl elution buffers showed 22, 12, and 2 µg/10 mL, calcitriol production, respectively. Ammonium sulfate was used for protein precipitation, and it did not affect the bioconversion process except at a concentration of 10%w/v. Secondary fractionation was carried out using 80 mL of the 50 mM NaCl elution buffer and the results showed the 80 mL eluent volume was enough for the complete elution of the active protein. The pH 7.8, temperature 28 °C, and 6 h reaction time were optimum for maximum calcitriol production (31 µg/10 mL). In conclusion, the transformation of vitamin D3 into calcitriol was successfully carried out within 6 h and at pH 7.8 and 28 °C using fractionated cell lysate. This process resulted in a 10-fold increase in calcitriol as compared to that produced in our previous study using a 14 L fermenter (32.8 µg/100 mL). Therefore, cell-free lysate should be considered for industrial and scaling up vitamin D3 bioconversion into calcitriol.

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Section: Discussionsupporting

confidence: 93%

Section: Discussionsupporting

confidence: 77%

Section: Introductionmentioning

confidence: 99%

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

Abbas,

Elkhatib,

Aboulwafa

et al. 2024

AMB Expr

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“…Unfortunately, the chemical synthesis of calcitriol is a high-cost and laborious procedure that needs a lot of reaction steps (Kametani and Furuyama 1987 ). The microbial conversion of vitamin D 3 has a lower cost than chemical synthesis; nevertheless, the order Actinomycetales (well known to establish vitamin D 3 bioconversion), namely the two genera Streptomyces and Amycolata , showed very few microorganisms capable of conducting this conversion (Sasaki et al 1991 , 1992 ; Sawada et al 2004 ; Kang et al 2006 ; Takeda et al 2006 ). Furthermore, traditional organic synthesis of the hydroxylated vitamin D3 derivatives is highly complex, which results in low yield (Andrews et al 1986 ; Jin et al 2018 ; Ryznar et al 2002 ).…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2023

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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.

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“…33,34 However, synthetic calcitriol is industrially produced through a multistep and expensive reaction making it more expensive than the precursor cholecalciferol. 35,36 Recently, it was suggested that some cell types including hMSCs, possess the enzymatic toolbox for VD3 metabolism and action, including the enzymes for cholecalciferol hydroxylation into calcitriol. 37 In an attempt to overcome the thermal instability of PLA and its inherent biological inertness, we explored the effect of VD3 as a plasticizer.…”

Section: Introductionmentioning

confidence: 99%

Cholecalciferol as Bioactive Plasticizer of High Molecular Weight Poly(D,L-Lactic Acid) Scaffolds for Bone Regeneration

Calore

Hadavi

Honing

et al. 2022

Tissue Engineering Part C: Methods

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Optimization of culture conditions for the bioconversion of vitamin D3 to 1α,25-dihydroxyvitamin D3 usingPseudonocardia autotrophica ID 9302

Cited by 14 publications

References 21 publications

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

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

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

Cholecalciferol as Bioactive Plasticizer of High Molecular Weight Poly(D,L-Lactic Acid) Scaffolds for Bone Regeneration

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