Abstract
25-hydroxyvitamin D3 has attracted considerable attention due to its great medical value and huge market demand in animal husbandry. Microbial production of 25-hydroxyvitamin D3 has been recognized as an alternative superior to traditional chemical synthesis. In this study, a Gram-positive bacteria zju 4-2 (CCTCC M 2019385) was isolated from the soil using vitamin D3 as the sole carbon source and was identified as Bacillus cereus according to its physiological characteristics and 16S rRNA analysis, which also showed a relatively high capacity for 25-hydroxyvitamin D3 production. Through systematic optimization of different catalytic conditions, the optimal solvent system of vitamin D3, vitamin D3 addition time and concentration, temperature, and pH were shown to be propylene glycol/ethanol (v/v = 9:1), early stationary phase, 2 g/L, 37 °C, and pH 7.2, respectively. With these optimal conditions, 796 mg/L of 25-hydroxyvitamin D3 was achieved after 48 h bioconversion with zju 4-2 at the shake flask level. Finally, up to 830 mg/L 25-hydroxyvitamin D3 with a yield of 41.5% was obtained in a 5 L fermentation tank. Our developed biotransformation process with this newly isolated strain provides a platform to produce 25-hydroxyvitamin D3 efficiently at industrialization scale.
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References
Abbas A, Aboulwafa MM, Aboshanab K, Hassouna N (2011) Optimization of culture conditions for transformation of vitamin D3 to calcitriol by Actinomyces hyovaginalis isolate A11-2. Arch Clinical Microb 2:1–15
Abdulmughni A, Jóźwik IK, Brill E, Hannemann F, Thunnissen A, Bernhardt R (2017) Biochemical and structural characterization of CYP109A2, a vitamin D3 25-hydroxylase from Bacillus megaterium. FEBS J 284:3881–3894. https://doi.org/10.1111/febs.14276
Andrews DR, Barton DHR, Hesse RH, Pechet MM (1986) Synthesis of 25-hydroxy- and 1.alpha., 25-dihydroxy vitamin D3 from vitamin D2 (calciferol). J Organomet Chem 51(25):4819–4828. https://doi.org/10.1021/jo00375a013
Ban J-G, Kim H-B, Lee M-J, Anbu P, Kim E-SJ (2014) Identification of a vitamin D3-specific hydroxylase genes through actinomycetes genome mining. J Ind Microbiol Biotechnol 41:265–273. https://doi.org/10.1007/s10295-013-1336-9
Carlberg C (2019) Nutrigenomics of vitamin D. Nutrients 11(3):676. https://doi.org/10.3390/nu11030676
Carlberg C, Haq A (2018) The concept of the personal vitamin D response index. J Steroid Biochem 175:12–17. https://doi.org/10.1016/j.jsbmb.2016.12.011
Cheng CYS, Kim TK, Jeayeng S, Slominski AT, Tuckey RC (2018) Properties of purified CYP2R1 in a reconstituted membrane environment and its 25-hydroxylation of 20-hydroxyvitamin D3. J Steroid Biochem 177:59–69. https://doi.org/10.1016/j.jsbmb.2017.07.011
Chun RF, Liu NQ, Lee T, Schall JI, Denburg MR, Rutstein RM, Adam JS, Zemel BS, Stallings VA, Hewison M (2015) Vitamin D supplementation and antibacterial immune responses in adolescents and young adults with HIV/AIDS. J Steroid Biochem 148:290–297. https://doi.org/10.1016/j.jsbmb.2014.07.013
Dhas Y, Banerjee J, Damle G, Mishra N (2019) Association of vitamin D deficiency with insulin resistance in middle-aged type 2 diabetics. Clin Chim Acta 492:95–101. https://doi.org/10.1016/j.cca.2019.02.014
Ehrhardt M, Gerber A, Hannemann F, Bernhardt R (2016) Expression of human CYP27A1 in B. megaterium for the efficient hydroxylation of cholesterol, vitamin D3 and 7-dehydrocholesterol. J Biotechnol 218:34–40. https://doi.org/10.1016/j.jbiotec.2015.11.021
Fujii Y, Tamura T (2010) Hydroxylase e and use thereof. US Patent 20100297712A1
Hayashi K, Yasuda K, Sugimoto H, Ikushiro S, Kamakura M, Kittaka A, Horst RL, Chen TC, Ohta M, Shiro Y, Sakaki T (2010) Three-step hydroxylation of vitamin D3 by a genetically engineered CYP105A1. FEBS J 277:3999–4009. https://doi.org/10.1111/j.1742-4658.2010.07791.x
Huang J, Du Y, Xu G, Zhang H, Zhu F, Huang L, Xu Z (2011) High yield and cost-effective production of poly (γ-glutamic acid) with Bacillus subtilis. Eng Life Sci 11:291–297. https://doi.org/10.1002/elsc.201000133
Imoto N, Nishioka T, Tamura T (2011) Permeabilization induced by lipid II-targeting lantibiotic nisin and its effect on the bioconversion of vitamin D3 to 25-hydroxyvitamin D3 by Rhodococcus erythropolis. Biochemi Bioph Res Co 405:393–398. https://doi.org/10.1016/j.bbrc.2011.01.038
Jacoby C, Eipper J, Warnke M, Tiedt O, Mergelsberg M, Stärk H-J, Daus B, Martín-Moldes Z, Zamarro MT, Díaz E, Boll M (2018) Four molybdenum-dependent steroid C-25 hydroxylases: heterologous overproduction, role in steroid degradation, and application for 25-hydroxyvitamin D3 synthesis. Mbio 9(3):e0694–e0618. https://doi.org/10.1128/mBio.00694-18
Janocha S, Schmitz D, Bernhardt R (2015) Terpene hydroxylation with microbial cytochrome P450 Monooxygenases. Adv Biochem Eng Biotechnol 148:215–250. https://doi.org/10.1007/10_2014_296
Jin C, Wang Y, Sun B, Su W (2018) A concise synthesis of 25-hydroxycholesterol from hyodesoxycholic acid. J Chem Res 42(2):96–99. https://doi.org/10.3184/174751918X15184426915885
Kakhki RAM, Heuthorst T, Mills A, Neijat M, Kiarie E (2018) Interactive effects of calcium and top-dressed 25-hydroxy vitamin D3 on egg production, egg shell quality, and bones attributes in aged Lohmann LSL-lite layers. Poult Sci 98:1254–1262. https://doi.org/10.3382/ps/pey446
Kalimuthu P, Wojtkiewicz AM, Szaleniec M, Bernhardt PV (2018) Electrocatalytic hydroxylation of sterols by steroid C25 dehydrogenase from Sterolibacterium denitrificans. Chem Eur J 24(30):7710–7717. https://doi.org/10.1002/chem.201800616
Kang D-J, Lee H-S, Park J-T, Bang JS, Hong S-K, Kim T-Y (2006) Optimization of culture conditions for the bioconversion of vitamin D3 to 1α,25-dihydroxyvitamin D3 using Pseudonocardia autotrophica ID 9302. Biotechnol Bioprocess Eng 11:408–413. https://doi.org/10.1007/BF02932307
Kang D-J, Im J-H, Kang J-H, Kim KH (2015a) Whole cell bioconversion of vitamin D3 to calcitriol using Pseudonocardia sp. KCTC 1029BP. Bioprocess Biosyst Eng 38:1281–1290. https://doi.org/10.1007/s00449-015-1368-9
Kang D-J, Im J-H, Kang J-H, Kim KH (2015b) Bioconversion of vitamin D3 to calcifediol by using resting cells of Pseudonocardia sp. Biotechnol Lett 37:1895–1904. https://doi.org/10.1007/s10529-015-1862-9
Kurek-Tyrlik A, Michalak K, Wicha J (2005) Synthesis of 17-epi-calcitriol from a common androstane derivative, involving the ring B photochemical opening and the intermediate triene ozonolysis. J Organomet Chem 70(21):8513–8521. https://doi.org/10.1021/jo051357u
Liang J, Xu Z, Liu T, Lin J, Cen P (2008) Effects of cultivation conditions on the production of natamycin with Streptomyces gilvosporeus LK-196. Enzyme Microb Tech 42:145–150. https://doi.org/10.1016/j.enzmictec.2007.08.012
Luo JQ, Jiang F, Fang WZ, Lu Q (2017) Optimization of bioconversion conditions for vitamin D3 to 25-hydroxyvitamin D using Pseudonocardia autotrophica CGMCC5098. Biocatal Biotransfor 35(1):11–18. https://doi.org/10.1080/10242422.2016.1268130
Manchand PS, Yiannikouros GP, Belica PS, Madan P (1995) Nickel-mediated conjugate addition. elaboration of calcitriol from ergocalciferol. J Organomet Chem 60(20):6574–6581. https://doi.org/10.1021/jo00125a051
Ogawa S, Kakiyama G, Muto A, Hosoda A, Mitamura K, Ikegawa S, Hofmann AF, Iida T (2009) A facile synthesis of C-24 and C-25 oxysterols by in situ generated ethyl(trifluoromethyl)dioxirane. Steroids 74(1):81–87. https://doi.org/10.1016/j.steroids.2008.09.015
Ohmura K, Kato M, Watanabe T, Oku K, Bohgaki T, Horita T, Yasuda S, Ito YM, Sato N, Atsumi T (2018) Effect of combined treatment with bisphosphonate and vitamin D on atherosclerosis in patients with systemic lupus erythematosus: a propensity score-based analysis. Arthritis Res Ther 20:72. https://doi.org/10.1186/s13075-018-1589-9
Putkaradze N, Litzenburger M, Abdulmughni A, Milhim M, Brill E, Hannemann F, Bernhardt RJAM (2017) CYP109E1 is a novel versatile statin and terpene oxidase from Bacillus megaterium. Appl Microbiol Biotechol 101:8379–8393. https://doi.org/10.1007/s00253-017-8552-6
Roth DE, Perumal N, Al Mahmud A, Baqui AH (2013) Maternal vitamin D3 supplementation during the third trimester of pregnancy: effects on infant growth in a longitudinal follow-up study in bangladesh. J Pediatr-Us 163:1605–1611. https://doi.org/10.1016/j.jpeds.2013.07.030
Rugor A, Tataruch M, Staroń J, Dudzik A, Niedzialkowska E, Nowak P, Hogendorf A, Michalik-Zym A, Napruszewska DB, Jarzębski A, Szymańska K, Białas W, Szaleniec MJAM (2017a) Regioselective hydroxylation of cholecalciferol, cholesterol and other sterol derivatives by steroid C25 dehydrogenase. Appl Microbiol Biotechnol 101(3):1163–1174. https://doi.org/10.1007/s00253-016-7880-2
Rugor A, Wójcik-Augustyn A, Niedzialkowska E, Mordalski S, Staroń J, Bojarski A, Szaleniec M (2017b) Reaction mechanism of sterol hydroxylation by steroid C25 dehydrogenase – homology model, reactivity and isoenzymatic diversity. J Inorg Biochem 173:28–43. https://doi.org/10.1016/j.jinorgbio.2017.04.027
Ryznar T, Krupa M, Kutner A (2002) Syntheses of vitamin D metabolites and analogs. Retrospect and prospects. Przem Chem 81(5):300–310
Sasaki J, Mikami A, Mizoue K, Omura S (1991) Transformation of 25- and 1 alpha-hydroxyvitamin D3 to 1 alpha, 25-dihydroxyvitamin D3 by using Streptomyces sp. strains. Appl Environ Microbiol 57:2841–2846
Sasaki J, Miyazaki A, Saito M, Adachi T, Mizoue K, Hanada K, Omura S (1992) Transformation of vitamin D3 to 1α,25-dihydroxyvitamin D3 via 25-hydroxyvitamin D3 using Amycolata sp. strains. Appl Microbiol Biotechnol 38(2):152–157. https://doi.org/10.1007/BF00174460
Shen Y, Yu Z, Yang X, Wang F, Luo J, Wang M (2017) A new technique for promoting cyclic utilization of cyclodextrins in biotransformation. J Ind Microbiol Biotechnol 44:1–7. https://doi.org/10.1007/s10295-016-1856-1
Shi Z, Wei P, Zhu X, Cai J, Huang L, Xu Z (2012) Efficient production of l-lactic acid from hydrolysate of Jerusalem artichoke with immobilized cells of Lactococcus lactis in fibrous bed bioreactors. Enzyme Microb Techn 51:263–268. https://doi.org/10.1016/j.enzmictec.2012.07.007
Szaleniec M, Wojtkiewicz AM, Bernhardt R, Borowski T, Donova MJAM, Biotechnology (2018) Bacterial steroid hydroxylases: enzyme classes, their functions and comparison of their catalytic mechanisms. Appl Microbiol Biotechnol 102(19):8153-8171. https://doi.org/10.1007/s00253-018-9239-3
Tamura K, Filipski A, Peterson D, Stecher G, Kumar S (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30:2725–2729. https://doi.org/10.1093/molbev/mst197
Tuckey RC, Li W, Ma D, Cheng CYS, Wang KM, Kim T-K, Jeayeng S, Slominski AT (2018) CYP27A1 acts on the pre-vitamin D3 photoproduct, lumisterol, producing biologically active hydroxy-metabolites. J Steroid Biochem 181:1–10. https://doi.org/10.1016/j.jsbmb.2018.02.008
Warnke M, Jung T, Dermer J, Hipp K, Jehmlich N, von Bergen M, Ferlaino S, Fries A, Müller M, Boll M (2016) 25-hydroxyvitamin D3 synthesis by enzymatic steroid side-chain hydroxylation with water. Angew Chem Int Ed Eng 55(5):1881–1884. https://doi.org/10.1002/anie.201510331
Yamamoto E, Jørgensen TN (2019) Immunological effects of vitamin D and their relations to autoimmunity. J Autoimmun 100:7–16. https://doi.org/10.1016/j.jaut.2019.03.002
Yasuda K, Endo M, Ikushiro S, Kamakura M, Ohta M, Sakaki T (2013) UV-dependent production of 25-hydroxyvitamin D2 in the recombinant yeast cells expressing human CYP2R1. Biochem Bioph Res Co 434(2):311–315. https://doi.org/10.1016/j.bbrc.2013.02.124
Yasutake Y, Nishioka T, Imoto N, Tamura T (2013) A single mutation at the ferredoxin binding site of p450 Vdh enables efficient biocatalytic production of 25-hydroxyvitamin D3. Chembiochem 14:2284–2291. https://doi.org/10.1002/cbic.201300386
Yoon S-H, Ha S-M, Kwon S, Lim J, Kim Y, Seo H, Chun J (2017) Introducing EzBioCloud: a taxonomically united database of 16S rRNA gene sequences and whole-genome assemblies. Int J Syst Evol Microbiol 67:1613–1617. https://doi.org/10.1099/ijsem.0.001755
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This work was financially supported by National Natural Science Foundation of China (No. 21576232, 21606205 & 21808199) and the National Key Research and Development Program of China (No. 2018YFC1603900).
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Tang, D., Liu, W., Huang, L. et al. Efficient biotransformation of vitamin D3 to 25-hydroxyvitamin D3 by a newly isolated Bacillus cereus strain. Appl Microbiol Biotechnol 104, 765–774 (2020). https://doi.org/10.1007/s00253-019-10250-1
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DOI: https://doi.org/10.1007/s00253-019-10250-1