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Production of β-apo-10′-carotenal from β-carotene by human β-carotene-9′,10′-oxygenase expressed in E. coli

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Abstract

The gene encoding human β-carotene-9′,10′-oxygenase, which cleaves the 9′,10′ double bond in β-carotene into β-apo-10′-carotenal, was cloned and expressed in Escherichia coli. Under aqueous conditions, the optimum organic solvent for the formation of detergent micelles was toluene. The optimum pH, temperature, detergent type, and the optimum concentrations of detergent, substrate, and enzyme for β-apo-10′-carotenal production were 8.0, 37°C, Tween 40, 2.4%, 300 mg β-carotene/l, and 0.25 U/ml, respectively. Under the optimum conditions, 43 mg β-apo-10′-carotenal/l was produced after 21 h with a conversion of 14%. This is the first report to describe the enzymatic production of β-apo-10′carotenal.

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References

  • Berry SD, Davis SR, Beattie EM, Thomas NL, Burrett AK, Ward HE, Stanfield AM, Biswas M, Ankersmit-Udy AE, Oxley PE, Barnett JL, Pearson JF, van der Does Y, Macgibbon AH, Spelman RJ, Lehnert K, Snell RG (2009) Mutation in bovine beta-carotene oxygenase 2 affects milk color. Genetics 182:923–926

    Article  PubMed  CAS  Google Scholar 

  • During A, Harrison EH (2004) Intestinal absorption and metabolism of carotenoids: insights from cell culture. Arch Biochem Biophys 430:77–88

    Article  PubMed  CAS  Google Scholar 

  • During A, Nagao A, Hoshino C, Terao J (1996) Assay of beta-carotene 15,15′-dioxygenase activity by reverse-phase high-pressure liquid chromatography. Anal Biochem 241:199–205

    Article  PubMed  CAS  Google Scholar 

  • Fidge N, Smith F, Goodman D (1969) Vitamin A and carotenoids. The enzymic conversion of beta-carotene into retinal in hog intestinal mucosa. Biochem J 114:689–694

    PubMed  CAS  Google Scholar 

  • Hu KQ, Liu C, Ernst H, Krinsky NI, Russell RM, Wang XD (2006) The biochemical characterization of ferret carotene-9′,10′-monooxygenase catalyzing cleavage of carotenoids in vitro and in vivo. J Biol Chem 281:19327–19338

    Article  PubMed  CAS  Google Scholar 

  • Kiefer C, Hessel S, Lampert JM, Vogt K, Lederer MO, Breithaupt DE, von Lintig J (2001) Identification and characterization of a mammalian enzyme catalyzing the asymmetric oxidative cleavage of provitamin A. J Biol Chem 276:14110–14116

    PubMed  CAS  Google Scholar 

  • Kim YS, Kim NH, Kim HJ, Lee JK, Kim SW, Oh DK (2007) Effective production of retinal from beta-carotene using recombinant mouse beta-carotene 15,15′-monooxygenase. Appl Microbiol Biotechnol 76:1339–1345

    Article  PubMed  CAS  Google Scholar 

  • Kim NH, Kim YS, Kim HJ, Oh DK (2008) Optimized formation of detergent micelles of beta-carotene and retinal production using recombinant human beta, beta-carotene 15,15′-monooxygenase. Biotechnol Prog 24:227–231

    Article  PubMed  CAS  Google Scholar 

  • Kim YS, Kim NH, Yeom SJ, Kim SW, Oh DK (2009) In vitro characterization of a recombinant Blh protein from an uncultured marine bacterium as a beta-carotene 15,15′-dioxygenase. J Biol Chem 284:15781–15793

    Article  PubMed  CAS  Google Scholar 

  • Kim YS, Park CS, Oh DK (2010) Retinal production from beta-carotene by beta-carotene 15,15′-dioxygenase from an unculturable marine bacterium. Biotechnol Lett

  • Lakshmanan M, Chansang H, Olson J (1972) Purification and properties of carotene 15,15′-dioxygenase of rabbit intestine. J Lipid Res 13:477–482

    PubMed  CAS  Google Scholar 

  • Lindqvist A, Andersson S (2002) Biochemical properties of purified recombinant human beta-carotene 15,15′-monooxygenase. J Biol Chem 277:23942–23948

    Article  PubMed  CAS  Google Scholar 

  • Lindqvist A, He YG, Andersson S (2005) Cell type-specific expression of beta-carotene 9′,10′-monooxygenase in human tissues. J Histochem Cytochem 53:1403–1412

    Article  PubMed  CAS  Google Scholar 

  • Nagao A, During A, Hoshino C, Terao J, Olson JA (1996) Stoichiometric conversion of all trans-beta-carotene to retinal by pig intestinal extract. Arch Biochem Biophys 328:57–63

    Article  PubMed  CAS  Google Scholar 

  • Redmond T, Gentleman S, Duncan T, Yu S, Wiggert B, Gantt E, Cunningham FJ (2001) Identification, expression, and substrate specificity of a mammalian beta-carotene 15,15′-dioxygenase. J Biol Chem 276:6560–6565

    Article  PubMed  CAS  Google Scholar 

  • Scherzinger D, Al-Babili S (2008) In vitro characterization of a carotenoid cleavage dioxygenase from Nostoc sp. PCC 7120 reveals a novel cleavage pattern, cytosolic localization and induction by highlight. Mol Microbiol 69:231–244

    Article  PubMed  CAS  Google Scholar 

  • Shikama K (2006) Nature of the FeO2 bonding in myoglobin and hemoglobin: a new molecular paradigm. Prog Biophys Mol Biol 91:83–162

    Article  PubMed  CAS  Google Scholar 

  • Tian R, Pitchford WS, Morris CA, Cullen NG, Bottema CD (2009) Genetic variation in the beta, beta-carotene-9′,10′-dioxygenase gene and association with fat colour in bovine adipose tissue and milk. Anim Genet 41:253–259

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

This study was supported by a grant (2009-0084490) from the Basic Research Program, Ministry of Education, Science and Technology.

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Correspondence to Deok-Kun Oh.

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Kim, YS., Yeom, SJ. & Oh, DK. Production of β-apo-10′-carotenal from β-carotene by human β-carotene-9′,10′-oxygenase expressed in E. coli . Biotechnol Lett 33, 1195–1200 (2011). https://doi.org/10.1007/s10529-011-0556-1

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  • DOI: https://doi.org/10.1007/s10529-011-0556-1

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