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.
Similar content being viewed by others
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
During A, Harrison EH (2004) Intestinal absorption and metabolism of carotenoids: insights from cell culture. Arch Biochem Biophys 430:77–88
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
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
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
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
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
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
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
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
Lindqvist A, Andersson S (2002) Biochemical properties of purified recombinant human beta-carotene 15,15′-monooxygenase. J Biol Chem 277:23942–23948
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
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
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
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
Shikama K (2006) Nature of the FeO2 bonding in myoglobin and hemoglobin: a new molecular paradigm. Prog Biophys Mol Biol 91:83–162
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
Acknowledgments
This study was supported by a grant (2009-0084490) from the Basic Research Program, Ministry of Education, Science and Technology.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
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
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10529-011-0556-1