Browse > Article

Characterization of Human ${\beta}-Carotene$ 15,15-dioxygenase Isolated from Recombinant Escherichia coli  

Shin, Won-Phil (Department of Food Science and Technology, Seoul National University of Technology)
Chang, Pahn-Shick (Department of Food Science and Technology, Seoul National University of Technology)
Publication Information
Korean Journal of Food Science and Technology / v.36, no.3, 2004 , pp. 440-447 More about this Journal
Abstract
Characteristics of human ${\beta}-carotene$ 15,15'-dioxygenase isolated by recombinant DNA technology was elucidated. Optimal pH and temperature were 9.0 and $40^{\circ}C$, respectively. Enzyme activity was temperature-sensitive. Enzyme was stable at pH 6.0-9.0 for 24 hr and under $5^{\circ}C$. Half-life of enzyme at $35^{\circ}C$ was 40 min. Crude preparations of enzyme were inhibited by ferrous ion-chelating agent and sulfhydryl-binding agent. GSH offsets inhibitory effect of PCMB. With increasing substrate concentrations, enzyme activity gave typical Michaelis-Menten curve, Based on Hanes-Woolf plot of data, $K_{m}\;and\;V_{max$ were $3.39{\times}10^{6}\;M\;and\;1.2\;pmol/mg$ protein/min, respectively.
Keywords
${\beta}-carotene$ 15,15'-dioxygenase; recombinant DNA technology; optimal pH and temperature; $K_m$; $V_{max}$;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Wang XD, Krinsky NI, Tang G, Russell RM. Retinoic acid can be produced from excentric cleavage of $\beta$-carotene in human intestinal mucosa. Arch. Biochm. Biophys. 293: 298-304 (1992)   DOI   ScienceOn
2 Goodman DS, Huang HS, Kanai M, Shiratori T. Mechanism of biosynthesis of vitamin A from $\beta$-carotene. J. Biol. Chem. 241: 1929-1932 (1966)
3 Sklan D. Carotene-cleavage activity in chick intestinal mucosa cytosol: association with a high-molecular-weight lipid-protein aggregate fraction and partial characterization of the activity. Br. J. Nutr. 50: 417-425 (1983)   DOI   ScienceOn
4 Goodman DS, Huang HS, Kanai M, Shiratori TJ. The enzymatic conversion of all-trans $\beta$-carotene into retinal. Biol. Chem. 242: 3543-3554 (1967)
5 Devery J, Milborrow BV. $\beta-Carotene$ 15,15'-dioxygenase (EC 1.13.11.21) isolation reaction mechanism and improved assay procedure. Br. J. Nutr. 72: 397-414 (1994)   DOI   ScienceOn
6 During A, Nagao A, Hoshino C, Terao J. Assay of $\beta-Carotene$ 15,15'-dioxygenase activity by reverse phase high pressure liquid chromatography. Anal. Biochem. 241: 199-205 (1996)   DOI   PUBMED   ScienceOn
7 Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, Fujimoto EK, Goeke NM, Olson BJ, Klenk DC. Measurement of protein using bicinchoninic acid. Anal. Biochem. 150: 76-85 (1985)   DOI   ScienceOn
8 Dmitrovskii AA, Gessler NN, Gomboeva SB, Ershov YU. Enzymatic oxidation of $\beta-apo-8-carotenal$ to $\beta-apo-14-carotenal$by an enzyme different from $\beta-carotene$ 15,15'-dioxygenase. Biochemistry 62: 787-792 (1997)
9 Nagao A, During A, Hoshino C, Terao J, Olson JA. Stoichiometric conversion of all $trans-\beta-carotene$ to retinal by pig intestinal extract. Arch. Biochem. Biophys. 328: 57-63 (1996)   DOI   PUBMED   ScienceOn
10 Burton GW, Ingold KU. $\beta$-Carotene: an unusual type of lipid antioxidant. Science 244: 569-573 (1984)   DOI
11 Edes TE, Thornton W. Jr., Shan J. $\beta-Carotene $ and aryl hydrocarbon hydroxylase in the rat: an effect of $\beta-Carotene $ independent of vitaman A activity. J. Nutr. 119: 796-799 (1989)
12 Ziegler RG. A review of epidemiologic evidence that carotenoids reduce the risk of cancer. J. Nutr. 119: 116-122 (1989)   PUBMED
13 Ganguly J, Sastry PS. Mechanism of conversion of $\beta$-carotene Fig. 15. Effect of temperature on the heat inactivation cofficient of $\beta$-carotene into vitamin A-central cleavage versus random cleavage. Rev. Nutr. Diet. 45: 198-220 (1993)
14 Olson JA, Hayaishi O. The enzymatic cleavage of $\beta$-carotene into vitamin A by soluble enzymes of rat liver and intestine. Proc. Natl. Acad. Sci. USA 54: 1364-1370 (1965)   DOI   ScienceOn
15 Krinsky NI. Actions of carotenoids in biological systems. Ann. Rev. Nutr. 13: 561-587 (1993)   DOI   PUBMED   ScienceOn
16 van Vliet T, van Schaik F, van den Berg H. Beta-carotene metabolism: the enzymatic cleavage to retinal. Neth. J. Nutr. 53: 186-190 (1992)
17 Weisburger JH. Nutritional approach to cancer prevention with emphasis on vitamins, antioxidants, and carotenoids. Am. J. Clin. Nutr. 53: 226S-237S (1991)   PUBMED
18 Hansen S, Maret W. Retinal is not formed in vitro by enzymatic central cleavage of $\beta$-carotene. Biochemistry 27: 200-206 (1988)   DOI   ScienceOn
19 Nagao A, Olson JA. Enzymatic formation of 9-cis, 13-cis, and all-trans retinals from isomers of $\beta-Carotene$ 15,15'-dioxygenase. FASEB J. 8: 968-973 (1994)
20 During A, Nagao A, Terao J. Beta-carotene 15,15'-dioxygenase activity and cellular retinol-binding protein type II level are enhanced by dietery unsaturated triacylglycerols in rat intestines. J. Nutr. 128: 1614-1619 (1998)
21 Gawienowski AM, Stacewicz-Sapuncakis M, Longley R. Biosynthesis of retinal in bovine corpus luteum. J. Lipid Res. 15: 375-379 (1974)
22 Goodman DS, Olson JA. The conversion of all $trans-\beta-Carotene $ into retinal. Methods Enzymol. 15: 462-475 (1969)   DOI
23 During AD, Fields M, Lewis CG, Smith JC. Intestinal $\beta$-carotene 15,15'-dioxygenase activity is markedly enhanced in copper-deficient rats fed on high-iron diets and fructose. Br. J. Nutr. 84: 117-124 (2000)
24 Goodman DS, Huang HS. Biosynthesis of vitamin A with rat intrstinal enzymes. Science 149: 879-880 (1965)   DOI   PUBMED   ScienceOn
25 Zhang LX, Cooney RV, Bertman JS. Carotenoids enhance gap junctional communication and inhibit lipid peroxidation in C3H/ 10T1/2 cell: relationship to their cancer chemopreventive action. Carcinogenesis 12: 2109-2114 (1991)   DOI   ScienceOn
26 Gronowska-Senger A, Wolf G. Effect of dietary protein on the enzyme from rat and human intestine which converts beta-carotene to retinal. J. Nutr. 100: 300-308 (1970)
27 Yang A, Tume, RK. A comparison of beta-carotene-splitting activity isolated from intestinal mucosa of pasture-grazed sheep, goats and cattle. Biochem. Mol. Biol. Int. 30: 209-217 (1993)
28 van Poppel G. Carotenoids and cancer-an update with emphasis on human intervention studies. Eur. J. Cancer 29A: 1335-1344 (1993)   DOI   ScienceOn
29 Glover J. The conversion of $\beta$-carotene into vitamin A. Vitam. Horm. 18: 371-386 (1960)   DOI   PUBMED
30 Wang XD. Absorption and metabolism of $\beta$-carotene. J. Am. Coll. Nutr. 13: 314-325 (1994)   DOI   PUBMED
31 Krinsky NI, Wang XD, Tang G, Russell RM. Conversion of carotenoids to retinoids. pp. 1-16. In: Retinoids: Progress in Research and Clinical Applications. Liverea MA, Packer L (eds). Marcel Dekker, New York, NY, USA (1993)
32 Bendich A. Carotenoids and the immune responses. J. Nutr. 116: 2254-2262 (1986)   PUBMED
33 Olson JA. Provitamin A function of carotenoids: conversion of $\beta$- carotene into vitamin A. J. Nutr. 119: 105-108 (1989)   PUBMED
34 Olson JA, Lakshman MR. Carotenoid conversion. Methods Enzymol. 189: 425-432 (1990)   DOI   PUBMED
35 Lakshman MR, Mychkovsky I, Attlesey M. Enzymatic conversion of $all-trans-\beta-Carotene$ by a cytosolic enzyme from rabbit and rat intestinal mucosa. Proc. Natl. Acad. Sci. USA 86: 9124-9128 (1989)   DOI   ScienceOn
36 Singh H, Cama HR. Enzymatic cleavage of carotenoids. Biochem. Biophys. Acta 370: 49-61 (1974)   DOI   ScienceOn
37 Straub O. Key to carotenoids. pp. 1-196. In: Key to Carotenoids. Pfander H (ed). Basel Birkhauser Verlag, Basel, Switzerland (1987)
38 Krinsky NI. Photobiology of carotenoid protection. pp. 397-403. In: The Science of Photomedicine. Regan JD, Parrish JA (eds). Plenum, New York, NY, USA (1982)
39 Ziegler RG. Vegetables, fruits, carotenoids and the risk of cancer. Am. J. Clin. Nutr. 53: 238S-246S (1991)
40 Moon RC, Mehta RG. Anticarcinogenic effects of retinoids in animals. Adv. Exp. Med. Biol. 206: 399-411 (1986)
41 Fidge NH, Smith FR, Goodman WS. Vitamin A and carotenoids: the enzymic conversion of $\beta$-carotene into retinal in hog intestinal mucosa. Biochem. J. 114: 689-694 (1969)
42 Krinsky NI. Effects of carotenoids in cellular and animal systems. Am. J. Clin. Nutr. 53: 238S-246S (1991)   PUBMED
43 Bertram JS, Pung A, Churley M, Kappock IVTJ, Wilkins LR, Cooney RV. Diverse carotenoid protect against chemically induced neoplastic transformation. Carcinogenesis 2: 671-678 (1991)
44 Yeum KJ, dos Anjos Ferreira AL, Smith D, Krinsky NI, Russell RM. The effect of $\alpha$-tocopherol on the oxidative cleavage of $\beta$-carotene. Free Radic. Biol. Med. 29: 105-114 (2000)   DOI   ScienceOn
45 van Vliet T, van Schaik F, Schreurs WHP, van den Berg H. In vitro measurement of $\beta-carotene$ cleavage activity: methodological considerations and the effect of other carotenoids on $\beta-carotene$ cleavage. Int. J. Vit. Nutr. Res. 66: 77-85 (1995)
46 Moon RC. Comparative aspects of carotenoids and retinoids as chemopreventive agents for cancer. J. Nutr. 119: 127-134 (1989)   PUBMED
47 Lakshmanan MR, Chansang H, Olson JA. Purification and properties of $\beta$-carotene 15,15'-dioxygenase of rabbit intestine. J. Lipid Res. 13: 477-482 (1972)
48 Villard L, Bates CJ. Carotene dioxygenase(EC 1.13.11.21) activity in rat intestine: effects of vitamin A deficiency and of pregnancy. Br. J. Nutr. 56: 115-122 (1986)   DOI   ScienceOn
49 Napoli JL, Race KR. Biogenesis of retinoic acid from $\beta-Carotene$: differences between the metabolism $\beta-Carotene$ and retinal. J. Biol. Chem. 263: 17372-17377 (1988)