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Identification and assessment of markers of biotin status in healthy adults

  • Wei Kay Eng (a1), David Giraud (a1), Vicki L. Schlegel (a2), Dong Wang (a3), Bo Hyun Lee (a2) and Janos Zempleni (a1)...


Human biotin requirements are unknown and the identification of reliable markers of biotin status is necessary to fill this knowledge gap. Here, we used an outpatient feeding protocol to create states of biotin deficiency, sufficiency and supplementation in sixteen healthy men and women. A total of twenty possible markers of biotin status were assessed, including the abundance of biotinylated carboxylases in lymphocytes, the expression of genes from biotin metabolism and the urinary excretion of biotin and organic acids. Only the abundance of biotinylated 3-methylcrotonyl-CoA carboxylase (holo-MCC) and propionyl-CoA carboxylase (holo-PCC) allowed for distinguishing biotin-deficient and biotin-sufficient individuals. The urinary excretion of biotin reliably identified biotin-supplemented subjects, but did not distinguish between biotin-depleted and biotin-sufficient individuals. The urinary excretion of 3-hydroxyisovaleric acid detected some biotin-deficient subjects, but produced a meaningful number of false-negative results and did not distinguish between biotin-sufficient and biotin-supplemented individuals. None of the other organic acids that were tested were useful markers of biotin status. Likewise, the abundance of mRNA coding for biotin transporters, holocarboxylase synthetase and biotin-dependent carboxylases in lymphocytes were not different among the treatment groups. Generally, datasets were characterised by variations that exceeded those seen in studies in cell cultures. We conclude that holo-MCC and holo-PCC are the most reliable, single markers of biotin status tested in the present study.

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Corresponding author

*Corresponding author: J. Zempleni, fax +1 402 472 1587, email


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1Zempleni, J, Wijeratne, SS & Hassan, YI (2009) Biotin. Biofactors 35, 3646.
2Camporeale, G, Shubert, EE, Sarath, G, et al. (2004) K8 and K12 are biotinylated in human histone H4. Eur J Biochem 271, 22572263.
3Kobza, K, Camporeale, G, Rueckert, B, et al. (2005) K4, K9, and K18 in human histone H3 are targets for biotinylation by biotinidase. FEBS J 272, 42494259.
4Pestinger, V, Wijeratne, SSK, Rodriguez-Melendez, R, et al. (2011) Novel histone biotinylation marks are enriched in repeat regions and participate in repression of transcriptionally competent genes. J Nutr Biochem 22, 328333.
5Rios-Avila, L, Pestinger, V & Zempleni, J (2012) K16-biotinylated histone H4 is overrepresented in repeat regions and participates in the repression of transcriptionally competent genes in human Jurkat lymphoid cells. J Nutr Biochem 23, 15591564.
6Stanley, JS, Griffin, JB & Zempleni, J (2001) Biotinylation of histones in human cells: effects of cell proliferation. Eur J Biochem 268, 54245429.
7Bailey, LM, Ivanov, RA, Wallace, JC, et al. (2008) Artifactual detection of biotin on histones by streptavidin. Anal Biochem 373, 7177.
8Kuroishi, T, Rios-Avila, L, Pestinger, V, et al. (2011) Biotinylation is a natural, albeit rare, modification of human histones. Mol Genet Metab 104, 537545.
9Camporeale, G, Oommen, AM, Griffin, JB, et al. (2007) K12-biotinylated histone H4 marks heterochromatin in human lymphoblastoma cells. J Nutr Biochem 18, 760768.
10Filenko, NA, Kolar, C, West, JT, et al. (2011) The role of histone H4 biotinylation in the structure and dynamics of nucleosomes. PLoS One 6, e16299.
11Bao, B, Pestinger, V, HY, I, et al. (2011) Holocarboxylase synthetase is a chromatin protein and interacts directly with histone H3 to mediate biotinylation of K9 and K18. J Nutr Biochem 22, 470475.
12National Research Council (1998) Dietary Reference Intakes for Thiamin, Riboflavin, Niacin, Vitamin B6, Folate, Vitamin B12, Pantothenic Acid, Biotin, and Choline. Washington, DC: National Academy Press.
13Zempleni, J & Mock, DM (2000) Biotin. In Modern Analytical Methodologies on Fat and Water-Soluble Vitamins, pp. 389409 [Song, WO and Beecher, GR, editors]. New York, NY: Wiley & Sons, Inc.
14Green, NM (1975) Avidin. Adv Protein Chem 29, 85133.
15Mock, N, Malik, M, Stumbo, P, et al. (1997) Increased urinary excretion of 3-hydroxyisovaleric acid and decreased urinary excretion of biotin are sensitive early indicators of decreased status in experimental biotin deficiency. Am J Clin Nutr 65, 951958.
16Sealey, WM, Teague, AM, Stratton, SL, et al. (2004) Smoking accelerates biotin catabolism in women. Am J Clin Nutr 80, 932935.
17Weiner, DL, Grier, RE & Wolf, B (1985) A bioassay for determining biotinidase activity and for discriminating biocytin from biotin using holocarboxylase synthetase-deficient cultured fibroblasts. J Inherit Metab Dis 8, 101102.
18Wolf, B, Heard, GS, Weissbecker, KA, et al. (1985) Biotinidase deficiency: initial clinical features and rapid diagnosis. Ann Neurol 18, 614617.
19Mock, DM & Dyken, ME (1997) Biotin catabolism is accelerated in adults receiving long-term therapy with anticonvulsants. Neurology 49, 14441447.
20Rathman, SC, Eisenschenk, S & McMahon, RJ (2002) The abundance and function of biotin-dependent enzymes are reduced in rats chronically administered carbamazepine. J Nutr 132, 34053410.
21Mock, DM & Stadler, DD (1997) Conflicting indicators of biotin status from a cross-sectional study of normal pregnancy. J Am Coll Nutr 16, 252257.
22Mock, DM, Stadler, D, Stratton, S, et al. (1997) Biotin status assessed longitudinally in pregnant women. J Nutr 127, 710716.
23Mock, DM, Mock, NI & Dankle, JA (1992) Secretory patterns of biotin in human milk. J Nutr 122, 546552.
24Camporeale, G & Zempleni, J (2006) Biotin. In Present Knowledge in Nutrition, 9th ed., pp. 314326 [Bowman, BA and Russell, RM, editors]. Washington, DC: International Life Sciences Institute.
25Stratton, SL, Henrich, CL, Matthews, NI, et al. (2012) Marginal biotin deficiency can be induced experimentally in humans using a cost-effective outpatient design. J Nutr 142, 2226.
26Baugh, CM, Malone, JH & Butterworth, CE Jr (1968) Human biotin deficiency: a case history of biotin deficiency induced by raw egg consumption in a cirrhotic patient. Am J Clin Nutr 21, 173182.
27Wolf, B & Heard, GS (1991) Biotinidase deficiency. In Advances in Pediatrics, pp. 121 [Barness, L and Oski, F, editors]. Chicago, IL: Medical Book Publishers.
28Zempleni, J, Helm, RM & Mock, DM (2001) In vivo biotin supplementation at a pharmacologic dose decreases proliferation rates of human peripheral blood mononuclear cells and cytokine release. J Nutr 131, 14791484.
29Manthey, KC, Griffin, JB & Zempleni, J (2002) Biotin supply affects expression of biotin transporters, biotinylation of carboxylases, and metabolism of interleukin-2 in Jurkat cells. J Nutr 132, 887892.
30Kaur Mall, G, Chew, YC & Zempleni, J (2010) Biotin requirements are lower in human Jurkat lymphoid cells but homeostatic mechanisms are similar to those of HepG2 liver cells. J Nutr 140, 10861092.
31Green, NM (1965) A spectrophotometric assay for avidin and biotin based on binding of dyes by avidin. Biochem J 94, 23c24c.
32Durance, TD (1991) Residual avidin activity in cooked egg white assayed with improved sensitivity. J Food Sci 56, 707709.
33Zempleni, J & Mock, DM (1998) Uptake and metabolism of biotin by human peripheral blood mononuclear cells. Am J Physiol Cell Physiol 275, C382C388.
34Livak, KJ & Schmittgen, TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2( − Delta Delta C(T)) method. Methods 25, 402408.
35O'Brien, D, Ibbott, FA & Rodgerson, DO (1968) Laboratory Manual of Pediatric Micro-biochemical Techniques, 4th ed.New York: Hoeber Medical Division, Harper & Row, Publishers, Inc.
36Mock, DM, Henrich, CL, Carnell, N, et al. (2002) Indicators of marginal biotin deficiency and repletion in humans: validation of 3-hydroxyisovaleric acid excretion and a leucine challenge. Am J Clin Nutr 76, 10611068.
37SAS Institute (1999) StatView Reference, 3rd ed.Cary, NC: SAS Publishing.
38Staggs, CG, Sealey, WM, McCabe, BJ, et al. (2004) Determination of the biotin content of select foods using accurate and sensitive HPLC/avidin binding. J Food Compost Anal 17, 767776.
39 Food Standards Australia New Zealand (2010) NUTTAB 2010 Online Searchable Database. = search (accessed 25 June 2012).
40Velazquez, A, Zamudio, S, Baez, A, et al. (1990) Indicators of biotin status: a study of patients on prolonged total parenteral nutrition. Eur J Clin Nutr 44, 1116.
41Mock, DM, Henrich, CL, Carnell, N, et al. (2002) Lymphocyte propionyl-CoA carboxylase and accumulation of odd-chain fatty acid in plasma and erythrocytes are useful indicators of marginal biotin deficiency. J Nutr Biochem 13, 462470.
42Mock, DM & Mock, NI (2002) Lymphocyte propionyl-CoA carboxylase is an early and sensitive indicator of biotin deficiency in rats, but urinary excretion of 3-hydroxypropionic acid is not. J Nutr 132, 19451950.
43Zempleni, J, Trusty, TA & Mock, DM (1997) Lipoic acid reduces the activities of biotin-dependent carboxylases in rat liver. J Nutr 127, 17761781.
44Mock, NI, Mock, DM, Malik, M, et al. (1995) Urinary excretion of biotin and 3-hydroxyisovaleric acid (3-HIA) are early indicators of biotin deficiency. FASEB J 9, A985 [abstract].
45Stratton, SL, Horvath, TD, Bogusiewicz, A, et al. (2011) Urinary excretion of 3-hydroxyisovaleryl carnitine is an early and sensitive indicator of marginal biotin deficiency in humans. J Nutr 141, 353358.
46Horvath, TD, Stratton, SL, Bogusiewicz, A, et al. (2010) Quantitative measurement of urinary excretion of 3-hydroxyisovaleryl carnitine by LC–MS/MS as an indicator of biotin status in humans. Anal Chem 82, 95439548.
47Mock, DM, Stratton, SL, Horvath, TD, et al. (2011) Urinary excretion of 3-hydroxyisovaleric acid and 3-hydroxyisovaleryl carnitine increases in response to a leucine challenge in marginally biotin-deficient humans. J Nutr 141, 19251930.
48Mock, DM, Lankford, GL & Cazin, J Jr (1993) Biotin and biotin analogs in human urine: biotin accounts for only half of the total. J Nutr 123, 18441851.
49Zempleni, J, McCormick, DB & Mock, DM (1997) Identification of biotin sulfone, bisnorbiotin methyl ketone, and tetranorbiotin-l-sulfoxide in human urine. Am J Clin Nutr 65, 508511.
50Mock, DM & Mock, NI (1997) Serum concentrations of bisnorbiotin and biotin sulfoxide increase during both acute and chronic biotin supplementation. J Lab Clin Med 129, 384388.
51Rodriguez-Melendez, R, Cano, S, Mendez, ST, et al. (2001) Biotin regulates the genetic expression of holocarboxylase synthetase and mitochondrial carboxylases in rats. J Nutr 131, 19091913.
52Solorzano-Vargas, RS, Pacheco-Alvarez, D & Leon-Del-Rio, A (2002) Holocarboxylase synthetase is an obligate participant in biotin-mediated regulation of its own expression and of biotin-dependent carboxylases mRNA levels in human cells. Proc Natl Acad Sci U S A 99, 53255330.



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