Assessment of lactase activity in humans by measurement of galactitol and galactonate in serum and urine after milk intake

Nathalie Vionnet, Linda H Münger, Carola Freiburghaus, Kathryn J Burton, Grégory Pimentel, François P Pralong, René Badertscher, Guy Vergères, Nathalie Vionnet, Linda H Münger, Carola Freiburghaus, Kathryn J Burton, Grégory Pimentel, François P Pralong, René Badertscher, Guy Vergères

Abstract

Background: Lactase is an enzyme that hydrolyzes lactose into glucose and galactose in the small intestine, where they are absorbed. Hypolactasia is a common condition, primarily caused by genetic programming, that leads to lactose maldigestion and, in certain cases, lactose intolerance. Galactitol and galactonate are 2 products of hepatic galactose metabolism that are candidate markers for the intake of lactose-containing foods.

Objectives: The primary objective of the study was to explore the changes in serum and urine metabolomes during postprandial dairy product tests through the association between lactase persistence genotype and the postprandial dynamics of lactose-derived metabolites.

Methods: We characterized the 6-h postprandial serum kinetics and urinary excretion of lactose, galactose, galactitol, and galactonate in 14 healthy men who had consumed a single dose of acidified milk (800 g) which contained 38.8 g lactose. Genotyping of LCT-13910 C/T (rs4988235) was performed to assess primary lactase persistence.

Results: There were 2 distinct postprandial responses, classified as high and low metabolite responses, observed for galactose, and its metabolites galactitol and galactonate, in serum and urine. In all but 1 subject, there was a concordance between the high metabolite responses and genetic lactase persistence and between the low metabolite responses and genetic lactase nonpersistence (accuracy 0.92), galactitol and galactonate being more discriminative than galactose.

Conclusions: Postprandial galactitol and galactonate after lactose overload appear to be good proxies for genetically determined lactase activity. The development of a noninvasive lactose digestion test based on the measurement of these metabolites in urine could be clinically useful. This trial was registered at clinicaltrials.gov as NCT02230345.

Figures

FIGURE 1
FIGURE 1
Genetic dependency of the postprandial behavior of lactose-derived metabolites after intake of acidified milk. Serum lactose (A), galactose (B), galactitol (C), and galactonate (D) after milk intake in 14 healthy subjects. Participants with CC (solid grey, n = 8) and CT or TT (solid black, n = 5) or nondetermined (black dashed, n = 1) genotype at rs4988235. Comparison of postprandial serum (iAUC) lactose (E), galactose (F), galactitol (G), and galactonate (H) after milk intake between CC (grey circles, n = 8), CT (black circles, n = 2), and TT (black triangles, n = 3) carriers in 13 healthy subjects. Comparison of postprandial urine (6-h pool) lactose (I), galactose (J), galactitol (K), and galactonate (L) after milk intake between CC (grey circles, n = 8), CT (black circles, n = 2), and TT (black triangles, n = 3) carriers in 13 healthy subjects. Boxplots indicate medians and interquartile ranges. The Box-Whiskers plots indicate medians, interquartile ranges, and minimum and maximum values excluding outliers. *P < 0.05, Wilcoxon's signed-rank test between CC and CT or TT genotypes. Subject F3_009, who was of African origin, was excluded from the statistical analyses of panels E–L because genotypes at rs4988235 were not relevant for diagnosis of lactase persistence in this population. iAUC, incremental area under the curve.
FIGURE 2
FIGURE 2
Correlation between postprandial serum and urine lactose (A), galactose (B), galactitol (C), and galactonate (D) after milk intake in 14 subjects. Partial Spearman's correlation test controlling for lactase persistence. Participants with CC (grey circles, n = 8), CT (black circles, n = 2), TT (black triangles, n = 3), or nondetermined (unfilled circle, n = 1) genotype at rs4988235. Subject F3_015 is indicated on the figure. iAUC, incremental area under the curve.
FIGURE 3
FIGURE 3
Correlation between postprandial serum galactose and galactitol (A), galactose and galactonate (B), and galactonate and galactitol (C), and between postprandial urine galactose and galactitol (D), galactose and galactonate (E), and galactonate and galactitol (F), after milk intake in 14 subjects. Partial Spearman's correlation test controlling for lactase persistence. Participants with CC (grey circles, n = 8), CT (black circles, n = 2), TT (black triangles, n = 3), or nondetermined (unfilled circle, n = 1) genotype at rs4988235. Subject F3_015 is indicated on the figure. iAUC, incremental area under the curve.

References

    1. Johnson JM, Conforti FD. Lactose A2. In: Caballero B.editor. Encyclopedia of Food Sciences and Nutrition. 2nd ed Oxford: Academic Press; 2003:3472–6.
    1. Labrie V, Buske OJ, Oh E, Jeremian R, Ptak C, Gasiunas G, Maleckas A, Petereit R, Zvirbliene A, Adamonis K et al. .. Lactase nonpersistence is directed by DNA-variation-dependent epigenetic aging. Nat Struct Mol Biol. 2016;23(6):566–73.
    1. He T, Priebe MG, Harmsen HJ, Stellaard F, Sun X, Welling GW, Vonk RJ. Colonic fermentation may play a role in lactose intolerance in humans. J Nutr. 2006;136(1):58–63.
    1. Gerbault P, Liebert A, Itan Y, Powell A, Currat M, Burger J, Swallow DM, Thomas MG. Evolution of lactase persistence: an example of human niche construction. Philos Trans R Soc Lond B Biol Sci. 2011;366(1566):863–77.
    1. Misselwitz B, Pohl D, Fruhauf H, Fried M, Vavricka SR, Fox M. Lactose malabsorption and intolerance: pathogenesis, diagnosis and treatment. United European Gastroenterol J. 2013;1(3):151–9.
    1. Ingram CJ, Mulcare CA, Itan Y, Thomas MG, Swallow DM. Lactose digestion and the evolutionary genetics of lactase persistence. Hum Genet. 2009;124(6):579–91.
    1. Aragon JJ, Hermida C, Martinez-Costa OH, Sanchez V, Martin I, Sanchez JJ, Codoceo R, Cano JM, Cano A, Crespo L et al. .. Noninvasive diagnosis of hypolactasia with 4-galactosylxylose (gaxilose): a multicentre, open-label, phase IIB–III nonrandomized trial. J Clin Gastroenterol. 2014;48(1):29–36.
    1. Simren M, Stotzer PO. Use and abuse of hydrogen breath tests. Gut. 2006;55(3):297–303.
    1. Gasbarrini A, Corazza GR, Gasbarrini G, Montalto M, Di Stefano M, Basilisco G, Parodi A, Usai-Satta P, Vernia P, Anania C et al. .. Methodology and indications of H2-breath testing in gastrointestinal diseases: the Rome Consensus Conference. Aliment Pharmacol Ther. 2009;29 Suppl 1:1–49.
    1. Perets TT, Hamouda D, Layfer O, Ashorov O, Boltin D, Levy S, Niv Y, Dickman R. Small intestinal bacterial overgrowth may increase the likelihood of lactose and sorbitol but not fructose intolerance false positive diagnosis. Ann Clin Lab Sci. 2017;47(4):447–51.
    1. Pimentel G, Burton KJ, Rosikiewicz M, Freiburghaus C, von Ah U, Munger LH, Pralong FP, Vionnet N, Greub G, Badertscher R et al. .. Blood lactose after dairy product intake in healthy men. Br J Nutr. 2017;118(12):1070–7.
    1. Coelho AI, Berry GT, Rubio-Gozalbo ME. Galactose metabolism and health. Curr Opin Clin Nutr Metab Care. 2015;18(4):422–7.
    1. Trimigno A, Münger L, Picone G, Freiburghaus C, Pimentel G, Vionnet N, Pralong FP, Capozzi F, Badertscher R, Vergères G. GC-MS based metabolomics and NMR spectroscopy investigation of food intake biomarkers for milk and cheese in serum of healthy humans. Metabolites. 2018;8(2):26.
    1. Münger LH, Trimigno A, Picone G, Freiburghaus C, Pimentel G, Burton KJ, Pralong FP, Vionnet N, Capozzi F, Badertscher R et al. .. Identification of urinary food intake biomarkers for milk, cheese, and soy-based drink by untargeted GC-MS and NMR in healthy humans. J Proteome Res. 2017;16(9):3321–35.
    1. Burton KJ, Rosikiewicz M, Pimentel G, Butikofer U, von Ah U, Voirol MJ, Croxatto A, Aeby S, Drai J, McTernan PG et al. .. Probiotic yogurt and acidified milk similarly reduce postprandial inflammation and both alter the gut microbiota of healthy, young men. Br J Nutr. 2017;117(9):1312–22.
    1. Pimentel G, Burton KJ, von Ah U, Bütikofer U, Pralong FP, Vionnet N, Portmann R, Vergères G. Metabolic footprinting of fermented milk consumption in serum of healthy men. J Nutr. 2018;148(6):851–60.
    1. Burton KJ, Pimentel G, Zangger N, Vionnet N, Drai J, McTernan PG, Pralong FP, Delorenzi M, Vergeres G. Modulation of the peripheral blood transcriptome by the ingestion of probiotic yoghurt and acidified milk in healthy, young men. PLoS One. 2018;13(2):e0192947.
    1. Dunn WB, Broadhurst D, Begley P, Zelena E, Francis-McIntyre S, Anderson N, Brown M, Knowles JD, Halsall A, Haselden JN et al. .. Procedures for large-scale metabolic profiling of serum and plasma using gas chromatography and liquid chromatography coupled to mass spectrometry. Nat Protoc. 2011;6(7):1060–83.
    1. Sumner LW, Amberg A, Barrett D, Beale MH, Beger R, Daykin CA, Fan TW, Fiehn O, Goodacre R, Griffin JL et al. .. Proposed minimum reporting standards for chemical analysis Chemical Analysis Working Group (CAWG) Metabolomics Standards Initiative (MSI). Metabolomics. 2007;3(3):211–21.
    1. Ekstrom C. MESS: Miscellaneous Esoteric Statistical Scripts. R package version 0.3-2[Internet] 2014. Available from: .
    1. Noguchi K, Gel YR, Brunner E, Konietschke F. nparLD: an R software package for the nonparametric analysis of longitudinal data in factorial experiments. J Stat Softw. 2012;50:1–12.
    1. Kim S. ppcor: an R package for a fast calculation to semi-partial correlation coefficients. Commun Stat Appl Methods. 2015;22(6):665–74.
    1. Dzialanski Z, Barany M, Engfeldt P, Magnuson A, Olsson LA, Nilsson TK. Lactase persistence versus lactose intolerance: is there an intermediate phenotype?. Clin Biochem. 2016;49(3):248–52.
    1. Enattah NS, Kuokkanen M, Forsblom C, Natah S, Oksanen A, Jarvela I, Peltonen L, Savilahti E. Correlation of intestinal disaccharidase activities with the C/T-13910 variant and age. World J Gastroenterol. 2007;13(25):3508–12.
    1. Ingram CJ, Raga TO, Tarekegn A, Browning SL, Elamin MF, Bekele E, Thomas MG, Weale ME, Bradman N, Swallow DM. Multiple rare variants as a cause of a common phenotype: several different lactase persistence associated alleles in a single ethnic group. J Mol Evol. 2009;69(6):579–88.
    1. Itan Y, Jones BL, Ingram CJ, Swallow DM, Thomas MG. A worldwide correlation of lactase persistence phenotype and genotypes. BMC Evol Biol. 2010;10:36.
    1. Rezaie A, Buresi M, Lembo A, Lin H, McCallum R, Rao S, Schmulson M, Valdovinos M, Zakko S, Pimentel M. Hydrogen and methane-based breath testing in gastrointestinal disorders: the North American Consensus. Am J Gastroenterol. 2017;112(5):775–84.
    1. Keiding S, Johansen S, Winkler K, Tonnesen K, Tygstrup N. Michaelis-Menten kinetics of galactose elimination by the isolated perfused pig liver. Am J Physiol. 1976;230(5):1302–13.
    1. Merkel C, Marchesini G, Fabbri A, Bianco S, Bianchi G, Enzo E, Sacerdoti D, Zoli M, Gatta A. The course of galactose elimination capacity in patients with alcoholic cirrhosis: possible use as a surrogate marker for death. Hepatology. 1996;24(4):820–3.
    1. Ficicioglu C, Hussa C, Gallagher PR, Thomas N, Yager C. Monitoring of biochemical status in children with Duarte galactosemia: utility of galactose, galactitol, galactonate, and galactose 1-phosphate. Clin Chem. 2010;56(7):1177–82.
    1. Szumilo T. Purification and properties of D-galactonate dehydratase from Mycobacterium butyricum. Biochim Biophys Acta. 1981;661(2):240–6.

Source: PubMed

Sponsors en medewerkers

Medische omstandigheden

Geneesmiddelinterventies

3
Abonneren