Identification of a Saccharomyces Cerevisiae Glucosidase That Hydrolyzes Flavonoid Glucosides

Overview

Journal Appl Environ Microbiol

Specialties Environmental Health
Microbiology

Date 2011 Jan 11

PMID 21216897

Citations 18

Authors

Sabine Schmidt

Sandra Rainieri

Simone Witte

Ulrich Matern

Stefan Martens

Affiliations

Soon will be listed here.

Abstract

Baker's yeast (Saccharomyces cerevisiae) whole-cell bioconversions of naringenin 7-O-β-glucoside revealed considerable β-glucosidase activity, which impairs any strategy to generate or modify flavonoid glucosides in yeast transformants. Up to 10 putative glycoside hydrolases annotated in the S. cerevisiae genome database were overexpressed with His tags in yeast cells. Examination of these recombinant, partially purified polypeptides for hydrolytic activity with synthetic chromogenic α- or β-glucosides identified three efficient β-glucosidases (EXG1, SPR1, and YIR007W), which were further assayed with natural flavonoid β-glucoside substrates and product verification by thin-layer chromatography (TLC) or high-performance liquid chromatography (HPLC). Preferential hydrolysis of 7- or 4'-O-glucosides of isoflavones, flavonols, flavones, and flavanones was observed in vitro with all three glucosidases, while anthocyanins were also accepted as substrates. The glucosidase activities of EXG1 and SPR1 were completely abolished by Val168Tyr mutation, which confirmed the relevance of this residue, as reported for other glucosidases. Most importantly, biotransformation experiments with knockout yeast strains revealed that only EXG1 knockout strains lost the capability to hydrolyze flavonoid glucosides.

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References

Martens S, Forkmann G, Britsch L, Wellmann F, Matern U, Lukacin R . Divergent evolution of flavonoid 2-oxoglutarate-dependent dioxygenases in parsley. FEBS Lett. 2003; 544(1-3):93-8. DOI: 10.1016/s0014-5793(03)00479-4. View

Marotti I, Bonetti A, Biavati B, Catizone P, Dinelli G . Biotransformation of common bean (Phaseolus vulgaris L.) flavonoid glycosides by bifidobacterium species from human intestinal origin. J Agric Food Chem. 2007; 55(10):3913-9. DOI: 10.1021/jf062997g. View

Hempel J, Pforte H, Raab B, Engst W, Bohm H, Jacobasch G . Flavonols and flavones of parsley cell suspension culture change the antioxidative capacity of plasma in rats. Nahrung. 1999; 43(3):201-4. DOI: 10.1002/(SICI)1521-3803(19990601)43:3<201::AID-FOOD201>3.0.CO;2-1. View

Bouarab K, Melton R, Peart J, Baulcombe D, Osbourn A . A saponin-detoxifying enzyme mediates suppression of plant defences. Nature. 2002; 418(6900):889-92. DOI: 10.1038/nature00950. View

Lesage G, Bussey H . Cell wall assembly in Saccharomyces cerevisiae. Microbiol Mol Biol Rev. 2006; 70(2):317-43. PMC: 1489534. DOI: 10.1128/MMBR.00038-05. View

Morant A, Jorgensen K, Jorgensen C, Paquette S, Sanchez-Perez R, Moller B . beta-Glucosidases as detonators of plant chemical defense. Phytochemistry. 2008; 69(9):1795-813. DOI: 10.1016/j.phytochem.2008.03.006. View

Ebel J, Hahlbrock K . Enzymes of flavone and flavonol-glycoside biosynthesis. Coordinated and selective induction in cell-suspension cultures of Petroselinum hortense. Eur J Biochem. 1977; 75(1):201-9. DOI: 10.1111/j.1432-1033.1977.tb11518.x. View

Tahara S . A journey of twenty-five years through the ecological biochemistry of flavonoids. Biosci Biotechnol Biochem. 2007; 71(6):1387-404. DOI: 10.1271/bbb.70028. View

Henstrand J, McCue K, Brink K, Handa A, Herrmann K, Conn E . Light and Fungal Elicitor Induce 3-Deoxy-d-arabino-Heptulosonate 7-Phosphate Synthase mRNA in Suspension Cultured Cells of Parsley (Petroselinum crispum L.). Plant Physiol. 1992; 98(2):761-3. PMC: 1080257. DOI: 10.1104/pp.98.2.761. View

10.

Muthukumar G, Suhng S, Magee P, Jewell R, Primerano D . The Saccharomyces cerevisiae SPR1 gene encodes a sporulation-specific exo-1,3-beta-glucanase which contributes to ascospore thermoresistance. J Bacteriol. 1993; 175(2):386-94. PMC: 196152. DOI: 10.1128/jb.175.2.386-394.1993. View

11.

Huh W, Falvo J, Gerke L, Carroll A, Howson R, Weissman J . Global analysis of protein localization in budding yeast. Nature. 2003; 425(6959):686-91. DOI: 10.1038/nature02026. View

12.

Hahlbrock K, Knobloch K, Kreuzaler F, Potts J, Wellmann E . Coordinated induction and subsequent activity changes of two groups of metabolically interrelated enzymes. Light-induced synthesis of flavonoid glycosides in cell suspension cultures of Petroselinum hortense. Eur J Biochem. 1976; 61(1):199-206. DOI: 10.1111/j.1432-1033.1976.tb10012.x. View

13.

Ruan C, Zhang H, Zhang L, Liu Z, Sun G, Lei J . Biotransformation of ginsenoside Rf to Rh1 by recombinant beta-glucosidase. Molecules. 2009; 14(6):2043-8. PMC: 6254368. DOI: 10.3390/molecules14062043. View

14.

Martens S, Forkmann G, Matern U, Lukacin R . Cloning of parsley flavone synthase I. Phytochemistry. 2001; 58(1):43-6. DOI: 10.1016/s0031-9422(01)00191-1. View

15.

Ramirez M, Hernandez L, Larriba G . A similar protein portion for two exoglucanases secreted by Saccharomyces cerevisiae. Arch Microbiol. 1989; 151(5):391-8. DOI: 10.1007/BF00416596. View

16.

Moehs C, ALLEN P, Friedman M, Belknap W . Cloning and expression of solanidine UDP-glucose glucosyltransferase from potato. Plant J. 1997; 11(2):227-36. DOI: 10.1046/j.1365-313x.1997.11020227.x. View

17.

Ververidis F, Trantas E, Douglas C, Vollmer G, Kretzschmar G, Panopoulos N . Biotechnology of flavonoids and other phenylpropanoid-derived natural products. Part II: Reconstruction of multienzyme pathways in plants and microbes. Biotechnol J. 2007; 2(10):1235-49. DOI: 10.1002/biot.200700184. View

18.

Bradford M . A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976; 72:248-54. DOI: 10.1016/0003-2697(76)90527-3. View

19.

Suzuki K, Yabe T, Maruyama Y, Abe K, Nakajima T . Characterization of recombinant yeast exo-beta-1,3-glucanase (Exg 1p) expressed in Escherichia coli cells. Biosci Biotechnol Biochem. 2001; 65(6):1310-4. DOI: 10.1271/bbb.65.1310. View

20.

Kotaka A, Bando H, Kaya M, Kato-Murai M, Kuroda K, Sahara H . Direct ethanol production from barley beta-glucan by sake yeast displaying Aspergillus oryzae beta-glucosidase and endoglucanase. J Biosci Bioeng. 2008; 105(6):622-7. DOI: 10.1263/jbb.105.622. View