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Targeting of mitochondrial Saccharomyces cerevisiae Ilv5p to the cytosol and its effect on vicinal diketone formation in brewing

  • Applied Genetics and Molecular Biotechnology
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Abstract

Vicinal diketones (VDK) cause butter-like off-flavors in beer and are formed by a non-enzymatic oxidative decarboxylation of α-aceto-α-hydroxybutyrate and α-acetolactate, which are intermediates in isoleucine and valine biosynthesis taking place in the mitochondria. On the assumption that part of α-acetolactate can be formed also in the cytosol due to a mislocalization of the responsible acetohydroxyacid synthase encoded by ILV2 and ILV6, functional expression in the cytosol of acetohydroxyacid reductoisomerase (Ilv5p) was explored. Using the cytosolic Ilv5p, I aimed to metabolize the cytosolically formed α-aetolactate, thereby lowering the total VDK production. Among mutant Ilv5p enzymes with varying degrees of N-terminal truncation, one with a 46-residue deletion (Ilv5pΔ46) exhibited an unequivocal localization in the cytosol judged from microscopy of the Ilv5pΔ46-green fluorescent protein fusion protein and the inability of Ilv5pΔ46 to remedy the isoleucine/valine requirement of an ilv5Δ strain. When introduced into an industrial lager brewing strain, a robust expression of Ilv5pΔ46 was as effective as that of a wild-type Ilv5p in lowering the total VDK production in a 2-l scale fermentation trial. Unlike the case of the wild-type Ilv5p, an additional expression of Ilv5pΔ46 did not alter the quality of the resultant beer in terms of contents of aromatic compounds and organic acids.

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Acknowledgments

The author thanks Morten C Kielland-Brandt and Kjeld Olesen for critical reading of the manuscript. The expert technical assistance of Yumiko Itokui, Yachiyo Wada, Kaoru Nakagawa, and Hiromi Hata is gratefully acknowledged.

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  1. Suntory Research Center, 1-1-1 Wakayamadai, Shimamoto-cho, Mishima-gun, Osaka, 618-8503, Japan

    Fumihiko Omura

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  1. Fumihiko Omura
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Omura, F. Targeting of mitochondrial Saccharomyces cerevisiae Ilv5p to the cytosol and its effect on vicinal diketone formation in brewing. Appl Microbiol Biotechnol 78, 503–513 (2008). https://doi.org/10.1007/s00253-007-1333-x

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  • DOI: https://doi.org/10.1007/s00253-007-1333-x

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