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The alternative d-galactose degrading pathway of Aspergillus nidulans proceeds via l-sorbose

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Abstract

The catabolism of d-galactose in yeast depends on the enzymes of the Leloir pathway. In contrast, Aspergillus nidulans mutants in galactokinase (galE) can still grow on d-galactose in the presence of ammonium—but not nitrate—ions as nitrogen source. A. nidulans galE mutants transiently accumulate high (400 mM) intracellular concentrations of galactitol, indicating that the alternative d-galactose degrading pathway may proceed via this intermediate. The enzyme degrading galactitol was identified as l-arabitol dehydrogenase, because an A. nidulans loss-of-function mutant in this enzyme (araA1) did not show NAD+-dependent galactitol dehydrogenase activity, still accumulated galactitol but was unable to catabolize it thereafter, and a double galE/araA1 mutant was unable to grow on d-galactose or galactitol. The product of galactitol oxidation was identified as l-sorbose, which is a substrate for hexokinase, as evidenced by a loss of l-sorbose phosphorylating activity in an A. nidulans hexokinase (frA1) mutant. l-Sorbose catabolism involves a hexokinase step, indicated by the inability of the frA1 mutant to grow on galactitol or l-sorbose, and by the fact that a galE/frA1 double mutant of A. nidulans was unable to grow on d-galactose. The results therefore provide evidence for an alternative pathway of d-galactose catabolism in A. nidulans that involves reduction of the d-galactose to galactitol and NAD+-dependent oxidation of galactitol by l-arabitol dehydrogenase to l-sorbose.

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Acknowledgements

We are grateful to Professor Axel A. Brakhage (University of Hannover, Germany), Dr. Michel J. A. Flipphi (University of Valencia, Spain) and Dr. George J. Ruijter (Leiden University, The Netherlands) for the A. nidulans R21, frA1 and araA1 strains, respectively. The project was carried out in the framework of an Austrian-Hungarian Intergovernmental Science and Technology Cooperation Programme (A-26/2000, and was grant-aided by the OTKA (Hungarian Scientific Research Fund; grants F 031985 and F 042602) and the FKFP (Fund for Research and Development in Higher Education; grant 0009/2001). Dr. Erzsébet Sándor is a recipient of an OTKA Postdoctoral Fellowship (grant D 37975).

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Authors and Affiliations

  1. Department of Microbiology and Biotechnology, University of Debrecen, Faculty of Sciences, P.O.Box 63, 4010, Debrecen, Hungary

    Erzsébet Fekete, Levente Karaffa, Adél Sepsi & Attila Szentirmai

  2. Department of Plant Protection, University of Debrecen, Faculty of Agriculture, P.O.Box 36, 4015, Debrecen, Hungary

    Erzsébet Sándor

  3. Department of Physical Chemistry, University of Debrecen, Faculty of Sciences, P.O. Box 7, 4010, Debrecen, Hungary

    István Bányai

  4. Division of Gene Technology and Applied Biochemistry, TU Wien, Getreidemarkt 9/1665, Institute of Chemical Engineering, 1060, Vienna, Austria

    Bernhard Seiboth & Christian P. Kubicek

  5. Department of Biochemistry, University of Debrecen, Faculty of Sciences, P.O.Box 55, 4010, Debrecen, Hungary

    Gyöngyi Gyémánt

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  1. Erzsébet Fekete
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  2. Levente Karaffa
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Correspondence to Levente Karaffa.

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Fekete, E., Karaffa, L., Sándor, E. et al. The alternative d-galactose degrading pathway of Aspergillus nidulans proceeds via l-sorbose. Arch Microbiol 181, 35–44 (2004). https://doi.org/10.1007/s00203-003-0622-8

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  • DOI: https://doi.org/10.1007/s00203-003-0622-8

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