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A Role of Formate Dehydrogenase in the Oxalate Title in the Wood-destroying Basidiomycete Ceriporiopsis subvermispora

WATANABE, Tomoki; SABRINA, Tengku; HATTORI, Author(s) Takefumi; SHIMADA, Mikio

Wood research : bulletin of the Wood Research Institute Kyoto Citation University (2003), 90: 7-8

Issue Date 2003-09-30

URL http://hdl.handle.net/2433/53104

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Type Departmental Bulletin Paper

Textversion publisher

Kyoto University Preliminary

A Role of Formate Dehydrogenase in the Oxalate Metabolism in the Wood-destroying Basidiomycete Ceriporiopsis subvermispora*l

Tomoki WATANABE*2, Tengku SABRINA *3, Takefumi HATTORI*2 and Mikio SHIMADA*2 (Received June 7, 2003)

Keywords: oxalate metabolism, NAD +-dependent formate dehydrogenase, white-rot fungi, Ceriporiopsis subvermispora

metabolism that oxalate decarboxylase (ODO; EO 4.1.1.2) Introduction converts oxalate to formate and , and the It is a common physiological trait that brown-rot fungi, formate thus produced is converted to carbon dioxide by including Fomitopsis palustris accumulate in large formate dehydrogenase (FDH; EO 1.2.1.2), yielding quantities in the cultures. Oxalic acid serves as an acid NADH. However, recently, Aguilar et al. successfully catalyst for the hydrolytic breakdown of wood purified oxalate oxidase (OXO ; EO 1.2.3.4) from white­ polysaccharides during brown-rot wood decay processes. rot fungus Ceriporiopsis subvermispora and they proposed a Furthermore, F. palustris has been reported to acquire novel pathway in which oxalate is metabolized by OXO to energy for growth by "oxalate-"l). An carbon dioxide, accompanied with the production of 11 oxalate-producing , glyoxylate dehydrogenase H 20 2 ) (GLOXDH) linked with oxalate biosynthesis, and Thus, we were motivated to investigate whether C. isocitrate (IOL) as a key enzyme of the glyoxylate subvermispora has the oxalate-metabolizing systems with ll cycle, have been successfully purified and characterized ODO, FDH and OXO ) We report here preliminary from F. palustrii,3). Furthermore, another oxalate­ results for the purification and characterization of FDH producing enzyme, oxaloacetase has been detected from and the detection of ODO activity from C. subvermispora. 4 wood-rotting fungi ). The results are discussed in relation to oxalate metabolism On the contrary, white-rot fungi accumulate much by this fungus. smaller amounts of oxalic acid because they have oxalate­ 5 8 decomposing systems - ). Under the extracellular condi­ Results and Discussion tion, the two biochemical mediators, including veratryl alcohol cation radicals and Mn3 + produced by lignin Ceriporiopsis subvermispora OS 105 that was kindly provided peroxidase and manganese peroxidase, respectively, have from Dr. Vicuna was cultivated at 2rO in the Kirk's basal l2 been reported to catalyze the decomposition of oxalic acid medium ) containing 2.5% glucose as a carbon source, 3.0 to carbon dioxide9,lO). As a result, oxalic acid seemingly mM ammonium tartrate as a nitrogen source, which was inhibits ligninolytic enzymes4). Furthermore, it has been supplemented with 7-fold minerals. We have purified proposed as a general mechanism for intracellular oxalate FDH from C. subvermispora by varIOUS column

o Route A !JJr2CO, + H,O, TCA a ~COOH) Isocitrate Glyoxylate NAD+ NADH+H+ cycle V .A 2 CO: "-.HCOOH U CO, ® RouteS @

Figure A possible biochemical mechanism for oxalate metabolism in C. subvermispora. Notes: CD Oxalate oxidase, CV Oxalate decarboxylase, ® Formate dehydrogenase.

*1 Laboratory of Biochemical Control, Wood Research Institute, Kyoto University, Uji, Kyoto 611-0011, Japan. *2 North Sumatra University, Indonesia. *3 A part of this work was presented at the 55th Annual Meeting of the Wood Research Society in Fukuoka, March 2003.

-7- WOOD RESEARCH No. 90 (2003) chromatographies. The purified FDH was found to be and the role in white-rot wood decay (Figure). electrophoreticallya single band on SDS-PAGE gel. The purified FDH was similar in molecular mass to the FDHs I3 I4 References purified from ) and plants ) But the K m value for formate was about one twentieth that of the 1) E. MUNIR, J.J. YOON, T. TOKIMATSU, T. HATTORI and M. SHIMADA: Proc. Nat!. Acad. Sci. USA.) 98(20), 11126-11130 enzyme, although the K m value for NAD+ was almost the I3 same ). The enzyme showed greater activities at the (2001). neutral pH range. 2) E. MUNIR, T. HATTORI and M. SHIMADA: Arch. Biochem. Biophys.) 399(2), 225-231 (2002). The optimum temperature for native FDH was at a 3) T. TOKIMATSU, Y. NAGAI, T. HATTORI and M. SHIMADA: room temperature. Formate was the best FEBS Lett.) 437(1-2), 117-121 (1998). among various intermediate organic acids tested. The 4) Y. AKAMATSU, M. TAKAHASHI and M. SHIMADA: Mokuzai FDH activity was inhibited by NADH (60 pM), ATP (10 Gakkaishi) 39, 352-356 (1993). mM), and ADP (10 mM). Interestingly, 2-oxoglutarate 5) H. SHIMAZONO: J. Biolchem.) 42, 321-340 (1955). and oxaloacetate also inhibited the . These 6) H. SHIMAZONO: Bull. Forest Expt. Station) 33, 393-397 results suggest that these a-ketoacids may control the (1951). enzyme activity intrftcellularly. 7) S. TAKAO: Appl. Microbiol.) 13, 727-732 (1965). The ODe activity was detected from the cell-free 8) M.V. DUTTON, C.S. EVANS, P.T. ATKEY and D.A. WOOD: extracts of C. subvermispora. Thus, the results strongly Appl. Microbiol. Biotechnol.) 39(1), 5-10 (1993). 9) Y. AKAMATSU, D.E. MA, T. HIGUCHI and M. SHIMADA: suggest that C. subvermispora decomposes oxalate to CO2 via formate (Figure, Route B), besides another oxalate FEBS Lett.) 269, 261-263 (1990). metabolizing pathway which was reported by Aguilar et at. 10) D.B. MA, T. HATTORI, Y. AKAMATSU, M. ADACHI and M. (Route A) 11). SHIMADA: Biosci. Biochem. Biotechnol.) 56,1378-1381 (1992). 11) C. AGUILAR, U. URZUA, C. KOENING and R. VICUNA: Arch. We suspect that NADH produced as the results of the Biochem. Biophys.) 366, 275-282 (1999). oxidation of formate may serve as an electron donor for I5 12) T.K. KIRK, E. SCHULTZ, W.]. CONNORS, L.F. LORENZ and ATP generation as in the case ofyeasts ). Alternatively, J.G. ZEICUS: Arch. Microbiol.) 117, 277-285 (1978). NADH may be used as a cosubstrate for several enzymes to 13) T.V. AVILOVA, O.A. EGOROVA, L.S. LOANESYAN and A.M. reduce quinones derived from lignin. It is speculated that EGOROV: Eur. J. Biochem.) 152, 657-662 (1985). white-rot fungi are superior to brown-rot ones in 14) D. PEACOCK and D. BOULTER: Biochem. J.) 120, 763-769 biochemical evolution on conversion of oxalate to an (1970). energy source. However, further research is needed to 15) N. KATO, H. SAHM and F. WANER: Biochim. Biophys. Acta) elucidate the reaction mechanisms for oxalate metabolism 566(1), 12-20 (1979).

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