Molecular cloning and characterization of the alcohol dehydrogenase ADH1 gene of Candida utilis ATCC 9950

Oxford University Press (OUP) - Tập 33 - Trang 1032-1036 - 2006
Yong-Cheol Park1, Na-Rae Yun1, Ka-Yiu San2, George N. Bennett1
1Department of Biochemistry and Cell Biology, Rice University, Houston, USA
2Department of Bioengineering, Rice University, Houston, USA

Tóm tắt

The alcohol dehydrogenase gene (ADH1) of Candida utilis ATCC9950 was cloned and expressed in recombinant Escherichia coli. C. utilis ADH1 was obtained by PCR amplification of C. utilis genomic DNA using two degenerate primers. Amino acid sequence analysis of C. utilis ADH1 indicated that it contained a zinc-binding consensus region and a NAD(P)+-binding site, and lacked a mitochondrial targeting peptide. It has a 98 and 73% identity with ADH1s of C. albicans and Saccharomyces cerevisiae, respectively. Amino acid sequence analysis and enzyme characterization with various aliphatic and branched alcohols suggested that C. utilis ADH1 might be a primary alcohol dehydrogenase existing in the cytoplasm and requiring zinc ion and NAD(P)+ for reaction.

Tài liệu tham khảo

Benetzen JL, Hall BD (1982) The primary structure of the Saccharomyces cerevisiae gene for alcohol dehydrogenase I. J Biol Chem 257:3018–3025 Cho JY, Jeffries TW (1998) Pichia stipitis genes for alcohol dehydrogenase with fermentative and respiratory functions. Appl Environ Microbiol 64:1350–1358 Leskovac V, Trivić S, Peričin D (2002) The three zinc-containing alcohol dehydrogenases from baker’s yeast, Saccharomyces cerevisiae. FEMS Yeast Res 2:481–494 Russell DW, Smith M, Williamson VM, Young ET (1983) Nucleotide sequence of the yeast alcohol dehydrogenase II. J Biol Chem 258:2674–2682 Saliola M, Shuster JR, Falcone C (1990) The alcohol dehydrogenase system in the yeast, Kluyveromyces lactis. Yeast 6:193–204 Saliola M, Gonnell R, Mazzoni C, Falcone C (1991) Two genes encoding putative mitochondrial alcohol dehydrogenases are present in the yeast Kluyveromyces lactis. Yeast 7:391–400 Young ET, Pilgrim D (1985) Isolation and DNA sequence of ADH3, a nuclear gene encoding the mitochondrial isozyme of alcohol dehydrogenase in Saccharomyces cerevisiae. Mol Cell Biol 5:3024–3034 Yurimoto H, Lee B, Yasuda F, Sakai Y, Kato N (2004) Alcohol dehydrogenases that catalyze methyl formate synthesis participate in formaldehyde detoxification in the methylotrophic yeast Candida boidinii. Yeast 21:341–350 Murdanoto AP, Sakai Y, Sembiring L, Tani Y, Kato N (1997) Ester synthesis by NAD+-dependent dehydrogenation of hemiacetal: production of methyl formate by cells of methylotrophic yeasts. Biosci Biotech Biochem 61:1391–1393 Bertram G, Swoboda RK, Gooday GW, Gow NAR, Brown AJ (1996) Structure and regulation of the Candida albicans ADH1 gene encoding an immunogenic alcohol dehydrogenase. Yeast 12:115–127 Jones T, Federspiel NA, Chibana H, Dungan J, Kalman S, Magee BB, Newport G, Thrstenson YR, Agabian N, Magee PT, Davis RW, Scherer S (2004) The diploid genome sequence of Candida albicans. Proc Nat Acad Sci 101:7329–7334 Verduyn C, Breedveld GJ, Acheffers WA, van Dijken JP (1988) Substrate specificity of alcohol dehydrogenase from the yeasts Hansenula polymorpha CBS 4732 and Candida utilis CBS 621. Yeast 4:143–148 Kusano M, Sakai Y, Kato N, Yoshimoto H, Tamai Y (1999) A novel hemiacetal dehydrogenase activity involved in ethyl acetate synthesis in Candida utilis. J Biosci Bioeng 87:690–692 Park YC, Kim SJ, Choi JH, Lee WH, Park KM, Kawamukai M, Ryu YW, Seo JH (2005) Batch and fed-batch production of coenzyme Q10 in recombinant Escherichia coli containing the decaprenyl diphosphate synthase gene from Gluconobacter subdoxydans. Appl Microbiol Biotechnol 67:192–196 Reid MR, Fewson CA (1994) Molecular characterization of microbial alcohol dehydrogenases. Crit Rev Microbiol 20:13–56 Fan F, Lorenzen JA, Plapp BV (1991) An aspartate residue in yeast alcohol dehydrogenase I determines the specificity for coenzyme. Biochemistry 30:6397–6401 van Iersel MFM, Eppink MHM, van Berkel WJH, Rombouts FM, Abee T (1997) Purification and characterization of a novel NADP-dependent branched-chain alcohol dehydrogenase from Saccharomyces cerevisiae. Appl Environ Microbiol 63:4079–4082 Green DW, Sun HW, Plapp BV (1993) Inversion of the substrate specificity of yeast alcohol dehydrogenase. J Biol Chem 268:7792–7798 Kotani T, Yamamoto T, Yurimoto H, Sakai Y, Kato N (2003) Propane monooxygenase and NAD+-dependent secondary alcohol dehydrogenase in propane metabolism by Gordonia sp. strain TY-5. J Bacteriol 185:7120–7128 Kusano M, Sakai Y, Kato N, Yoshimoto H, Sone H, Tamai Y (1998) Hemiacetal dehydrogenase activity of alcohol dehydrogenases in Saccharomyces cerevisiae. Biosci Biotechnol Biochem 62:1956–1961 Kondo K, Miura Y, Sone H, Kobayashi K, Iijima H (1997) High-level expression of a sweet protein, monellin, in the food yeast Candida utilis. Nat Biotechnol 15:453–457 Fan F, Plapp BV (1999) Probing the affinity and specificity of yeast alcohol dehydrogenase I for coenzymes. Arch Biochem Biophy 367:240–249 Sambrook J, Fritsch EF, Maniatis T (1989) Molecular coning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor