Starch debranching enzyme (R-enzyme or pullulanase) from developing rice endosperm: purification, cDNA and chromosomal localization of the gene

Planta - Tập 199 - Trang 209-218 - 1996
Yasunori Nakamura1, Takayuki Umemoto2, Naoki Ogata1, Yoshihide Kuboki3, Masahiro Yano3, Takuji Sasaki3
1Department of Applied Physiology, National Institute of Agrobiological Resources (NIAR), Ibaraki, Japan
2Department of Lowland Farming, Tohoku National Agricultural Experiment Station, Akita, Japan
3Rice Genome Research Program, NIAR/STAFF, Ibaraki, Japan

Tóm tắt

Starch debranching enzyme (R-enzyme or pullulanase) was purified to homogeneity from developing endosperm of rice (Oryza sativa L. cv. Fujihikari) using a variety of high-performance liquid chromatography columns, and characterized. A cDNA clone encoding the full length of the rice endosperm debranching enzyme was isolated and its nucleotide sequence was determined. The cDNA contains an open reading frame of 2958 bp. The mature debranching enzyme of rice appears to be composed of 912 amino acids with a predicted relative molecular mass (Mr) of 102069 Da, similar in size to its Mr of about 100 000 Da estimated by polyacrylamide gel electrophoresis in sodium dodecyl sulfate. The amino acid sequence of rice debranching enzyme is substantially similar to that of bacterial pullulanase, while it bears little similarity to that of bacterial isoamylase or to glycogen debranching enzymes from human muscle and rabbit muscle. Southern blot analyses strongly suggest that the debranching enzyme gene is present as a single copy in the rice genome. Analysis by restriction fragment length polymorphism with a probe including the 3′-untranslated region of cDNA for rice debranching enzyme confirmed that the debranching enzyme gene is located on chromosome 4.

Tài liệu tham khảo

Amemura A, Chakraborty R, Fujita M, Noumi T, Futai (1988) Cloning and nucleotide sequence of the isoamylase gene from Pseudomonas amyloderamosa SB-15. J Biol Chem 263: 9271–9275 Baecker PA, Greenberg E, Preiss J (1986) Biosynthesis of bacterial glycogen. J Biol Chem 261: 8738–8743 Cleveland DW, Fisher GS, Krischer MW, Laemmli UK (1977) Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem 252: 1102–1106 Doehlert DC, Knutson CA (1991) Two classes of starch debranching enzymes from developing maize kernels. J Plant Physiol 138: 566–572 Hobson PN, Whelan WJ, Peat S (1951) The enzymic synthesis and degradation of starch. Part XIV. R-Enzyme. J Chem Soc 1451–1459 James MG, Robertson DS, Myers AM (1995) Characterization of the gene sugaryl, a determinant of starch composition in kernels. Plant Cell 7: 417–429 Jespersen HM, MacGregor EA, Henrissat B, Sierks MR, Svensson B (1993) Starch-and glycogen-debranching and branching enzymes: Prediction of structural features of the catalytic (β/α)8-barrel domain and evolutionary relationship to other amylolytic enzymes. J Protein Chem 12: 791–805 Katsuragi N, Takizawa N, Murooka Y (1987) Entire nucleotide sequence of the pullulanase gene of Klebsiella aerogenes W70. J Bacteriol 169: 2301–2306 Kornacker MG, Pugsley AP (1990) Molecular characterization of pulA and its product, pullulanase, a secreted enzyme of Klebsiella pneumoniae UNF5023. Mol Microbiol 4: 73–85 Kurata N, Nagamura Y, Yamamoto K, Harushima Y, Sue N, Wu J, Antonio BA, Shomura A, Shimizu T, Lin S-Y, Inoue T, Fukuda A, Shimano T, Kuboki Y, Toyama T, Miyamoto Y, Kirihara T, Hayasaka K, Miyao A, Monna L, Zhong HS, Tamura Y, Wang Z-X, Momma T, Umehara Y, Yano M, Sasaki T, Minobe Y (1994) A 300 kilobase interval genetic map of rice including 883 expressed sequences. Nature Genet 8: 365–372 Kuriki T, Imanaka T (1989) Nucleotide sequence of the neopullulanase gene from Bacillus stearothermophilus. J Gen Microbiol 135: 1512–1528 Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 80–685 Lander ES, Green P, Abrahamson J, Barlow A, Daly MJ, Lincoln SE, Newburg L (1987) MAPMAKER: An interactive computer package for constructing primary genetic linkage map of experimental and natural populations. Genomics 1: 174–181 Lee EYC, Whelan WJ (1971) Glycogen and starch debranching enzymes. In: Boyer PD (ed) The enzymes, vol 4. Academic Press, New York, pp 191–234 Liu W, Castro ML, Takrama J, Bilous PT, Vinayagamoorthy T, Madsen NB, Bleackley RC (1993) Molecular cloning, sequencing, and analysis of the cDNA for rabbit muscle glycogen debranching enzyme. Arch Biochem Biophys 306: 232–239 Maniatis T, Fritsch EF, Sambrook J (1982) In: Molecular cloning: A laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y. Martin C, Smith AM (1995) Starch biosynthesis. Plant Cell 7: 971–985 Nakamura Y, Yamanouchi H (1992) Nucleotide sequence of a cDNA encoding starch-branching enzyme, or Q-enzyme I, from rice endosperm. Plant Physiol 99: 1265–1266 Nakamura Y, Yuki K, Park S-Y, Ohya T (1989) Carbohydrate metabolism in the developing endosperm of rice grains. Plant Cell Physiol 30: 833–839 Nelson N (1944) A photometric adaptation of the Somogyi method for the determination of glucose. J Biol Chem 153: 375–380 Pan D, Nelson ON (1984) A debranching enzyme deficiency in endosperms of the sugary-1 mutants of maize. Plant Physiol 74: 324–328 Romeo T, Kumar A, Preiss J (1988) Analysis of the Escherichia coli glycogen gene cluster suggests that catabolic enzymes are encoded among the biosynthetic genes. Gene 70: 363–376 Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 Somogyi M (1952) Notes on sugar determination. J Biol Chem 195: 19–23 Svensson B (1994) Protein engineering in the α-amylase family: catalytic mechanism, substrate specificity, and stability. Plant Mol Biol 25: 141–157 Takata H, Kuriki T, Okada S, Takesada Y, Iizuka M, Minamiura N, Imanaka T (1992) Action of neopullulanase. J Biol Chem 267: 18447–18452 Thon VJ, Khalil M, Cannon JF (1993) Isolation of human glycogen branching enzyme cDNAs by screening complementation in yeast. J Biol Chem 268: 7509–7513 Toguri T (1991) Changes of a rice debranching enzyme during seed formation and germination. J Plant Physiol 137: 541–546 Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedures and some applications. Proc Natl Acad Sci USA 76: 4350–4354 Yamada J, Izawa M (1979) A debranching enzyme of rice seeds at milky stage. Its purification and substrate specificities. Agric Biol Chem 43: 37–44 Yang BZ, Ding JH, Enghild JJ, Bao Y, Chen YT (1992) Molecular cloning and nucleotide sequence of cDNA encoding human muscle glycogen debranching enzyme. J Biol Chem 267: 9294–9299 Yano M, Isono Y, Satoh H, Omura T (1984) Gene analysis of sugary and shrunken mutants of rice, Oryza sativa L. Jpn J Breed 34: 43–49