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The α-Amylase-Pullulanase (apu) Gene from Clostridium thermohydrosulfuricum: Nucleotide Sequence and Expression in Escherichia coli

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Genetics and Molecular Biology of Anaerobic Bacteria

Part of the book series: Brock/Springer Series in Contemporary Bioscience ((BROCK/SPRINGER))

Abstract

Starch consists of two high molecular mass fractions, amylose and amylopectin, both composed of anhydro glucose units connected to each other by α-glycosidic linkages. In the straight-chain fraction, amylose, there are only α-1,4 linkages, whereas in the branched-chain fraction, amylopectin, short α-1,4 chains are linked to each other by α-1,6 linkages at the branch points. The relative amount of α-1,6 linkages depends on the source of the starch but is approximately 3%–4% in most common starches (Fogarty and Kelly, 1979). α-Amylases (EC 3.2.1.1) hydrolyze α-1,4 linkages in amylose and amylopectin in an endo fashion, producing dextrins and maltooligosaccharides and causing a rapid decrease in the viscosity and iodine-binding capacity of the starch.

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References

  • Alff-Steinberger, C. 1984. Evidence for a coding pattern on the non-coding strand of the E. coli genome. Nucleic Acids Res. 12:2235–2241.

    Article  PubMed  CAS  Google Scholar 

  • Amemura, A., R. Chakraborty, M. Fujita, T. Noumi, and M. Futai. 1988. Cloning and nucleotide sequence of the isoamylase gene from Pseudomonas amyloderamosa SB-15. J. Biol Chem. 263:9271–9275.

    PubMed  CAS  Google Scholar 

  • Antranikian, G. 1990. Physiology and enzymology of thermophilic anaerobic bacteria degrading starch. FEMS Microbiol Rev. 75:201–218.

    Article  CAS  Google Scholar 

  • Aymerich, S., G. Gonzy-Tréboul, and M. Steinmetz. 1986. 5’-Noncoding region sacR is the target of all identified regulation affecting the levansucrase gene in Bacillus subtilis. J. Bacteriol. 166:993–998.

    PubMed  CAS  Google Scholar 

  • Béguin, P., P. Cornet, and J.-P. Aubert. 1985. Sequence of a cellulase gene of the thermophilic bacterium Clostridium thermocellum. J. Bacteriol. 162:102–105.

    PubMed  Google Scholar 

  • Binder, F., O. Huber, and A. Bock. 1986. Cyclodextrin-glycosyltransferase from Klebsiella pneumoniae M5al: cloning, nucleotide sequence and expression. Gene (Amst.) 47:269–277.

    Article  CAS  Google Scholar 

  • Brennan, R.G. and B.W. Matthews. 1989. Structural basis of DNΑ-protein recognition. Trends Biochem. Sci. 14:286–290.

    Article  PubMed  CAS  Google Scholar 

  • Buisson, G., E. Duée, R. Haser, and F. Payan. 1987. Three dimensional structure of porcine pancreatic α-amylase at 2.9 Å resolution. Role of calcium in structure and activity. EMBO J. 6:3909–3916.

    PubMed  CAS  Google Scholar 

  • Coleman, R.D., S.-S. Yang, and M.P. McAlister. 1987. Cloning of the debranching-enzyme gene from Thermoanaerobium brockii into Escherichia coli and Bacillus subtilis. J. Bacteriol. 169:4302–4307.

    PubMed  CAS  Google Scholar 

  • Crombrugghe, B., S. Busby, and H. Buc. 1984. Cyclic AMP receptor protein: role in transcription activation. Science 224:831–838.

    Article  PubMed  Google Scholar 

  • Crutz, A.-M., M. Steinmetz, S. Aymerich, R. Richter, and D. Le Coq. 1990. Induction of levansucrase in Bacillus subtilis: an antitermination mechanism negatively controlled by the phosphotransferase system. J. Bacteriol. 172:1043–1050.

    PubMed  CAS  Google Scholar 

  • Ebright, R.H., Y.W. Ebright, and A. Gunasekera. 1989. Consensus DNA site for the Escherichia coli catabolite gene activator protein (CAP): CAP exhibits a 450-fold higher affinity for the consensus DNA site than for the E. coli lac DNA site. Nucleic Acids Res. 17:10295–10305.

    Article  PubMed  CAS  Google Scholar 

  • d’Enfert, C, C. Chapon, and A.P. Pugsley. 1987. Export and secretion of the lipoprotein pullulα-nase by Klebsiella pneumoniae. Mol Microbiol. 1:107–116.

    Article  PubMed  Google Scholar 

  • Fogarty, W.M. and C.T. Kelly. 1979. Starch-degrading enzymes of microbial origin. In: Progress in Industrial Microbiology, Vol. 15, M.J. Bull, ed., pp. 87–150. Amsterdam: Elsevier.

    Google Scholar 

  • Gamier, J., D.J. Osguthorpe, and B. Robson. 1978. Analysis of the accuracy and implications of simple methods for predicting the secondary structure of globular proteins. J. Mol Biol 120:97–120.

    Article  Google Scholar 

  • Grépinet, O., M.-C. Chebrou, and P. Béguin. 1988. Nucleotide sequence and deletion analysis of the xylanase gene (xynZ) of Clostridium thermocellum. J. Bacteriol. 170:4582–4588.

    PubMed  Google Scholar 

  • Grosjean, H. and W. Fiers. 1982. Preferential codon usage in procaryotic genes: the optimal codon-anticodon interaction energy and the selective codon usage in efficiently expressed genes. Gene (Amst.) 18:199–209.

    Article  CAS  Google Scholar 

  • Hershey, J.W.B. 1987. Protein synthesis. In:Escherichia coli and Salmonella typhimurium, Cellular and Molecular Biology, Vol. 1, F.C. Neidhardt, ed., pp. 613–647. Washington, D.C: American Society for Microbiology.

    Google Scholar 

  • Hoshiko, S., O. Makabe, C. Nojiri, K. Katsumata, E. Satoh, and K. Nagaoka. 1987. Molecular cloning and characterization of the Streptomyces hygroscopicus α-amylase gene. J. Bacteriol. 169: 1029–1036.

    PubMed  CAS  Google Scholar 

  • Hyun, H.H. and J.G. Zeikus. 1985a. General biochemical characterization of thermostable pullulanase and glucoamylase from Clostridium thermohydrosulfuricum. Appl. Environ. Microbiol. 49:1168–1173.

    PubMed  CAS  Google Scholar 

  • Hyun, H.H. and J.G. Zeikus. 1985b. Regulation and genetic enhancement of glucoamylase and pullulanase production in Clostridium thermohydrosulfuricum. J. Bacteriol. 164:1146–1152.

    PubMed  CAS  Google Scholar 

  • Ihara, H., T. Sasaki, A. Tsuboi, H. Yamagata, N. Tsukagoshi, and S. Udaka. 1985. Complete nucleotide sequence of a thermophilic flamylase gene: homology between procaryotic and eucaryotic α-amylases at the active sites. J. Biochem. 98:95–103.

    PubMed  CAS  Google Scholar 

  • Joliff, G., P. Béguin, and J.-A. Aubert. 1986. Nucleotide sequence of the cellulase gene celD encoding endoglucanase D of Clostridium thermo-cellum. Nucleic Acids Res. 14:8605–8613.

    Article  PubMed  CAS  Google Scholar 

  • Katsuragi, N., N. Takizawa, and Y. Murooka. 1987. Entire nucleotide sequence of the pullulanase gene of Klebsiella aerogenes W70. J. Bacteriol. 169:2301–2306.

    PubMed  CAS  Google Scholar 

  • Klingeberg, M., H. Hippe, and G. Antranikian. 1990. Production of novel pullulanases at high concentrations by two newly isolated thermophilic Clostridia. FEMS Microbiol. Lett. 69:145–152.

    Article  CAS  Google Scholar 

  • Kreuzer, P., D. Gärtner, R. Allmansberger, and W. Hillen. 1989. Identification and sequence analysis of the Bacillus subtilis W23 xylR gene and xyl operator. J. Bacteriol. 171:3840–3845.

    PubMed  CAS  Google Scholar 

  • Kuriki, T. and T. Imanaka. 1989. Nucleotide sequence of the neopullulanase gene from Bacillus stearothermophilus. J. Gen. Microbiol. 135: 1521–1528.

    PubMed  CAS  Google Scholar 

  • Kyte, J. and R.F. Doolittle. 1982. A simple method for displaying the hydropathic character of a protein. J. Mol. Biol. 157:105–132.

    Article  PubMed  CAS  Google Scholar 

  • Laoide, B.M., G.H. Chambliss, and DJ. McCon-nell. 1989. Bacillus licheniformis α-amylase gene, amyh, is subject to promoter-independent catabolite repression in Bacillus subtilis. J. Bacteriol. 171:2435–2442.

    PubMed  CAS  Google Scholar 

  • Long, C.M., M.-J. Virolle, S.-Y. Chang, S. Chang, and M.J. Bibb. 1987. α-Amylase gene of Streptomyces limosus: nucleotide sequence, expression motifs, and amino acid sequence homology to mammalian and invertebrate α-amylases. J. Bacteriol. 169:5745–5754.

    PubMed  CAS  Google Scholar 

  • Mathupala, S., B.C. Saha, and J.G. Zeikus. 1990. Substrate competition and specificity at the active site of amylopullulanase from Clostridium thermohydrosulfuricum. Biochem. Biophys. Res. Commun. 166:126–132.

    Article  PubMed  CAS  Google Scholar 

  • Matsuura, Y., M. Kusunoki, W. Harada, and M. Kakudo. 1984. Structure and possible catalytic residues of Takα-amylase A. J. Biochem. (Tokyo) 95:697–702.

    CAS  Google Scholar 

  • McConnell, D.J., B.A. Cantwell, K.M. Devine, A.J. Forage, B.M. Laoide, C. O’Kane, J.F. Ollington, and P.M. Sharp. 1986. Genetic engineering of extracellular enzyme systems of Bacilli. In: Biochemical Engineering IV, C.H. Lim, and K. Venkatasubramanian, eds., Ann. N.Y. Acad. Sci. 469:1–17.

    Google Scholar 

  • Melasniemi, H. 1987a. Effect of carbon source on production of thermostable α-amylase, pullulanase and α-glucosidase by Clostridium thermohydrosulfuricum. J. Gen. Microbiol. 133:883–890.

    CAS  Google Scholar 

  • Melasniemi, H. 1987b. Characterization of α-amylase and pullulanase activities of Clostridium thermohydrosulfuricum. Evidence for a novel thermostable amylase. Biochem. J. 246: 193–197.

    PubMed  CAS  Google Scholar 

  • Melasniemi, H. 1988. Purification and some properties the extracellular α-amylase-pullulanase produced by Clostridium thermohydrosulfuricum. Biochem. J. 250:813–818.

    PubMed  CAS  Google Scholar 

  • Melasniemi, H. and M. Paloheimo. 1989. Cloning and expression of the Clostridium thermohydrosulfuricum α-amylase-pullulanase gene in Escherichia coli. J. Gen. Microbiol. 135:1755–1762.

    PubMed  CAS  Google Scholar 

  • Melasniemi, H., M. Paloheimo, and L. Hemiö. 1990. Nucleotide sequence of the α-amylase-pullulanase gene from Clostridium thermohydrosulfuricum. J. Gen. Microbiol. 136:447–454.

    PubMed  CAS  Google Scholar 

  • Nakajima, R., T. Imanaka, and S. Aiba. 1985. Nucleotide sequence of the Bacillus stearothermophilus α-amylase gene. J. Bacteriol. 163:401–406.

    PubMed  CAS  Google Scholar 

  • Nakajima, R., T. Imanaka, and S. Aiba. 1986. Comparison of amino acid sequences of eleven different α-amylases. Appl. Microbiol. Biotechnol. 23:355–360.

    Article  CAS  Google Scholar 

  • Nakamura, N., N. Sashihara, H. Nagayama, and K. Horikoshi. 1989. Characterization of pullulanase and α-amylase activities of a Thermus sp. AMD33. Starch Stärke 41:112–117.

    Article  Google Scholar 

  • Odibo, F.J.C. and S.K.C. Obi. 1988. Purification and characterization of a thermostable pullulanase from Thermoactinomyces thalpophilus. J. Ind. Microbiol. 3:343–350.

    Article  CAS  Google Scholar 

  • Plant, A.R., R.M. Clemens, H.W. Morgan, and R.M. Daniel. 1987. Active-site- and substrate-specificity of Thermoanaerobium Tok6-Bl pullulanase. Biochem. J. 246:537–541.

    PubMed  CAS  Google Scholar 

  • Pugsley, A.P., C. Chapon, and M. Schwartz. 1986. Extracellular pullulanase of Klebsiella pneumoniae is a lipoprotein. J. Bacteriol. 166:1083–1088.

    PubMed  CAS  Google Scholar 

  • Rogers, J.C. 1985. Two barley α-amylase gene families are regulated differently in aleurone cells. J. Biol Chem. 260:3731–3738.

    PubMed  CAS  Google Scholar 

  • Rosenberg, M. and D. Court. 1979. Regulatory sequences involved in the promotion and termination of RNA transcription. Annu. Rev. Genet. 13:319–353.

    Article  PubMed  CAS  Google Scholar 

  • Saha, B.C. and J.G. Zeikus. 1989. Novel highly thermostable pullulanase from thermophiles. Trends Biotechnol. 7:234–239.

    Article  CAS  Google Scholar 

  • Saha, B.C., S.P. Mathupala, and J.G. Zeikus. 1988. Purification and characterization of a highly thermostable novel pullulanase from Clostridium thermohydrosulfuricum. Biochem. J. 252:343–348.

    PubMed  CAS  Google Scholar 

  • Saha, B.C., R. Lamed, C.-Y. Lee, S.P. Mathupala, and J.G. Zeikus. 1990. Characterization of an endo-acting amylopullulanase from Thermo-anaerobacter strain B6A. Appl Environ. Microbiol. 56:881–886.

    PubMed  CAS  Google Scholar 

  • Saha, B.C., G.-J. Shen, K.C. Srivastava, L.W. LeCureux, and J.G. Zeikus. 1989. New thermostable α-amylase-like pullulanase from thermophilic Bacillus sp. 3183. Enzyme Microb. Technol 11:760–764.

    Article  CAS  Google Scholar 

  • Sata, H., M. Umeda, C.-H. Kim, H. Taniguchi, and Y. Maruyama. 1989. Amylase-pullulanase enzyme produced by B. circulans F-2. Biochim. Biophys. Acta 991:388–394.

    Article  CAS  Google Scholar 

  • Schwarz, W.H., S. Schimming, K.P. Rücknagel, S. Burgschwaiger, G. Kreil, and W.L. Staudenbauer. 1988. Nucleotide sequence of the celC gene encoding endoglucanase C of Clostridium thermocellum. Gene (Amst.) 63:23–30.

    Article  CAS  Google Scholar 

  • Sharon, N. and H. Lis. 1981. Glycoproteins: research booming on long-ignored, ubiquitous compounds. Chem. Eng. News 30:21–44.

    Article  Google Scholar 

  • Spreinat, A. and G. Antranikian. 1990. Purification and properties of a thermostable pullulanase from Clostridium thermosulfurogenes EMI which hydrolyses both α-1,6 and α-1,4-glycosidic linkages. Appl. Microbiol. Biotechnol 33:511–518.

    Article  CAS  Google Scholar 

  • Takano, T., M. Fukuda, M. Monma, S. Kobayashi, K. Kainuma, and K. Yamane. 1986. Molecular cloning, DNA nucleotide sequencing, and expression in Bacillus subtilis cells of the Bacillus macerons cyclodextrin glucanotransferase gene. J. Bacteriol. 166:1118–1122.

    PubMed  CAS  Google Scholar 

  • Takasaki, Y. 1987. Pullulanase-amylase complex enzyme from Bacillus subtilis. Agric. Biol. Chem. 51:9–16.

    Article  CAS  Google Scholar 

  • Takkinen, K., R.F. Pettersson, N. Kalkkinen, I. Palva, H. Söderlund, and L. Kääriäinen. 1983. Amino acid sequence of α-amylase from Bacillus amyloliquefaciens deduced from the nucleotide sequence of the cloned gene. J. Biol. Chem. 258:1007–1013.

    PubMed  CAS  Google Scholar 

  • Vihinen, M. and P. Mäntsälä. 1990. Conserved residues of liquefying α-amylases are concentrated in the vicinity of active site. Biochem. Biophys. Res. Commun. 166:61–65.

    Article  PubMed  CAS  Google Scholar 

  • Wiegel, J., L.G. Ljungdahl, and J.R. Rawson. 1979. Isolation from soil and properties of the extreme thermophile Clostridium thermohydrosulfuricum. J. Bacteriol. 139:800–810.

    PubMed  CAS  Google Scholar 

  • Wu, H.C. 1987. Posttranslational modification and processing of membrane proteins in bacteria. In: Bacterial Outer Membrane as Model System, M. Inouye, ed., pp. 37–71. New York: Academic Press.

    Google Scholar 

  • Yang, M., A. Galizzi, and D. Henner. 1983. Nucleotide sequence of the amylase gene from Bacillus subtilis. Nucleic Acids Res. 11:237–249.

    Article  CAS  Google Scholar 

  • Yuuki, T., T. Nomura, H. Tezuka, A. Tsuboi, H. Yamagata, N. Tsukagoshi, and S. Udaka. 1985. Complete nucleotide sequence of a gene coding for heat- and pH-stable α-amylase of Bacillus licheniformis: comparison of the amino acid sequences of three bacterial liquefying α-amylases deduced from the DNA sequences. J. Biochem. 98:1147–1156.

    PubMed  CAS  Google Scholar 

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  1. Hannes Melasniemi
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  1. Unité des Anaérobies, Institut Pasteur, 75724, Paris, France

    Madaleine Sebald

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Melasniemi, H. (1993). The α-Amylase-Pullulanase (apu) Gene from Clostridium thermohydrosulfuricum: Nucleotide Sequence and Expression in Escherichia coli . In: Sebald, M. (eds) Genetics and Molecular Biology of Anaerobic Bacteria. Brock/Springer Series in Contemporary Bioscience. Springer, New York, NY. https://doi.org/10.1007/978-1-4615-7087-5_32

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  • DOI: https://doi.org/10.1007/978-1-4615-7087-5_32

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