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Partial purification and comparative characterization of α-amylase secreted byLactobacillus amylovorus

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Abstract

An α-amylase (E.C. 3.2.1.1.) secreted byLactobacillus amylovorus was partially purified and characterized. This high-molecular-weight enzyme [Imam SH, Burgess-Cassler A, Côté GL, Gordon SH, Baker FL (1991) Curr Microbiol 22:365–370] was quantified with a clinical α-amylase assay adapted to a microplate format. It was isolated from concentrated cell-free culture medium by ammonium sulfate precipitation, ion exchange, and hydrophobic interaction chromatographies. The enzyme was not particularly thermostable, but like three other microbial α-amylases tested for comparison, was renaturable following treatment with SDS and heat. The pH optimum and pI were 5.5±0.5 and 5.0, respectively; its temperature optimum was 60–65°C, and the molecular weight on SDS gels was 140±10 kDa.

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Literature Cited

  1. Adler-Nissen J (1987) Newer uses of microbial enzymes in food processing. Tibtech 5:170–174

    Google Scholar 

  2. Brumm PJ, Hebeda RE, Teague WM (1988) Purification and properties of a new, commercial, thermostableBacillus stear-othermophilus α-amylase. Food Biotechnol 2:67–80

    Google Scholar 

  3. Burgess-Cassler A, Imam SH, Gould JM (1991) High-molecular-weight amylase activities from bacteria degrading starchplastic films. Appl Environ Microbiol 57:612–614

    Google Scholar 

  4. Candussio A, Schmid G, Böck A (1990) Biochemical and genetic analysis of a maltopentaose-producing amylase from an alkaliphilic gram-positive bacterium. Eur J Biochem 191:177–185

    Google Scholar 

  5. Champ M, Szylit O, Raibaud P, Nadra A-A (1983) Amylase production by threeLactobacillus strains isolated from chicken crop. J Appl Bacteriol 55:487–493

    Google Scholar 

  6. Cheng P, Mueller RE, Jaeger S, Bajpai R, Iannotti EL (1991) Lactic acid production from enzyme-thinned cornstarch usingLactobacillus amylovorus. J Indust Microbiol 7:27–34

    Google Scholar 

  7. Chiang JP, Alter JE, Sternberg-Elkhart M (1979) Purification and characterization of a thermostable α-amylase fromBacillus licheniformis. Starch/Stärke 31:86–92

    Google Scholar 

  8. de Man JC, Rogosa M, Sharpe ME (1960) A medium for the cultivation of lactobacilli. J Appl Bacteriol 23:130–135

    Google Scholar 

  9. de Mot R, Verachtert H (1987) Purification and characterization of extra-cellular α-amylase and glucoamylase from the yeastCandida antarctica CBS 6678. Eur J Biochem 164:643–654

    Google Scholar 

  10. Giulian GG, Moss RL, Greaser M (1983) Improved methodology for analysis and quantitation of proteins on one-dimensional silver-stained slab gels. Anal Bioch 129:277–287

    Google Scholar 

  11. Hanzawa S, Odagawa A, Sakata H, Takeuchi M, Ichishima E (1986) Degradation of α-amylase fromAspergillus oryzae with firmly bound proteinases at pH 3.0. Curr Microbiol 14:235–240

    Google Scholar 

  12. Heeb MJ, Gabriel O (1984) Enzyme localization in gels. Methods Enzymol 104:416–439

    Google Scholar 

  13. Imam SH, Burgess-Cassler A, Côté GL, Gordon SH, Baker FL (1991) A study of cornstarch granule digestion by an unusually high molecular weight α-amylase secreted byLactobacillus amylovorus: identification of an unusually high molecular weight α-amylase. Curr Microbiol, 22:365–370

    Google Scholar 

  14. Kennedy JF, Cabalda VM, White CA (1988) Enzymatic starch utilization and genetic engineering. Tibtech 6:184–189

    Google Scholar 

  15. Koch H, Röper H (1988) New industrial products from starch. Starch/Stärke 40:121–131

    Google Scholar 

  16. Lacks SA, Springhorn SS (1980) Renaturation of enzymes after polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. J Biol Chem 255:7467–7473

    Google Scholar 

  17. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Google Scholar 

  18. McCleary BV, Codd R (1989) Measurement of β-amylase in cereal flours and commercial enzyme preparations. J Cereal Sci 9:17–33

    Google Scholar 

  19. McCleary BV, Sheehan H (1987) Measurement of cereal α-amylase: a new assay procedure. J Cereal Sci 6:237–251

    Google Scholar 

  20. Morgan FJ, Priest FG (1981) Characterization of a thermostable α-amylase fromBacillus licheniformis NCIB 6346. J Appl Bacteriol 50:107–114

    Google Scholar 

  21. Nakamura LK (1981)Lactobacillus amylovorus, a new starch-hydrolyzing species from cattle waste-corn fermentations. Int J Syst Bacteriol 31:56–63

    Google Scholar 

  22. Nakamura LK, Crowell CD (1979)Lactobacillus amylophilus, a new starch-hydrolyzing species from swine waste-corn fermentation. Dev Ind Microbiol 20:531–540

    Google Scholar 

  23. Nosoh Y, Sekiguchi T (1990) Protein engineering for thermostability. Tibtech 8:16–20

    Google Scholar 

  24. Olympia M, Takano M (1989) Isolation of an amylolytic lactic acid bacterial from “Burong Bangus,” In: Ghee AH, Hen NB, and Kong LK (eds), Trends in food biotechnology. Singapore: Singapore Institute of Food Science and Technology. pp 114–120

    Google Scholar 

  25. Pace CN (1990) Measuring and increasing protein stability. Tibtech 8:93–98

    Google Scholar 

  26. Ramesh MV, Lonsane BK (1990) Characteristics and novel features of thermostable α-amylase produced byBacillus licheniformis M27 under solid state fermentation. Starch/Stärke 42:233–238

    Google Scholar 

  27. Sen S, Chakrabarty SL (1986) Amylase fromLactobacillus cellobiosus D-39 isolated from vegetable wastes: purification and characterization. J Appl Bacteriol 60:419–423

    Google Scholar 

  28. Soor SK, Hincke MT (1990) Microplate reader-based kinetic determination of α-amylase activity: application to quantitation of secretion from rat parotid acini. Anal Biochem 188:187–191

    Google Scholar 

  29. Vihinen M, Mäntsälä P (1989) Microbial amylolytic enzymes. Crit Rev Biochem Molec Biol 24:329–418

    Google Scholar 

  30. Whitaker JR (ed.) (1972) Principles of enzymology for the food sciences, New York: Marcel Dekker Inc., pp 1–636

    Google Scholar 

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

    Google Scholar 

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Authors and Affiliations

  1. Agricultural Research Service, Biopolymer Research Unit, National Center for Agricultural Utilization Research, USDA, 61604-3999, Peoria, IL, USA

    Anthony Burgess-Cassler & Syed Imam

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  1. Anthony Burgess-Cassler
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  2. Syed Imam
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Burgess-Cassler, A., Imam, S. Partial purification and comparative characterization of α-amylase secreted byLactobacillus amylovorus . Current Microbiology 23, 207–213 (1991). https://doi.org/10.1007/BF02092280

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