Characteristics of a Bifidobacterium longum LL04 ${\beta}$-Galactosidase (recombinant) Produced in Escherichia coli

  • Lim, Seong-Il (Division of Fermented Food Research, Korea Food Research Institute) ;
  • Kim, Geun-Bae (Department of Animal Science and Technology, Chung-Ang University) ;
  • Yi, Sung-Hun (Department of Food Science and Agricultural Chemistry, McGill University) ;
  • Lee, Byong-Hoon (Department of Food Science and Agricultural Chemistry, McGill University)
  • Published : 2006.12.31

Abstract

Recombinant ${\beta}$-galactosidase from Bifidobacterium longum LL04 was expressed in Escherichia coli and partially purified by ammonium sulphate precipitation and anion-exchange chromatography (Mono-Q). The optimum temperature and pH of the partially purified enzyme were $50^{\circ}C$ and pH 7.0-8.0, respectively, when o-nitrophenyl-${\beta}$-D-galactopyranoside was used as a substrate. The enzyme was stable over the pH range of 5.0-9.0, and was active at $40^{\circ}C$ for more than 60 min at pH 7.0. The enzyme was significantly activated by $Na^+$ and $K^+$. Maximal activity was observed at the concentration of 10 mM for both $Na^+$ and $K^+$. The enzyme activity was strongly inhibited by most bivalent metal ions. The Km and Vmax on ONPG at 37 and $50^{\circ}C$ were 0.72, 167.9, and 0.507 mM, 310.9 U/mL, respectively.

Keywords

References

  1. Meghrous J, Euloge P, Junelles AM, Ballongue J, Petitdemange H. Screening of Bijidobacterium strains from bacteriocin production. Biotechnol. Lett. 12: 575-580 (1990) https://doi.org/10.1007/BF01030755
  2. Fujiwara S, Hashiba H, Hirota T, Forstner JF. Proteinaceous factor(s) in culture supernatant fluids of bifidobacteria which prevents the binding of enterotoxigenic Escherichia coli to gangliotetraosylceramide. Appl. Environ. Microbiol. 63: 506-512 (1997)
  3. Kim JW, Lee SJ, Park KH. Antimicrobial effect of Bifidobacterium breve and Bijidobacterium infantis against Salmonella typhimurium KCTC 1925 and E. coli O157:H7 ATCC 43895. Food Sci. Biotechnol. 11: 89-92 (2002)
  4. Toida T, Sekine K, Tatsuki T, Saito M, Kawashima T, Hashimoto Y, Sakurai Y. Biochemical characterization and antitumor activity of a new cell wall preparation, whole peptidoglycan (WPG) from Bijidobacterium infantis. J. Cancer Res. Clin. 116: 342-348 (1990)
  5. Bouhnik Y, Flourie B, Andrieux C, Bisetti N, Briet F, Rambaud rc. Effects of Bijidobacterium sp. fermented milk ingested with or without inulin on colonic bifidobacteria and enzymatic activities in healthy humans. Eur. J. Clin. Nutr. 50: 269-273 (1996)
  6. Singh J, Rivenson A, Tomita M, Shimamura S, Ishibashi N, Reddy BS. Bijidobacterium longum, a lactic acid-producing intestinal bacterium inhibits colon cancer and modulates the intermediate biomarkers of colon carcinogenesis. Carcinogenesis 18: 833-841 (1997) https://doi.org/10.1093/carcin/18.4.833
  7. Shin MS, Yu KW, Shin KS, Lee H. Enhancement of immunological activity in mice with oral administration of cell wall components of Bijidobacterium infidum. Food Sci. Biotechnol. 13: 85-89 (2004)
  8. Tahri K, Grill JP, Schneider F. Bifidobacteria strain behavior toward cholesterol: coprecipitation with bile salts and assimilation. Curr. Microbiol. 33: 187-193 (1996) https://doi.org/10.1007/s002849900098
  9. Tahri K, Grill JP, Schneider F. Involvement oftrihydroxyconjugated bile salts in cholesterol assimilation by bifidobacteria. Curr. Microbiol. 34: 79-84 (1997) https://doi.org/10.1007/s002849900148
  10. Deguchi Y, Morishita T, Mutai M. Comparative studies on systhesis of water-soluble vitamins among human species of bifidobacteria. Agr. Biol. Chem. Tokyo 49: 13-19 (1985) https://doi.org/10.1271/bbb1961.49.13
  11. Teraguchi S, Ono J, Kiyosawa I, Fukuwatari Y, Araki J, Okonogi S. Vitamin production by bifidobacteria originated from human intestine. J. Jpn. Soc. Nutr. Food Sci. 32: 157-169 (1986)
  12. Rossi M, Altomare L, Gohzalez A, Roodriguez V, Brigidi P, Matteuzzi D. Nucleotide sequence, expression, and transcriptional analysis of the Bifidobacterium longum MB 219 lacZ gene. Arch. Michrobiol. 174: 74-80 (2000) https://doi.org/10.1007/s002030000178
  13. Schell MA, Karrnirantzou M, Snel B, Vilanova D, Berger B, Pessi G, Zwahlen MC, Desiere F, Bork P, Delley M, Pridmore RD, Arigoni F. The genome sequence of Bifidobacterium longum reflects its adaptation to the human gastrointestinal tract. P. Natl. Acad. Sci. USA 99: 14422-14427 (2002)
  14. Hung MN, Lee BH. Cloning and expression of $\beta$-galactosidase genes from Bifidobacterium irfantis into Escherichia coli. Biotechnol. Lett. 20: 659-662 (1998) https://doi.org/10.1023/A:1005314422383
  15. Nunoura N, Ohdan K, Tanaka K, Tamaki H, Yano T, Inui M, Yukawa H, Yamamoto K, Kumagai H. Cloning and nucleotide sequence of the $\beta$-D-galactosidase gene from Bifidobacterium breve clb, and expression of $\beta$-D-galactosidase activity in Escherichia coli. Biosci. Biotech. Bioch. 60: 2011-2018 (1996) https://doi.org/10.1271/bbb.60.2011
  16. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of proteindye binding. Anal. Biochem. 72: 248-254 (1970) https://doi.org/10.1016/0003-2697(76)90527-3
  17. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685 (1970) https://doi.org/10.1038/227680a0
  18. Bhowmik T, Johnson MC, Ray B. Factors influencing synthesis and activity of $\beta$-galactosidase in Lactobacillus acidophilus. J. Ind. Microbiol. 2: 1-7 (1987) https://doi.org/10.1007/BF01569399
  19. Hung MN, Lee BH. Purification and characterization of a recombinant $\beta$-galactosidase with transgalactosylation activity from Bifidobacterium infantis HL96. Appl. Microbiol. Biochem. 58: 439-445 (2002) https://doi.org/10.1007/s00253-001-0911-6
  20. Huang DQ, Prevost H, Divies C. Principal characteristics of $\beta$-galactosidase from Leuconostoc spp. Int. Dairy J. 5: 29-43 (1995) https://doi.org/10.1016/0958-6946(94)P1597-7
  21. Greenberg NA, Mahoney RR. Production and characterization of $\beta$-galactosidase from Streptococcus thermophilus. J. Food Sci. 47: 1824-1828 (1982) https://doi.org/10.1111/j.1365-2621.1982.tb12891.x
  22. Trimbur DE, Gutshall KR, Prema P, Brenchley JE. Characterization of a psychrotrophic arthrobacter gene and its cold-active $\beta$-galactosidase. Appl. Environ. Microbiol. 60: 4544-4552 (1994)
  23. Huber RE, Kurz G, Wallenfels K. A quantitation of the factor which affect the hydrolase and transgalactosylase activities of $\beta$-galactosidase (E. coli) on lactose. Biochemistry-US 15: 1994-2001 (1976) https://doi.org/10.1021/bi00654a029
  24. Chakraborti S, Sani RK, Banerjee UC, Sobti RC. Purification and characterization of a novel $\beta$-galactosidase from Bacillus sp MTCC 3088. J. Ind. Microbiol. Biot. 24: 58-63 (2000) https://doi.org/10.1038/sj.jim.2900770
  25. Nakao M, Harada M, Kodama Y, Nakayama T, Shibano Y, Amachi T. Purification and characterization of a thermostable $\beta$-galactosidase with high transgalactosylation activity from Saccharopolyspora rectivirgula. Appl. Microbiol. Biot. 40: 657-663 (1994) https://doi.org/10.1007/BF00173325
  26. Greenberg NA, Mahoney RR. Immobilization of lactose ($\beta$galactosidase) for use in dairy processing: A Review. Process Biochem. 16: 2-8 (1981)
  27. Ulrich JT, McFeter GA, Temple KL. Induction and characterization of $\beta$-galactosidase in an extreme thermophile. J. Bacteriol. 110: 691-698 (1972)
  28. Bhowmik T, Marth EH. $\beta$-Galactosidase of Pedococcus species: induction, purification and partial characterization. Appl. Microbiol. Biot. 33: 317-323 (1990) https://doi.org/10.1007/BF00164529
  29. Mahoney RR, Nickerson TA, Whitaker JR. Selection of strain, growth conditions and extraction procedures for optimum production of lactase from Kluyveromyces fragilis. J. Dairy Sci. 58: 1620-1629 (1975) https://doi.org/10.3168/jds.S0022-0302(75)84760-6
  30. Abdelrahim KA, Lee BH. Production and characterization of $\beta$-galactosidase from psychrotrophic Bacillus subtilis KL88. Biotechnol. Appl. Bioc. 13: 246-256 (1991)
  31. Itoh T, Ohhashi M, Toba T, Adachi S. Purification and properties of $\beta$-galactosidase from Lactobacillus bulgaricus. Milchwissenschaft 35: 593-597 (1980)
  32. Garman J, Coolbear T, Smart J. The effect of cations on the hydrolysis of lactose and the transferase reactions catalysed by $\beta$-galactosidase from six strains of lactic acid bacteria. Appl. Microbiol. Biot. 46: 22-27 (1996) https://doi.org/10.1007/s002530050778
  33. Cavaille D, Combes D. Characterization of $\beta$-galactosidase from Kluyveromyces lactis. Biotechnol. Appl. Biochem. 22: 55-64 (1995)
  34. Smart J, Richardson B. Molecular properties and sensitivity to cations of $\beta$-galactosidase from Streptococcus thermophilus with four enzyme substrates. Appl. Microbiol. Biot. 26: 177-185 (1987) https://doi.org/10.1007/BF00253905
  35. Park YK, DeSanti MSS, Pastore OM. Production and characterization of $\beta$-galactosidase from Aspergillus oryzae. J. Food Sci. 44: 100-103 (1979) https://doi.org/10.1111/j.1365-2621.1979.tb10016.x
  36. Dumortier V, Brassart C, Bouquelet S. Purification and properties of a $\beta$-D-galactosidase from Bifidobacterium bifidum exhibiting a transgalactosylation reaction. Biotechnol. Appl. Bioc. 19: 341-354 (1994)
  37. Ohtsuka K, Tanoh A, Ozawa O, Kanematsu T, Uchida T, Shinke R. Purification and properties of a $\beta$-galactosidase with high galactosyl transfer activity from Cryptococcus laurentii. J. Ferment. Bioeng. 70: 301-307 (1990) https://doi.org/10.1016/0922-338X(90)90138-M