Bifidobacterium bifidum NCIMB41171 carries four genes encoding different β-galactosidases. One of them, named bbgIII, consisted of an open reading frame of 1,935 amino acid (a.a.) residues encoding a protein with a multidomain structure, commonly identified on cell wall bound enzymes, having a signal peptide, a membrane anchor, FIVAR domains, immunoglobulin Ig-like and discoidin-like domains. The other three genes, termed bbgI, bbgII and bbgIV, encoded proteins of 1,291, 689 and 1,052 a.a. residues, respectively, which were most probably intracellularly located. Two cases of protein evolution between strains of the same species were identified when the a.a. sequences of the BbgI and BbgIII were compared with homologous proteins from B. bifidum DSM20215. The homologous proteins were found to be differentiated at the C-terminal a.a. part either due to a single nucleotide insertion or to a whole DNA sequence insertion, respectively. The bbgIV gene was located in a gene organisation surrounded by divergently transcribed genes putatively for sugar transport (galactoside-symporter) and gene regulation (LacI-transcriptional regulator), a structure that was found to be highly conserved in B. longum, B. adolescentis and B. infantis, suggesting optimal organisation shared amongst those species.
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Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410
Baumgartner S, Hofmann K, Chiquet-Ehrismann R, Bucher P (1998) The discoidin domain family revisited: New members from prokaryotes and homology-based fold prediction. Protein Sci 7:1626–1631
Bezkorovainy A, Catchpole MR (1989) Biochemistry and Physiology of Bifidobacteria. CRC, Florida
Biery CM, Stewart JF, Stellwagen AE, Raleigh EA, Craig LN (2000) A simple in vitro Tn7-based transposition system with low target site selectivity for genome and gene analysis. Nucleic Acids Res 28(5):1067–1077
Derensy-Dron D, Krzewinski F, Brassart C, Bouquelet S, (1999). β-1,3-Galactosyl-N-acetylhexosamine phosphorylase from Bifidobacterium bifidum DSM 20082: characterisation, partial purification and relation to mucin degradation. Biotechnol Appl Biochem 29:3–10
Dumortier V, Brassart C, Bouquelet S, (1994). Purification and properties of a β-galactosidase from Bifidobacterium bifidum exhibiting transgalactosylation reaction. Biotechnol Appl Biochem 19:341–354
Fujita K, Oura F, Nagamine N, Katayama T, Hiratake J, Sakata K, Kumagai H, Yamamoto K (2005) Identification and molecular cloning of a novel glycoside hydrolase family of core 1 type O-glycan-specific endo-alpha-N-acetylgalactosaminidase from Bifidobacterium longum. J Biol Chem 280 (45):37415–37422
Gardy JL, Laird MR, Chen F, Rey S, Walsh SJ, Ester M, Brinkman FSL (2005) PSORTb v.2.0: Expanded prediction of bacterial protein subcellular localization and insights gained from comparative proteome analysis. Bioinformatics 21(5):617–23
Gascell A, Crennell S, Taylor G (1995) The three domains of a bacterial sialidase: A β-propeller, an immunoglobulin module and a galactose binding jelly-roll. Structure 3:1197–1205
Gibson GR, Roberfroid MB (1995) Dietary modulation of the human colonic microbiota: Introducing the concept of prebiotics. J Nutr 125:1401–1412
Goulas KA, Tzortzis G, Gibson RG (2006) Development of a process for the production and purification of α- and β-galactooligosaccharides from Bifidobacterium bifidum NCIMB 41171. Int Dairy J 17(6):648–656
Hanahan D (1983) Studies on transformation of Escherichia coli with plasmids. J Mol Biol 166:557–580
Henrissat B, Bairoch A (1993) New families in the classification of glycosyl hydrolases based on amino acid sequences similarities. Biochem J 293:781–788
Hofmann K, Stoffel W, (1993) TMbase—A database of membrane spanning proteins segments. Biol Chem Hoppe-Seyler 374:166
Hung MN, Xia Z, Hu NT, Lee B (2001) Molecular and biochemical analysis of two β-galactosidases from Bifidobacterium infantis HL96. Appl Environ Microbiol 67 (9):4256–4263
Katayama T, Sakuma A, Kimura T, Makimura Y, Hiratake J, Sakata K, Yamanoi T, Kumagai H, Yamamoto K (2004) Molecular cloning and characterization of Bifidobacterium bifidum 1,2-alpha-L-fucosidase (AfcA), a novel inverting glycosidase (glycoside hydrolase family 95). J Bacteriol 186(15):4885–4893
Kitaoka M, Tian J, Nishimoto M (2005) Novel putative galactose operon involving lacto-N-biose phosphorylase in Bifidobacterium longum. Appl Environ Microbiol 71(6):3158–3162
Lawson PA, Gharbia SE, Shatt HN, Clark DR (1989) Recognition of Fusobacterium nucleatum subgroups FN-1, FN-2 and FN-3 by ribosomal-RNA gene restriction patterns. FEMS Microbiol Lett 65(1–2):41–45
Moller PL, Jorgensen F, Hansen OC, Madsen SM, Stougaard P (2001) Intra-and extracellular β-galactosidases from Bifidobacterium bifidum and B. infantis: Molecular cloning, Heterologous Expression, and Comparative Characterisation. Appl Environ Microbiol 67(5):2276–2283
Navarre WW, Schneewind O (1999) Surface proteins in Gram-positive bacteria and mechanisms of their targeting to the cell wall envelope. Microbiol Mol Biol Review 63:174–229
Nielsen H, Engelbrecht J, Brunak S, Heijne G (1997) Identification of prokaryotic and eukaryotic signal peptides and prediction of their cleavage sites. Protein Eng 10:1–6
Nquyen CC, Saier MH Jr (1995) Phylogenetic, structural and functional analyses of the LacI-GalR family of bacterial transcription factors. FEBS Lett 377:98–102
Parkinson JS (1993) Signal transduction schemes in bacteria. Cell 73:857–871
Prenosil JE, Stuker E, Bourne JR (1987) Formation of oligosaccharides during enzymatic lactose: Part I: State of the art. Biotechnol Bioeng 30:1019–1025
Reese MG (2001) Application of a time-delay neural network to promoter annotation in the Drosomphila melanogaster genome. Comput Chem 26:51–56
Roger LC, McCartney AL (2005) Prebiotics and the infant microbiota. In: Tannock GW (ed) Probiotics and Prebiotics: Scientific aspects. Caister Academic, UK
Rossi M, Altomare L, Conzalez Vara y Rodriguez P, Brigidi P, Matteuzzi D (2000) Nucleotide sequence, expression and transcriptional analysis of the Bifidobacterium longum MB 219 lacZ gene. Arch Microbiol 174:74–80
Sambrook J, Russell WD (2001) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, New York
Satokari RM, Vaughan EE, Smidt H, Saarela M, Matto J, de Vos WM (2003) Molecular approaches for the detection and identification of bifidobacteria and lactobacilli in the human gastrointestinal tract. System Appl Microbiol 26:572–584
Schell MA, Karmiratzou M, Snel B, Vilanova D, Berger B, Pessi G, Zwahlen MC, Desiere F, Bork P, Delley M, Pridmore DR, Arigoni F (2002) The genome sequence of Bifidobacterium longum reflects its adaptation to the human gastrointestinal tract. Proc Natl Acad Sci USA 99(22):14422–14427
Sgorbati B, Biavati B, Palenzona D (1995) The genus of Bifidobacterium: In: Wood BJB and Holzapfel WH (eds) The Lactic Acid Bacteria. Blackie Academic and Professional, London
Suzuki T, Tsuda Y, Kanou N, Inoue T, Kumazaki K, Nagano S, Hirai S, Tanaka K, Watanabe K, (2006) Bifidobacterium adolescentis complete genome sequence. NCBI Database
Tzortzis G, Goulas AK, Gibson GR (2005a) Synthesis of prebiotic galactooligosaccharides using the whole cells of a novel strain Bifidobacterium bifidum NCIMB 41171. Appl Microbiol Biotechnol 68:412–416
Tzortzis G, Goulas AK, Gee MJ, Gibson RG (2005b) A novel galactooligosaccharide mixture increases the bifidobacterial population numbers in a continuous in vitro fermentation system and in the proximal colonic contents of pigs in vivo. J Nutr 135:1726–1731
Van Laere KMJ, Abee T, Schols HA, Beldman G, Voragen AGJ (2000) Characterisation of a novel β-galactosidase from Bifidobacterium adolescentis DSM 20083 active towards transgalactooligosaccharides. Appl Environ Microbiol 66(4):1379–1384
This work was supported by the Greek State Scholarship’s Foundation (I.K.Y.).
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Goulas, T.K., Goulas, A.K., Tzortzis, G. et al. Molecular cloning and comparative analysis of four β-galactosidase genes from Bifidobacterium bifidum NCIMB41171. Appl Microbiol Biotechnol 76, 1365–1372 (2007). https://doi.org/10.1007/s00253-007-1099-1
- B. bifidum
- Protein evolution