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Abstract
Strains of Lactococcus lactis ssp. lactis and Lactococcus lactis ssp. cremoris were cultured under aerobic and anaerobic conditions on plates of whey agar, Elliker agar, and M17L agar at 15, 20, and 30°C to determine the environmental conditions required for the expression of the ropy phenotype. Two strains, L. lactis ssp. cremoris Ropy 352 and L. lactis ssp. cremoris Hollandicus, exhibited two distinct polysaccharide phenotypes, ropy and mucoid. Expression of these phenotypes could be induced individually or simultaneously. The inducible nature of this response suggests that genetic regulators were present. Western blots were used to determine whether or not Lon protease and RcsA, two regulators of polysaccharide expression in Escherichia coli, were present in lactococci. Lon, a negative regulator, and RcsA, an unstable positive regulator, have been shown at the structural level to be conserved in a number of Gram-negative and Gram-positive microorganisms. The present study found evidence for structural conservation of Lon protease in lactococci. Less of the Lon-like protein was observed in the ropy strains than in the nonropy strains.
Key words
Abbreviation Key:
EPS (exopolysaccharide), LAB (lactic acid bacteria), TBST (Tris-buffered saline plus 1% Tween 20)References
- Sequence identification of 2,375 human brain genes.Nature (Lond.). 1992; 355: 632
- Isolation from Klebsiella and characterization of two rcs genes that activate colanic acid capsular biosynthesis in Escherichia coli.J. Gen. Microbiol. 1987; 133: 331
- Site-directed mutagenesis of La protease.FEBS Lett. 1991; 287: 211
- Characterization of heat shock in Bacillus subtilis.J. Bacteriol. 1986; 168: 1243
- Heat shock-induced protein synthesis in Lactococcus lactis ssp. lactis.Curr. Microbiol. 1992; 24: 281
- Improved enumeration of lactic acid streptococci on Elliker agar containing phosphate.Appl. Environ. Microbiol. 1979; 38: 173
- The rcsA gene from Erwinia amylovora: identification, nucleotide sequence analysis, and regulation of exopolysaccharide biosynthesis.Mol. Plant-Microbe Interact. 1990; 3: 429
- Is thermotolerance correlated to heatshock protein synthesis in Lactococcus lactis ssp. lactis? Int.J. Food Microbiol. 1992; 16: 227
- Exopolysaccharide production in milk by Lactobacillus delbrueckii ssp. bulgaricus CNRZ 1187 and by two colonial variants.J. Dairy Sci. 1996; 79: 205
- Exocellular polysaccharides produced by lactic acid bacteria.FEMS Microbiol. Lett. 1990; 87: 113
- Carbon source requirements for exopolysaccharide production by Lactobacillus casei CG11 and partial structure analysis of the polymer.J. Bacteriol. 1994; 60: 3914
- Isolation and characterization of an rcsA-like gene of Erwinia amylovora that activates extracellular polysaccharide production in Erwinia species. Escherichia coli, and Salmonella typhimurium.Mol. Plant-Microbe Interact. 1990; 3: 429
- Sequence of the lon gene in Escherichia coli: a heat shock gene which encodes the ATP-dependent protease La.J. Biol. Chem. 1988; 263: 11718
- Molecular cloning, expression and nucleotide sequence of the rcsA gene of Erwinia amylovora, encoding a positive regulator of capsule expression: evidence for a family of related capsule activator proteins.J. Gen. Microbiol. 1990; 136: 1799
- Evidence for structural conservation of Lon and RcsA.J. Bacteriol. 1994; 176: 5126
- Polysaccharide expression in lactococci. Genetics of streptococci, enterococci and lactococci. 85. Dev. Biol. Stand, Basel1995: 469
- Isolation and characterization of lon mutants in Salmonella typhimurium.J. Bacteriol. 1986; 165: 193
- Cloning, expression, and characterization of the lon gene of Erwinia amylovora: evidence for a heat shock response.J. Bacteriol. 1995; 177: 932
- An agar culture medium for lactic streptococci and lactobacilli.J. Dairy Sci. 1956; 39: 1611
- Exopolysaccharide production in different media by lactic acid bacteria.Cult. Dairy Products J. 1995; 30: 18
- Plasmid complements of Streptococcus lactis NCD0712 and other lactic streptococci after protoplastinduced curing.J. Bacteriol. 1983; 154: 1
- Myxococcus xanthus encodes an ATP-dependent protease which is required for developmental gene transcription and intercellular signaling.J. Bacteriol. 1993; 175: 4538
- Regulation of capsule synthesis: modification of the two-component paradigm by an accessory unstable regulator.in: Hoch J.A. Silhavy T.J. Two-Component Signal Transduction. Am. Soc. Microbiol, Washington, DC1995: 253
- Structure of the exopolysaccharide produced by Lactococcus lactis subspecies cremoris H414 grown in a defined medium or skimmed milk.Carbohydr. Res. 1992; 231: 273
- Antibodies: A Laboratory Manual.Cold Spring Harbor Lab, Cold Spring Harbor, NY1988
- Cloning, characterization, and inactivation of the Bacillus brevis lon gene.J. Bacteriol. 1992; 174: 2281
- The rcsA gene of Escherichia coli 09:K30:H12 is involved in the expression of the serotype specific Group 1 K (capsular) antigen.J. Bacteriol. 1992; 174: 8
- Finnish fermented milk “Viili”: involvement of two cell surface proteins in production of slime by Streptococcus lactis ssp. cremoris.J. Dairy Sci. 1988; 71: 3197
- Isolation of two different polysaccharides from halophilic Zoogloea sp.Biotechnol. Lett. 1994; 16: 783
- Cleavage of structural proteins during the assembly of the head of bacteriophage T4.Nature. 1970; 227: 680
- Scandinavian ropy milk—identification and characterization of endogenous ropy lactic streptococci and their extracellular excretion.J. Dairy Sci. 1984; 67: 735
- Structure of the extracellular polysaccharide from slimeforming Lactococcus lactis ssp. cremoris SBT 0495.Carbohydr. Res. 1992; 224: 245
- A novel phosphopolysaccharide from slime-forming Lactococcus lactis subspecies cremoris SBT 0495.J. Dairy Sci. 1990; 73: 1472
- Exopolysaccharides from Rhizobium meliloti YE-2 grown under different osmolarity conditions: viscoelastic properties.Carbohydr. Res. 1992; 223: 227
- Plasmid encoded functions of ropy lactic strains from Scandinavian fermented milk.Biochimie. 1988; 70: 437
- Cloning and nucleotide sequence of the Myxococcus xanthus lon gene: indispensability of lon for vegetative growth.J. Bacteriol. 1993; 175: 2271
- The lonD gene is homologous to the lon gene encoding an ATP-dependent protease and is essential for the development of Myxococcus xanthus.J. Bacteriol. 1993; 175: 4545
- Structural and functional analysis of the rcsA gene from Erwinia stewartii.Mol. Gen. Genet. 1991; 229: 155
- Asymmetric segregation of heat-shock proteins upon cell division in Caulobacter crescentus.J. Mol. Biol. 1987; 194: 653
- Cloning, nucleotide sequence, and expression of the Bacillus subtilus lon gene.J. Bacteriol. 1994; 176: 6518
- Molecular Cloning: A Laboratory Manual.2nd ed. Cold Spring Harbor Lab, Cold Spring Harbor, NY1989
- Ecology of the lactic streptococci.A review. J. Milk Food Technol. 1972; 35: 176
- Identification and characterization of the eps (exopolysaccharide) gene cluster from Streptococcus thermophilus Sfi6.J. Bacteriol. 1996; 178: 1680
- Biosynthesis of microbial exopolysaccharides.Adv. Microb. Physiol. 1982; 23: 79
- Requirement for the yeast gene lon in intramitochondrial proteolysis and maintenance of respiration.Science (Washington, DC). 1994; 264: 273
- A method for the efficient blotting of strongly basic proteins from sodium dodecyl sulfatepolyacrylamide gels to nitrocellulose.Anal. Biochem. 1985; 150: 403
- Improved medium for lactic streptococci and their bacteriophages.Appl. Microbiol. 1975; 29: 807
- Capsular polysaccharide of a slime-forming Lactococcus lactis ssp. cremoris LAPT 3001 isolated from Swedish fermented milk “langfil”.Int. J. Food Microbiol. 1991; 12: 167
- Cloning and nucleotide sequence of the Myxococcus xanthus lon gene: indispensability of lon for vegetative growth.J. Bacteriol. 1993; 175: 2271
- Capsule synthesis in Escherichia coli K-12 is regulated by proteolysis.J. Bacteriol. 1987; 169: 981
- Control of extracellular polysaccharide biosynthesis in Erwinia stewartii and Escherichia coli K12.J. Bacteriol. 1987; 160: 184
- Enhanced immunodetection of blotted house dust mite protein allergen on nitrocellulose following blocking with Tween 20.Electrophoresis. 1989; 10: 243
- Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications.Proc. Natl. Acad. Sci. USA. 1979; 76: 4350
- Involvement of a plasmid in production of ropiness (mucoidness) in milk cultures by Streptococcus cremoris MS.Appl. Microbiol. 1986; 51: 677
- Plasmidencoded ropiness production in Lactobacillus casei ssp. casei.Biotechnol. Lett. 1989; II: 709
- Characterization of the rcsA and rcsB genes from Salmonella typhi: rcsB through tviA is involved in regulation of Vi antigen synthesis.J. Bacteriol. 1996; 178: 1691
- Construction of Streptococcus lactis ssp. lactis strains with a single plasmid associated with mucoid phenotype.Appl. Environ. Microbiol. 1987; 53: 1385
- Yogurt production from reconstituted skim milk powders using different polymer and non-polymer forming starter cultures.J. Dairy Res. 1993; 60: 247
- A human mitochondrial ATPdependent protease that is highly homologous to bacterial Lon protease.Proc. Natl. Acad. Sci. USA. 1993; 90 (247): 11
- Characterization of the heat shock response in Lactococcus lactis ssp. lactis.Appl. Microbiol. 1991; 57: 1408
Article info
Publication history
Accepted:
January 27,
1997
Received:
July 8,
1996
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© 1997 American Dairy Science Association. Published by Elsevier Inc.
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