Heterologous expression and characterization of the exopolysaccharide from Streptococcus thermophilus Sfi39.
The genes responsible for exopolysaccharide (EPS) synthesis in Streptococcus thermophilus Sfi39 were identified on a 20-kb genomic fragment. The two genes, epsE and epsG, were shown to be involved in EPS synthesis as their disruption lead to the loss of the ropy phenotype. Several naturally selected nonropy mutants were isolated, one acquired an insertion sequence (IS)-element (IS905) in the middle of the eps gene cluster. The eps gene cluster was cloned and transferred into a nonEPS-producing heterologous host, Lactococcus lactis MG1363. The EPS produced was shown by chemical analysis and NMR spectroscopy to be identical to the EPS produced by S. thermophilus Sfi39. This demonstrated first that all genes needed for EPS production and export were present in the S. thermophilus Sfi39 eps gene cluster, and second that the heterologous production of an EPS was possible by transfer of the complete eps gene cluster alone, provided that the heterologous host possessed all necessary genetic information for precursor synthesis.
Germond
J E
JE
Nestlé Research Center, Vers-chez-les-Blanc, CH-1000 Lausanne 26, Switzerland. jacques-edouard.germond@rdls.nestle.com
Delley
M
M
D'Amico
N
N
Vincent
S J
SJ
D001483:Base Sequence; D017931:DNA Primers; D004269:DNA, Bacterial; D005798:Genes, Bacterial; D013294:Lactococcus lactis / Q000235:genetics; D005810:Multigene Family; D016296:Mutagenesis; D019906:Nuclear Magnetic Resonance, Biomolecular; D011135:Polysaccharides, Bacterial / Q000737:chemistry / Q000235:genetics*; D013291:Streptococcus / Q000737:chemistry / Q000235:genetics*
D017931:DNA Primers; D004269:DNA, Bacterial; D011135:Polysaccharides, Bacterial