ABSTRACT
This study evaluated the shelf life of fresh pasta filled with cheese subjected to modified atmosphere packaging (MAP) or air packaging (AP). After a pasteurization treatment, fresh pasta was packaged under a 50/50 N(2)/CO(2) ratio or in air (air batch). Changes in microbial growth, in-package gas composition, chemical-physical parameters and sensory attributes were monitored for 42 days at 4 (°)C. The pasteurization treatment resulted in suitable microbiological reduction. MAP allowed a mold-free shelf life of the fresh filled pasta of 42 days, whereas air-packaged samples got spoilt between 7 and 14 days. The hurdle approach used (MAP and low storage temperature) prevented the growth of pathogens and alterative microorganisms. MAP samples maintained a high microbiological standard throughout the storage period. The panel judged MAP fresh pasta above the acceptability threshold throughout the shelf life.
Subject(s)
Food Handling , Food Microbiology , Food Packaging/methods , Food Preservation , Hydrogen-Ion Concentration , Time Factors , WaterABSTRACT
Streptococcus thermophilus strain CNRZ 302 is unable to ferment galactose, neither that generated intracellularly by lactose hydrolysis nor the free sugar. Nevertheless, sequence analysis and complementation studies with Escherichia coli demonstrated that strain CNRZ 302 contained structurally intact genes for the Leloir pathway enzymes. These were organized into an operon in the order galKTE, which was preceded by a divergently transcribed regulator gene, galR, and followed by a galM gene and the lactose operon lacSZ. Results of Northern blot analysis showed that the structural gal genes were transcribed weakly, and only in medium containing lactose, by strain CNRZ 302. However, in a spontaneous galactose-fermenting mutant, designated NZ302G, the galKTE genes were well expressed in cells grown on lactose or galactose. In both CNRZ 302 and the Gal(+) mutant NZ302G, the transcription of the galR gene was induced by growth on lactose. Disruption of galR indicated that it functioned as a transcriptional activator of both the gal and lac operons while negatively regulating its own expression. Sequence analysis of the gal promoter regions of NZ302G and nine other independently isolated Gal(+) mutants of CNRZ 302 revealed mutations at three positions in the galK promoter region, which included substitutions at positions -9 and -15 as well as a single-base-pair insertion at position -37 with respect to the main transcription initiation point. Galactokinase activity measurements and analysis of gusA reporter gene fusions in strains containing the mutated promoters suggested that they were gal promoter-up mutations. We propose that poor expression of the gal genes in the galactose-negative S. thermophilus CNRZ 302 is caused by naturally occurring mutations in the galK promoter.
Subject(s)
Galactose/metabolism , Gene Expression Regulation, Bacterial , Genes, Bacterial , Lactose/metabolism , Regulon , Streptococcus/genetics , Base Sequence , Cloning, Molecular , DNA, Intergenic , Escherichia coli Proteins , Lac Operon , Molecular Sequence Data , Promoter Regions, Genetic , Repressor Proteins/genetics , Sequence Analysis, DNA , Transcription, Genetic , beta-Galactosidase/geneticsABSTRACT
Streptococci from different collections and dairy materials were characterized by conventional and molecular methods. After amplification of the 16S-23S rDNA spacer region, all the strains referable to the genus Streptococcus exhibited a single polymerase chain reaction (PCR) product, allowing their differentiation from enterococci. Cleaving this PCR product with Hae III, two different restriction patterns could be observed, allowing Streptococcus salivarius DSM 20560T, Strep. thermophilus NCDO 822 and two strains of Streptococcus spp. to be gathered in one group and all the other strains in another. In order to achieve strain typing, all the cultures were investigated by random amplified polymorphic DNA (RAPD)-PCR analysis employing two selected primers. The results were treated by cluster analysis, appearing significantly consistent with both the taxonomic position and the origin of the strains. Pulsed-field gel electrophoresis (PFGE) of Sma I digests of the genomic DNA from 11 representative strains with decreasing levels of RAPD similarity allowed their diversity to be confirmed, even though RAPD-PCR proved to be less discriminating than PFGE analysis. The results are discussed with reference to the capability of the analytical procedures used to aid both identification and strain typing of streptococci, as well as the taxonomic structure of the species Strep. thermophilus.
