Cloning, molecular characterization and expression analysis of two small heat shock genes isolated from wine Lactobacillus plantarum

Spano, Giuseppe; Capozzi, Vittorio; Vernile, A.; Massa, Salvatore

doi:10.1111/j.1365-2672.2004.02359.x

Cited by 35 publications

(34 citation statements)

References 32 publications

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“…CtsR negatively autoregulates its own synthesis by the same mechanism (14). The CtsR regulon was previously shown to be involved in ethanol and heat stress responses in B. subtilis (23) and L. plantarum (49). Analogously, our results show that the CtsR regulon was partially induced after 30 min and 24 h of ethanol exposure and included elevated expression of ClpP-, ClpE-, and Hsp1-encoding genes.…”

Section: Discussionsupporting

confidence: 72%

Short- and Long-Term Adaptation to Ethanol Stress and Its Cross-Protective Consequences in Lactobacillus plantarum

Veen

Abee

Tempelaars

et al. 2011

Appl Environ Microbiol

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This paper describes the molecular responses of Lactobacillus plantarum WCFS1 toward ethanol exposure. Global transcriptome profiling using DNA microarrays demonstrated adaptation of the microorganism to the presence of 8% ethanol over short (10-min and 30-min) and long (24-h) time intervals. A total of 57 genes were differentially expressed at all time points. Expression levels of an additional 859 and 873 genes were modulated after 30 min and 24 h of exposure to the solvent, respectively. Ethanol exposure led to induced expression of genes involved in citrate metabolism and cell envelope architecture, as well as canonical stress response pathways controlled by the central stress regulators HrcA and CtsR. Correspondingly, cells grown for 24 h in medium containing 8% ethanol exhibited higher levels of citrate consumption and modified cell membrane fatty acid composition and showed invaginating septa compared with cells grown in liquid medium without ethanol. In addition, these physiological changes resulted in cross-protection against high temperatures but not against several other stresses tested. To evaluate the role of HrcA and CtsR in ethanol tolerance, ctsR and hrcA gene deletion mutants were constructed. The growth rate of the L. plantarum ⌬ctsR::cat strain was impaired in de Man-Rogosa-Sharpe (MRS) medium containing 8% ethanol, whereas growth of the L. plantarum ⌬hrcA::cat and ⌬ctsR ⌬hrcA::cat mutants was indistinguishable from that of wild-type cells. Overall, these results suggest that the induction of CtsR class III stress responses provides cross-protection against heat stress.

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Section: Discussionsupporting

confidence: 72%

Short- and Long-Term Adaptation to Ethanol Stress and Its Cross-Protective Consequences in Lactobacillus plantarum

Veen

Abee

Tempelaars

et al. 2011

Appl Environ Microbiol

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“…Heat stress was previously shown to enhance the expression of the heat shock genes hsp18.5, hsp19.3, and hsp18.55, carried on the genome of Lb. plantarum NC8 (339,456). As mentioned earlier, CSPs are induced as a result of cold shock pretreatment, and they have been associated with the stabilization of mRNA (457).…”

Section: Stress Associated With Technological Productionmentioning

confidence: 97%

Stress Physiology of Lactic Acid Bacteria

Papadimitriou

Alegría

Bron

et al. 2016

Microbiol Mol Biol Rev

511

404

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SUMMARYLactic acid bacteria (LAB) are important starter, commensal, or pathogenic microorganisms. The stress physiology of LAB has been studied in depth for over 2 decades, fueled mostly by the technological implications of LAB robustness in the food industry. Survival of probiotic LAB in the host and the potential relatedness of LAB virulence to their stress resilience have intensified interest in the field. Thus, a wealth of information concerning stress responses exists today for strains as diverse as starter (e.g.,Lactococcus lactis), probiotic (e.g., severalLactobacillusspp.), and pathogenic (e.g.,EnterococcusandStreptococcusspp.) LAB. Here we present the state of the art for LAB stress behavior. We describe the multitude of stresses that LAB are confronted with, and we present the experimental context used to study the stress responses of LAB, focusing on adaptation, habituation, and cross-protection as well as on self-induced multistress resistance in stationary phase, biofilms, and dormancy. We also consider stress responses at the population and single-cell levels. Subsequently, we concentrate on the stress defense mechanisms that have been reported to date, grouping them according to their direct participation in preserving cell energy, defending macromolecules, and protecting the cell envelope. Stress-induced responses of probiotic LAB and commensal/pathogenic LAB are highlighted separately due to the complexity of the peculiar multistress conditions to which these bacteria are subjected in their hosts. Induction of prophages under environmental stresses is then discussed. Finally, we present systems-based strategies to characterize the “stressome” of LAB and to engineer new food-related and probiotic LAB with improved stress tolerance.

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“…Similar results were obtained by Walker et al (28) when Lactobacillus johnsonii was exposed to a heat shock at a temperature at which GroESL expression was highest (55°C for 45 min) prior to freezing at Ϫ20°C for 7 days. Interestingly, expression of small heat shock proteins during exposure to cold has been reported for Lactobacillus plantarum, suggesting that there is a link between chaperone induction and cold stress (21,22). It has been reported that GroESL is responsible for low-temperature growth and mRNA stability in Escherichia coli (7,9), which may explain the protective functions associated with GroESL overproduction during freeze-drying.…”

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confidence: 99%

Enhanced Survival of GroESL-Overproducing Lactobacillus paracasei NFBC 338 under Stressful Conditions Induced by Drying

Corcoran

Ross

Fitzgerald

et al. 2006

Appl Environ Microbiol

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GroESL-overproducing Lactobacillus paracasei NFBC 338 was dried, and its viability was compared with that of controls. Spray-and freeze-dried cultures overproducing GroESL exhibited ϳ10-fold and 2-fold better survival, respectively, demonstrating the importance of GroESL in stress tolerance, which can be exploited to enhance the technological performance of sensitive probiotic cultures.The health benefits associated with consumption of probiotic bacteria have been well characterized (13,18). From a processing perspective, these microorganisms must be suitable for large-scale industrial production, so that up to 10 7 CFU g Ϫ1 are present in a food product at the end of its shelf-life. Spraydrying is an effective method for producing stable powders at low operating costs; however, the survival rates of lactic acid bacteria are often low (12), and the loss of viability is caused principally by cell membrane damage (1). Freeze-drying exposes cells to attenuating effects of freezing and rehydration. In addition, the secondary structures of RNA and DNA stabilize, resulting in reduced efficacy of DNA replication, transcription, and translation (27). Lactobacillus paracasei NFBC 338 is a suitable probiotic candidate for spray-drying (8), and heat or salt adaptation increased its tolerance to spray drying (6). As GroESL was upregulated following heat stress of L. paracasei NFBC 338, we homologously overexpressed the GroESL operon in L. paracasei NFBC 338, thereby conferring protection during heat, salt, and butanol stresses (5). The chaperone protein GroESL refolds denatured proteins during heat stress (17). The functions of the chaperone do not appear to be limited to heat stress but involve a wider role in cellular processes, such as growth (7), mRNA stability (9), and cytoplasmic protein folding (10). The studies described in this paper built on these findings and demonstrated that L. paracasei NFBC 338 overproducing GroESL exhibited enhanced survival during drying, thus demonstrating a novel use for an overproduced chaperone protein.In order to evaluate the significance of overexpression of GroESL on the technological performance of a probiotic strain during drying, L. paracasei NFBC 338 containing either pGRO2 (an overproducer of GroESL) or pMSP3535 (control) was grown to the late exponential phase as previously described (5), centrifuged, suspended in reconstituted skim milk (RSM) (20%, wt/vol) to a final density of 2.0 ϫ 10 9 to 2.5 ϫ 10 9 CFU g Ϫ1 , and spray-dried. A laboratory-scale spray-dryer (model B191 Buchi mini spray-dryer; Flawil, Switzerland) was used to process samples at a constant air inlet temperature of 180°C and an outlet temperature of 95 to 100°C, and the percentage of surviving bacteria was determined as described previously (8). Following spray-drying, cultures overproducing GroESL exhibited approximately 10-fold better survival (P Ͻ 0.05) than controls; this treatment resulted in powders that contained 8.03 ϫ 10 7 CFU g Ϫ1 and exhibited 5.15% survival, while cultures of L. paracasei NFBC 338 ...

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Cloning, molecular characterization and expression analysis of two small heat shock genes isolated from wine Lactobacillus plantarum

Cited by 35 publications

References 32 publications

Short- and Long-Term Adaptation to Ethanol Stress and Its Cross-Protective Consequences in Lactobacillus plantarum

Short- and Long-Term Adaptation to Ethanol Stress and Its Cross-Protective Consequences in Lactobacillus plantarum

Stress Physiology of Lactic Acid Bacteria

Enhanced Survival of GroESL-Overproducing Lactobacillus paracasei NFBC 338 under Stressful Conditions Induced by Drying

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