scholarly journals Transcriptome analysis revealed growth phase-associated changes of a centenarian-originated probiotic Bifidobacterium animalis subsp. lactis A6

2021 ◽  
Author(s):  
Hui Wang ◽  
Jieran An ◽  
Chengfei Fan ◽  
Zhengyuan Zhai ◽  
Hongxing Zhang ◽  
...  
Keyword(s):  
Carbon Source ◽  
Stationary Phase ◽  
Gene Cluster ◽  
Growth Phase ◽  
Differential Expression Analysis ◽  
Acid Tolerance ◽  
Glucose Consumption ◽  
Exponential Phase ◽  
Putative Gene ◽  
Bifidobacterium Animalis

Abstract Background The physiology and application characteristics of probiotics are closely associated with the growth phase. Bifidobacterium animalis subsp. lactis A6 is a promising probiotic strain isolated from the feces of a healthy centenarian in China. In this study, RNA-seq was carried out to investigate the metabolic mechanism between the exponential and the stationary phase in B. lactis A6. Results Differential expression analysis showed that a total of 810 genes were significantly changed in the stationary phase compared to the exponential phase, which consisted of 392 up-regulated and 418 down-regulated genes. The results showed that the transport and metabolism of cellobiose, xylooligosaccharides and raffinose were enhanced at the stationary phase, which expanded carbon source utilizing profile to confront with glucose consumption. Meanwhile, genes involved in NH3 production were up-regulated at the stationary phase to enhance acid tolerance during fermentation. In addition, peptidoglycan biosynthesis was significantly repressed, which is comparable with the decreased growth rate during the stationary phase. Remarkably, a putative gene cluster encoding Tad pili was up-regulated 6.5~12.1-fold, which is consistent with the significantly increased adhesion rate to mucin from 2.38–4.90% during the transition from the exponential phase to the stationary phase. Conclusions This study reported growth phase-associated changes of B. lactis A6 during fermentation, including expanded carbon source utilizing profile, enhanced acid tolerance, and up-regulated Tad pili gene cluster responsible for bacterial adhesion in the stationary phase. These findings provide a novel insight into the growth phase associated characteristics in B. lactis A6 and provide valuable information for further application in the food industry.

Morphological Characterization of Mycobacteriophage R1

1974 ◽  
Vol 32 ◽  
pp. 254-255
Author(s):  
B. L. Soloff ◽  
T. A. Rado
Keyword(s):  
Carbon Source ◽  
Stationary Phase ◽  
Growth Phase ◽  
Minimal Medium ◽  
Morphological Characterization ◽  
Logarithmic Growth Phase ◽  
Complete Lysis ◽  
Lysogenic Culture ◽  
Logarithmic Growth

Mycobacteriophage R1 was originally isolated from a lysogenic culture of M. butyricum. The virus was propagated on a leucine-requiring derivative of M. smegmatis, 607 leu−, isolated by nitrosoguanidine mutagenesis of typestrain ATCC 607. Growth was accomplished in a minimal medium containing glycerol and glucose as carbon source and enriched by the addition of 80 μg/ ml L-leucine. Bacteria in early logarithmic growth phase were infected with virus at a multiplicity of 5, and incubated with aeration for 8 hours. The partially lysed suspension was diluted 1:10 in growth medium and incubated for a further 8 hours. This permitted stationary phase cells to re-enter logarithmic growth and resulted in complete lysis of the culture.

scholarly journals RocA Regulates Phosphatase Activity of Virulence Sensor CovS of Group A Streptococcus in Growth Phase- and pH-Dependent Manners

mSphere ◽  
2020 ◽  
Vol 5 (3) ◽  
Author(s):  
Chuan Chiang-Ni ◽  
He-Jing Chiou ◽  
Huei-Chuan Tseng ◽  
Chih-Yun Hsu ◽  
Cheng-Hsun Chiu
Keyword(s):  
Stationary Phase ◽  
Phosphatase Activity ◽  
Virulence Factor ◽  
Growth Phase ◽  
Exponential Phase ◽  
Acidic Stress ◽  
Phosphorylation Level ◽  
Factor Expression ◽  
Group A ◽  
Ph Dependent

ABSTRACT The control of the virulence response regulator and sensor (CovR-CovS) two-component regulatory system in group A Streptococcus (GAS) strains regulates more than 15% of gene expression and has critical roles in invasive GAS infection. The membrane-embedded CovS has kinase and phosphatase activities, and both are required for modulating the phosphorylation level of CovR. Regulator of Cov (RocA) is a positive regulator of covR and also been shown to be a pseudokinase that interacts with CovS to enhance the phosphorylation level of CovR; however, how RocA modulates the activity of CovS has not been determined conclusively. Although the phosphorylation level of CovR was decreased in the rocA mutant in the exponential phase, the present study shows that phosphorylated CovR in the rocA mutant increased to levels similar to those in the wild-type strain in the stationary phase of growth. In addition, acidic stress, which is generally present in the stationary phase, enhanced the phosphorylation level of CovR in the rocA mutant. The phosphorylation levels of CovR in the CovS phosphatase-inactivated mutant and its rocA mutant were similar under acidic stress and Mg2+ (the signal that inhibits CovS phosphatase activity) treatments, suggesting that the phosphatase activity, but not the kinase activity, of CovS is required for RocA to modulate CovR phosphorylation. The phosphorylation level of CovR is crucial for GAS strains to regulate virulence factor expression; therefore, the growth phase- and pH-dependent RocA activity would contribute significantly to GAS pathogenesis. IMPORTANCE The emergence of invasive group A streptococcal infections has been reported worldwide. Clinical isolates that have spontaneous mutations or a truncated allele of the rocA gene (e.g., emm3-type isolates) are considered to be more virulent than isolates with the intact rocA gene (e.g., emm1-type isolates). RocA is a positive regulator of covR and has been shown to enhance the phosphorylation level of intracellular CovR regulator through the functional CovS protein. CovS is the membrane-embedded sensor and modulates the phosphorylation level of CovR by its kinase and phosphatase activities. The present study shows that the enhancement of CovR phosphorylation is mediated via the repression of CovS’s phosphatase activity by RocA. In addition, we found that RocA acts dominantly on modulating CovR phosphorylation under neutral pH conditions and in the exponential phase of growth. The phosphorylation level of CovR is crucial for group A Streptococcus species to regulate virulence factor expression and is highly related to bacterial invasiveness; therefore, growth phase- and pH-dependent RocA activity and the sequence polymorphisms of rocA gene would contribute significantly to bacterial phenotype variations and pathogenesis.

scholarly journals The Nature of Antibacterial Adaptive Immune Responses against Staphylococcus aureus Is Dependent on the Growth Phase and Extracellular Peptidoglycan

2019 ◽  
Vol 88 (1) ◽  
Author(s):  
Payal P. Balraadjsing ◽  
Lisbeth D. Lund ◽  
Yuri Souwer ◽  
Sebastian A. J. Zaat ◽  
Hanne Frøkiær ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Cell Wall ◽  
T Cell ◽  
Stationary Phase ◽  
Growth Phase ◽  
Cell Response ◽  
Th17 Cell ◽  
Exponential Phase ◽  
Th1 Cell ◽  
Content Type

ABSTRACT Staphylococcus aureus has evolved different strategies to evade the immune response, which play an important role in its pathogenesis. The bacteria express and shed various cell wall components and toxins during different stages of growth that may affect the protective T cell responses to extracellular and intracellular S. aureus. However, if and how the dendritic cell (DC)-mediated T cell response against S. aureus changes during growth of the bacterium remain elusive. In this study, we show that exponential-phase (EP) S. aureus bacteria were endocytosed very efficiently by human DCs, and these DCs strongly promoted production of the T cell polarizing factor interleukin-12 (IL-12). In contrast, stationary-phase (SP) S. aureus bacteria were endocytosed less efficiently by DCs, and these DCs produced small amounts of IL-12. The high level of IL-12 production induced by EP S. aureus led to the development of a T helper 1 (Th1) cell response, which was inhibited after neutralization of IL-12. Furthermore, preincubation with the staphylococcal cell wall component peptidoglycan (PGN), characteristically shed during the exponential growth phase, modulated the DC response to EP S. aureus. PGN preincubation appeared to inhibit IL-12p35 expression, leading to downregulation of IL-12 and an increase of IL-23 production by DCs, enhancing Th17 cell development. Taken together, our data indicate that exponential-phase S. aureus bacteria induce a stronger IL-12-dependent Th1 cell response than stationary-phase S. aureus and that this Th1 cell response shifted toward a Th17 cell response in the presence of PGN.

scholarly journals Unraveling the regulation of sophorolipid biosynthesis in Starmerella bombicola

2020 ◽  
Vol 20 (3) ◽  
Author(s):  
Sofie Lodens ◽  
Sophie L K W Roelants ◽  
Goedele Luyten ◽  
Robin Geys ◽  
Pieter Coussement ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stationary Phase ◽  
Gene Cluster ◽  
Biosynthetic Gene Cluster ◽  
Exponential Phase ◽  
Biosynthetic Gene ◽  
Reporter System ◽  
Gfp Expression ◽  
Qpcr Analysis ◽  
Starmerella Bombicola

ABSTRACT Starmerella bombicola very efficiently produces the secondary metabolites sophorolipids (SLs). Their biosynthesis is not-growth associated and highly upregulated in the stationary phase. Despite high industrial and academic interest, the underlying regulation of SL biosynthesis remains unknown. In this paper, potential regulation of SL biosynthesis through the telomere positioning effect (TPE) was investigated, as the SL gene cluster is located adjacent to a telomere. An additional copy of this gene cluster was introduced elsewhere in the genome to investigate if this results in a decoy of regulation. Indeed, for the new strain, the onset of SL production was shifted to the exponential phase. This result was confirmed by RT-qPCR analysis. The TPE effect was further investigated by developing and applying a suitable reporter system for this non-conventional yeast, enabling non-biased comparison of gene expression between the subtelomeric CYP52M1- and the URA3 locus. This was done with a constitutive endogenous promotor (pGAPD) and one of the endogenous promotors of the SL biosynthetic gene cluster (pCYP52M1). A clear positioning effect was observed for both promotors with significantly higher GFP expression levels at the URA3 locus. No clear GFP upregulation was observed in the stationary phase for any of the new strains.

scholarly journals Genetic Evidence that Legionella pneumophila RpoS Modulates Expression of the Transmission Phenotype in Both the Exponential Phase and the Stationary Phase

2004 ◽  
Vol 72 (5) ◽  
pp. 2468-2476 ◽  
Author(s):  
Michael A. Bachman ◽  
Michele S. Swanson
Keyword(s):  
Life Cycle ◽  
Rna Polymerase ◽  
Stationary Phase ◽  
Growth Phase ◽  
Legionella Pneumophila ◽  
Opportunistic Pathogen ◽  
Sigma Factors ◽  
Exponential Phase ◽  
Posttranscriptional Control ◽  
Multiple Copies

ABSTRACT The opportunistic pathogen Legionella pneumophila alternates between two states: replication within phagocytes and transmission between host amoebae or macrophages. In broth cultures that model this life cycle, during the replication period, CsrA inhibits expression of transmission traits. When nutrients become limiting, the alarmone (p)ppGpp accumulates and the sigma factors RpoS and FliA and the positive activators LetA/S and LetE promote differentiation to the transmissible form. Here we show that when cells enter the postexponential growth phase, RpoS increases expression of the transmission genes fliA, flaA, and mip, factors L. pneumophila needs to establish a new replication niche. In contrast, in exponential (E)-phase cells whose (p)ppGpp levels are low, rpoS inhibits expression of transmission traits, on the basis of three separate observations. First, rpoS RNA levels peak in the E phase, suggestive of a role for RpoS during replication. Second, in multiple copies, rpoS decreases the amounts of csrA, letE, fliA, and flaA transcripts and inhibits the transmission traits of motility, infectivity, and cytotoxicity. Third, rpoS blocks expression of cytotoxicity and motility by E-phase bacteria that have been induced to express the LetA activator ectopically. The data are discussed in the context of a model in which the alarmone (p)ppGpp enables RpoS to outcompete other sigma factors for binding to RNA polymerase to promote transcription of transmission genes, while LetA/S acts in parallel to relieve CsrA posttranscriptional repression of the transmission regulon. By coupling transcriptional and posttranscriptional control pathways, intracellular L. pneumophila could respond to stress by rapidly differentiating to a transmissible form.

scholarly journals The expression of the acarbose biosynthesis gene cluster in Actinoplanes sp. SE50/110 is dependent on the growth phase

2021 ◽  
Author(s):  
Julian Droste ◽  
Vera Ortseifen ◽  
Lena Schaffert ◽  
Marcus Persicke ◽  
Susanne Schneiker-Bekel ◽  
...  
Keyword(s):  
Transcriptional Regulation ◽  
Stationary Phase ◽  
Gene Cluster ◽  
Growth Phase ◽  
Formation Rate ◽  
High Temporal Resolution ◽  
Biosynthesis Gene Cluster ◽  
Metabolic Switch ◽  
A Genome ◽  
Post Transcriptional Regulation

Abstract Background: Actinoplanes sp. SE50/110 is the natural producer of the diabetes mellitus drug acarbose, which is highly produced during the growth phase and ceases during the stationary phase. In previous works, the growth-dependency of acarbose formation was assumed to be caused by a decreasing transcription of the acarbose biosynthesis genes during transition and stationary growth phase. Results: In this study, transcriptomic data using RNA-seq and state-of-the-art proteomic data from seven time points of controlled bioreactor cultivations were used to analyze expression dynamics during growth of Actinoplanes sp. SE50/110. A hierarchical cluster analysis revealed co-regulated genes, which display similar transcription dynamics over the cultivation time. Aside from an expected metabolic switch from primary to secondary metabolism during transition phase, we observed a continuously decreasing transcript abundance of all acarbose biosynthetic genes from the early growth phase until stationary phase, with the strongest decrease for the monocistronically transcribed genes acbA, acbB, acbD and acbE. Our data confirm a similar trend for acb gene transcription and acarbose formation rate. Surprisingly, the proteome dynamics does not follow the respective transcription for all acb genes. This suggests different protein stabilities or post-transcriptional regulation of the Acb proteins, which in turn could indicate bottlenecks in the acarbose biosynthesis. Furthermore, several genes are co-expressed with the acb gene cluster over the course of the cultivation, including eleven transcriptional regulators (e.g. ACSP50_0424), two sigma factors (ACSP50_0644, ACSP50_6006) and further genes, which have not previously been in focus of acarbose research in Actinoplanes sp. SE50/110. Conclusion: In conclusion, we have demonstrated, that a genome wide transcriptome and proteome analysis in a high temporal resolution is well suited to study the acarbose biosynthesis and the transcriptional and post-transcriptional regulation thereof.

Augmentation of Killing of Escherichia coli O157 by Combinations of Lactate, Ethanol, and Low-pH Conditions

1999 ◽  
Vol 65 (3) ◽  
pp. 1308-1311 ◽  
Author(s):  
Sarah L. Jordan ◽  
Jayne Glover ◽  
Laura Malcolm ◽  
Fiona M. Thomson-Carter ◽  
Ian R. Booth ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Stationary Phase ◽  
Acid Tolerance ◽  
Low Ph ◽  
Exponential Phase ◽  
Escherichia Coli O157 ◽  
Cytoplasmic Ph ◽  
Ph Homeostasis ◽  
Acid Tolerant ◽  
Ph Conditions

ABSTRACT The acid tolerance of Escherichia coli O157:H7 strains can be overcome by addition of lactate, ethanol, or a combination of the two agents. Killing can be increased by as much as 4 log units in the first 5 min of incubation at pH 3 even for the most acid-tolerant isolates. Exponential-phase, habituated, and stationary-phase cells are all sensitive to incubation with lactate and ethanol. Killing correlates with disruption of the capacity for pH homeostasis. Habituated and stationary-phase cells can partially offset the effects of the lowering of cytoplasmic pH.

scholarly journals Growth-phase dependence of susceptibility to antimicrobial peptides in Staphylococcus aureus

Microbiology ◽  
2011 ◽  
Vol 157 (6) ◽  
pp. 1786-1797 ◽  
Author(s):  
Miki Matsuo ◽  
Yuichi Oogai ◽  
Fuminori Kato ◽  
Motoyuki Sugai ◽  
Hitoshi Komatsuzawa
Keyword(s):  
Staphylococcus Aureus ◽  
Cell Surface ◽  
Antimicrobial Peptides ◽  
Stationary Phase ◽  
Surface Charge ◽  
Growth Phase ◽  
Exponential Phase ◽  
Dependent Manner ◽  
Phase Dependence ◽  
Cell Surface Charge

Bacterial cell surface charge is responsible for susceptibility to cationic antimicrobial peptides. Previously, Staphylococcus aureus dlt and mprF were identified as factors conferring a positive charge upon cell surfaces. In this study, we investigated the regulation of cell surface charge during growth. Using a group of S. aureus MW2 mutants, which are gene-inactivated in 15 types of two-component systems (TCSs), we tested dltC and mprF expression and found that two TCSs, aps and agr, were associated with dltC and mprF expression in a growth phase-dependent manner. The first of these, aps, which had already been identified as a sensor of antimicrobial peptides and a positive regulator of dlt and mprF expression, was expressed strongly in the exponential phase, while its expression was significantly suppressed by agr in the stationary phase, resulting in higher expression of dltC and mprF in the exponential phase and lower expression in the stationary phase. Since both types of expression affected the cell surface charge, the susceptibility to antimicrobial peptides and cationic antibiotics was changed during growth. Furthermore, we found that the ability to sense antimicrobial peptides only functioned in the exponential phase. These results suggest that cell surface charge is tightly regulated during growth in S. aureus.

scholarly journals Taxon- and Growth Phase-Specific Antioxidant Production by Chlorophyte, Bacillariophyte, and Haptophyte Strains Isolated From Tropical Waters

2020 ◽  
Vol 8 ◽  
Author(s):  
Norazira Abdu Rahman ◽  
Tomoyo Katayama ◽  
Mohd Effendy Abd Wahid ◽  
Nor Azman Kasan ◽  
Helena Khatoon ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Stationary Phase ◽  
Antioxidant Capacity ◽  
Growth Phase ◽  
Stationary Phases ◽  
Exponential Phase ◽  
Antioxidant Compounds ◽  
Antioxidant Compound ◽  
Scavenging Capacity ◽  
Β Carotene

Antioxidants found in microalgae play an essential role in both animals and humans, against various diseases and aging processes by protecting cells from oxidative damage. In this study, 26 indigenous tropical marine microalgae were screened. Out of the 26 screened strains, 10 were selected and were further investigated for their natural antioxidant compounds which include carotenoids, phenolics, and fatty acids collected in their exponential and stationary phases. The antioxidant capacity was also evaluated by a total of four assays, which include ABTS, DPPH, superoxide radical (O2•–) scavenging capacity, and nitric oxide (•NO–) scavenging capacity. This study revealed that the antioxidant capacity of the microalgae varied between divisions, strains, and growth phase and was also related to the content of antioxidant compounds present in the cells. Carotenoids and phenolics were found to be the major contributors to the antioxidant capacity, followed by polyunsaturated fatty acids linoleic acid (LA), eicosapentaenoic acid (EPA), arachidonic acid (ARA), and docosahexaenoic acid (DHA) compared to other fatty acids. The antioxidant capacity of the selected bacillariophytes and haptophytes was found to be positively correlated to phenolic (R2-value = 0.623, 0.714, and 0.786 with ABTS, DPPH, and •NO–) under exponential phase, and to carotenoid fucoxanthin and β-carotene (R2 value = 0.530, 0.581 with ABTS, and 0.710, 0.795 with O2•–) under stationary phase. Meanwhile, antioxidant capacity of chlorophyte strains was positively correlated with lutein, β-carotene and zeaxanthin under the exponential phase (R2 value = 0.615, 0.615, 0.507 with ABTS, and R2 value = 0.794, 0.659, and 0.509 with •NO–). In the stationary phase, chlorophyte strains were positively correlated with violaxanthin (0.755 with •NO–), neoxanthin (0.623 with DPPH, 0.610 with •NO–), and lutein (0.582 with •NO–). This study showed that antioxidant capacity and related antioxidant compound production of tropical microalgae strains are growth phase-dependent. The results can be used to improve the microalgal antioxidant compound production for application in pharmaceutical, nutraceutical, food, and feed industry.

Influence of Bacterial Detachment on the Bioflotation of Malachite

2015 ◽  
Vol 1130 ◽  
pp. 485-488
Author(s):  
Ga Hee Kim ◽  
So Yeon Park ◽  
Kyuh Yeong Park ◽  
Junh Yun Choi ◽  
Si Young Q. Choi ◽  
...  
Keyword(s):  
Fluid Flow ◽  
Stationary Phase ◽  
Surface Properties ◽  
Growth Phase ◽  
Morphological Characteristics ◽  
Exponential Phase ◽  
Rhodococcus Opacus ◽  
Cell Detachment ◽  
Flotation Process ◽  
Flotation Behavior

The effects of the bacterial growth phase on the malachite flotation were investigated in a well-controlled Hallimond tube system. Rhodococcus opacus, which is one of representative hydrophobic bacteria, was employed for this study. The test results showed that the bacteria in the stationary phase exhibit two-fold greater floatability than those in mid-exponential phase. To understand the observed flotation behavior, complementary cell characterization tests (e.g., zeta potential and contact angle measurements) and cell attachment tests were conducted. Interestingly, the bacteria at both phases exhibited similar surface properties as well as almost identical amount of cells attached onto the malachite, suggesting that the growth phase dependent flotation behavior cannot be attributed to the variation of cell surface properties and the extent of cell adsorption. On the other hand, cell detachment tests revealed that the amount of cells detached from the malachite surface is greater for the mid-exponential phase than the stationary phase due to the higher fluid drag applied to the cells at the mid-exponential phase, which was explained by the differences in the size and shape of attached bacteria onto the malachite surface. Specifically, the bacteria in the mid-exponential phase had a larger size and formed loosely-packed structures like an end-to end on the malachite surface. These morphological characteristics were found to cause the bacteria of mid-exponential phase to be separated highly sensitive and easy from malachite surface due to the fluid flow. The findings from this study suggest that in the case of bioflotation using a relatively large bacteria size than the collector, it is important to consider the cell detachment by the fluid flow that occurs during a flotation process.

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