Lysogeny Broth (LB) v1

protocols.io ◽  
2016 ◽  
Author(s):  
Steven Burgess
Keyword(s):  
protocols.io ◽  
2017 ◽  
Author(s):  
Haven Himmighoefer ◽  
Rachel Ancar
Keyword(s):  

2014 ◽  
Vol 89 (1-2) ◽  
pp. 384-389 ◽  
Author(s):  
Tian Li ◽  
Xinping Deng ◽  
Jinjun Wang ◽  
Yucheng Chen ◽  
Lin He ◽  
...  

protocols.io ◽  
2019 ◽  
Author(s):  
Brian Smith ◽  
baltrus email
Keyword(s):  

2016 ◽  
Vol 62 (6) ◽  
pp. 464-474 ◽  
Author(s):  
Che O’May ◽  
Olivier Amzallag ◽  
Karim Bechir ◽  
Nathalie Tufenkji

Proteus mirabilis is a major cause of catheter-associated urinary tract infection (CAUTI), emphasizing that novel strategies for targeting this bacterium are needed. Potential targets are P. mirabilis surface-associated swarming motility and the propensity of these bacteria to form biofilms that may lead to catheter blockage. We previously showed that the addition of cranberry powder (CP) to lysogeny broth (LB) medium resulted in impaired P. mirabilis swarming motility over short time periods (up to 16 h). Herein, we significantly expanded on those findings by exploring (i) the effects of cranberry derivatives on biofilm formation of P. mirabilis, (ii) whether swarming inhibition occurred transiently or over longer periods more relevant to real infections (∼3 days), (iii) whether swarming was also blocked by commercially available cranberry juices, (iv) whether CP or cranberry juices exhibited effects under natural urine conditions, and (v) the effects of cranberry on medium pH, which is an indirect indicator of urease activity. At short time scales (24 h), CP and commercially available pure cranberry juice impaired swarming motility and repelled actively swarming bacteria in LB medium. Over longer time periods more representative of infections (∼3 days), the capacity of the cranberry material to impair swarming diminished and bacteria would start to migrate across the surface, albeit by exhibiting a different motility phenotype to the regular “bull’s-eye” swarming phenotype of P. mirabilis. This bacterium did not swarm on urine agar or LB agar supplemented with urea, suggesting that any potential application of anti-swarming compounds may be better suited to settings external to the urine environment. Anti-swarming effects were confounded by the ability of cranberry products to enhance biofilm formation in both LB and urine conditions. These findings provide key insights into the long-term strategy of targeting P. mirabilis CAUTIs.


2019 ◽  
Vol 8 (1) ◽  
pp. 62 ◽  
Author(s):  
Sarah Gingichashvili ◽  
Danielle Duanis-Assaf ◽  
Moshe Shemesh ◽  
John D. B. Featherstone ◽  
Osnat Feuerstein ◽  
...  

Biofilms are commonly defined as accumulations of microbes, embedded in a self-secreted, polysaccharide-rich extra-cellular matrix. This study aimed to characterize specific morphological changes that occur in Bacillus subtilis biofilms under nutrient-limiting growth conditions. Under varying levels of nutrient depletion, colony-type biofilms were found to exhibit different rates of spatial expansion and green fluorescent protein production. Specifically, colony-type biofilms grown on media with decreased lysogeny broth content exhibited increased spatial expansion and more stable GFP production over the entire growth period. By modeling the surface morphology of colony-type biofilms using confocal and multiphoton microscopy, we analyzed the appearance of distinctive folds or “wrinkles” that form as a result of lysogeny broth content reduction in the solid agar growth media. When subjected to varying nutritional conditions, the channel-like folds were shown to alter their morphology; growth on nutrient-depleted media was found to trigger the formation of large and straight wrinkles connecting the colony core to its periphery. To test a possible functional role of the formed channels, a fluorescent analogue of glucose was used to demonstrate preferential native uptake of the molecules into the channels’ interiors which supports their possible role in the transport of molecules throughout biofilm structures.


2014 ◽  
Vol 4 ◽  
pp. 1-9 ◽  
Author(s):  
Antigoni Nikolaki ◽  
Anastasia Papadioti ◽  
Katerina Arvaniti ◽  
Eleni Kassotaki ◽  
Julian D. Langer ◽  
...  
Keyword(s):  

2017 ◽  
Author(s):  
Savita Chib ◽  
Farhan Ali ◽  
Aswin Sai Narain Seshasayee

AbstractProlonged stationary-phase is an approximation of natural environments presenting a range of stresses. Survival in prolonged stationary-phase requires alternative metabolic pathways for survival. This study describes the repertoire of mutations accumulating in starvingE. colipopulations in lysogeny broth. A wide range of mutations accumulate over the course of one month in stationary-phase. SNPs constitute 64% of all mutations. A majority of these mutations are non-synonymous and are located at conserved loci. There is an increase in genetic diversity in the evolving populations over time. Computer simulations of evolution in stationary phase suggest that the maximum frequency obtained by mutations in our experimental populations can not be explained by neutral drift. Moreover there is frequent genetic parallelism across populations suggesting that these mutations are under positive selection. Finally functional analysis of mutations suggests that regulatory mutations are frequent targets of selection.


Author(s):  
EDITHA RENESTEEN ◽  
FURQON DWI CAHYO ◽  
AMARILA MALIK

Objective: Sucrose phosphorylase (SPase) is an enzyme that catalyzes the transfer of glucosyl to various acceptor molecules. Distinct types of SPaseshave been reported, and their transglycosylase activities have been shown to differ. In general, glycosylation is a process that is used to modifybioactive compounds. As such, glycosylation can increase the chemical stability of compounds and improve their characteristics such as reduce strongsmell and sour taste. We previously cloned recombinant SPase (SPaseWRS-3[1]) from Leuconostoc mesenteroides MBFWRS-3[1] in Escherichia coli.In the current study, we aimed to characterize SPaseWRS-3 and determine its transglycosylation activity using benzoic acid (BA), ascorbic acid, andkojic acid (KA).Methods: Expression analyses were conducted in lysogeny broth (LB) medium supplemented with tetracycline and expression was induced usingisopropyl-β-d-thiogalactopyranoside. The characteristics of the 56 kDa recombinant SPase (rec-SPase) were confirmed using sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE). Rec-SPase activity was determined spectrophotometrically using sucrose as the substrate and NADPHas the end-product at 340 nm. Transglycosylation activity was evaluated using thin-layer chromatography (TLC) on silica gel plates.Results: Our results demonstrated that the rec-SPase had an activity of 98.52% relative to the reference SPase (ref-SPase). BA and KA were determinedto undergo glucosyl transfer by rec-SPase using ref-SPase, as observed with TLC. Our findings are consistent with those reported previously for theSPase isolated from L. mesenteroides.Conclusion: Recombinant SPase activity is comparable to reference SPase activity. Our study could be the initial study to deeply observe SPase activityin other substrates as well.


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