scholarly journals Aspartate-90 and arginine-269 of hamster aspartate transcarbamylase affect the oligomeric state of a chimaeric protein with an Escherichia coli maltose-binding domain

1998 ◽  
Vol 329 (2) ◽  
pp. 243-247 ◽  
Author(s):  
Yu QIU ◽  
N. Jeffrey DAVIDSON
Keyword(s):  
Escherichia Coli ◽  
Structural Integrity ◽  
Quaternary Structure ◽  
Binding Domain ◽  
Carbamoyl Phosphate ◽  
Oligomeric State ◽  
Aspartate Transcarbamylase ◽  
Catalytic Subunits ◽  
E Coli ◽  
Kinetics Of

Residues Asp-90 and Arg-269 of Escherichia coli aspartate transcarbamylase seem to interact at the interface of adjacent catalytic subunits. Alanine substitutions at the analogous positions in the hamster aspartate transcarbamylase of a chimaeric protein carrying an E. coli maltose-binding domain lead to changes in both the kinetics of the enzyme and the quaternary structure of the protein. The Vmax for the Asp-90 → Ala and Arg-269 → Ala substitutions is decreased to 1/21 and 1/50 respectively, the [S]0.5 for aspartate is increased 540-fold and 826-fold respectively, and the [S]0.5 for carbamoyl phosphate is increased 60-fold for both. These substitutions decrease the oligomeric size of the protein. Whereas the native chimaeric protein behaves as a pentamer, the Asp-90 variant is a trimer and the Arg-269 variant is a dimer. The altered enzymes also exhibit marked decreases in thermal stability and are inactivated at much lower concentrations of urea than is the unaltered enzyme. Taken together, these results are consistent with the hypothesis that both Asp-90 and Arg-269 have a role in the enzymic function and structural integrity of hamster aspartate transcarbamylase.

Inactivation of Escherichia coli by Ultrasound Combined with Nisin

2018 ◽  
Vol 81 (6) ◽  
pp. 993-1000 ◽  
Author(s):  
ZUWEN WANG ◽  
XIUFANG BI ◽  
RUI XIANG ◽  
LIYI CHEN ◽  
XIAOPING FENG ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Synergistic Effect ◽  
Linear Models ◽  
Treatment Time ◽  
Nutrient Broth ◽  
Inactivation Kinetics ◽  
Growth Phases ◽  
E Coli ◽  
Kinetics Of ◽  
Inactivation Effect

ABSTRACT The aim of this study was to investigate the inactivation of nonpathogenic Escherichia coli in nutrient broth and milk through the use of either ultrasound (US) alone or US combined with nisin (US + nisin) treatments. The E. coli cells were treated at 0 to 55°C, 242.04 to 968.16 W/cm2 for 0 to 15 min. The results showed that the inactivation of E. coli by US and US + nisin increased when the temperature, US power density, and treatment time were increased. The inactivation kinetics of E. coli in nutrient broth by US and US + nisin both conformed to linear models. The largest reductions of 2.89 and 2.93 log cycles by US and US + nisin, respectively, were achieved at 968.16 W/cm2 and at 25°C for 15 min. The suspension media of the E. coli cells influenced the inactivation effect of US, while the growth phases of E. coli cells did not affect their resistance to US. Under all experiment conditions of this study, the differences between US and US + nisin in their respective inactivation effects on E. coli were not obvious. The results suggested that nisin had either no effect at all or a weak synergistic effect with US and that the E. coli cells were inactivated mainly by US, thus indicating that the inactivation of E. coli by US is an “all or nothing” event.

Kinetics of the Quaternary Structure Change of Aspartate Transcarbamylase Triggered by Succinate, a Competitive Inhibitor

Biochemistry ◽  
1994 ◽  
Vol 33 (33) ◽  
pp. 10007-10012 ◽  
Author(s):  
Hirotsugu Tsuruta ◽  
Patrice Vachette ◽  
Takayuki Sano ◽  
Michael F. Moody ◽  
Yoshiyuki Amemiya ◽  
...  
Keyword(s):  
Quaternary Structure ◽  
Competitive Inhibitor ◽  
Structure Change ◽  
Aspartate Transcarbamylase ◽  
Kinetics Of

scholarly journals Bacterial protein translocation requires only one copy of the SecY complex in vivo

2012 ◽  
Vol 198 (5) ◽  
pp. 881-893 ◽  
Author(s):  
Eunyong Park ◽  
Tom A. Rapoport
Keyword(s):  
Escherichia Coli ◽  
Small Molecules ◽  
Protein Translocation ◽  
Cross Linking ◽  
Bacterial Protein ◽  
Oligomeric State ◽  
E Coli ◽  
Cotranslational Translocation ◽  
Escherichia Coli Cells

The transport of proteins across the plasma membrane in bacteria requires a channel formed from the SecY complex, which cooperates with either a translating ribosome in cotranslational translocation or the SecA ATPase in post-translational translocation. Whether translocation requires oligomers of the SecY complex is an important but controversial issue: it determines channel size, how the permeation of small molecules is prevented, and how the channel interacts with the ribosome and SecA. Here, we probe in vivo the oligomeric state of SecY by cross-linking, using defined co- and post-translational translocation intermediates in intact Escherichia coli cells. We show that nontranslocating SecY associated transiently through different interaction surfaces with other SecY molecules inside the membrane. These interactions were significantly reduced when a translocating polypeptide inserted into the SecY channel co- or post-translationally. Mutations that abolish the interaction between SecY molecules still supported viability of E. coli. These results show that a single SecY molecule is sufficient for protein translocation.

Viability of Salmonella, Escherichia coli O157:H7, and Listeria monocytogenes in Yellow Fat Spreads as Affected by Storage Temperature

2003 ◽  
Vol 66 (4) ◽  
pp. 549-558 ◽  
Author(s):  
SARAH L. HOLLIDAY ◽  
LARRY R. BEUCHAT
Keyword(s):  
Escherichia Coli ◽  
Listeria Monocytogenes ◽  
Storage Temperature ◽  
Salt Content ◽  
Inactivation Kinetics ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Inhibition Of Growth ◽  
Time Required ◽  
Kinetics Of

A study was conducted to characterize the survival and inactivation kinetics of a five-serotype mixture of Salmonella (6.23 to 6.55 log10 CFU per 3.5-ml or 4-g sample), a five-strain mixture of Escherichia coli O157:H7 (5.36 to 6.14 log10 CFU per 3.5-ml or 4-g sample), and a six-strain mixture of Listeria monocytogenes (5.91 to 6.18 log10 CFU per 3.5-ml or 4-g sample) inoculated into seven yellow fat spreads (one margarine, one butter-margarine blend, and five dairy and nondairy spreads and toppings) after formulation and processing and stored at 4.4, 10, and 21°C for up to 94 days. Neither Salmonella nor E. coli O157:H7 grew in any of the test products. The time required for the elimination of each pathogen depended on the product and the storage temperature. Death was more rapid at 21°C than at 4.4 or 10°C. Depending on the product, the time required for the elimination of viable cells at 21°C ranged from 5 to 7 days to >94 days for Salmonella, from 3 to 5 days to 28 to 42 days for E. coli O157:H7, and from 10 to 14 days to >94 days for L. monocytogenes. Death was most rapid in a water-continuous spray product (pH 3.66, 4.12% salt) and least rapid in a butter-margarine blend (pH 6.66, 1.88% salt). E. coli O157:H7 died more rapidly than did Salmonella or L. monocytogenes regardless of storage temperature. Salmonella survived longer in high-fat (≥61%) products than in products with lower fat contents. The inhibition of growth is attributed to factors such as acidic pH, salt content, the presence of preservatives, emulsion characteristics, and nutrient deprivation. L. monocytogenes did not grow in six of the test products, but its population increased between 42 and 63 days in a butter-margarine blend stored at 10°C and between 3 and 7 days when the blend was stored at 21°C. On the basis of the experimental parameters examined in this study, traditional margarine and spreads not containing butter are not “potentially hazardous foods” in that they do not support the growth of Salmonella, E. coli O157:H7, or L. monocytogenes.

scholarly journals Environmental inactivation and irrigation-mediated regrowth of Escherichia coli O157:H7 on romaine lettuce when inoculated in a fecal slurry matrix

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e6591 ◽  
Author(s):  
Jennifer A. Chase ◽  
Melissa L. Partyka ◽  
Ronald F. Bond ◽  
Edward R. Atwill
Keyword(s):  
Escherichia Coli ◽  
Field Trials ◽  
Inactivation Rate ◽  
Bacterial Inactivation ◽  
Romaine Lettuce ◽  
E Coli ◽  
Log Reduction ◽  
Salinas Valley ◽  
Kinetics Of ◽  
Insight Into

Field trials were conducted in July–August and October 2012 to quantify the inactivation rate of Escherichia coli O157:H7 when mixed with fecal slurry and applied to romaine lettuce leaves. Lettuce was grown under commercial conditions in Salinas Valley, California. One-half milliliter of rabbit, chicken, or pig fecal slurry, containing an average of 4.05 × 107 CFU E. coli O157:H7 (C0), was inoculated onto the upper (adaxial) surface of a lower leaf on 288 heads of lettuce per trial immediately following a 2.5 h irrigation event. To estimate the bacterial inactivation rate as a function of time, fecal matrix, irrigation and seasonal climate effects, sets of lettuce heads (n = 28) were sampled each day over 10 days and the concentration of E. coli O157:H7 (Ct) determined. E. coli O157:H7 was detected on 100% of heads during the 10-day duration, with concentrations ranging from ≤340 MPN/head (∼5-log reduction) to >3.45 × 1012 MPN/head (∼5-log growth). Relative to C0, on day 10 (Ct = 12) we observed an overall 2.6-log and 3.2-log mean reduction of E. coli O157:H7 in July and October, respectively. However, we observed relative maximum concentrations due to bacterial growth on day 6 (maximum Ct = 8) apparently stimulated by foliar irrigation on day 5. From this maximum there was a mean 5.3-log and 5.1-log reduction by day 10 (Ct = 12) for the July and October trials, respectively. This study provides insight into the inactivation and growth kinetics of E. coli O157:H7 on romaine lettuce leaves under natural field conditions. This study provides evidence that harvesting within 24 h post irrigation has the potential to increase the concentration of E. coli O157:H7 contamination, if present on heads of romaine lettuce; foliar irrigation can temporarily stimulate substantial regrowth of E. coli O157:H7.

scholarly journals Escherichia coli Survival in, and Release from, White-Tailed Deer Feces

2014 ◽  
Vol 81 (3) ◽  
pp. 1168-1176 ◽  
Author(s):  
Andrey K. Guber ◽  
Jessica Fry ◽  
Rebecca L. Ives ◽  
Joan B. Rose
Keyword(s):  
Escherichia Coli ◽  
Fate And Transport ◽  
Exponential Model ◽  
Temperature Correction ◽  
Content Type ◽  
E Coli ◽  
Naturally Occurring ◽  
Different Temperatures ◽  
Kinetics Of ◽  
Source Materials

ABSTRACTWhite-tailed deer are an important reservoir for pathogens that can contribute a large portion of microbial pollution in fragmented agricultural and forest landscapes. The scarcity of experimental data on survival of microorganisms in and release from deer feces makes prediction of their fate and transport less reliable and development of efficient strategies for environment protection more difficult. The goal of this study was to estimate parameters for modelingEscherichia colisurvival in and release from deer (Odocoileus virginianus) feces. Our objectives were as follows: (i) to measure survival ofE. coliin deer pellets at different temperatures, (ii) to measure kinetics ofE. colirelease from deer pellets at different rainfall intensities, and (iii) to estimate parameters of models describing survival and release of microorganisms from deer feces. Laboratory experiments were conducted to studyE. colisurvival in deer pellets at three temperatures and to estimate parameters of Chick's exponential model with temperature correction based on the Arrhenius equation. Kinetics ofE. colirelease from deer pellets were measured at two rainfall intensities and used to derive the parameters of Bradford-Schijven model of bacterial release. The results showed that parameters of the survival and release models obtained forE. coliin this study substantially differed from those obtained by using other source materials, e.g., feces of domestic animals and manures. This emphasizes the necessity of comprehensive studies of survival of naturally occurring populations of microorganisms in and release from wildlife animal feces in order to achieve better predictions of microbial fate and transport in fragmented agricultural and forest landscapes.

scholarly journals Engineering a trifunctional proline utilization A chimaera by fusing a DNA-binding domain to a bifunctional PutA

2016 ◽  
Vol 36 (6) ◽  
Author(s):  
Benjamin W. Arentson ◽  
Erin L. Hayes ◽  
Weidong Zhu ◽  
Harkewal Singh ◽  
John J. Tanner ◽  
...  
Keyword(s):  
Dna Binding ◽  
Modular Design ◽  
Quaternary Structure ◽  
Lipid Binding ◽  
Binding Domain ◽  
Dna Binding Domain ◽  
Oligomeric State ◽  
Proline Utilization ◽  
Functional Switching ◽  
Proline Utilization A

Proline utilization A (PutA) is a bifunctional flavoenzyme with proline dehydrogenase (PRODH) and Δ1-pyrroline-5-carboxylate (P5C) dehydrogenase (P5CDH) domains that catalyses the two-step oxidation of proline to glutamate. Trifunctional PutAs also have an N-terminal ribbon–helix–helix (RHH) DNA-binding domain and moonlight as autogenous transcriptional repressors of the put regulon. A unique property of trifunctional PutA is the ability to switch functions from DNA-bound repressor to membrane-associated enzyme in response to cellular nutritional needs and proline availability. In the present study, we attempt to construct a trifunctional PutA by fusing the RHH domain of Escherichia coli PutA (EcRHH) to the bifunctional Rhodobacter capsulatus PutA (RcPutA) in order to explore the modular design of functional switching in trifunctional PutAs. The EcRHH–RcPutA chimaera retains the catalytic properties of RcPutA while acquiring the oligomeric state, quaternary structure and DNA-binding properties of EcPutA. Furthermore, the EcRHH–RcPutA chimaera exhibits proline-induced lipid association, which is a fundamental characteristic of functional switching. Unexpectedly, RcPutA lipid binding is also activated by proline, which shows for the first time that bifunctional PutAs exhibit a limited form of functional switching. Altogether, these results suggest that the C-terminal domain (CTD), which is conserved by trifunctional PutAs and certain bifunctional PutAs, is essential for functional switching in trifunctional PutAs.

scholarly journals Disinfection of Escherichia coli and Pseudomonas aeruginosa by copper in water

2016 ◽  
Vol 14 (3) ◽  
pp. 424-432 ◽  
Author(s):  
Andrew M. Armstrong ◽  
Mark D. Sobsey ◽  
Lisa M. Casanova
Keyword(s):  
Escherichia Coli ◽  
Pseudomonas Aeruginosa ◽  
Enteric Bacteria ◽  
Inactivation Kinetics ◽  
Log10 Reduction ◽  
E Coli ◽  
Piped Water ◽  
Ionic Copper ◽  
Household Water ◽  
Kinetics Of

When households lack access to continuous piped water, water storage in the home creates opportunities for contamination. Storage in copper vessels has been shown to reduce microbes, but inactivation kinetics of enteric bacteria in water by copper alone needs to be understood. This work characterized inactivation kinetics of Escherichia coli and Pseudomonas aeruginosa by dissolved ionic copper in water. Reductions of E. coli and P. aeruginosa increase with increasing dose. At 0.3 mg/L, there was a 2.5 log10 reduction of E. coli within 6 hours. At 1 and 3 mg/L, the detection limit was reached between 3 and 6 hours; maximum reduction measured was 8.5 log10. For P. aeruginosa, at 6 hours there was 1 log10 reduction at 0.3 mg/L, 3.0 log10 at 1 mg/L, and 3.6 log10 at 3 mg/L. There was no significant decline in copper concentration. Copper inactivates bacteria under controlled conditions at doses between 0.3 and 1 mg/L. E. coli was inactivated more rapidly than P. aeruginosa. Copper at 1 mg/L can achieve 99.9% inactivation of P. aeruginosa and 99.9999997% inactivation of E. coli over 6 hours, making it a candidate treatment for stored household water.

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