142,472 research outputs found
Heterologous expression and site-directed mutagenesis of soluble methane monoxygenase
No full textThe purpose of this investigation was to study the heterologous expression of
soluble methane monooxygenase (sMMO) genes from Methylococcus capsulatus
(Bath) and Methylosinus trichosporium OB3b. Using the T7-RNA polymerase
expression system, the entire sMMO operon and subclones (constructed using the
polymerase chain reaction) were over-expressed in E. coli. Results obtained using the
Me. capsulatus (Bath) sMMO operon confirmed previous reports (C. West, G. P. C.
Salmond, H. Dalton and 1. C. Murrell (1992). J Gen. Microbial. 138, 1301-1307)
that functional expression of protein B and the reductase occurred but the hydroxylase
was inactive. Similar results were obtained by expressing the sMMO operon of Ms.
trichosporium OB3b in E. coli, using plasmids previously described (D. Jahng and T.
K. Wood (1994). Appl. Environ. Microbiol. 60, 2473-2482). Protein B, the reductase
and orfY were over-expressed and purified from E. coli using glutathione-Stransferase
fusion proteins and affinity chromatography.
The expression of sMMO genes from Mc. capsulatus (Bath) and Ms.
trichosporium OB3b was studied in Pseudomonas putida. A previous report (D.
Jahng and T. K. Wood (1994). Appl. Environ. Microbial. 60, 2473-2482) had
suggested that functional expression of sMMO from Ms. trichosporium OB3b was
achieved in P. putida Fl. Attempts to repeat this work proved that protein B and the
reductase were functionally expressed, but the hydroxylase was inactive. Similar
results were obtained for the heterologous expression of the sMMO operon from Mc.
capsulatus (Bath) in P. putida.
Methanotrophs were used for the heterologous expression of sMMO via two
strategies. (1) The expression of sMMO from Mc. capsulatus (Bath) and Ms.
trichosporium OB3b was studied in Methylomonas album B08 and Methylocystis
parvus OBBP. These are methanotrophs that do not express sMMO, but express
particulate MMO (pMMO) only, to utilise methane as a sole carbon and energy
source. Functional expression of the sMMO operon of Ms. trichosporium OB3b was
achieved in Mm. album BG8, however, recombinant sMMO enzyme activity was poor
and problems were encountered with the growth of the sMMO positive transconjugant
methanotrophs. (2) sMMO-minus marker exchange mutants of Ms. trichosporium
OB3b (H. Martin and 1. C. Murrell (1995). FEMS Microbiol. Letts. 127, 243-248)
were complemented with plasmid encoded genes and functional sMMO expression
was obtained. Southern hybridisation analysis revealed that the plasmid DNA had
integrated into the chromosome of the Ms. trichosporium OB3b sMMO-minus mutant
via a single homologous recombination event between the mmoX genes.
Protein B from Mc. capsulatus (Bath) is inactivated by proteolysis to give rise
to a truncated form designated protein B'. The Met 12-Gly 13 cleavage site was
modified by site-directed mutagenesis to Met12-Gln13 which improved the stability
of the protein when incubated at room temperature. Only after prolonged incubation
was protein B' formed. Recombinant protein B from Ms. trichosporium OB3b also
appears to be unstable, and readily degraded when incubated at room temperature.
The cleavage of protein B to inactive protein B' may be a general regulatory
mechanism that occurs within the cell to regulate sMMO activity
The particulate methane monooxygenase from Methylococcus capsulatus (Bath)
No full textThe isolation procedure for the pMMO complex has been optimised to obtain a high
specific activity enzyme from Methylococcus capsulatus (Bath). The enzyme is
comprised of the pMMO hydroxylase (pMMOH) consisting of polypeptides 47,26 and
23kDa molecular mass. In addition to this, a putative pMMO reductase (pMMOR) was
also found to be necessary to maintain propylene oxidising activity. This component
was found to consist of two polypeptides of approximately 63 and 8kDa. Preliminary Nterminal
sequence data of the large subunit ofpMMOR indicates that the sequence bears
70% similarity to the methanol dehydrogenase (MDH) from Methylococcus capsulatus
(Bath). Therefore, we tentatively propose that the" MDH can act as a reductase
component to the pMMOH.
The significance of this result prompted investigations into the previous published
proposals that electrons derived from the methanol oxidation reaction can be channelled
back into the methane oxidation reaction by the methanol dehydrogenase, independent
of NADH. Any effect of methanol to act as a reductant to pMMO in membrane
preparations was lost upon isolation of the pMMO complex, indicating the necessity to
maintain a fully functional methanol dehydrogenase (MDH) upon isolation. In addition
to this, the in vitro electron donors of pMMO, NADH and duroquinol were found to act
via distinct pathways of electron transfer (electron transport inhibitor studies).
Electron paramagnetic resonance (EPR) spectroscopy data provided evidence that the
copper in the active site of pMMO existed as a mononuclear copper (II) centre not a
trinuclear copper centre suggested by Chan and coworkers (Chan et al., 1993; Nguyen et
al., 1994, 1996a, 1996b, 1998). In addition to this preliminary data also indicates the
presence of an iron centre which is only EPR visible after reduction of the complex
suggesting the majority of iron in the complex is EPR invisible. The exact nature of this
iron centre is still unclear.
A structural study of the pMMO complex has also been undertaken using electron
microscopy studies in conjunction with single particle analysis. This allowed low
resolution projection maps of different views of the pMMO complex to be generated.
The complex appears to exist in a polymeric state of at least a dimer, possibly a tetramer
if the molecular weight analysis calculated by sedimentation equilibrium analysis is
taken into account.
This study has provided some insight into basic characteristics and the structure of a
duroquinol-driven pMMO complex and its interaction with other electron transfer
proteins
Dataset for "Pioneering Net Zero Carbon Construction Policy in Bath & North East Somerset"
No full textThis data was collected as part of a collaborative project between the University of Bath and Bath and North East Somerset Council, which studied the implementation of new sustainable construction planning policies, the first of their kind in the UK to require that: • All new residential and major non-residential building developments must achieve net zero operational energy, conforming to ambitious energy consumption targets, matching this with on-site renewables, and only offsetting the difference in exceptional circumstances. • Major residential and non-residential developments must demonstrate an embodied carbon lower than a threshold value, including the substructure, superstructure and finishes, with no offsetting permitted. The data includes analysis of incoming planning application, relating to the characteristics of proposed buildings and key parameters submitted to comply with the net zero energy requirements. A questionnaire was also sent out to applicants, with both written and numerical responses included in this dataset
Dataset for "Pioneering Net Zero Carbon Construction Policy in Bath & North East Somerset: Evaluating the effectiveness of novel planning policies over time"
No full textThis data was collected as part of a continuing collaboration between the University of Bath and Bath and North East Somerset Council, exploring the impacts of (and reception to) pioneering sustainable planning policies for new buildings which were first introduced in January 2023. This project evaluates the success of the policies two years on, establishing long-term trends, opportunities for refinement, and the national policy implications of this unique policy case study. The deposited data relates to two parts of the methodology. The first is an analysis of incoming planning application, relating to the characteristics of proposed buildings and key parameters submitted to comply with the net zero energy requirements. The second is the results of a questionnaire sent out to applicants
bath n
No full textbath (n)Then we'd put 'em in bath,see,an' bath 'em for a couple o' hours after that,you know,an' preserve 'em.... (receptacle in which cans of lobster or salmon are boiled to preserve the meat/fish)YesDNE-citPhrase is: _to put in bath_J. D. A. WIDDOWSON AUG 1975Used IUsed INot use
Characterisation of a putative N-terminal GLUT4 kinase
No full textSIGLEAvailable from British Library Document Supply Centre- DSC:DXN059680 / BLDSC - British Library Document Supply CentreGBUnited Kingdo
bath n
No full textbath (n)The bath is boilin' water,you know,fresh water,an'...and boil 'em for about _ we'd give 'em two baths. We'd _ we'd give 'em one bath, a hour, a hour an' a quarter, an' then we'd blow 'em off, let the wind out of 'em an' stop 'em again. Put 'em in again an' boil 'em an' take 'em out... (receptacle in which cans of lobster or salmon are boiled)receptacle in which cans of lobster or salmon areYesDNE-citJ. D. A. WIDDOWSON AUG 1975Used IUsed IUsed ISource appears in DNE as: T 455/6-67
Properties of Al-doped ZnS films grown by chemical bath deposition
No full textZinc sulphide (ZnS) buffer layers are a cadmium free, wider energy band gap, alternative to the cadmium sulphide(CdS) buffer layers commonly used in copper indium gallium diselenide (CuInGaSe2)-based solar cells. However extrinsic doping of the ZnS is important to lower the resistivity of the layers and to improve flexibility of device design. In this work, Al-doped ZnS nanocrystalline films have been produced on glass substrates using a chemical bath deposition (CBD) method. The Al- concentration was varied from 0 at. % to 10 at. %, keeping other deposition parameters constant. The elemental composition of a typical sample with 6 at. % ‘Al’ in ZnS was Zn=44.9 at. %, S=49.8 at. % and Al=5.3 at.%. The X-ray diffraction data taken on these samples showed a broad peak corresponding to the (111) plane of ZnS while the crystallite size varied in the range, 8 – 15 nm, depending on the concentration of Al in the layers. The films with a Al-doping content of 6 at. % had an optical transmittance of 75 % in the visible range and the energy band gap evaluated from the data was 3.66 eV. The films n-type electrical conductivities and the electrical resistivity varied in the range, 107-103 Ωcm, it decreasing with an increase of the Al-concentration in the solution
Late Gothic architecture in South West England : four major centres of building activity at Wells, Bristol, Sherbourne and Bath
No full textBy 1360 the Perpendicular style was established as the successor to Decorated
architecture. During the subsequent one hundred and eighty years, until the Reformation,
major building work was carried out at four great churches in the south west of England.
The complete reconstructions of St Mary Redcliffe, Sherborne Abbey and Bath Abbey, and
considerable work to the precinct at Wells Cathedral during this period, form the basis for
this thesis. Through a study of each of these major centres, the issues of workshop identity
and stylistic trendsetters are considered.
It is shown how the interpretation of documentary evidence has impeded an
understanding of these buildings, which can be revealed by an analysis of the fabric. Based
primarily on a methodology of buildings archaeology and assessment of moulding profiles,
traditional assumptions concerning the chronology and patronage are challenged.
The new chronology for works at Sherborne Abbey, and the redating of the
commencement of Bath Abbey further our understanding of the nature of masons'
workshops, patronage and stylistic development within a regional context. Introspection in
masons' workshops during the 15th century, and retrospection in later design in the region,
demonstrates a reliance on the innovations of the 14th century, and the significance of the
parish church tradition in the region, respectively. The thesis concludes with a discussion on
the influence of major church workshops on domestic architecture, and the impact of the
dissemination of the lodges in the early 16th century
Dataset for “Ketogenic diet but not free-sugar restriction alters glucose tolerance, lipid metabolism, peripheral tissue phenotype, and gut microbiome: RCT”
No full textThe dataset includes demographic data of participants (n=60) randomised to one of three groups: 1. CONTROL (moderate sugar) - reflecting the composition of a typical European diet; 2. Low sugar - the same composition of a typical European diet but with <5% energy intake from sugar; 3. Low carbohydrate - low carbohydrate diet with <5% energy intake from sugar, replacing carbohydrate energy with fat. All laboratory trials took place at the University of Bath. Participants consumed these diets for period of 12 weeks, with laboratory visits at baseline, at week 4, and at week 12. The types of data that were collected during the trials are as follows: - Height; - Body mass; - Body composition (by Dual-energy x-ray absorptiometry); - Resting metabolic rate; - Postprandial substrate oxidation; - Exercise metabolism; - Physical activity energy expenditure; - Fasting and postprandial concentrations of circulating metabolites (e.g., glucose, lactate triacylglycerol) and hormones (e.g. insulin, C-peptide); - Nuclear magnetic resonance spectroscopy measures of plasma (e.g., lipoprotein size and particle number); - Skeletal muscle protein levels; - Adipose tissue mRNA levels; - Fasting haematology (e.g., white blood cell count, haematocrit, haemoglobin concentration); - Urinary acetoacetate concentrations; - Self-reported energy intake; - Visual analogue scales for appetite and mood; - Continuous glucose monitoring
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