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1.
Cytochrome P-450 isozymes represent a critical component of nature’s spectrum of detoxification catalysts that could be exploited
for bioremediation. The ethanol-inducible human cytochrome P-450 2E1 serves as a model eukaryotic P-450 that complements the
bacterial P-450 cam in dehalogenation and detoxification of environmental pollutants. We explored the construction of novel
chimeric P-450s using cytochrome P-450 camC and 2E1 genes. For construction of chimera 1 (478 amino acids, 55.14 kDa), 145 amino acids from the N-terminus of P-450 2E1 protein
(493 amino acids, 56.84 kDa) were replaced with 130 amino acids from the N-terminus of P-450 camC protein (415 amino acids,
46.66 kDa). In chimera 2 (525 amino acids, 60.24 kDa) the strategy involves replacement of 28 amino acids in the C-terminus
of chimera 1 with 75 amino acids from the C-terminus of P-450 camC gene. Homology models of both the chimeric proteins were
developed using SWISS-MODEL based on the known crystal structure of cytochrome P-450 camC, BM-3, 1DT6A, and 2C17A. The models
indicated that the proposed heme-binding site was intact, which is inevitable for catalytic activity of cytochrome P-450s.
The expression of chimera 1 and 2 genes in Escherichia coli DH5α was evident from light-pink cell pellets, protein band in sodium dodecyl sulfate polyacrylamide gel electrophoresis,
and diagnostic carbon monoxide-difference spectra. Our studies show that strategies can be developed to exploit the natural
diversity of the P-450 superfamily to generate chimeric biocatalysts that would provide new templates amenable to directed
evolution. 相似文献
2.
A technique is reported for measuring triplet spin-lattice relaxation times in fluid solution. It is based on the observation of chemically induced electron spin polarization, in the presence of a triplet quencher. Spin-lattice relaxation times of the order of 10 ns are reported for the duroquinone triplet in various solvents. 相似文献
3.
Philip A. Wheeler Michael Brungs Graham R. Atkins 《Journal of Sol-Gel Science and Technology》2003,27(3):321-326
Organically modified silicates (ormosils) have been viewed as possible materials for optical signal processing devices for communications networks. One significant problem obstructing their use in this application is the absorption of infrared light by hydroxy species (present for example in silanol groups (SiOH) and water) around the communications wavelengths (1310 and 1550 nm). We have investigated several ormosil compositions, synthesised via an aqueous sol-gel method, to determine if residual hydroxy groups can be removed more effectively via reducing the connectivity of the material by increasing the number of organic groups (such as Si—CH3). Furthermore, we have investigated the effect of different inert atmospheres on the drying process. It was found that reducing the connectivity does promote the removal of hydroxy absorption, mainly via the condensation of residual silanol groups, but also by removal of residual water, depending on the other precursors in the system. It was also found that the drying atmosphere used (nitrogen or helium) had no effect on the drying process. 相似文献
4.
Ram Raghava Robert M. Caddell Lynn Buege Anthony G. Atkins 《Journal of Macromolecular Science: Physics》2013,52(4):655-665
The through thickness total strains of hydrostatically bulged domes of materials are usually determined indirectly. The in-plane circumferential strains are readily measured, and then constancy of volume is invoked to determine the “missing” or unknown strain. Some of our recent work with polymers has shown that volume constancy in large-scale deformation does not hold so that the usual computations of stress and strain during bulging of sheet may be in error. We describe an electrical device that reduces such errors by directly measuring the through thickness (and, thus, the thickness “strain”) for nonmetallic materials. 相似文献
5.
The effect of a magnetic field on the intensity of fluorescence from chemiluminescent reactions occurring in fluid solutions is calculated. The calculation first considers pairs of triplet molecules rotating in each other's vicinity and calculates the effect of a magnetic field on the rate at which the overall singlet state is populated. The field diminishes this rate, as in the solid-state situation. The triplets are then allowed to diffuse apart, and the process of populating the overall singlet state of the pair is treated as a relaxation process occurring during the diffusive trajectory. In this case too at high fields the intensity diminishes. The calculations are repeated for triplet-doublet quenching, and, in accord with the solid-state results, the intensity is enhanced at high fields because the quenching rate for doublet-triplet collisions is diminished, and the resultant fluorescence arises via triplets which escape abortive D-T encounters. Explicit expressions involving the dynamical parameters of the fluid (e.g. the translational and rotational correlation times of the species in the solutions) are given for the fluorescence intensity. 相似文献
6.
Amanda J. Gregory Jonathan P. Atkins Daryl Burdon Michael Elliott 《European Journal of Operational Research》2013
The purpose of this paper is to learn from Complex Adaptive Systems (CAS) theory to inform the development of Problem Structuring Methods (PSMs) both in general and in the specific context of marine management. The focus on marine management is important because it is concerned with a CAS (formed through the interconnection between natural systems, designed systems and social systems) which exemplifies their particularly ‘wicked’ nature. Recognition of this compels us to take seriously the need to develop tools for knowledge elicitation and structuring which meet the demands of CAS. In marine management, chief among those tools is the DPSIR (Drivers – Pressures – State Changes – Impacts – Responses) model and, although widely applied, the extent to which it is appropriate for dealing with the demands of a CAS is questionable. Such questioning is particularly pertinent in the context of the marine environment where there is a need to not only recognise a broad range of stakeholders (a question of boundary critique) but also to manage competing knowledge (economic, local and scientific) and value claims. Hence this paper emphasises how a CAS perspective might add impetus to the development of a critical perspective on DPSIR and PSM theory and practice to promote a more systemic view of decision-making and policy development. 相似文献
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