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排序方式: 共有3195条查询结果,搜索用时 31 毫秒
31.
The effects of five polyethylene glycol (PEG) compounds of different molecular weight on the thermal stability of penicillin
G acylase (PGA) obtained from a mutant ofEscherichia coli ATCC 11105 have been investigated. The molecular weights of PEG compounds were 400, 4000, 6000, 10,000, and 15,000. The thermal
inactivation mechanisms of both native and PEG-containing PGA were considered to obey first order inactivation kinetics during
prolonged heat treatments. Optimal concentrations of PEGs at molecular weights of 400,4000, 6000,10,000, and 15,000 were found
to be 250,150,150,100, and 50 mM, respectively. The greatest enhancement of thermostability was observed with PEG 4000 and
PEG 6000, as a nearly 20-fold increase above 50°C. PGA showed almost the same temperature activity profile and optimal temperature
values both in the presence and absence of PEG. The addition of PEGs did not cause any change in the optimal temperature value
of PGA, but the parametersV
m
,K
m
, the activation energy, and thek
cat
values of enzyme were markedly decreased because of the mixed inhibition by PEG compounds. The type of inhibition was found
to be hyperbolic uncompetitive. 相似文献
32.
An optical biosensor for the determination of hydrogen peroxide based on immobilized horseradish peroxidase is described. The fluorescence of the dimeric product of the enzyme catalysed oxidation of homovanillic acid is utilized to determine the concentration of H2O2. The membrane-bound enzyme is attached to a bifurcated fibre bundle permitting excitation and detection of the fluorescence by a fluorometer. The response of the sensor is linear from 1 to 130 M hydrogen peroxide; the coefficient of variation is 3%. The sensor is stable for more than 10 weeks. The operating pH for maximal sensor response is 8.15. This allows the sensor to be used in combination with oxidase reactions producing hydrogen peroxide, as is demonstrated with a co-immobilized lactate oxidase-horseradish peroxidase optode for the determination of L-lactate. The fluorescence intensity of this sensor depends linearly on the concentration of lactate between 3 and 200 M and a throughput of 10 samples per hour is possible. The precision is in the same range as that of the monoenzyme optode. The lifetime of the bienzyme sensor for lactate is considerably shorter than that of the peroxidase sensor; it is limited by the stability of the immobilized lactate oxidase enzyme. The sensor has been applied to the determination of lactate in control serum. 相似文献
33.
34.
The exact residues within severe acute respiratory syndrome coronavirus (SARS-CoV) S1 protein and its receptor, human ACE2, involved in their interaction still remain largely undetermined. Identification of exact amino acid residues that are crucial for the interaction of S1 with ACE2 could provide working hypotheses for experimental studies and might be helpful for the development of antiviral inhibitor. In this paper, a molecular docking model of SARS-CoV S1 protein in complex with human ACE2 was constructed. The interacting residue pairs within this complex model and their contact types were also identified. Our model, supported by significant biochemical evidence, suggested receptor-binding residues were concentrated in two segments of S1 protein. In contrast, the interfacial residues in ACE2, though close to each other in tertiary structure, were found to be widely scattered in the primary sequence. In particular, the S1 residue ARG453 and ACE2 residue LYS341 might be the key residues in the complex formation. 相似文献
35.
以交联聚乙烯醇为载体的离子交换剂对蛋白质的分离性能 总被引:2,自引:0,他引:2
本文对阴离子交换剂DEA—PVT常压液相离子交换色谱分离蛋白质的性能、分离条件进行了探讨。结果表明其对蛋白质的分离性能良好,容易洗脱。与载体交联聚乙烯醇相比,DEA—PVT对蛋白质的非特异性吸附明显降低。 相似文献
36.
Surface plasmon resonance (SPR) has been successfully applied for the simple, rapid, and label-free assay of various biomolecules. This assay evaluates a novel wavelength modulation SPR biosensor for the detection of tetanus toxin. The wavelength modulation SPR biosensor is designed based on fixing the incident angle of light and measuring the reflected intensities in the resonance wavelength range spanning 400-800 nm simultaneously. Tetanus toxin (TeNT), one of the most potent toxins known, is synthesized as a 150 kDa single polypeptide chain. The SPR biosensor has been shown to be capable of directly detecting concentration of tetanus toxin as low as 0.028 Lf ml−1. Under selected experimental conditions, the SPR biosensor has a good reproducibility, sensitivity and reversibility. The results illustrate how wavelength modulation SPR biosensor can be used to detect biomolecular interactions. 相似文献
37.
J. Sereikaité D. Iljasevičiené G. Dienys H. Danilčenko V. Gavrilova 《Applied biochemistry and biotechnology》1993,43(2):153-160
Ascorbate oxidase fromCucurbita sp. was isolated by ammonium sulfate precipitation and DEAE-dextran-silochrome column chromatography. The thermal and pH stabilities
of the purified enzyme were investigated. TheK
M forl-ascorbic acid (1.5 mM) and chlorohydroquinone (0.37 mM) was determined. Substrate specificity of ascorbate oxidase was investigated and compared with those of laccases fromCoriolus hirsutus andCerrena maxima. Ascorbate oxidase was covalently bound to a polymeric membrane and used in an enzyme electrode for ascorbic acid. 相似文献
38.
A rapid microtiter plate assay for the detection of inhibitors of the Na+, K+-ATPase has been developed. The assay is based on the measurement of inorganic phosphate released from the substrate, ATP,
and has been designed to be carried out in the individual wells of a microtiter plate. Since the production of inorganic phosphate
is determined colorimetrically, multiple samples can be tested simultaneously using a microtiter plate reader. This microtiter
plate assay is particularly useful for screening large numbers of samples, such as microbial culture supernatants. 相似文献
39.
Toshiyuki Chikuma Yoko ShimabukuroTakeya Iguchi Akira TanakaKyoji Taguchi Takeshi KatoMitsune Yamaguchi Hiroshi Hojo 《Analytica chimica acta》2002,457(2):157-164
A rapid and sensitive assay for the determination of Dns-His-Lys-Arg-His-Lys cleaving enzyme activity is reported. This assay is based on fluorimetric detection of a dansylated dipeptide, 5-dimethylaminonaphthalene-1-sulfonyl-His-Lys, enzymatically formed from the substrate 5-dimethylaminonaphthalene-1-sulfonyl-His-Lys-Arg-His-d-Lys, after separation by high-performance liquid chromatography (HPLC) using a C-18 reversed-phase column by isocratic elution. This assay is sensitive enough to measure 5-dimethylaminonaphthalene-1-sulfonyl-His-Lys at concentrations as low as 7 pmol, and yields highly reproducible results and requires less than 9.0 min per sample for separation and quantitation. The optimum pH for Dns-His-Lys-Arg-His-Lys cleaving enzyme activity was 7.5-8.0. The Michaelis constant (Km) and the maximum velocity (Vmax) values were 33.3 μM and 47.07 pmol/(μg h), respectively with the use of enzyme extract obtained from bovine pituitary. By using this assay, axonal transport of this enzyme activity was observed 48 h after double ligations of rat sciatic nerves. The high sensitivity and selectivity of this assay would be useful for clarification of the physiological role of this enzyme. 相似文献
40.
InteractionbetweenMetalinMetalloEnzymeandSmallBiologicalMolecules¥HuJie-Han;ShuZan-Yong;TaoLi-Mei;ChengGuo-Bao(DalianInstitut... 相似文献