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Isolation of amylolytic strains of Thermoactinomyces vulgaris and production of thermophilic actinomycete amylases 总被引:6,自引:1,他引:5
Kuo, M. J. (Iowa State University, Ames), and P. A. Hartman. Isolation of amylolytic strains of Thermoactinomyces vulgaris and production of thermophilic actinomycete amylases. J. Bacteriol. 92:723-726. 1966.-Of 759 isolates obtained from dung, compost, and soil samples, a culture of Thermoactinomyces vulgaris (strain 5) was selected for further study on the basis of quantities of amylase produced in synthetic and nonsynthetic media, rapid growth and sporulation, culture stability upon prolonged storage at 5 C, and growth temperature range. Inoculum preparation, temperature optimum for amylase formation, and the effects of various kinds and levels of carbon and nitrogen sources on amylase production were studied with T. vulgaris strain 5. An optimal procedure for production of T. vulgaris amylases is proposed. 相似文献
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The Saccharomyces cerevisiae SOC8-1 gene and its relationship to a nucleotide kinase 总被引:1,自引:0,他引:1
W J Choi J L Campbell C L Kuo A Y Jong 《The Journal of biological chemistry》1989,264(26):15593-15599
The yeast SOC8-1 gene was originally identified by partial complementation of cdc8 mutant strains. We have carried out Bal31 deletion analysis of the SOC8-1 gene to define the minimal size which is required for the complementation of the cdc8 mutation. When the SOC8-1 gene is cloned in a multicopy plasmid, it enables temperature-resistant growth in the cdc8 mutant strain, while the SOC8-1 gene in a single copy plasmid does not. Thus, its suppression of the cdc8 mutant is dosage dependent. The high copy number vector carrying the SOC8-1 gene can complement five different cdc8 alleles, indicating that the suppression is not allele specific. Since CDC8 encodes thymidylate kinase, cells bearing a high copy number plasmid containing SOC8-1 gene were tested for the ability to phosphorylate several nucleoside monophosphates, including UMP, GMP and dTMP. Significantly increased phosphorylation activity was observed, suggesting that SOC8-1 encodes a nucleotide kinase. Both restriction enzyme analysis of the SOC8-1 gene and partial purification of the overproduced kinase in SOC8-1 overproducing strains suggest that SOC8-1 may be allelic with URA6. Consistent with these results, both SOC8-1 and URA6 are located on chromosome XI. Thus, one possible suppression mechanism is that SOC8-1 may provide a trans-acting dTMP kinase activity, bypassing the cdc8 gene defect. 相似文献
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X-ray diffraction analysis on single crystals of recombinant Escherichia coli ornithine transcarbamoylase 总被引:3,自引:0,他引:3
Single crystals of recombinant Escherichia coli ornithine transcarbamoylase suitable for x-ray analysis have been grown from polyethylene glycol and 2-methyl-2,4-pentanediol. The space group has been determined as P3(1) or P3(2), with one protein trimer of three identical 36.8-kDa subunits in the asymmetric unit. The unit cell dimensions are a = b = 105.1 A and c = 87.8 A. The crystals diffract well to 3-A resolution and are quite resistant to radiation damage. Single crystals have also been grown of a genetically engineered site-specific mutant for which the replacement of an arginine (Arg-57) to a glycine has been shown to not only drastically affect the enzyme activity but also its kinetic mechanism (Kuo, L. C., Miller, A. W., Lee, S., and Kozuma, C. (1988) Biochemistry 27, 8823-8832). The crystals of the Arg-57----Gly mutant protein are isomorphous to those of the wild type. Crystal soaking experiments using both wild-type and Arg-57----Gly crystals in the presence of various ligands have provided evidence of specific conformational changes upon substrate binding which supports our previous kinetic and spectroscopic observations. 相似文献
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Using 3T[14C]malate it was possible to show intermolecular T-transfer to unlabeled fumarate. The rate of dissociation of ET derived from the malate was not rapid, only about as fast as required for KMcat. Because of the slow dissociation of ET derived from T-malate, the awkward complex ET-malate is readily formed. As shown by the effect of added malate on the partition of ET, otherwise captured by fumarate, ET.malate must be functional. Its rate of dissociation to E.M determines the V/Km value of malate. Hydrogen dissociation of the complex ET.F was linearly related to the concentration and basicity of the buffer provided, varying from < 10% to > 60% of the overall rate with alkyl phosphonates. Partition of EH.F to free malate or fumarate occurs in a ratio approximately 2:1 at both low and high buffer. This agrees well with the comparison of the equilibrium exchange rates: malate with [18O]water to malate with [14C]-fumarate [Hansen, J.N., Dinovo, E.C., & Boyer, P.D. (1969) J. Biol. Chem. 244, 6270-6279]. Therefore, the abstracted hydroxyl group is fully exchanged from the enzyme when the bound hydrogen and fumarate return to malate and must be much more accessible to the medium than the abstracted proton. The fact that buffer increases the rate of proton transfer to the medium in the central complex makes it appear that a proton relay connects the active site donor with a remote site that interfaces with the ultimate proton source, water. 相似文献
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Relationship between receptor/ligand binding affinity and adhesion strength. 总被引:10,自引:7,他引:3 下载免费PDF全文
Receptor-mediated cell adhesion is a central phenomenon in many physiological and biotechnological processes. Mechanical strength of adhesion is generally presumed to be related to chemical affinity of receptor/ligand bonds, but no experimental study has been previously directed toward this issue. Here we investigate the dependence of receptor/ligand adhesion strength on bond affinity using a radial fluid flow chamber assay to measure the force needed to detach polystyrene beads covalently coated with immunoglobulin G from glass surfaces covalently coated with protein A. A spectrum of animal species sources for immunoglobulin G permits examination of three decades of protein A/immunoglobulin G binding affinity. Our results for this model system demonstrate that adhesion strength varies with the logarithm of the binding affinity, consistent with a prediction from the theoretical model by Dembo et al. (Dembo, M., D.C. Torney, K. Saxman, and D. Hammer. 1988. Proc. R. Soc. Lond. Ser. B 234:55-83). 相似文献
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Jin-Cherng Lien Li-Jiau Huang Jih-Pyang Wang Che-Ming Teng Kuo-Hsiung Lee Sheng-Chu Kuo 《Bioorganic & medicinal chemistry》1997,5(12):2111-2120
A series of 2-substituted 3-chloro-1,4-naphthoquinones was synthesized, and the antiplatelet, antiinflammatory, and antiallergic activities of these compounds were evaluated. The structure-activity relationships in this series were also examined. Most of the 2-alkyl/arylcarboxamido derivatives of 3-chloro-1,4-naphthoquinone showed potent activities with similar trends in each of the activities evaluated. 相似文献