Home About us Contact
|
Home About us Contact | ||
|
|
||
Low Stability (low + stability)
Selected AbstractsCatalytic Hydroxylation in Biphasic Systems using CYP102A1 MutantsADVANCED SYNTHESIS & CATALYSIS (PREVIOUSLY: JOURNAL FUER PRAKTISCHE CHEMIE), Issue 7-8 2005Steffen Abstract Cytochrome P450 monooxygenases are biocatalysts that hydroxylate or epoxidise a wide range of hydrophobic organic substrates. Their technical application is, however, limited to a small number of whole-cell processes. The use of the isolated P450 enzymes is believed to be impractical due to their low stability, stoichiometric need of the expensive cofactor NAD(P)H and low solubility of most substrates in aqueous media. We investigated the behaviour of an isolated bacterial monooxygenase (mutants of CYP102A1) in a biphasic reaction system supported by cofactor recycling with the NADP+ -dependent formate dehydrogenase from Pseudomonas sp 101. Using this experimental set-up cyclohexane, octane and myristic acid were hydroxylated. To reduce the process costs a novel NADH-dependent mutant of CYP102A1 was designed. For recycling of NADH an NAD+ -dependent FDH was used. The stability of the monooxygenase mutants under the reaction conditions in the biphasic system was quite high as revealed by total turnover numbers of up to 12,850 in the NADPH-dependent cyclohexane hydroxylation and up to 30,000 in the NADH-dependent myristic acid oxidation. [source] Analysis of interaction modes in calix[4]arene,levofloxacin complexes by quantum methodsJOURNAL OF PHYSICAL ORGANIC CHEMISTRY, Issue 3 2006Alexandrine Lambert Abstract Host,guest interactions between chiral calix[4]arenes and the antibiotic levofloxacin are analyzed on the basis of quantum mechanical calculations at the density functional (for model systems) and semi-empirical levels. The calix[4]arene macrocycle carries two (+)-isomenthyl groups attached to opposing phenyl groups at the lower rim and different substituents (R,=,H, CH3, tBu, CH2CHCH2, COCH3 and NO2) are considered at the upper rim. Nitro derivatives are expected to form ionized complexes whereas the other derivatives should form neutral complexes with a very low stability. Copyright © 2006 John Wiley & Sons, Ltd. [source] Multiple bacteria encode metallothioneins and SmtA-like zinc fingersMOLECULAR MICROBIOLOGY, Issue 5 2002Claudia A. Blindauer Summary Zinc is essential but toxic in excess. Bacterial metallothionein, SmtA from Synechococcus PCC 7942, sequesters and detoxifies four zinc ions per molecule and contains a zinc finger structurally similar to eukaryotic GATA. The dearth of other reported bacterial metallothioneins has been surprising. Here we describe related bacterial metallothioneins (BmtA) from Anabaena PCC 7120, Pseudomonas aeruginosa and Pseudomonas putida that bind multiple zinc ions with high stability towards protons. Thiol modification demonstrates that cysteine coordinates zinc in all of these proteins. Additionally, 111Cd-NMR, and 111Cd-edited 1H-NMR, identified histidine ligands in Anabaena PCC 7120 BmtA, analogous to SmtA. A related Escherichia coli protein bound only a single zinc ion, via four cysteine residues, with low stability towards protons; 111Cd-NMR and 111Cd-edited 1H-NMR confirmed exclusive cysteine-coordination, and these cysteine residues reacted rapidly with 5,5,-dithiobis-(2-nitrobenzoic acid). 1H-NMR of proteins from P. aeruginosa, Anabaena PCC 7120 and E. coli generated fingerprints diagnostic for the GATA-like zinc finger fold of SmtA. These studies reveal first the existence of multiple bacterial metallothioneins, and second proteins with SmtA-like lone zinc fingers, devoid of a cluster, and designated GatA. We have identified 12 smtA -like genes in sequence databases including four of the gatA type. [source] New attempts to quantify concentric needle electromyographyMUSCLE AND NERVE, Issue S11 2002Masahiro Sonoo MD, PhDArticle first published online: 4 JUN 200 Abstract Quantitative motor unit potential (MUP) analysis, which is a leading method of quantitative evaluation of concentric needle electromyography, has several inherent limitations. First, the most essential features of neurogenic or myogenic changes manifest as recruitment abnormalities, rather than as changes in MUP morphology. Second, two factors related to MUP sampling, focusing and level of contraction, greatly influence the parameters of sampled MUPs. Third, the MUP duration, considered to be the cardinal parameter in MUP analysis, has several drawbacks, including low stability and low discriminant sensitivity. We developed a new MUP parameter, the size index (SI), which is calculated from the MUP amplitude and area/amplitude ratio (thickness). The SI remained almost constant during electrode movements, as demonstrated by manual scanning of MUPs. It is a stable and robust parameter and achieved an extremely high ability to discriminate between normal and large neurogenic MUPs. It identifies features related to the sound produced by the MUP on the audio monitor, which is often used by trained electromyographers for qualitative assessments of MUPs. © 2002 Wiley Periodicals, Inc. Muscle Nerve Supplement 11: S98,S102, 2002 [source] Structure of a highly stable mutant of human fibroblast growth factor 1ACTA CRYSTALLOGRAPHICA SECTION D, Issue 1 2009Anna Szlachcic Fibroblast growth factors (FGFs) are involved in diverse cellular processes such as cell migration, angiogenesis, osteogenesis, wound healing and embryonic and foetal development. Human acidic fibroblast growth factor (FGF-1) is the only member of the FGF family that binds with high affinity to all four FGF receptors and thus is considered to be the human mitogen with the broadest specificity. However, pharmacological applications of FGF-1 are limited owing to its low stability. It has previously been reported that the introduction of single mutations can significantly improve the stability of FGF-1 and its resistance to proteolytic degradation. Here, the structure of the Q40P/S47I/H93G triple mutant of FGF-1, which exhibits much higher stability, a prolonged half-life and enhanced mitogenic activity, is presented. Compared with the wild-type structure, three localized conformational changes in the stable triple mutant were observed, which is in agreement with the perfect energetic additivity of the single mutations described in a previous study. The huge change in FGF-1 stability (the denaturation temperature increased by 21.5,K, equivalent to ,,Gden = 24.3,kJ,mol,1) seems to result from the formation of a short 310 -helix (position 40), an improvement in the propensity of amino acids to form ,-sheets (position 47) and the rearrangement of a local hydrogen-bond network (positions 47 and 93). [source] Improving the Industrial Production of 6-APA: Enzymatic Hydrolysis of Penicillin G in the Presence of Organic SolventsBIOTECHNOLOGY PROGRESS, Issue 6 2003Olga Abian The hydrolysis of penicillin G in the presence of an organic solvent, used with the purpose of extracting it from the culture medium, may greatly simplify the industrial preparation of 6-APA. However, under these conditions, PGA immobilized onto Eupergit displays very low stability (half-life of 5 h in butanone-saturated water) and a significant degree of inhibition by the organic solvent (30%). The negative effect of the organic solvent strongly depended on the type of solvent utilized: water saturated with butanone (around 28% v/v) had a much more pronounced negative effect than that of methylisobutyl ketone (MIBK) (solubility in water was only 2%). These problems were sorted out by using a new penicillin G acylase derivative designed to work in the presence of organic solvents (with each enzyme molecule surrounded by an hydrophilic artificial environment) and a suitable organic solvent (MIBK). Using such solvent, this derivative kept its activity unaltered for 1 week at 32 °C. Moreover, the enzyme activity was hardly inhibited by the presence of the organic solvent. In this way, the new enzyme derivative thus prepared enables simplification of the industrial hydrolysis of penicillin G. [source] Mechanism of tyrosinase inhibition by deoxyArbutin and its second-generation derivativesBRITISH JOURNAL OF DERMATOLOGY, Issue 6 2008S. Chawla Summary Background, Disorders, such as age spots, melasma and hyperpigmentation at sites of actinic damage, emanate from the augmentation of an increased amount of epidermal melanin. Objectives, The ineptness of current therapies in treating these conditions, as well as high cytotoxicity, mutagenicity, poor skin penetration and low stability of skin-depigmenting formulations led us to investigate new compounds that meet the medical requirements for depigmentation agents. We have shown previously that the tyrosinase inhibitor deoxyArbutin (dA) is a more effective and less toxic skin lightener than hydroquinone (HQ). Methods, The efficacy and reversibility of dA and its derivatives on inhibiting tyrosine hydroxylase and DOPAoxidase was assessed using standard assays. Results, dA and its second-generation derivatives inhibit tyrosine hydroxylase and DOPAoxidase activities of tyrosinase dose dependently thereby inhibiting melanin synthesis in intact melanocytes, when used at concentrations that retain 95% cell viability in culture. This depigmenting effect was completely reversible when the compounds were removed. Tyrosinase inhibition was also observed in vitro when tested using human and purified mushroom tyrosinase, establishing that they are direct enzyme inhibitors. Lineweaver,Burk reciprocal plot analysis using mushroom tyrosinase illustrated that dA and its derivatives are more robust competitive inhibitors than HQ, when tyrosine is used as substrate. Conclusions, Thus, dA and its second-generation derivatives, which inhibit melanogenesis at safe concentrations by specifically acting on the tyrosinase enzyme at a post-translational level, are promising agents to ameliorate hyperpigmented lesions or lighten skin. [source] | ||||||||||||||||