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Hydrolyzing Activity (hydrolyzing + activity)
Selected AbstractsGenes for an alkaline d -stereospecific endopeptidase and its homolog are located in tandem on Bacillus cereus genomeFEMS MICROBIOLOGY LETTERS, Issue 1 2003Hidenobu Komeda Abstract Alkaline d -peptidase (Adp) from Bacillus cereus DF4-B is a d -stereospecific endopeptidase acting on oligopeptides composed of d -phenylalanine and the primary structure deduced from its gene, adp, shows a similarity with d -stereospecific hydrolases from Ochrobactrum anthropi strains. We have isolated DNA fragments covering the flanking region of adp from DF4-B genome and found an additional gene, adp2, located upstream of adp. The deduced amino acid sequence of Adp2 showed 96% and 85% identity with those of Adp from B. cereus strains AH559 and DF4-B, respectively. The recombinant Adp2 expressed in Escherichia coli was purified to homogeneity and characterized. It had hydrolyzing activity toward (d -Phe)3, (d -Phe)4, and (d -Phe)6 but did not act on (l -Phe)4, d -Phe-NH2, and l -Phe-NH2, some characteristics that are closely related to those of Adp from strain DF4-B. These results indicate that highly homologous genes encoding d -stereospecific endopeptidases are arranged in a tandem manner on the genomic DNA of B. cereus DF4-B. [source] Affinity and catalytic heterogeneity of polyclonal myelin basic protein-hydrolyzing IgGs from sera of patients with multiple sclerosisJOURNAL OF CELLULAR AND MOLECULAR MEDICINE, Issue 3 2010Galina A. Legostaeva Abstract Human myelin basic protein (hMBP)-hydrolyzing activity was recently shown to be an intrinsic property of antibodies (Abs) from multiple sclerosis (MS) patients. Here, we present the first evidence demonstrating a significant diversity of different fractions of polyclonal IgGs (pIgGs) from MS patients in their affinity for hMBP and in the ability of pIgGs to hydrolyze hBMP at different optimal pHs (3,10.5). IgGs containing ,- and ,-types of light chains demonstrated comparable relative activities in the hydrolysis of hMBP. IgGs of IgG1,IgG4 sub-classes were analyzed for catalytic activity. IgGs of all four sub-classes were catalytically active, with their contribution to the total activity of Abzs in the hydrolysis of hMBP and its 19-mer oligopeptide increasing in the order: IgG1 (1.5,2.1%) < IgG2 (4.9,12.8%) < IgG3 (14.7,25.0%) < IgG4 (71,78%). Our findings suggest that the immune systems of individual MS patients generate a variety of anti-hMBP abzymes with different catalytic properties, which can attack hMBP of myelin-proteolipid shell of axons, playing an important role in MS pathogenesis. [source] Variation of Fructooligosaccharides and their Metabolizing Enzymes in Onion Bulb (Allium cepa L. cv. Tenshin) During Long-term StorageJOURNAL OF FOOD SCIENCE, Issue 3 2005Noureddine Benkeblia ABSTRACT: The objective of this study was to assess the status of fructooligosaccharides (FOS) in onion bulbs (Allium cepa L. cv. Tenshin) and their metabolizing-enzymes,1-fructoexohydrolase (1-FEH), 1-kestose hydrolyzing enzyme (1-KH), fructan:fructan 1F -fructosyltransferase (1-FFT) and fructan:fructan 6G -fructosyltransferase (6G-FFT),during storage at 15°C. Fructose varies slightly, whereas 1-kestose peaked after 6 wk and then decreased progressively during the last 18 wk of storage. Lower degree of polymerization (DP) 3 to 6) FOS, higher (DP 7 to 12) FOS, total FOS, and total carbohydrates showed similar and close patterns during 24 wk. They varied slightly at the beginning of the storage period; afterward they decreased progressively and regularly during the last 20 wk of storage. 1-FEH and 1-KH activities were low but peaked abruptly after 12 and 16 wk, respectively, after which they decreased to levels higher (1-FEH) or similar (1-KH) to those observed at the beginning of the storage. Surprisingly, 1-FFT activity showed similar pattern to the variation of 1-KH hydrolyzing activity; on the other hand, 6G-FFT, although higher, was stable during 16 wk but decreased after that. The results allowed us to associate FOS to the dormancy and sprouting states, and the peaks of the degrading enzymes were shown to signal the release of dormancy of onion bulb. [source] Workshop 5: NAAG and NAALADase: Functional Properties in the Central and Peripheral Nervous SystemJOURNAL OF NEUROCHEMISTRY, Issue 2002D. Bacich Glutamate carboxypeptidase II (GCPII, also known as N-acetylated-alpha-linked acidic dipeptidase or NAALADase) knockout (KO) mice were generated by inserting a GCPII targeting cassette containing a PGK-Neo resistance marker and stop codons in exons 1 and 2, and removal of exons 1 and 2 intron/exon boundary sequence. Embryonic stem cells were injected into C57BL6 blastocysts, and chimeric offspring born. Germline transmission was confirmed by mating the chimeras to generate heterozygous KO mice. Crossing heterozygous mice generated F2 generation mice homozygous for the null mutant, as confirmed by loss of GCPII protein. NAAG hydrolyzing activity was minimal (0.07 pmol/mg/min) in KO tissue, with normal levels (4.82 pmol/mg/min) in wild types and intermediate levels (1.73 pmol/mg/min) in heterozygotes. Preliminary neuropathy experiments showed KO mice are less affected by nerve-crush and recover faster from the damage-induced neuropathy, as indicated by EMG recording and nerve morphology. Similarly, GCPII KO mice subjected to high dose vitamin B6 displayed less severe neuropathy than wild types, as indicated by reduced sensory nerve conduction velocity and morphological deficits. Also, in a transient middle cerebral artery occlusion model, GCPII KO mice were significantly more resistant to the effects of cerebral ischemia than their wildtype littermates. Findings support GCPII involvement in stroke and in mediating chronic neuropathic conditions and suggest GCPII inhibitors may be useful in treatment of brain ischemia as well as peripheral neuropathies. [source] | ||||||||||