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Swi See Ang Abu Bakar Salleh Leow Thean Chor Yahaya M. Normi Bimo Ario Tejo Mohd Basyaruddin Abdul Rahman Mariam-Aisha Fatima 《The protein journal》2018,37(2):180-193
The bioconversion of vitamin D3 catalyzed by cytochrome P450 (CYP) requires 25-hydroxylation and subsequent 1α-hydroxylation to produce the hormonal activated 1α,25-dihydroxyvitamin D3. Vitamin D3 25-hydroxylase catalyses the first step in the vitamin D3 biosynthetic pathway, essential in the de novo activation of vitamin D3. A CYP known as CYP107CB2 has been identified as a novel vitamin D hydroxylase in Bacillus lehensis G1. In order to deepen the understanding of this bacterial origin CYP107CB2, its detailed biological functions as well as biochemical characteristics were defined. CYP107CB2 was characterized through the absorption spectral analysis and accordingly, the enzyme was assayed for vitamin D3 hydroxylation activity. CYP-ligand characterization and catalysis optimization were conducted to increase the turnover of hydroxylated products in an NADPH-regenerating system. Results revealed that the over-expressed CYP107CB2 protein was dominantly cytosolic and the purified fraction showed a protein band at approximately 62 kDa on SDS–PAGE, indicative of CYP107CB2. Spectral analysis indicated that CYP107CB2 protein was properly folded and it was in the active form to catalyze vitamin D3 reaction at C25. HPLC and MS analysis from a reconstituted enzymatic reaction confirmed the hydroxylated products were 25-hydroxyitamin D3 and 1α,25-dihydroxyvitamin D3 when the substrates vitamin D3 and 1α-hydroxyvitamin D3 were used. Biochemical characterization shows that CYP107CB2 performed hydroxylation activity at 25 °C in pH 8 and successfully increased the production of 1α,25-dihydroxyvitamin D3 up to four fold. These findings show that CYP107CB2 has a biologically relevant vitamin D3 25-hydroxylase activity and further suggest the contribution of CYP family to the metabolism of vitamin D3. 相似文献
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The F420S substitution enhances the specific activity of Ralstonia eutropha PHA synthase (PhaCRe). We have now carried out site-directed saturation mutagenesis of F420 of PhaCRe and, amongst the F420 mutants, the F420S mutant gave the highest poly(3-hydroxybutyrate) (PHB) content. In vitro activity assay showed that the F420S enzyme had a significant decrease in its lag phase compared to that of the wild-type enzyme. Enhancement of PHB accumulation was achieved by combination of the F420S mutation with a G4D mutation, which conferred high PHB content and high in vivo concentration of PhaCRe enzyme. The G4D/F420S mutant gave a higher PHB content and in vivo concentration of PhaCRe enzyme than the F420S mutant, while the molecular weight of the PHB polymer of the double mutant was similar to that of the F420S mutant. 相似文献
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Sohrab?P?Shah David?YM?He Jessica?N?Sawkins Jeffrey?C?Druce Gerald?Quon Drew?Lett Grace?XY?Zheng Tao?Xu BF?Francis?OuelletteEmail author 《BMC bioinformatics》2004,5(1):40
Background
We present Pegasys – a flexible, modular and customizable software system that facilitates the execution and data integration from heterogeneous biological sequence analysis tools. 相似文献5.
Saadi Bayat Bimo A. Tejo Abu Bakar Salleh Emilia Abdmalek Yahaya M. Normi Mohd Basyaruddin Abdul Rahman 《Chirality》2013,25(11):726-734
A series of tripeptide organocatalysts containing a secondary amine group and two amino acids with polar side chain units were developed and evaluated in the direct asymmetric intermolecular aldol reaction of 4‐nitrobenzaldehyde and cyclohexanone. The effectiveness of short polar peptides as asymmetric catalysts in aldol reactions to attain high yields of enantio‐ and diastereoselective isomers were investigated. In a comparison, glutamic acid and histidine produced higher % ee and yields when they were applied as the second amino acid in short trimeric peptides. These short polar peptides were found to be efficient organocatalysts for the asymmetric aldol addition reaction in aqueous media. Chirality 25:726–734, 2013. © 2013 Wiley Periodicals, Inc 相似文献
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Hernández Pérez A E Cerna Chávez JC Delgado Ortiz M Beltrán Beache LM Tapia Vargas YM Ochoa Fuentes 《Phyton》2019,88(1):11-13
Mexico is the main producer, consumer and exporter
of avocado in the world, being Michoacan the main producer state
contributing more than 80% of the national production. There
are phytopathogens that decimate the production causing the
death of the tree. Root samples were collected in avocado trees
that showed the characteristic symptomatology of the disease
known as avocado sadness, the sampling was carried out in four
of the main avocado producing towns, in the state of Michoacan,
Mexico. The isolation consisted in sowing root tissue in Petri
dishes with V8®-PARPH culture medium, subsequently they were
identified morphologically and for species level it was determined
by molecular biology, with the PCR-ITS technique. Pathogenicity
tests were performed in triplicate with avocado seedlings with more
than six leaves. After 24 hours, the inoculated plants expressed
decay in the apical part, after 120 hours the leaves showed yellowing
and after 15 days there was a generalized wilt on the stem and
leaves, re-isolating the phytopathogen Phytopythium vexans.
This study confirms the first report of the oomycete P. vexans
affecting avocado trees in the most important producing region of
the Mexican Republic. 相似文献
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Rauda A. Mohamed Abu Bakar Salleh Adam Thean Chor Leow Normi M. Yahaya Mohd Basyaruddin Abdul Rahman 《Molecular biotechnology》2017,59(7):284-293
An enzyme with broad substrate specificity would be an asset for industrial application. T1 lipase apparently has the same active site residues as polyhydroxyalkanoates (PHA) depolymerase. Sequences of both enzymes were studied and compared, and a conserved lipase box pentapeptide region around the nucleophilic serine was detected. The alignment of 3-D structures for both enzymes showed their active site residues were well aligned with an RMSD value of 1.981 Å despite their sequence similarity of only 53.8%. Docking of T1 lipase with P(3HB) gave forth high binding energy of 5.4 kcal/mol, with the distance of 4.05 Å between serine hydroxyl (OH) group of TI lipase to the carbonyl carbon of the substrate, similar to the native PhaZ7 Pl . This suggests the possible ability of T1 lipase to bind P(3HB) in its active site. The ability of T1 lipase in degrading amorphous P(3HB) was investigated on 0.2% (w/v) P(3HB) plate. Halo zone was observed around the colony containing the enzyme which confirms that T1 lipase is indeed able to degrade amorphous P(3HB). Results obtained in this study highlight the fact that T1 lipase is a versatile hydrolase enzyme which does not only record triglyceride degradation activity but amorphous P(3HB) degradation activity as well. 相似文献
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