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- Abu-Bakar, A'edah, et al.
(author)
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Metabolism of bilirubin by human cytochrome P450 2A6
- 2012
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In: Toxicology and Applied Pharmacology. - : Elsevier BV. - 0041-008X .- 1096-0333. ; 261:1, s. 50-58
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Journal article (peer-reviewed)abstract
- The mouse cytochrome P450 (CYP) 2A5 has recently been shown to function as hepatic "Bilirubin Oxidase" (Abu-Bakar, A., et al., 2011. Toxicol. Appl. Pharmacol. 257, 14-22). To date, no information is available on human CYP isoforms involvement in bilirubin metabolism. In this paper we provide novel evidence for human CYP2A6 metabolising the tetrapyrrole bilirubin. Incubation of bilirubin with recombinant yeast microsomes expressing the CYP2A6 showed that bilirubin inhibited CYP2A6-dependent coumarin 7-hydroxylase activity to almost 100% with an estimated K-i of 2.231 mu M. Metabolite screening by a high-performance liquid chromatography/electrospray ionisation mass spectrometry indicated that CYP2A6 oxidised bilirubin to biliverdin and to three other smaller products with m/z values of 301,315 and 333. Molecular docking analyses indicated that bilirubin and its positively charged intermediate interacted with key amino acid residues at the enzyme's active site. They were stabilised at the site in a conformation favouring biliverdin formation. By contrast, the end product, biliverdin was less fitting to the active site with the critical central methylene bridge distanced from the CYP2A6 haem iron facilitating its release. Furthermore, bilirubin treatment of HepG2 cells increased the CYP2A6 protein and activity levels with no effect on the corresponding mRNA. Co-treatment with cycloheximide (CHX), a protein synthesis inhibitor, resulted in increased half-life of the CYP2A6 compared to cells treated only with CHX. Collectively, the observations indicate that the CYP2A6 may function as human "Bilirubin Oxidase" where bilirubin is potentially a substrate and a regulator of the enzyme.
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| 2. |
- Verkasalo, Erkki, et al.
(author)
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Extractives of Stemwood and Sawmill Residues of Scots Pine (Pinus sylvestris L.) for Biorefining in Four Climatic Regions in Finland-Phenolic and Resin Acid Compounds
- 2021
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In: Forests. - : MDPI. - 1999-4907. ; 12:2, s. 1-24
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Journal article (peer-reviewed)abstract
- This study aimed to identify and quantify phenolic and resin acid extractive compounds in Scots pine stemwood and sawmill residues in four climatic regions of Finland to evaluate their most optimal sources for bio-based chemical biorefining and bioenergy products. The sample consisted of 140 trees from 28 stands, and sawdust lots from 11 log stands. NMR for the overall extractive analysis and HPLC for the quantitative estimation of phenolic and resin acid compounds were employed. Correlation analysis, multivariate factor analysis, principle component analysis and multiple linear regression modelling were applied for statistical analysis. HPLC identified 12 extractive compounds and NMR five more resin acids. Pinosylvin (PS), pinosylvin monomethyl ether (PSMME), and partly neolignans/lignans occurred in the largest concentrations. Wood type caused the most variation, heartwood having larger concentrations than sapwood (sawdust between them). Regional differences in the concentrations were smaller, but factor analysis distinguished the northern and the southern regions into their own groups. The results indicated higher concentrations of PS, PSMME, and vanillic acid in southern regions and those of, e.g., PSMME glycoside, lignan 2, and neolignan 1 in northern regions. The rather low concentrations of extractives in stemwood and sawdust imply value-added products, efficient sorting and/or large raw material volumes.
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