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Title: | 放線菌 Nonomuraea angiospora 中 Cytochrome P450 Monooxygenase 之選殖 Molecular Cloning of Cytochrome P450 Monooxygenase Gene from Nonomuraea angiospora |
Authors: | Chih-Ying Chen 陳芝瑩 |
Advisor: | 王愛玉(Ai-Yu Wang) |
Keyword: | pravastatin,compactin,HMG-CoA reductase,cytochrome P450 monooxygenase (CYP450),Nonomuraea angiospora, |
Publication Year : | 2009 |
Degree: | 碩士 |
Abstract: | 普遍存在於生物體中的 Cytochrome P450 monooxygenases (CYP450s) 是一種血基質蛋白質 (hemoprotein),由一群大家族 (superfamily) 基因所表現。CYP450 參與許多的催化反應,包括 carbon hydroxylation、heteroatom oxygenation、dealkylation、epoxidation、aromatic hydroxylation、reduction 和 dehalogenation 等反應。市面上已有許多產品是應用 CYP450s的特性所開發,例如抗生素。
本論文利用不同的 PCR 方法,由 Nonomuraea angiospora 中選殖出 CYP450 基因。此菌株已被發現具有將 compactin 轉化成 pravastatin 的能力。Pravastatin為 HMG-CoA reductase 活性抑制因子,被用來降低血液中的膽固醇含量。Nonomuraea angiospora CYP450 基因的編碼區域 (open reading frame) 全長為 1224 bp,可轉譯成 407 個胺基酸,預估分子量為 45 kDa。Nonomuraea angiospora CYP450 之胺基酸序列與 Nonomuraea recticatena、Salinispora arenicola CNS-205 以及Saccharopolyspora erythraea NRRL2338 的 CYP450 分別具有 90%、66% 和 61 % 的同質性。由序列分析、二級結構預測及三級結構模擬分析結果,推測 Nonomuraea angiospora CYP450 隸屬於 CYP105 family,可以使用 mitochondrial/bacterial type 的電子傳遞系統,催化 compactin 轉化為 pravastatin 的反應。 Cytochrome P450 monooxygenases (CYP450s), which are ubiquitously distributed in organism, are hemoproteins encoded by a superfamily of genes. They catalyze a variety of chemical reactions including carbon hydroxylation, heteroatom oxygenation, dealkylation, epoxidation, aromatic hydroxylation, reduction and dehalogenation. The enzyme family has been used for production of a lot commercial products such as antibiotics. In this study, CYP450 gene was cloned from Nonomuraea angiospora, which has been found to be capable of converting compactin to pravastatin, by different PCR methods. Pravastatin is an inhibitor of HMG-CoA reductase and is used in reducing the cholesterol levels in blood. The open reading frame of Nonomuraea angiospora CYP450 gene consists of 1224 bp and is predicted to encode 407 amino acids with a molecular mass of 45 kDa. The amino acid sequence of Nonomuraea angiospora CYP450 shows 90%, 66% and 61% identities with CYP450s from Nonomuraea recticatena, Salinispora arenicola and Saccharopolyspora erythraea, respectively. The results of sequence analysis、secondary structure prediction and 3D structure modeling suggest that Nonomuraea angiospora CYP450 belongs to the CYP105 family and may catalyze the hydroxylation of compactin to pravastatin by using the mitochondrial/bacterial type electronic transport system. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/23013 |
Fulltext Rights: | 未授權 |
Appears in Collections: | 微生物學科所 |
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ntu-98-1.pdf Restricted Access | 2.89 MB | Adobe PDF |
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