Biodegradation of Bacterial Polyentens in Anerobic Conditions and is mechanisms (2002)

厌氧条件下细菌多聚体的生物降解及其机制 (2002)

• 批准号: 11217214
• 负责人: SAITO Terumi
• 金额: $ 22.85万
• 依托单位: Kanagawa University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research on Priority Areas
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11217214/
• 关键词: poly (3-hydroxybatyrate); polyhydroxy alkanoates; PHB depolymerase; PHB; inclusion bodies; intracellular PHB depolymerase; biodegradation; bacterial polyesters; 生分解性; 生分解性ポリエステル; ポリ-3-ヒドロキシ酪酸; ポリヒドロキシアルカノエート; PHA; 嫌気; ボリヒドロキシアルカノエート

Several bacteria which degrade bacterial polyester (PHB) and also perform denitrification were isolated. We attempted the isolation of extracellular PHB depolymerases and cloning of their genes, but failed. When small fish (gold fish) were kept in the presence of PHB, a considerable reduction of nigrogen compounds in water was observed. These results indicate that the important role of the bacteria with ability of both PHB-degradation and denitrification in environment. On the other hand, we coned the gene of an intracellular PHB depolymerase (phaZl) from Ralstonia eutropha H16 and charaterization its product (PhaZ1). The amino acid sequence of PhaZ1 was novel. The molecular mass of PhaZ1 was 47 Kda. PhaZ1, which localized solely in PHB inclusions, degraded artificial amorphous PHB granules, but not crystalline PHB granules. We further cloned several intracellular 3-hydroxybutyrate-oligomer hydrolases. One of them, an intracellular 3HB-oligomer hydrolase (PhaZ2) from R. eutropha, showed both 3HB-oligomer hydrolase and PHB depolymerase activities, PhaZ2 localizedd both in cytosol and PHB inclusions, The amounts of PhaZ1 and PhaZ2 increased with the content of PHB in cells. The null mutant lacking both PhaZ1and PhaZ2 revealed the increase PHB deposition in cells. These results indicate that PhaZ1 and PhazZ2 cooperate in intracellular degradation of PHB

分离出了几株能降解细菌聚酯(PHB)并能进行反硝化的细菌。我们尝试了胞外PHB解聚酶的分离和基因的克隆，但失败了。当小鱼(金鱼)在PHB存在下饲养时，观察到水中黑素化合物的相当大的减少。这些结果表明，兼具PHB降解和反硝化能力的细菌在环境中起着重要的作用。另一方面，我们克隆了真核细菌H16胞内PHB解聚酶(PhaZ1)的基因，并对其产物(PhaZ1)进行了鉴定。PhaZ1的氨基酸序列为新的。PhaZ1的相对分子质量为47kDa。PhaZ1仅定位在PHB包裹体中，降解人工无定形PHB颗粒，但不降解结晶PHB颗粒。我们进一步克隆了几个胞内3-羟基丁酸齐聚物水解酶。其中一种是富营养红曲霉的胞内3HB寡聚体水解酶(PhaZ2)，它同时具有3HB寡聚体水解酶和PHB解聚酶的活性，PhaZ2定位于胞浆和PHB包涵体中，PhaZ1和PhaZ2的含量随着细胞中PHB含量的增加而增加。缺失PhaZ1和PhaZ2的缺失突变体显示PHB在细胞内沉积增加。这些结果表明，PhaZ1和PhazZ2在PHB的胞内降解中起协同作用

项目成果

Y.Inagawa et al.: "Identification and characterization of poly-3-hydroxy butyrate granule-associated protein PGA12 and PGA16"Int. J. Biol. Macromol.. 30. 55-61 (2002)

Y.Inakawa 等人：“聚 3-羟基丁酸酯颗粒相关蛋白 PGA12 和 PGA16 的鉴定和表征”Int。

S.Miyazaki他: "roperties of a Poly(3-hydroxybutyrate) Depolymerase from Penicillium funiculosum"J.Polym.Environ.. 8. 175-182 (2000)

S. Miyazaki 等人：“来自绳状青霉的聚（3-羟基丁酸酯）解聚酶的特性”J. Polym. 8. 175-182 (2000)

Y.Suzuki他: "Involvement of Catalytic Amino Acid Residues in Enzyme Catalyzed Polymerization for the Synthesis of Polyesters"Biomacromolecules. 2. 541-544 (2001)

Y.Suzuki等人：“催化氨基酸残基参与聚酯合成的酶催化聚合”生物大分子2.541-544(2001)。

T.Kobayashi, A.Sugiyama, Y.Kawase, T.Saito, J.Mergaert, J.Swings: "Biochemical and Genetic Characterization of an Extracellular an Extracellular Poly (3-hydroxybutyrate) Depolymerase from Acidovorax sp. Strain TP4"J.Environ. Polym. Degrad.. 7. 9-17 (1999)

T.Kobayashi、A.Sugiyama、Y.Kawase、T.Saito、J.Mergaert、J.Swings：“来自 Acidovorax sp. 菌株 TP4 的细胞外聚（3-羟基丁酸酯）解聚酶的生化和遗传特征”J。

H.Yamane, K.Terao, S.Hiki, Y.Kawahara, Y.Kimura, T.Saito: "Enzymatic Degradation of Bacterial Homo-poly (3-hydroxybutyrate) Melt Spun Fibers"Polymer. 42. 7873-7878 (2001)

H.Yamane、K.Terao、S.Hiki、Y.Kawahara、Y.Kimura、T.Saito：“细菌均聚（3-羟基丁酸酯）熔纺纤维的酶降解”聚合物。