Production of 6-nylon monomer by metabolic engineering
| Project/Area Number |
16K14493
|
|---|
| Research Category |
Grant-in-Aid for Challenging Exploratory Research
|
| Allocation Type | Multi-year Fund |
|---|
| Research Field |
Biofunction/Bioprocess
|
|---|
| Research Institution | University of Hyogo |
|---|
Principal Investigator |
Negoro Seiji 兵庫県立大学, 工学研究科, 教授 (90156159)
|
|---|
| Research Collaborator |
TAKEO masahiro
KATO daiichiro
TAKEHARA ikki
FUJII tsubasa
|
|---|
| Project Period (FY) |
2016-04-01 – 2019-03-31
|
| Project Status |
Completed (Fiscal Year 2018)
|
| Budget Amount *help |
¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000)
Fiscal Year 2018: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2017: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2016: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
|
|---|
| Keywords | ナイロン / 代謝工学 / ６アミノヘキサン酸 / 6-アミノヘキサン酸 / アジピン酸 / アミノトランスフェラーゼ / ピリドキサルリン酸 / ４-アミノ酪酸 / ６-アミノヘキサン酸 / 代謝経路 / Arthrobacter / 廃棄物再資源化 / 環境材料 / バイオテクノロジー |
|---|
| Outline of Final Research Achievements |
Arthrobacter sp. strain KI72 grows on a 6-aminohexanoate oligomer as a sole source of carbon and nitrogen. We cloned the two genes, nylD1 and nylE1, responsible for 6-aminohexanoate metabolism. We amplified the DNA fragments that encode these genes by polymerase chain reaction using a synthetic primer DNA homologous to the 4-aminobutyrate metabolic enzymes. We inserted the amplified DNA fragments into the expression vector pColdI in Escherichia coli, purified the His-tagged enzymes to homogeneity, and performed biochemical studies. We confirmed that 6-aminohexanoate aminotransferase (NylD1) catalyzes the reaction of 6-aminohexanoate to adipate semialdehyde using alpha-ketoglutarate, pyruvate and glyoxylate as amino acceptors, generating glutamate, alanine and glycine, respectively. For further metabolism, adipate semialdehyde dehydrogenase (NylE1) catalyzes the oxidative reaction of adipate semialdehyde to adipate using NADP+ as a cofactor.
|
| Academic Significance and Societal Importance of the Research Achievements |
本研究は、繊維・プラスチックとして広く用いられているナイロンについて、６ナイロンの構成単位である6-アミノヘキサン酸の微生物代謝経路の解明、関連酵素群の取得と機能改良について、検討を行ったものである。特に、6-アミノヘキサン酸の代謝については、以前の研究例がなかったが、モノマー代謝の初段階に関与する２種類の酵素（アミノ基転移酵素と脱水素酵素）を用いて、6-アミノヘキサン酸が、90%以上の収率でアジピン酸へ変換できることが明らかとなった。本研究からNylA-E酵素群を統合的に用いることで、高収率で、６ナイロンを、６６ナイロン原料であるアジピン酸まで変換可能である可能性を示すことができた。
|
Report
(4 results)
Research Products
(27 results)
-
[Journal Article] Stability and subunit assembly of nylon hydrolase: structural basis of correct subunit assembly, aggregation, and intracellular degradation of the precursor protein2018
Author(s)
Seiji Negoro, Naoki Shibata, Young-Ho Lee, Ikki Takehara, Ryo Kinugasa, Keisuke Nagai, Yusuke Tanaka, Dai-ichiro Kato, Masahiro Takeo, Yuji Goto and Yoshiki Higuchi
-
Journal Title
Scientific Reports
Volume: 8
Issue: 1
Pages: 9725-9725
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research
-
-
[Journal Article] Biosynthetic Pathway and Genes of Chitin/Chitosan-Like Bioflocculant in the Genus Citrobacter.2018
Author(s)
Takeo M, Kimura K, Mayilraj S, Inoue T, Tada S, Miyamoto K, Kashiwa M, Ikemoto K, Baranwal P, Kato D, Negoro S.
-
Journal Title
Polymers (Basel).
Volume: 10
Issue: 3
Pages: 237-237
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research
-
-
-
-
[Journal Article] Mutations affecting the internal equilibrium of the reaction catalyzed by 6-aminohexanoate-dimer hydrolase2016
Author(s)
Negoro S, Kawashima Y, Shibata N, Kobayashi T, Baba T, Lee YH, Kamiya K, Shigeta Y, Nagai K, Takehara I, Kato D, Takeo M, Higuchi Y
-
Journal Title
FEBS Lett
Volume: 590
Pages: 3133-3143
Related Report
Peer Reviewed / Acknowledgement Compliant
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
