The first thermophilic α-oxoamine synthase family enzyme that has activities of 2-amino-3-ketobutyrate CoA ligase and 7-keto-8-aminopelargonic acid synthase: cloning and overexpression of the gene from an extreme thermophile, Thermus thermophilus and characterization of its gene product
-
- KUBOTA Takaaki
- Department of Biotechnology, Tottori University
-
- SHIMONO Jyunpei
- Department of Biotechnology, Tottori University
-
- KANAMEDA Chie
- Department of Biotechnology, Tottori University
-
- IZUMI Yoshikazu
- Department of Biotechnology, Tottori University
書誌事項
- タイトル別名
-
- The First Thermophilic .ALPHA.-Oxoamine Synthase Family Enzyme That Has Activities of 2-Amino-3-ketobutyrate CoA Ligase and 7-Keto-8-aminopelargonic Acid Synthase: Cloning and Overexpression of the Gene from an Extreme Thermophile, Thermus thermophilus, and Characterization of Its Gene Product
- The first thermophilic a oxoamine synthase family enzyme that has activities of 2 amino 3 ketobutyrate CoA ligase and 7 keto 8 aminopelargonic acid synthase cloning and overexpression of the gene from an extreme thermophile Thermus thermophilus and characterization of its gene product
- The First Thermophilic α-Oxoamine Synthase Family Enzyme That Has Activities of 2-Amino-3-ketobutyrate CoA Ligase and 7-Keto-8-aminopelargonic Acid Synthase: Cloning and Overexpression of the Gene from an Extreme Thermophile,<i>Thermus thermophilus</i>, and Characterization of Its Gene Product
この論文をさがす
抄録
The first thermophilic α-oxoamine synthase family enzyme was identified. The gene (ORF TTHA1582), which is annotated to code putative α-oxoamine synthase family enzymes, 7-keto-8-aminopelargonic acid (KAPA) synthase (BioF, 8-amino-7-oxononanoate synthase, EC 2.3.1.47) and 2-amino-3-ketobutyrate CoA ligase (KBL, EC 2.3.1.29), in a genomic database, was cloned from an extreme thermophile, Thermus thermophilus, and overexpressed in Escherichia coli. The recombinant TTHA1582 protein was purified and characterized. It exhibited activity of BioF, which catalyzes the condensation of pimeloyl-CoA and L-alanine to produce a biotin intermediate KAPA, CoASH, and CO2 with pyridoxal 5′-phosphate as a cofactor. The protein is a dimer with a subunit of 43 kDa that shows an amino acid sequence identity of 35% with E. coli BioF. The optimum temperature and pH were about 70 °C and about 6.0. The enzyme showed high thermostability at temperatures of up to 70 °C for 1 h, and a half-life of 1 h at 80 °C. Thus the TTHA1582 protein was found to have the highest optimum temperature and thermostablility of the α-oxoamine synthase family enzymes so far reported. Substrate specificity experiments revealed that it was also able to catalyze the KBL reaction, which used acetyl-CoA and glycine as substrates, and that enzyme activity was seen with the following combinations of substrates: acetyl-CoA and glycine, L-alanine, or L-serine; pimeloyl-CoA and L-alanine, glycine, or L-serine; palmitoyl-CoA and L-alanine. This suggests that the recombinant TTHA1582 protein has broad substrate specificity, unlike the reported mesophilic enzymes of the α-oxoamine synthase family.
収録刊行物
-
- Bioscience, Biotechnology, and Biochemistry
-
Bioscience, Biotechnology, and Biochemistry 71 (12), 3033-3040, 2007
公益社団法人 日本農芸化学会
- Tweet
キーワード
詳細情報 詳細情報について
-
- CRID
- 1390001206478023680
-
- NII論文ID
- 10027522246
-
- NII書誌ID
- AA10824164
-
- ISSN
- 13476947
- 09168451
-
- NDL書誌ID
- 9327018
-
- 本文言語コード
- en
-
- データソース種別
-
- JaLC
- NDL
- Crossref
- CiNii Articles
-
- 抄録ライセンスフラグ
- 使用不可