Deletion of uncharacterized domain from alpha-1,3-glucanase of Bacillus circulans KA-304 enhances heterologous enzyme production in Escherichia coli
-
- Yano Shigekazu
- Department of Biochemical Engineering, Graduate School of Sciences and Engineering, Yamagata University
-
- Suyotha Wasana
- Department of Industrial Biotechnology, Faculty of Agro-industry, Prince of Songkla University
-
- Zanma Sumika
- Department of Biochemical Engineering, Graduate School of Sciences and Engineering, Yamagata University
-
- Konno Hiroyuki
- Department of Biochemical Engineering, Graduate School of Sciences and Engineering, Yamagata University
-
- Cherdvorapong Vipavee
- Department of Biotechnology, Faculty of Life Sciences, Ritsumeikan University
-
- Wakayama Mamoru
- Department of Biotechnology, Faculty of Life Sciences, Ritsumeikan University
書誌事項
- タイトル別名
-
- Deletion of uncharacterized domain from α-1,3-glucanase of <i>Bacillus circulans</i> KA-304 enhances heterologous enzyme production in <i>Escherichia coli</i>
- Deletion of uncharacterized domain from α-1,3-glucanase of Bacillus circulans KA-304 enhances heterologous enzyme production in Escherichia coli
この論文をさがす
抄録
<p>α-1,3-Glucanase (Agl-KA) of Bacillus circulans KA-304 consists of an N-terminal discoidin domain (DS1), a carbohydrate binding module family 6 (CBM6), threonine and proline repeats (TP), a second discoidin domain (DS2), an uncharacterized conserved domain (UCD), and a C-terminal catalytic domain. Previously, we reported that DS1, CBM6, and DS2 have α-1,3-glucan-binding activity and contribute to α-1,3-glucan hydrolysis. In this study, UCD deletion mutant (AglΔUCD) was constructed, and its properties were compared with those of Agl-KA. α-1,3-Glucan hydrolyzing, α-1,3-glucan binding, and protoplast-forming activities of AglΔUCD were almost the same as those of Agl-KA. kcat/Km values of AgΔUCD and Agl-KA were 11.4 and 11.1 s–1 mg–1 mL, respectively. AglΔUCD and Agl-KA exhibited similar characteristics, such as optimal pH, pH stability, optimal temperature, and thermostability. These results suggest that UCD is not α-1,3-glucan-binding and flexible linker domain, and that deletion of UCD does not affect the affinity of N-terminal binding domains and the catalytic action of the C-terminal domain. Subsequently, heterologous UCenzyme productivity of AglΔD in Escherichia coli was compared with that of Agl-KA. The productivity of AglΔUCD was about 4-fold larger than that of Agl-KA after an 8-h induction at 30°C. In the case of induction at 20°C, the productivity of AglΔUCD was also larger than that of Agl-KA. These findings indicate that deletion of only UCD enhances the enzyme productivity in E. coli.</p>
収録刊行物
-
- The Journal of General and Applied Microbiology
-
The Journal of General and Applied Microbiology 64 (5), 212-220, 2018
公益財団法人 応用微生物学・分子細胞生物学研究奨励会