Bibliographic Details
| Title: |
Biochemical characterization, structure-guided mutagenesis, and application of a recombinant D-allulose 3-epimerase from Christensenellaceae bacterium for the biocatalytic production of D-allulose |
| Authors: |
Lijun Guan, Ling Zhu, Kunlun Wang, Yang Gao, Jialei Li, Song Yan, Xindi Zhang, Nina Ji, Jing Fan, Ye Zhou, Xinmiao Yao, Bo Li |
| Source: |
Frontiers in Bioengineering and Biotechnology, Vol 12 (2024) |
| Publisher Information: |
Frontiers Media S.A., 2024. |
| Publication Year: |
2024 |
| Collection: |
LCC:Biotechnology |
| Subject Terms: |
D-allulose, D-allulose 3-epimerase, bioconversion, site-directed iteration mutagenesis, apple juice, Biotechnology, TP248.13-248.65 |
| More Details: |
D-Allulose has become a promising alternative sweetener due to its unique properties of low caloric content, moderate sweetness, and physiological effects. D-Allulose 3-epimerase (DAEase) is a promising enzyme for D-Allulose production. However, the low catalytic efficiency limited its large-scale industrial applications. To obtain a more effective biocatalyst, a putative DAEase from Christensenellaceae bacterium (CbDAE) was identified and characterized. The recombinant CbDAE exhibited optimum activity at pH 7.5°C and 55°C, retaining more than 60% relative activity from 40°C to 70°C, and the catalytic activity could be significantly increased by Co2+ supplementation. These enzymatic properties of purified CbDAE were compared with other DAEases. CbDAE was also found to possess desirable thermal stability at 55°C with a half-life of 12.4 h. CbDAE performed the highest relative activity towards D-allulose and strong affinity for D-fructose but relatively low catalytic efficiency towards D-fructose. Based on the structure-guided design, the best double-mutation variant G36N/W112E was obtained which reached up to 4.21-fold enhancement of catalytic activity compared with wild-type (WT) CbDAE. The catalytic production of G36N/W112E with 500 g/L D-fructose was at a medium to a higher level among the DAEases in 3.5 h, reducing 40% catalytic reaction time compared to the WT CbDAE. In addition, the G36N/W112E variant was also applied in honey and apple juice for D-allulose conversion. Our research offers an extra biocatalyst for D-allulose production, and the comprehensive report of this enzyme makes it potentially interesting for industrial applications and will aid the development of industrial biocatalysts for D-allulose. |
| Document Type: |
article |
| File Description: |
electronic resource |
| Language: |
English |
| ISSN: |
2296-4185 |
| Relation: |
https://www.frontiersin.org/articles/10.3389/fbioe.2024.1365814/full; https://doaj.org/toc/2296-4185 |
| DOI: |
10.3389/fbioe.2024.1365814 |
| Access URL: |
https://doaj.org/article/ba95ee8d2aa142e1a13391fa82a03729 |
| Accession Number: |
edsdoj.ba95ee8d2aa142e1a13391fa82a03729 |
| Database: |
Directory of Open Access Journals |
