Identification of hyperthermophilic D-allulose 3-epimerase from Thermotoga sp. and its application as a high-performance biocatalyst for D-allulose synthesis

IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Bioprocess and Biosystems Engineering Pub Date : 2024-04-27 DOI:10.1007/s00449-024-02989-3
Ji-Dong Shen, Bao-Ping Xu, Te-Li Yu, Yong-Xiang Fei, Xue Cai, Liang-Gang Huang, Li-Qun Jin, Zhi-Qiang Liu, Yu-Guo Zheng
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Abstract

D-Allulose 3-epimerase (DAE) is a vital biocatalyst for the industrial synthesis of D-allulose, an ultra-low calorie rare sugar. However, limited thermostability of DAEs hinders their use at high-temperature production. In this research, hyperthermophilic TI-DAE (Tm = 98.4 ± 0.7 ℃) from Thermotoga sp. was identified via in silico screening. A comparative study of the structure and function of site-directed saturation mutagenesis mutants pinpointed the residue I100 as pivotal in maintaining the high-temperature activity and thermostability of TI-DAE. Employing TI-DAE as a biocatalyst, D-allulose was produced from D-fructose with a conversion rate of 32.5%. Moreover, TI-DAE demonstrated excellent catalytic synergy with glucose isomerase CAGI, enabling the one-step conversion of D-glucose to D-allulose with a conversion rate of 21.6%. This study offers a promising resource for the enzyme engineering of DAEs and a high-performance biocatalyst for industrial D-allulose production.

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嗜热菌 D-allulose 3-epimerase 的鉴定及其在 D-allulose 合成中作为高性能生物催化剂的应用
D- Allulose 3-epimerase(DAE)是工业合成超低卡路里稀有糖类 D- Allulose 的重要生物催化剂。然而,DAE 的热稳定性有限,阻碍了其在高温生产中的应用。在这项研究中,通过硅学筛选,从Thermotoga sp.中鉴定出了超嗜热TI-DAE(Tm = 98.4 ± 0.7 ℃)。通过对定点饱和突变突变体的结构和功能进行比较研究,发现残基 I100 是维持 TI-DAE 高温活性和耐热性的关键。使用 TI-DAE 作为生物催化剂,可以从 D-果糖生产出 D-阿洛酮糖,转化率高达 32.5%。此外,TI-DAE 与葡萄糖异构酶 CAGI 表现出了极佳的催化协同作用,可将 D-葡萄糖一步转化为 D-阿洛糖,转化率为 21.6%。这项研究为 DAE 的酶工程提供了一种前景广阔的资源,也为工业化生产 D-阿洛糖提供了一种高性能生物催化剂。
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来源期刊
Bioprocess and Biosystems Engineering
Bioprocess and Biosystems Engineering 工程技术-工程:化工
CiteScore
7.90
自引率
2.60%
发文量
147
审稿时长
2.6 months
期刊介绍: Bioprocess and Biosystems Engineering provides an international peer-reviewed forum to facilitate the discussion between engineering and biological science to find efficient solutions in the development and improvement of bioprocesses. The aim of the journal is to focus more attention on the multidisciplinary approaches for integrative bioprocess design. Of special interest are the rational manipulation of biosystems through metabolic engineering techniques to provide new biocatalysts as well as the model based design of bioprocesses (up-stream processing, bioreactor operation and downstream processing) that will lead to new and sustainable production processes. Contributions are targeted at new approaches for rational and evolutive design of cellular systems by taking into account the environment and constraints of technical production processes, integration of recombinant technology and process design, as well as new hybrid intersections such as bioinformatics and process systems engineering. Manuscripts concerning the design, simulation, experimental validation, control, and economic as well as ecological evaluation of novel processes using biosystems or parts thereof (e.g., enzymes, microorganisms, mammalian cells, plant cells, or tissue), their related products, or technical devices are also encouraged. The Editors will consider papers for publication based on novelty, their impact on biotechnological production and their contribution to the advancement of bioprocess and biosystems engineering science. Submission of papers dealing with routine aspects of bioprocess engineering (e.g., routine application of established methodologies, and description of established equipment) are discouraged.
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