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Preparation of combined cross-linked enzyme aggregates containing galactitol dehydrogenase and NADH oxidase for l-tagatose synthesis via in situ cofactor regeneration

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Abstract

The combined cross-linked enzyme aggregates (combi-CLEAs) containing galactitol dehydrogenase (Gdh) and NADH oxidase (Nox) were prepared for l-tagatose synthesis. To prevent the excess consumption of cofactor, Nox in the combi-CLEAs was used to in situ regenerate NAD+. In the immobilization process, ammonia sulfate and glutaraldehyde were used as the precipitant and cross-linking reagent, respectively. The preparation conditions were optimized as follows: 60% ammonium sulfate, 1:1 (molar ratio) of Gdh to Nox, 20:1 (molar ratio) of protein to glutaraldehyde, and 6 h of cross-linking time at 35 °C. Under these conditions, the activity of the combi-CLEAs was 210 U g−1. The combi-CLEAs exhibited higher thermostability and preserved 51.5% of the original activity after eight cycles of reuses at 45 °C. The combi-CLEAs were utilized for the preparation of l-tagatose without by-products. Therefore, the combi-CLEAs have the industrial potential for the bioconversion of galactitol to l-tagatose.

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Data availability

The datasets used or analyzed in this study are available from the corresponding author upon reasonable request.

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Funding

The authors appreciated the financial support from the Natural Science Foundation of Guangxi Province (No. 2019GXNSFAA185059).

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Authors and Affiliations

  1. School of Pharmacy, Jiangsu University, Zhenjiang, 212013, People’s Republic of China

    Xue-Yong Li, Meng-Qiu Xu, Hui Liu, Qiang Zhou & Ye-Wang Zhang

  2. College of Petroleum and Chemical Engineering, Qinzhou, 535100, People’s Republic of China

    Jian Gao

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  1. Xue-Yong Li
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Contributions

Design of experiments: Y-WZ, X-YL, and M-QX; performance of experiment and drawing of graphs: M-QX, X-YL, and HL; manuscript draft and compilation: X-YL and Y-WZ; correction and revision of the manuscript: Y-WZ, QZ, and JG.

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Correspondence to Ye-Wang Zhang.

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Li, XY., Xu, MQ., Liu, H. et al. Preparation of combined cross-linked enzyme aggregates containing galactitol dehydrogenase and NADH oxidase for l-tagatose synthesis via in situ cofactor regeneration. Bioprocess Biosyst Eng 45, 353–364 (2022). https://doi.org/10.1007/s00449-021-02665-w

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