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Efficient conversion of xylan to l-arabinose by multi-enzymatic cascade reaction including d-xylulose 4-epimerase as a new stereoselectivity-exchange enzyme

[Display omitted] •d-Xylulose 4-epimerase (DX4E) was engineered from d-tagaturonate 3-epimerase.•The kcat/Km of DX4E for xylulose increased 11-fold compared to the wild-type.•The equilibrium of d-xylose isomerase was overcome by sugar kinase and phosphatase.•An efficient pathway for converting d-xyl...

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Bibliographic Details
Published in:Bioresource technology 2024-12, Vol.413, p.131502, Article 131502
Main Authors: Lee, Tae-Eui, Shin, Kyung-Chul, Oh, Deok-Kun
Format: Article
Language:English
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Summary:[Display omitted] •d-Xylulose 4-epimerase (DX4E) was engineered from d-tagaturonate 3-epimerase.•The kcat/Km of DX4E for xylulose increased 11-fold compared to the wild-type.•The equilibrium of d-xylose isomerase was overcome by sugar kinase and phosphatase.•An efficient pathway for converting d-xylose to l-arabinose was established.•l-Arabinose is produced from xylan via multi-enzymatic cascade reactions. l-Arabinose has been produced by hydrolyzing arabinan, a component of hemicellulose. However, l-arabinose has limitations in industrial applications owing to its relatively high cost. Here, d-xylulose 4-epimerase as a new-type enzyme was developed from d-tagaturonate 3-epimerase from Thermotoga petrophila using structure-guided enzyme engineering. d-Xylulose 4-epimerase, which epimerized d-xylulose to l-ribulose, d-xylulokinase and sugar phosphatase, which overcame the equilibrium of d-xylose isomerase, were included to establish a new efficient conversion pathway from d-xylose to l-arabinose. l-Arabinose at 34 g/L was produced from 100 g/L xylan in 45 h by multi-enzymatic cascade reaction using xylanase and enzymes involved in the established conversion pathway. As l-ribulokinase was used instead of d-xylulokinase in the established conversion pathway, an efficient reverse-directed conversion pathway from l-arabinose to d-xylose and the production of d-xylose from arabinan using arabinanase and enzymes involved in the proposed pathway are proposed.
ISSN:0960-8524
1873-2976
1873-2976
DOI:10.1016/j.biortech.2024.131502