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Metabolic Engineering of Fungal Strains for Conversion of d-Galacturonate to meso-Galactarate

机译:真菌菌株的代谢工程将d-半乳糖醛酸酯转化为内消旋半乳糖酸酯

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摘要

d-Galacturonic acid can be obtained by hydrolyzing pectin, which is an abundant and low value raw material. By means of metabolic engineering, we constructed fungal strains for the conversion of d-galacturonate to meso-galactarate (mucate). Galactarate has applications in food, cosmetics, and pharmaceuticals and as a platform chemical. In fungi d-galacturonate is catabolized through a reductive pathway with a d-galacturonate reductase as the first enzyme. Deleting the corresponding gene in the fungi Hypocrea jecorina and Aspergillus niger resulted in strains unable to grow on d-galacturonate. The genes of the pathway for d-galacturonate catabolism were upregulated in the presence of d-galacturonate in A. niger, even when the gene for d-galacturonate reductase was deleted, indicating that d-galacturonate itself is an inducer for the pathway. A bacterial gene coding for a d-galacturonate dehydrogenase catalyzing the NAD-dependent oxidation of d-galacturonate to galactarate was introduced to both strains with disrupted d-galacturonate catabolism. Both strains converted d-galacturonate to galactarate. The resulting H. jecorina strain produced galactarate at high yield. The A. niger strain regained the ability to grow on d-galacturonate when the d-galacturonate dehydrogenase was introduced, suggesting that it has a pathway for galactarate catabolism.
机译:d-半乳糖醛酸可通过水解果胶而获得,果胶是一种丰富而低价值的原料。通过代谢工程,我们构建了用于将d-半乳糖醛酸酯转化为半乳糖半乳糖(粘酸盐)的真菌菌株。半乳酸盐在食品,化妆品和药品中以及作为平台化学品都有应用。在真菌中,d-半乳糖醛酸酯通过还原途径分解代谢,其中以d-半乳糖醛酸酯还原酶为第一酶。删除真菌Hypocrea jecorina和黑曲霉中的相应基因导致菌株无法在d-半乳糖醛酸上生长。即使在d-半乳糖醛酸还原酶基因缺失的情况下,黑曲霉中d-半乳糖醛酸分解代谢途径的基因也被上调,表明d-半乳糖醛酸本身是该途径的诱导物。将编码d-半乳糖醛酸脱氢酶的细菌基因催化d-半乳糖醛酸分解代谢的NAD依赖氧化成半乳糖酸酯。两种菌株都将d-半乳糖醛酸酯转化为半乳糖酸酯。所得的红褐肉座菌菌株以高产率产生半乳酸盐。当引入d-半乳糖醛酸脱氢酶时,黑曲霉菌株恢复了在d-半乳糖醛酸上生长的能力,这表明其具有半乳糖酸酯分解代谢的途径。

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