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Effect of particle size and enzyme load on the simultaneous reactions of lactose hydrolysis and transgalactosylation with glyoxyl-agarose immobilized β-galactosidase from Aspergillus oryzae

机译:粒径和酶负荷对米曲霉乳糖固定的β-半乳糖苷酶与乳糖水解和反式半乳糖同时反应的影响

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摘要

The simultaneous reactions of lactose hydrolysis and transgalactosylation in the production of galacto-oligosaccharides (GOS) were evaluated with immobilizedAspergillus oryzaeβ-galactosidase in glyoxyl-agarose of different particle sizes (fine and macro) and enzyme loads (1, 10 and 30 mgprotein/gsupport) to produce biocatalysts subjected to different magnitudes of internal diffusional restrictions. The ratio of initial reaction rates depended on the initial concentration of lactose: at values higher than 800 mM, the ratio of hydrolysis to total reaction rate(rhydr/rtotal), which reflects the hydrolytic potential, had values of 16 and 30%, and the ratio of transgalactosylation to total reaction rates(rtransgal/rtotal), which reflects the transgalactosylation potential, had values of 84 and 70% with the biocatalysts of smaller size and enzyme load, and bigger size and enzyme load, respectively. Results obtained highlight the fact that the biocatalyst should be optimized with regard to its intended use; β-galactosidase biocatalysts that have been optimized for lactose hydrolysis can be quite inadequate for performing GOS synthesis.
机译:用固定化米曲霉β-半乳糖苷酶在不同粒径(细颗粒和大颗粒)和酶负荷(1、10和30 mg蛋白/ g支持物)的固定化米曲霉β-半乳糖苷酶中评估乳糖水解和半乳糖基化在生产半乳糖寡糖(GOS)中的同时反应)以产生受到不同程度的内部扩散限制的生物催化剂。初始反应速率的比率取决于乳糖的初始浓度:在高于800µmM的值时,反映水解潜力的水解与总反应速率的比率(rhydr / rtotal)分别为16%和30%,并且反式半乳糖基化反应与总反应速率之比(rtransgal / rtotal)反映了反式半乳糖基化反应的潜力,其大小分别为84和70%,生物催化剂的尺寸和酶负载较小。获得的结果突出了这样一个事实,即应根据其预期用途优化生物催化剂。针对乳糖水解进行了优化的β-半乳糖苷酶生物催化剂可能不足以进行GOS合成。

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