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Characterization of a GH20 β-N-Acetylhexosaminidase from Flavobacterium algicola Suitable to Synthesize Lacto-N-triose II
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2024-02-22 , DOI: 10.1021/acs.jafc.3c07919
Chengqiang Li 1, 2, 3 , Zhuoning Cao 1, 2, 3 , Hong Jiang 1, 2, 3, 4 , Francesco Secundo 5 , Xiangzhao Mao 1, 2, 3, 4, 6
Affiliation  

β-N-Acetylhexosaminidases have attracted much attention in the enzymatic synthesis of lacto-N-triose II (LNT2) as a backbone precursor of human milk oligosaccharides (HMOs). In this study, a novel glycoside hydrolase (GH) 20 family β-N-acetylhexosaminidase, FlaNag2353, from Flavobacterium algicola was biochemically characterized and applied to synthesize LNT2. FlaNag2353 displayed optimal activity to p-nitrophenyl N-acetyl-β-d-glucosaminide (pNP-GlcNAc) at 40 °C and pH 8.0. In addition to its excellent hydrolysis activity toward pNP-GlcNAc and chitooligosaccharides, FlaNag2353 showed trans-glycosylation activity. Under conditions of pH 9.0 and 55 °C for 2 h and utilizing 200 mM lactose and 10 mM pNP-GlcNAc, FlaNag2353 synthesized LNT2 with a conversion ratio of 4.15% calculated from pNP-GlcNAc. Moreover, when applied to LNT2 synthesis with 10 mM pNP-GlcNAc and 9.7% (w/v) industrial waste whey powder, FlaNag2353 achieved a conversion ratio of 2.39%. This study has significant implications for broadening the applications of GH20 β-N-acetylhexosaminidases and promoting the high-value utilization of whey powder.

中文翻译:


适用于合成乳-N-丙糖 II 的海藻黄杆菌 GH20 β-N-乙酰己糖胺酶的表征



β- N-乙酰己糖胺酶作为人乳低聚糖(HMO)的主链前体,在乳糖-N-丙糖 II(LNT2)的酶促合成中引起了广泛关注。在本研究中,对来自黄杆菌的一种新型糖苷水解酶 (GH) 20 家族 β- N-乙酰己糖胺酶 FlaNag2353 进行了生化表征,并将其应用于合成 LNT2。 FlaNag2353 在 40 °C 和 pH 8.0 条件下对对硝基苯N-乙酰基-β- d-氨基葡萄糖 ( p NP-GlcNAc) 显示出最佳活性。 FlaNag2353 除了对p NP-GlcNAc 和壳寡糖具有优异的水解活性外,还表现出转糖基化活性。在pH 9.0和55℃条件下,利用200 mM乳糖和10 mM p NP-GlcNAc,2小时,FlaNag2353合成了LNT2,根据p NP-GlcNAc计算,转化率为4.15%。此外,当使用 10 mM p NP-GlcNAc 和 9.7% (w/v) 工业废乳清粉合成 LNT2 时,FlaNag2353 的转化率为 2.39%。该研究对于拓宽GH20β- N-乙酰氨基己糖苷酶的应用,促进乳清粉的高值化利用具有重要意义。
更新日期:2024-02-22
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