Characterization of a GH20 β-N-Acetylhexosaminidase from Flavobacterium algicola Suitable to Synthesize Lacto-N-triose II,Journal of Agricultural and Food Chemistry

当前位置： X-MOL 学术 › J. Agric. Food Chem. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

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 条件下对对硝基苯基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

点击分享查看原文

点击收藏

阅读更多本刊新发论文本刊介绍/投稿指南