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- Title
Enhancing lactose recognition of a key enzyme in 2'-fucosyllactose synthesis: α-1,2-fucosyltransferase.
- Authors
Wenxian Liu; Shizhe Tang; Jing Peng; Yuling Zhu; Lina Pan; Jiaqi Wang; Xiaoyu Peng; Haina Cheng; Zhu Chen; Yuguang Wang; Hongbo Zhou
- Abstract
BACKGROUND: 2'-Fucosyllactose, a representative oligosaccharide in human milk, is an emerging and promising food and pharmaceutical ingredient due to its powerful health benefits, such as participating in immune regulation, regulation of intestinal flora, etc. To enable economically viable production of 2'-fucosyllactose, different biosynthesis strategies using precursors and pathway enzymes have been developed. The α-1,2-fucosyltransferases are an essential part involved in these strategies, but their strict substrate selectivity and unsatisfactory substrate tolerance are one of the key roadblocks limiting biosynthesis. RESULTS: To tackle this issue, a semi-rational manipulation combining computer-aided designing and screening with biochemical experiments were adopted. The mutant had a 100-fold increase in catalytic efficiency compared to the wild-type. The highest 2'-fucosyllactose yield was up to 0.65 mol mol-1 lactose with a productivity of 2.56 g mL-1 h-1 performed by enzymatic catalysis in vitro. Further analysis revealed that the interactions between the mutant and substrates were reduced. The crucial contributions of wild-type and mutant to substrate recognition ability were closely related to their distinct phylotypes in terms of amino acid preference. CONCLUSION: It is envisioned that the engineered α-1,2-fucosyltransferase could be harnessed to relieve constraints imposed on the bioproduction of 2'-fucosyllactose and lay a theoretical foundation for elucidating the substrate recognition mechanisms of fucosyltransferases.
- Subjects
LACTOSE; BREAST milk; RECOGNITION (Psychology); FUCOSYLTRANSFERASES; ENZYMES; BIOSYNTHESIS
- Publication
Journal of the Science of Food & Agriculture, 2023, Vol 103, Issue 3, p1303
- ISSN
0022-5142
- Publication type
Article
- DOI
10.1002/jsfa.12224