| 초록 |
2'-Fucosyllactose(2’-FL) accounts for the largest portion of Human Milk Oligosaccharide compounds, which are functional materialswith prebiotic effects. Compared to its industrial importance, the efficiency of the current 2'-FL production enzyme, α-1,2-fucosyltransferase, is low. Directed evolution through random mutagenesis is a powerful tool forenhancing enzyme properties, but it takes a lot of time and money without asimple screening method. Here we report High Throughput screening system, whole cell colorimetric analysis method for measure the activity of mutant library. Calfintestine alkaline phosphatase(CIAP) separated GDP generated by the reactionof α-1,2-fucosyltransferase to single phosphate and GMP, and the amount of single phosphate was detected with Malachite Green. Through comparison with HPLC, it was confirmed that the activity of α-1,2-fucosyltransferase can be measured by this method. |
