Institut National de la Recherche Scientifique (INRS), EMT Research Center, Varennes, Quebec, Canada.
Biotechnol Bioeng. 2020 Mar;117(3):868-872. doi: 10.1002/bit.27229. Epub 2019 Dec 2.
Screening libraries of mutant proteins by phage display is now relatively common. However, one unknown factor is how the bacteriophage scaffold itself influences the properties of the displayed protein. This communication evaluates the effect of solution parameters on the catalytic activity of phage displayed Bacillus subtilis Lipase A (BSLA), compared to the free enzyme in solution. While the pH- and temperature-activity profiles of BSLA were not intrinsically affected by phage display, the nanoscale distribution of BSLA within the micellar assay buffer was. This lead to a pronounced increase of activity of phage-BSLA relative to the free enzyme, owing to the accumulation of phage-BSLA at the substrate-rich micelles. Considering this result obtained for BSLA, caution is warranted and similar effects should be considered when selecting other enzymes/proteins by phage display, as the activity of the displayed protein may differ from that of the free protein.
通过噬菌体展示筛选突变蛋白现在已经相对普遍。然而，有一个未知因素是噬菌体支架本身如何影响所展示蛋白质的性质。本通讯评估了溶液参数对噬菌体展示枯草芽孢杆菌脂肪酶 A（BSLA）的催化活性的影响，与溶液中的游离酶进行了比较。虽然 pH 和温度对 BSLA 的活性曲线没有内在影响，但 BSLA 在胶束测定缓冲液中的纳米级分布受到了影响。这导致噬菌体-BSLA 的活性相对于游离酶显著增加，这是由于噬菌体-BSLA在富含底物的胶束处聚集所致。考虑到BSLA 得到的这个结果，在通过噬菌体展示选择其他酶/蛋白质时需要谨慎，因为所展示蛋白质的活性可能与游离蛋白质不同。
