Lowering energy consumption for fermentable sugar production from Ramulus mori: Engineered xylanase synergy and improved pretreatment strategy

Engineering of xylanases for the development of biotechnologically important characteristics

“…In xylanase engineering with multiple methods, epPCR has been commonly performed to increase the xylanase gene diversity at first. The xylanase variants with a higher improved trait have been selected for the application of the next method(s) (Damis et al, 2019;Xing et al, 2021;You et al, 2021You et al, , 2022. For instance, Bispora sp.…”

“…MEY‐1 GH10 xylanase, which was truncated N‐terminal (You et al, 2019), has been randomly mutagenized by epPCR. Among these, two variants I133E and Q143M have had higher activity than the WT and thus these substitutions have been then saturated to further develop their catalytic activity (You et al, 2022). In some works, the existence of common mutations in more than one mutant can be a criterion in the selection of the variants for the application of the next methods.…”

“…Recent investigations have reported new strategies with the site‐saturation mutagenesis method, which has been applied alone or together with the other methods. If site‐saturation mutagenesis was applied after the directed evolution method (e.g., epPCR), only sequence data derived from epPCR can be sufficient to determine the residues of the xylanase for saturation (You et al, 2022). Even so, sometimes, computational tools could be mostly helpful for the prediction of the effect of the mutations produced by several rounds of multiple methods.…”