초록
<P><B>Abstract</B></P> <P>Pectinases represent about one fifth of the enzyme worldwide market due their wide range of biotechnological applications. Current commercial pectinases are exclusively obtained from microbial sources, but here we report a pectin methylesterase (<I>Sl</I>-PME) and an <I>endo</I>-polygalacturonase (<I>Sl</I>-EPG) bioprospected from the sugarcane weevil, <I>Sphenophorus levis</I>, which revealed good potential for industrial applications. <I>Sl</I>-PME and <I>Sl</I>-EPG were overexpressed in <I>Pichia pastoris</I>, purified and enzymatically characterized. <I>Sl</I>-EPG presents optimal activity at pH 4–5 and 50 °C, showing that it can be used for juice extraction and clarification. On the other hand, <I>Sl</I>-PME presents optimal activity at pH 6–8 and 40 °C, and thus, suitable for both acidic and alkaline processing, such as coffee and tea fermentation. <I>Sl</I>-EPG shows V<SUB> <I>max</I> </SUB> = 3.23 mM/min, K<SUB> <I>M</I> </SUB> = 2.4 g/L and k<SUB> <I>cat</I> </SUB> = 418.6 s<SUP>−1</SUP>. While <I>Sl</I>-PME shows V<SUB> <I>max</I> </SUB> = 0.14 mM/min, K<SUB> <I>M</I> </SUB> = 4.1 g/L and k<SUB> <I>cat</I> </SUB> = 1.7 s<SUP>−1</SUP>. A PG inhibitor (PGIP2) weakly interfered in the <I>Sl</I>-EPG activity and <I>Sl</I>-PME was not affected by a usual PME inhibitor. Moreover, these enzymes manifested synergistic action towards methylesterified pectin. Here, we propose these enzymes as novel alternative tools for the current commercial pectinases.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Pectinases represent about one fifth of the enzymes in the worldwide market. </LI> <LI> Currently, commercial pectinases are exclusively from microbial sources. </LI> <LI> Could insect pectinases also be appropriate candidates for industrial pectin processing? </LI> <LI> Here, the pectinases from the Sugarcane Weevil were over expressed, purified and biochemically characterized. </LI> <LI> Our data suggest these enzymes as innovative alternative to the current commercial pectinases from microbial sources. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>