Accessory active site residues of Streptomyces sp. N174 chitosanase: variations on a common theme in the lysozyme superfamily

The FEBS Journal
Marie-Eve Lacombe-HarveyRyszard Brzezinski

Abstract

The chitosanase from Streptomyces sp. N174 (CsnN174) is an inverting glycoside hydrolase belonging to family 46. Previous studies identified Asp40 as the general base residue. Mutation of Asp40 into glycine revealed an unexpectedly high residual activity. D40G mutation did not affect the stereochemical mechanism of catalysis or the mode of interaction with substrate. To explain the D40G residual activity, putative accessory catalytic residues were examined. Mutation of Glu36 was highly deleterious in a D40G background. Possibly, the D40G mutation reconfigured the catalytic center in a way that allowed Glu36 to be positioned favorably to perform catalysis. Thr45 was also found to be essential. Thr45 is thought to orientate the nucleophilic water molecule in a position to attack the glycosidic link. The finding that expression of heterologous CsnN174 in Escherichia coli protects cells against the antimicrobial effect of chitosan, allowed the selection of active chitosanase variants after saturation mutagenesis. Thr45 could be replaced only by serine, indicating the importance of the hydroxyl group. The newly identified accessory catalytic residues, Glu36 and Thr45 are located on a three-strand beta sheet highly conserved in GH19,...Continue Reading

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Citations

Nov 26, 2010·Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire·Mariana Gabriela GhinetRyszard Brzezinski
Jun 19, 2010·Marine Drugs·Berit B AamVincent G H Eijsink
Nov 1, 2015·Marine Drugs·Pascal ViensRyszard Brzezinski
Apr 29, 2014·The Biochemical Journal·Qianqian LyuWeizhi Liu
Jul 7, 2015·Biochimica Et Biophysica Acta·Qianqian LyuWeizhi Liu
Nov 5, 2016·Applied Biochemistry and Biotechnology·Zhanping ZhouJiangning Song
Sep 20, 2017·Applied Microbiology and Biotechnology·Mina ZitouniRyszard Brzezinski
May 26, 2018·Glycobiology·Yuichi NishitaniKunio Miki
Jun 23, 2018·Applied Microbiology and Biotechnology·Marie-Ève Lacombe-HarveyCarole Beaulieu
Oct 28, 2020·International Journal of Molecular Sciences·Martin BoninBruno M Moerschbacher
Aug 11, 2011·Bioengineered Bugs·Ryszard Brzezinski
Sep 14, 2010·Biomacromolecules·Ellinor B HeggsetKjell M Vårum
Jan 5, 2021·Journal of Agricultural and Food Chemistry·Bhoopal BhuvanachandraAppa Rao Podile

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