title: How the rice weevil breaks down the pectin network: Enzymatic synergism and sub-functionalization.
abstract: Pectin is the most complex polysaccharide in nature and highly abundant in plant cell walls and middle lamellae, where it functions in plant growth and development. Phytopathogens utilize plant pectin as an energy source through enzyme-mediated degradation. These pectolytic enzymes include polygalacturonases (PGs) of the GH28 family and pectin methylesterases (PMEs) of the CE8 family. Recently, PGs were also identified in herbivorous insects of the distantly related plant bug, stick insect and Phytophaga beetle lineages. Unlike all other insects, weevils possess PMEs in addition to PGs. To investigate pectin digestion in insects and the role of PMEs in weevils, all PME and PG family members of the rice weevil Sitophilus oryzae were heterologously expressed and functionally characterized. Enzymatically active and inactive PG and PME family members were identified. The loss of activity can be explained by a lack of substrate binding correlating with substitutions of functionally important amino acid residues. We found subfunctionalization in both enzyme families, supported by expression pattern and substrate specificities as well as evidence for synergistic pectin breakdown. Our data suggest that the rice weevil might be able to use pectin as an energy source, and illustrates the potential of both PG and PME enzyme families to functionally diversify after horizontal gene transfer.
authors.affiliation: Department of Entomology, Max Planck Institute for Chemical Ecology, Hans-Knoell-Str. 8, Jena, 07745, Germany. Electronic address: rkirsch@ice.mpg.de.
authors.forename: Roy
authors.initials: R
authors.lastname: Kirsch
authors.affiliation: Department of Entomology, Max Planck Institute for Chemical Ecology, Hans-Knoell-Str. 8, Jena, 07745, Germany.
authors.forename: David G
authors.initials: DG
authors.lastname: Heckel
authors.affiliation: Department of Entomology, Max Planck Institute for Chemical Ecology, Hans-Knoell-Str. 8, Jena, 07745, Germany. Electronic address: ypauchet@ice.mpg.de.
authors.forename: Yannick
authors.initials: Y
authors.lastname: Pauchet
mesh_terms: D000818:Animals; D002265:Carboxylic Ester Hydrolases / Q000235:genetics / Q000378:metabolism*; D019476:Insect Proteins / Q000235:genetics / Q000378:metabolism*; D005810:Multigene Family; D012275:Oryza / Q000378:metabolism / Q000469:parasitology*; D010368:Pectins / Q000378:metabolism*; D010802:Phylogeny; D011096:Polygalacturonase / Q000235:genetics / Q000378:metabolism*; D048889:Weevils / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*
chemical_list: D019476:Insect Proteins; D010368:Pectins; D002265:Carboxylic Ester Hydrolases; C022323:pectinesterase; D011096:Polygalacturonase
keywords: Horizontal gene transfer; Pectin methylesterase; Plant cell wall degradation; Polygalacturonase; Weevils