biostudies-literatureAl-Hinai TZSMinistry of Higher EducationBiotechnology and Biological Sciences Research Council547-558https://www.ebi.ac.uk/biostudies/studies/S-EPMC11037484biostudies-literatureUnknown133(4)<h4>Background and aims</h4>The softening of ripening fruit involves partial depolymerization of cell-wall pectin by three types of reaction: enzymic hydrolysis, enzymic elimination (lyase-catalysed) and non-enzymic oxidative scission. Two known lyase activities are pectate lyase and rhamnogalacturonan lyase (RGL), potentially causing mid-chain cleavage of homogalacturonan and rhamnogalacturonan-I (RG-I) domains of pectin respectively. However, the important biological question of whether RGL exhibits action in vivo had not been tested.<h4>Methods</h4>We developed a method for specifically and sensitively detecting in-vivo RGL products, based on Driselase digestion of cell walls and detection of a characteristic unsaturated 'fingerprint' product (tetrasaccharide) of RGL action.<h4>Key resuAnnals of botanyFruit softening: evidence for rhamnogalacturonan lyase action in vivo in ripe fruit cell walls.PMC11037484BB/N002458/1Mackay CLFry SCAl-Hinai TZSfalseFruit softening: evidence for rhamnogalacturonan lyase action in vivo in ripe fruit cell walls.<h4>Background and aims</h4>The softening of ripening fruit involves partial depolymerization of cell-wall pectin by three types of reaction: enzymic hydrolysis, enzymic elimination (lyase-catalysed) and non-enzymic oxidative scission. Two known lyase activities are pectate lyase and rhamnogalacturonan lyase (RGL), potentially causing mid-chain cleavage of homogalacturonan and rhamnogalacturonan-I (RG-I) domains of pectin respectively. However, the important biological question of whether RGL exhibits action in vivo had not been tested.<h4>Methods</h4>We developed a method for specifically and sensitively detecting in-vivo RGL products, based on Driselase digestion of cell walls and detection of a characteristic unsaturated 'fingerprint' product (tetrasaccharide) of RGL action.<h4>Key resu2024-01-01T00:00:00Z2024 Apr2025-04-26T23:12:36.54Z2025-04-06T17:31:15.582ZS-EPMC110374843818046010.1093/aob/mcad197