title: Listeria monocytogenes has a functional chitinolytic system and an active lytic polysaccharide monooxygenase.
abstract: Chitinases and chitin-active lytic polysaccharide monooxygenases (LPMOs) are most commonly associated with chitin metabolism, but are also reported as virulence factors in pathogenic bacteria. Listeria monocytogenes, a well-known virulent bacterium, possesses two chitinases (ChiA and ChiB) and a multi-modular lytic polysaccharide monooxygenase (LmLPMO10). These enzymes have been related to virulence and their role in chitin metabolism is poorly understood. It is thus of interest to functionally characterize the individual enzymes in order to shed light on their roles in vivo. Our results demonstrate that L. monocytogenes has a fully functional chitinolytic system. Both chitinases show substrate degradation rates similar to those of the nonprocessive endo-chitinase SmChiC from Serratia marcescens. Compared to the S. marcescens LPMO chitin-binding protein CBP21, LmLPMO10 shows a similar rate but different product profiles depending on the substrate. In LPMO-chitinase synergy experiments, CBP21 is able to boost the activity of both ChiA and ChiB more than LmLPMO10. Product analysis of the synergy assays revealed that the chitinases were unable to efficiently hydrolyse the LPMO products (chitooligosaccharide aldonic acids) with a degree of polymerization below four (ChiA and SmChiC) or three (ChiB). Gene transcription and protein expression analysis showed that LmLPMO10 is neither highly transcribed, nor abundantly secreted during the growth of L. monocytogenes in a chitin-containing medium. The chitinases on the other hand are both abundantly secreted in the presence of chitin. Although LmLPMO10 is shown to promote chitin degradation in tandem with the chitinases in vitro, the secretome and transcription data question whether this is the primary role of LmLPMO10 in vivo.
authors.affiliation: Department of Veterinary Disease Biology, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark.
authors.forename: Dafni K
authors.initials: DK
authors.lastname: Paspaliari
authors.forename: Jennifer S M
authors.initials: JS
authors.lastname: Loose
authors.forename: Marianne H
authors.initials: MH
authors.lastname: Larsen
authors.forename: Gustav
authors.initials: G
authors.lastname: Vaaje-Kolstad
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism; D020134:Catalytic Domain; D002686:Chitin / Q000378:metabolism; D002688:Chitinases / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D006868:Hydrolysis; D008089:Listeria monocytogenes / Q000235:genetics / Q000254:growth & development / Q000378:metabolism* / Q000472:pathogenicity; D006899:Mixed Function Oxygenases / Q000235:genetics / Q000378:metabolism*; D011134:Polysaccharides / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D037521:Virulence Factors / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D011134:Polysaccharides; D037521:Virulence Factors; D002686:Chitin; D006899:Mixed Function Oxygenases; D002688:Chitinases
keywords: AA10; CBM33; Listeria monocytogenes; chitin; chitinase