title: Characterization of genes for chitin catabolism in Haloferax mediterranei.
abstract: Chitin is the second most abundant natural polysaccharide after cellulose. But degradation of chitin has never been reported in haloarchaea. In this study, we revealed that Haloferax mediterranei, a metabolically versatile haloarchaeon, could utilize colloidal or powdered chitin for growth and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) accumulation, and the gene cluster (HFX_5025-5039) for the chitin catabolism pathway was experimentally identified. First, reverse transcription polymerase chain reaction results showed that the expression of the genes encoding the four putative chitinases (ChiAHme, ChiBHme, ChiCHme, and ChiDHme, HFX_5036-5039), the LmbE-like deacetylase (DacHme, HFX_5027), and the glycosidase (GlyAHme, HFX_5029) was induced by colloidal or powdered chitin, and chiA Hme, chiB Hme, and chiC Hme were cotranscribed. Knockout of chiABC Hme or chiD Hme had a significant effect on cell growth and PHBV production when chitin was used as the sole carbon source, and the chiABCD Hme knockout mutant lost the capability to utilize chitin. Knockout of dac Hme or glyA Hme also decreased PHBV accumulation on chitin. These results suggested that ChiABCDHme, DacHme, and GlyAHme were indeed involved in chitin degradation in H. mediterranei. Additionally, the chitinase assay showed that each chitinase possessed hydrolytic activity toward colloidal or powdered chitin, and the major product of colloidal chitin hydrolysis by ChiABCDHme was diacetylchitobiose, which was likely further degraded to monosaccharides by DacHme, GlyAHme, and other related enzymes for both cell growth and PHBV biosynthesis. Taken together, this study revealed the genes and enzymes involved in chitin catabolism in haloarchaea for the first time and indicated the potential of H. mediterranei as a whole-cell biocatalyst in chitin bioconversion.
authors.affiliation: State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, People's Republic of China.
authors.forename: Jing
authors.initials: J
authors.lastname: Hou
authors.forename: Jing
authors.initials: J
authors.lastname: Han
authors.forename: Lei
authors.initials: L
authors.lastname: Cai
authors.forename: Jian
authors.initials: J
authors.lastname: Zhou
authors.forename: Yang
authors.initials: Y
authors.lastname: Lü
authors.forename: Cheng
authors.initials: C
authors.lastname: Jin
authors.forename: Jingfang
authors.initials: J
authors.lastname: Liu
authors.forename: Hua
authors.initials: H
authors.lastname: Xiang
mesh_terms: D001711:Biotransformation; D002244:Carbon / Q000378:metabolism; D002686:Chitin / Q000378:metabolism*; D020869:Gene Expression Profiling; D055786:Gene Knockout Techniques; D019623:Haloferax mediterranei / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D053858:Metabolic Networks and Pathways / Q000235:genetics*; D005810:Multigene Family; D011091:Polyesters / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000094122:Polyhydroxybutyrates
chemical_list: D002244:Carbon; D002686:Chitin; D011091:Polyesters; C052620:poly(3-hydroxybutyrate)-co-(3-hydroxyvalerate); D000094122:Polyhydroxybutyrates