title: Genomic, proteomic and biochemical analysis of the chitinolytic machinery of Serratia marcescens BJL200.
abstract: The chitinolytic machinery of Serratia marcescens BJL200 has been studied in detail over the last couple of decades, however, the proteome secreted by this Gram-negative bacterium during growth on chitin has not been studied in depth. In addition, the genome of this most studied chitinolytic Serratia strain has until now, not been sequenced. We report a draft genome sequence for S. marcescens BJL200. Using label-free quantification (LFQ) proteomics and a recently developed plate-method for assessing secretomes during growth on solid substrates, we find that, as expected, the chitin-active enzymes (ChiA, B, C, and CBP21) are produced in high amounts when the bacterium grows on chitin. Other proteins produced in high amounts after bacterial growth on chitin provide interesting targets for further exploration of the proteins involved in degradation of chitin-rich biomasses. The genome encodes a fourth chitinase (ChiD), which is produced in low amounts during growth on chitin. Studies of chitin degradation with mixtures of recombinantly produced chitin-degrading enzymes showed that ChiD does not contribute to the overall efficiency of the process. ChiD is capable of converting N,N'-diacetyl chitobiose to N-acetyl glucosamine, but is less efficient than another enzyme produced for this purpose, the Chitobiase. Thus, the role of ChiD in chitin degradation, if any, remains unclear.
authors.affiliation: Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences (NMBU), P.O. Box 5003, 1432 Ås, Norway.
authors.forename: Tina R
authors.initials: TR
authors.lastname: Tuveng
authors.affiliation: Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences (NMBU), P.O. Box 5003, 1432 Ås, Norway.
authors.forename: Live Heldal
authors.initials: LH
authors.lastname: Hagen
authors.affiliation: Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences (NMBU), P.O. Box 5003, 1432 Ås, Norway.
authors.forename: Sophanit
authors.initials: S
authors.lastname: Mekasha
authors.affiliation: Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences (NMBU), P.O. Box 5003, 1432 Ås, Norway.
authors.forename: Jeremy
authors.initials: J
authors.lastname: Frank
authors.affiliation: Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences (NMBU), P.O. Box 5003, 1432 Ås, Norway.
authors.forename: Magnus Øverlie
authors.initials: MØ
authors.lastname: Arntzen
authors.affiliation: Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences (NMBU), P.O. Box 5003, 1432 Ås, Norway.
authors.forename: Gustav
authors.initials: G
authors.lastname: Vaaje-Kolstad
authors.affiliation: Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences (NMBU), P.O. Box 5003, 1432 Ås, Norway. Electronic address: vincent.eijsink@nmbu.no.
authors.forename: Vincent G H
authors.initials: VGH
authors.lastname: Eijsink
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002686:Chitin / Q000378:metabolism*; D020543:Proteome / Q000235:genetics / Q000378:metabolism; D040901:Proteomics; D012706:Serratia marcescens / Q000201:enzymology / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D020543:Proteome; D002686:Chitin
keywords: Biomass degradation; Chitinases; Proteomics; Secretome; Serratia marcescens