title: Modification of genetic regulation of a heterologous chitosanase gene in Streptomyces lividans TK24 leads to chitosanase production in the absence of chitosan.
abstract: BACKGROUND
  Chitosanases are enzymes hydrolysing chitosan, a β-1,4 linked D-glucosamine bio-polymer. Chitosan oligosaccharides have numerous emerging applications and chitosanases can be used for industrial enzymatic hydrolysis of chitosan. These extracellular enzymes, produced by many organisms including fungi and bacteria, are well studied at the biochemical and enzymatic level but very few works were dedicated to the regulation of their gene expression. This is the first study on the genetic regulation of a heterologous chitosanase gene (csnN106) in Streptomyces lividans.
  
  
  RESULTS
  Two S. lividans strains were used for induction experiments: the wild type strain and its mutant (ΔcsnR), harbouring an in-frame deletion of the csnR gene, encoding a negative transcriptional regulator. Comparison of chitosanase levels in various media indicated that CsnR regulates negatively the expression of the heterologous chitosanase gene csnN106. Using the ΔcsnR host and a mutated csnN106 gene with a modified transcription operator, substantial levels of chitosanase could be produced in the absence of chitosan, using inexpensive medium components. Furthermore, chitosanase production was of higher quality as lower levels of extracellular protease and protein contaminants were observed.
  
  
  CONCLUSIONS
  This new chitosanase production system is of interest for biotechnology as only common media components are used and enzyme of high degree of purity is obtained directly in the culture supernatant.
authors.affiliation: Centre d'Étude et de Valorisation de la Diversité Microbienne, Département de Biologie, Faculté des Sciences, Université de Sherbrooke, 2500 boulevard de l'Université, Sherbrooke, J1K 2R1, (Québec) Canada.
authors.forename: Marie-Pierre
authors.initials: MP
authors.lastname: Dubeau
authors.forename: Isabelle
authors.initials: I
authors.lastname: Guay
authors.forename: Ryszard
authors.initials: R
authors.lastname: Brzezinski
mesh_terms: D001483:Base Sequence; D048271:Chitosan / Q000737:chemistry; D006026:Glycoside Hydrolases / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D048170:Streptomyces lividans / Q000235:genetics* / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D048271:Chitosan; D006026:Glycoside Hydrolases; C019868:chitosanase