title: The secreted L-arabinose isomerase displays anti-hyperglycemic effects in mice.
abstract: BACKGROUND
  The L-arabinose isomerase is an intracellular enzyme which converts L-arabinose into L-ribulose in living systems and D-galactose into D-tagatose in industrial processes and at industrial scales. D-tagatose is a natural ketohexose with potential uses in pharmaceutical and food industries. The D-galactose isomerization reaction is thermodynamically equilibrated, and leads to secondary subproducts at high pH. Therefore, an attractive L-arabinose isomerase should be thermoactive and acidotolerant with high catalytic efficiency. While many reports focused on the set out of a low cost process for the industrial production of D-tagatose, these procedures remain costly. When compared to intracellular enzymes, the production of extracellular ones constitutes an interesting strategy to increase the suitability of the biocatalysts.
  
  
  RESULTS
  The L-arabinose isomerase (L-AI) from Lactobacillus sakei was expressed in Lactococcus lactis in fusion with the signal peptide of usp45 (SP(Usp45)). The L-AI protein and activity were detected only in the supernatant of the induced cultures of the recombinant L. lactis demonstrating the secretion in the medium of the intracellular L. sakei L-AI in an active form. Moreover, we showed an improvement in the enzyme secretion using either (1) L. lactis strains deficient for their two major proteases, ClpP and HtrA, or (2) an enhancer of protein secretion in L. lactis fused to the recombinant L-AI with the SP(Usp45). Th L-AI enzyme secreted by the recombinant L. lactis strains or produced intracellularly in E. coli, showed the same functional properties than the native enzyme. Furthermore, when mice are fed with the L. lactis strain secreting the L-AI and galactose, tagatose was produced in vivo and reduced the glycemia index.
  
  
  CONCLUSIONS
  We report for the first time the secretion of the intracellular L-arabinose isomerase in the supernatant of food grade L. lactis cultures with hardly display other secreted proteins. The secreted L-AI originated from the food grade L. sakei 23 K was active and showed the same catalytic and structural properties as the intracellular enzyme. The L. lactis strains secreting the L-arabinose isomerase has the ability to produce D-tagatose in vivo and conferred an anti-hyperglycemic effect to mice.
authors.affiliation: INRA, UMR 1319 Micalis, Interactions of Firmicutes With Their Environments, 78352, Jouy-en-Josas Cedex, France. moez.rhimi@jouy.inra.fr.|AgroParisTech, Micalis, Interactions of Firmicutes With Their Environments, 78352, Jouy-en-Josas Cedex, France. moez.rhimi@jouy.inra.fr.
authors.forename: Moez
authors.initials: M
authors.lastname: Rhimi
authors.affiliation: INRA, UMR 1319 Micalis, Commensal and Probiotics-Host Interactions Laboratory, 78352, Jouy-en-Josas Cedex, France. luis.bermudez@jouy.inra.fr.|AgroParisTech, Micalis, Commensal and Probiotics-Host Interactions Laboratory, 78352, Jouy-en-Josas Cedex, France. luis.bermudez@jouy.inra.fr.
authors.forename: Luis G
authors.initials: LG
authors.lastname: Bermudez-Humaran
authors.affiliation: INRA, UMR 1319 Micalis, Interactions of Firmicutes With Their Environments, 78352, Jouy-en-Josas Cedex, France. yuan.huang.cn@hotmail.com.|AgroParisTech, Micalis, Interactions of Firmicutes With Their Environments, 78352, Jouy-en-Josas Cedex, France. yuan.huang.cn@hotmail.com.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Huang
authors.affiliation: INRA, UMR 1319 Micalis, Interactions of Firmicutes With Their Environments, 78352, Jouy-en-Josas Cedex, France. samira.boudebbouze@jouy.inra.f.|AgroParisTech, Micalis, Interactions of Firmicutes With Their Environments, 78352, Jouy-en-Josas Cedex, France. samira.boudebbouze@jouy.inra.f.
authors.forename: Samira
authors.initials: S
authors.lastname: Boudebbouze
authors.affiliation: INRA, UMR 1319 Micalis, Interactions of Firmicutes With Their Environments, 78352, Jouy-en-Josas Cedex, France. nadia.gaci@etu.udamail.fr.|AgroParisTech, Micalis, Interactions of Firmicutes With Their Environments, 78352, Jouy-en-Josas Cedex, France. nadia.gaci@etu.udamail.fr.
authors.forename: Nadia
authors.initials: N
authors.lastname: Gaci
authors.affiliation: INRA, UMR 1319 Micalis, Interactions of Firmicutes With Their Environments, 78352, Jouy-en-Josas Cedex, France. alexandrine.garnier@gmail.com.|AgroParisTech, Micalis, Interactions of Firmicutes With Their Environments, 78352, Jouy-en-Josas Cedex, France. alexandrine.garnier@gmail.com.
authors.forename: Alexandrine
authors.initials: A
authors.lastname: Garnier
authors.affiliation: INRA, UMR 1319 Micalis, Commensal and Probiotics-Host Interactions Laboratory, 78352, Jouy-en-Josas Cedex, France. jean-jacques.gratadoux@jouy.inra.fr.|AgroParisTech, Micalis, Commensal and Probiotics-Host Interactions Laboratory, 78352, Jouy-en-Josas Cedex, France. jean-jacques.gratadoux@jouy.inra.fr.
authors.forename: Jean-Jacques
authors.initials: JJ
authors.lastname: Gratadoux
authors.affiliation: INRA, UMR 1319 Micalis, Interactions of Firmicutes With Their Environments, 78352, Jouy-en-Josas Cedex, France. hela.mkaouar@jouy.inra.fr.|AgroParisTech, Micalis, Interactions of Firmicutes With Their Environments, 78352, Jouy-en-Josas Cedex, France. hela.mkaouar@jouy.inra.fr.
authors.forename: Héla
authors.initials: H
authors.lastname: Mkaouar
authors.affiliation: INRA, UMR 1319 Micalis, Commensal and Probiotics-Host Interactions Laboratory, 78352, Jouy-en-Josas Cedex, France. philippe.langella@jouy.inra.fr.|AgroParisTech, Micalis, Commensal and Probiotics-Host Interactions Laboratory, 78352, Jouy-en-Josas Cedex, France. philippe.langella@jouy.inra.fr.
authors.forename: Philippe
authors.initials: P
authors.lastname: Langella
authors.affiliation: INRA, UMR 1319 Micalis, Interactions of Firmicutes With Their Environments, 78352, Jouy-en-Josas Cedex, France. emmanuelle.maguin@jouy.inra.fr.|AgroParisTech, Micalis, Interactions of Firmicutes With Their Environments, 78352, Jouy-en-Josas Cedex, France. emmanuelle.maguin@jouy.inra.fr.
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Maguin
mesh_terms: D019747:Aldose-Ketose Isomerases / Q000378:metabolism*; D000818:Animals; D006601:Hexoses / Q000378:metabolism*; D007004:Hypoglycemic Agents / Q000008:administration & dosage / Q000627:therapeutic use*; D051379:Mice
chemical_list: D006601:Hexoses; D007004:Hypoglycemic Agents; D019747:Aldose-Ketose Isomerases; C019715:L-arabinose isomerase; C030192:tagatose