title: In situ characterization of mycobacterial growth inhibition by lytic enzymes expressed in vectorized E. coli.
abstract: The emergence of extremely drug resistant Mycobacterium tuberculosis necessitates new strategies to combat the pathogen. Engineered bacteria may serve as vectors to deliver proteins to human cells, including mycobacteria-infected macrophages. In this work, we target Mycobacterium smegmatis, a nonpathogenic tuberculosis model, with E. coli modified to express trehalose dimycolate hydrolase (TDMH), a membrane-lysing serine esterase. We show that TDMH-expressing E. coli are capable of lysing mycobacteria in vitro and at low pH. Vectorized E. coli producing TDMH were found suppress the proliferation of mycobacteria in infected macrophages.
authors.affiliation: 2013 Paris Bettencourt iGEM team, Centre for Research and Interdisciplinarity, Paris Descartes University , 75014 Paris, France.
authors.forename: Iva
authors.initials: I
authors.lastname: Atanaskovic
authors.forename: Amel Camélia
authors.initials: AC
authors.lastname: Bencherif
authors.forename: Matthew
authors.initials: M
authors.lastname: Deyell
authors.forename: Sebastián
authors.initials: S
authors.lastname: Jaramillo-Riveri
authors.forename: Marguerite
authors.initials: M
authors.lastname: Benony
authors.forename: Aude G
authors.initials: AG
authors.lastname: Bernheim
authors.forename: Vincent K
authors.initials: VK
authors.lastname: Libis
authors.forename: Nicolas
authors.initials: N
authors.lastname: Koutsoubelis
authors.forename: Yonatan
authors.initials: Y
authors.lastname: Zegman
authors.forename: Anne C
authors.initials: AC
authors.lastname: Löchner
authors.forename: Clovis
authors.initials: C
authors.lastname: Basier
authors.forename: Idonnya
authors.initials: I
authors.lastname: Aghoghogbe
authors.forename: Zoran S
authors.initials: ZS
authors.lastname: Marinkovic
authors.forename: Sarah
authors.initials: S
authors.lastname: Zahra
authors.forename: Matthias
authors.initials: M
authors.lastname: Toulouze
authors.forename: Ariel B
authors.initials: AB
authors.lastname: Lindner
authors.forename: Edwin H
authors.initials: EH
authors.lastname: Wintermute
mesh_terms: D057005:Bioengineering / Q000379:methods*; D002478:Cells, Cultured; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D004950:Esterases / Q000235:genetics* / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008264:Macrophages / Q000382:microbiology; D020102:Mycobacterium smegmatis / Q000378:metabolism*
chemical_list: D004950:Esterases; C022635:serine esterase
keywords: LLO; TDMH; macrophages; mycobacteria; protein vectors; tuberculosis