Mycobacterial Lipolytic Enzymes: a Gold Mine for Tuberculosis Research Luc Dedieu, C

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Mycobacterial lipolytic enzymes: A gold mine for tuberculosis research Luc Dedieu, C. Serveau-Avesque, L. Kremer, Stéphane Canaan

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Luc Dedieu, C. Serveau-Avesque, L. Kremer, Stéphane Canaan. Mycobacterial lipolytic enzymes: A gold mine for tuberculosis research. Biochimie, Elsevier, 2013, 95 (1), pp.66-73. 10.1016/j.biochi.2012.07.008. hal-02646757

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Distributed under a Creative Commons Attribution - NonCommercial - NoDerivatives| 4.0 International License Manuscrit d’auteur / Author manuscript Manuscrit d’auteuVersionr / Author postprint manuscript Manuscrit d’auteur / Author manuscript Journal homepage B Ve http://dx.doi.org/10.1016/j.biochi.2012.07.008 0300-9084/$ control reliable of causa- lack the The of bacterium, strains tive world. resistant multi-drug the of emergence in measures, diseases deadly and [1] Introduction 1. Mycobacterium Diagnosis Immunogenicity Physiopathology wall Cell Localization Keywords: 2012 July 20 online Available 2012 July 10 Accepted 2012 April 4 Received history: Article info article c b research a tuberculosis for Dedieu L. mine gold A enzymes: lipolytic Mycobacterial Review nrclua iii nlso;T,tbruoi;PD rti puri protein PPD, tuberculosis; TB, inclusion; lipidic intracellular 49 otele ee 5 France 05, Cedex Montpellier 34095 rhtcueo h neoermiseimtc Characteristic intricate enigmatic. the remains lipids, envelope wall cell the the of of architecture functions, bacteria pathways biological other chemistry, biosynthetic the of from and on envelope it studies cell numerous distinguishes Despite the which with tuberculosis, associated M. are characteristics unique of disease. the evidence of increasing physiopathology the the in of role alternative their because gener- TB interest have for considerable metabolism global need lipid context,ated mycobacterial this the urgent in In involved for treatment. an enzymes and reasons vaccination, diagnosis, is principal of there methods indi- the infected Therefore, of in stage epidemic. some dormant are a in viduals, undetected remain to latter iochimie NEM IN,PaeEgn aalo,305MnplirCdx0,France 05, Cedex Montpellier 34095 Bataillon, Eugène Place DIMNP, INSERM, NSUR22AxMreleUniversité UMR7282-Aix-Marseille CNRS aoaor eDnmqedsItrcin ebaarsNrae tPtooius nvrié eMnplir2e ,CR M 25 ae17 lc uèeBataillon, Eugène Place 107, case 5235, UMR CNRS 1, et 2 Montpellier de Universités Pathologiques, et Normales Membranaires Interactions des Dynamique de Laboratoire * r si ueclss(B otne ob n ftems threatening most the of one be to continues (TB) tuberculosis , orsodn uhr Tel.: author. Corresponding -aladdress: E-mail ih88mlinnwifcin n . ilo etsi 2010 in deaths million 1.5 and infections new million 8.8 With Abbreviations: ned ag ubro hmclycmlxlpd with lipids complex chemically of number large a Indeed, on définitive du du m on définitive Mycobacterial lipolytic enzymes:Agold minefortuberculosis research. Biochimie, 95(1),66-73. , Dedieu, L.,Serveau-Avesque, C.,Kremer, L.,Canaan, S.(Auteurde correspondance) (2013).

e (201 e rn matter front see a .Serveau-Avesque C. , A,ticllcrl M om arpae B ii oy ILI, body; lipid LB, macrophage; foamy FM, triacylglycerol; TAG, [email protected] yoatru tuberculosis Mycobacterium 3 ),

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95, p. Ó 349 64 3 fax: 93; 40 16 91 4 33 02Esve asnSS l ihsreserved. rights All SAS. Masson Elsevier 2012 sc r S Canaan). (S. it e 66- p nyooi nefcaee hsooi el ioye D,3 hmnJsp iue,142MreleCdx2,France 20, Cedex Marseille 13402 Aiguier, Joseph chemin 31 SDV, Lipolyse, la de Physiologie et Interfaciale Enzymologie ublié DOI :10.1016/j.biochi.2012.07.008 73, Comment citer cedocument: DOI e e a .Kremer L. , n h blt fthe of ability the and , alascae,sraeepsdlpltcezmscaatrzdt ae tde ntelocalization, usefulness the the possible in on their markers highlighted Studies diagnostic enzymes cell date. new these or of as secreted to properties the characterized on immunological and emphasis enzymes activity an enzymatic with lipolytic review, exposed this in surface discussed wall-associated, be will disease the of physiopathology omnyadratvto.Terl flpltcezmsi h hsooyof physiology the in during roles enzymes physiological contagious. lipolytic highly important of be play role can to that The thought infection reactivation. are TB reac- and enzymes active During an dormancy lipolytic granulomas. to host and within leading lipids state again infection, Mycobacterial dormant replicating TB latent non-replicating start a a bacilli In in infection. tivation, latent present and are asymptomatic bacilli chronic, a persistent establish and system immune host ueclss(B soeo h edis netosdsae olwd ihasrn mati devel- in impact strong a with worldwide diseases infectious deadliest the countries. of oping one is (TB) Tuberculosis abstract dan þ : : 349 15 57. 58 71 91 4 33 10.1016/j.biochi.2012.07.008 s s / Fi ﬁ dderivative. ed l nal

b yoatru tuberculosis Mycobacterium ver , c .Canaan S. , si [2] on of theof on ma . nmcbceilseissc as bovis such Mycobacterium species mycobacterial bacteria in several in demonstrated been have consumption more occurs individuals. usually immunocompromised in infection frequently latent of reactivation this However, ramn.I eciaincss Asaeue ofe the fuel to used infection are active to an TAGs causing recalcitrant replication cases, particularly cellular of are reactivation resumption and In decades treatment. several for could bacilli persist stage, non-replicating this In dormancy. during energy eoos n ec h ug hr hyaepaoyoe by phagocytosed are they where lungs the reach and aerosols, smegmatis Mycobacterium raygyeo (TAG) triacylglycerol htaepoal eie rmlpd ftehs elmembrane cell host the of lipids from [5 derived probably are that ies a enivsiae.I a hw htdrn infection, during that the shown tuberculosis of was M. It physiopathology investigated. and been that lipids has decade disease last intracellular the between in only link is the It major neglected. largely been as have lipids considered are research extensive mycobacteria in of determinants have virulence that focus lipoarabinomannan, the and been lip- lipomannan as as well such as oglycans dimycocerosates, phthiocerol phosphatidyl mannosides, acids, inositol mycolic including (glyco)lipids mycobacterial e cuuaino iisdrvdfo otclsadtheir and cells host from derived lipids of Accumulation yial,tetbrl ail ne h oyb naainof inhalation by body the enter bacilli tubercle the Typically, ncnrs otecl alascae iis intracytoplasmic lipids, wall-associated cell the to contrast In ﬁ 8] h gis TB. against ght a yti way, this By . , * nu sc h tooia gn fT,hsahg aaiyt vd the evade to capacity high a has TB, of agent etiological the ,

ri pt publis cuuae nrclua ii nlsos(ILI) inclusions lipid intracellular accumulates .tuberculosis M. BCG [9 Ó e 02Esve asnSS l ihsreserved. rights All SAS. Masson Elsevier 2012 hed 12] [4,10] [13,18] in o s sasuc fcro and carbon of source a as use for . :

, trsftyaisi h omof form the in acids fatty stores yoatru leprae Mycobacterium .tuberculosis M. [3] . .tuberculosis M. [4,9,12,15 [14] [19] e and , [13] and 17] and [4] . , Manuscrit d’auteur / Author manuscript Manuscrit d’auteuVersionr / Author postprint manuscript Manuscrit d’auteur / Author manuscript Journal homepage B Ve iochimie r si eaoielpd ahrta carbohydrates than rather lipids preferentially metabolize bacteria phagocytosed macro- (FM), macrophages the foamy the gives ( appearance neutral that foamy layer, characteristic of their phospholipid phages composed a by mainly surrounded accumulate (LB), cellular lipids bacilli bodies organized containing lipid macrophages highly intracytoplasmic structure, a this In of of TB. hallmark formation histopathological major a the granuloma, termed structure, to leading dendritic cells, and lymphocytes response macrophages, of host recruitment subsequent of consists The macrophages. alveolar pulmonary yolsi ii nlsos(Ls.()Elre iwo B showing (B) of view Enlarged (C) (ILIs). inclusions lipid cytoplasmic 1. Fig. .tuberculosis M. with cells nuclear a lobe eotdin in and reported been dormant also the infection has for TB energy active lead an and can to that carbon reactivation aiding of thereby granulomas, sources within bacilli as serve ILI) an and in demonstrated As vitro infection. latent in a i.e., dormancy, to leading hgsmlbcei cur n cuuaeIIi hi cytoplasm their in ( ILI accumulate and acquire metabolism bacteria mycobacterial phagosomal lipid several in of involved up-regulation genes of evidence by supported fitaellrTG ek ntelt xoeta rwhphase growth exponential late the in [23] peaks TAGs intracellular synthesized or of fatty host free the from from derived imported [9] be are may latter that The acids TAGs. of composed mainly i.1 Fig. on définitive du du m on définitive Mycobacterial lipolytic enzymes:Agold minefortuberculosis research. Biochimie, 95(1),66-73. , Asacmlt uigmcbceilgot n h amount the and growth mycobacterial during accumulate TAGs . iceia nlsshv eeldthat revealed have analyses Biochemical Dedieu, L.,Serveau-Avesque, C.,Kremer, L.,Canaan, S.(Auteurde correspondance) (2013). n h o-elctn phase non-replicating the and nrclua ii ois(B n hgsmsin phagosomes and (LB) bodies lipid Intracellular n )adpriti o-elctn tt,ultimately state, non-replicating a in persist and C) and B .leprae M. oe fhmngranulomas human of model

(201 ne:arw niaetepaooa ebaeaon h atru.()L uruddb several by surrounded LB (B) bacterium. the around membrane phagosomal the indicate arrows Inset: . .tuberculosis M. 3 ), ifce arpae n cwn cells Schwann and macrophages -infected

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95, p. [21] 3R,adpoesdfreeto irsoya a 1ps-neto.()Atpclfaymcohg arigLsadpaooe containing phagosomes and LBs carrying macrophage foamy typical A (A) post-infection. 11 day at microscopy electron for processed and H37Rv, sc .tuberculosis- M. nadto oF,IIaccumulation ILI FM, to addition In . r it 66- p ublié DOI :10.1016/j.biochi.2012.07.008 73, [15] [17] Comment citer cedocument: [12] ute,i a enshown been has it Further, . DOI e e tti tg,teintra- the stage, this At . hs ii rpes(LB droplets lipid these , dan netdadipocytes infected : : .tuberculosis M. .tuberculosis M. 10.1016/j.biochi.2012.07.008 s s / Fi [20] i.1 Fig. iwta is that view a , A) l nal .Dde ta./Bohme9 21)66 (2013) 95 Biochimie / al. et Dedieu L.

.tuberculosis M. [12] ifce om arpae.Gauoa eeobtained were Granulomas macrophages. foamy -infected ver Within . [22] enovo de Lsare ILIs si on of theof on ma . [7] ail otiiglreII Aatdfo Ref. from (Adapted ILIs large containing bacilli nye,wihhv pndu xiigpsiiiiso their of possibilities exciting lipolytic up mycobacterial opened of have properties which biological enzymes, the of standing muoeiiy hc a rmtdteruea imresof biomarkers as use TB their active prompted their is has light, which to come immunogenicity, recently has of that aspect treatment interesting cystic an in lipases, in adjuvants or their as pancreatitis have against chronic or of agents enzymes diseases therapeutic Because cardiovascular as lipolytic health obesity, adipocytes. metabolism, human in in lipid attention received stored in fat role fundamental of as the well mobilization as in metabolism, in lipoprotein characterized digestion, fat well in roles been important have Lipases kingdom animal long- acids. containing de esters fatty be of can chain hydrolysis and the lipids for of biocatalysts turnover essential metabolic a provide that ae,hv nybe h ou fitnersac o h atfew last the for research phospholi- intense of and years focus lipases the been enzymes, only have associated pases, the ago, years many the to of contribute host. survival infected and of capacity functions extraordinary physiological lipase important TAG in serve increase an with coincides activity levels TAG in reduction a dormancy during elevated of expression that nti eiw ewl ics h eetavne nunder- in advances recent the discuss will we review, this In ept ioyi ciiishv endsrbdi mycobacteria in described been have activities lipolytic Despite nu [24 [13] sc e

[3 27] ri e 1 hs nye novdi ii erdto perto appear degradation lipid in involved enzymes Thus, . pt publis 73 ,32] iae n hshlpssaepcla molecules peculiar are phospholipases and Lipases . . [28e .tuberculosis-speciﬁ M. atclryi aml hr hyplay they where mammals in particularly 30], .tuberculosis M. hed [16] in : nvitro in [12]

n hti h eciae bacilli, reactivated the in that and , fi ). ail.Sm ftebcei ipa intra- display bacteria the of Some bacilli. rss ncs fmycobacterial of case In brosis. yifcino eihrlbodmono- blood peripheral of infection by .tuberculosis M. iaegnsi strongly is genes lipase c ihnthe within fi e as ned 67 Manuscrit d’auteur / Author manuscript Manuscrit d’auteuVersionr / Author postprint manuscript Manuscrit d’auteur / Author manuscript Journal homepage B Ve nerdta iYi epnil o h tlzto fTAGs of utilization the for responsible is LipY that inferred impeded severely facmltdTG nteasneo nte oreo abn In carbon. of source another of a absence the in TAGs accumulated of effi by accompanied TAGs induced, highly ILI-containing was expression hydrolyze to able [16,35] is LipY that demonstrated h Efml fprotein of family an of presence PE the quali while also the family, domain HSL PE the N-terminal of additional member a as protein hydrolytic and localization extracellular and activity intra dual LipY: 3.1. the in speci involved a date, trigger to to able infection. during and up response wall cell discovered response the enzymes of immune architecture three the the of on modulation in that [35,36] participate suggests also may localization Surface-exposed they lipids. cell host degradation enzymatic of in role potential of their components and/or structural membrane as the proteins these of possibility the raising several wall cell mycobacterial the architecture in enzymes Lipolytic 3. functions. physiological and oee,bohmclcaatrzto fteeezmsusing site Ser-His-Glu/Asp. enzymes active these triad an of speci possess catalytic characterization enzymes above biochemical characteristic other However, the all the to PLCs, comprising the belonging family. for (PLC) C not phospholipases Except the to 3 4 and family lipase family, mentioned (HSL) lipase Lipase the monoglyceride Hormone-Sensitive to human 7 the family, to 12 family, (BSSL) parapsilosis r classi are motn oei h yoatra iecceadprasin perhaps and cycle life mycobacterial the virulence. in role important tuberculosis M. 2. targets. and therapeutic TB putative active of as diagnosis of methods accurate more in applications 68 rmr eune,2 fte31 the of 28 sequences, primary ebr fthe of feature of characteristic a members is of which presence GXSXG, the sequence on consensus based the lipases or esterases putative as annotated so-called belonging the hydrolases, in to lipid/ester involved 24 including (enzymes degradation), enzymes lipids lipolytic encoding genes putative eoesz.Ti etr,cmie ihteeteeyhigh extremely 30 the representing lipids with lipids, of combined of degradation feature, amount This or in size. genes synthesis genome 50 the to in compared metabolism involved lipid enzymes in tuberculosis involved M. encoding enzymes genes of putative number large unusually an possesses bacterium tuberculosis M. iochimie r si lipY nsilico In h rmr euneo iY(v07)ceryidenti clearly (Rv3097c) LipY of sequence primary The of association the revealed have studies Immunolocalization h nlssof analysis The on définitive du du m on définitive Mycobacterial lipolytic enzymes:Agold minefortuberculosis research. Biochimie, 95(1),66-73. , fi dltdmtn of mutant -deleted Dedieu, L.,Serveau-Avesque, C.,Kremer, L.,Canaan, S.(Auteurde correspondance) (2013). usrtsi rrqiiet sals hi reactivities true their establish to prerequisite a is substrates c .tuberculosis.InM. n/ri naino otcells host of invasion in and/or .tuberculosis M. fi nsilico In dit ifrn aiis eogn othe to belonging 1 families: different into ed

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95, p. ( i.2 Fig. .tuberculosis M. shrci coli Escherichia sc [16] r it ) rmteersls tcnbe can it results, these From A). .tuberculosis M. 66- iceia characterization Biochemical . p fi uiae(u)fml,3t the to 3 family, (Cut) cutinase dtepeec faon 30 around of presence the ed e ublié DOI :10.1016/j.biochi.2012.07.008 73, eoervae htthis that revealed genome 0 ftedywih of weight dry the of 40% [5,8] Comment citer cedocument: DOI e e hscatrwl focus will chapter This . A yrlsswas hydrolysis TAG , hc a similar a has which , fi dan di samme of member a as it ed [34] : : 10.1016/j.biochi.2012.07.008 s s / Fi ioyi enzymes lipolytic ae ntheir on Based . in utilization cient .Dde ta./Bohme9 21)66 (2013) 95 Biochimie / al. et Dedieu L. [33] l nal fi

humoral c Indeed, . ﬁ Candida ver dthis ed si lipY on of theof on ma uigei rmdrac,laigt h reactivation. the and/or to leading dormancy dormancy, during from exit cytoplasm during bacterial the in accumulated eocl ihislclzto ntebceilsraea demon- as microscopy surface electron bacterial immunogold the by on strated localization its with reconcile erto fpoen arigP/P_GSdmisin domains PE/PPE_PGRS carrying proteins tuberculosis identiﬁ M. of ( been species also this has secretion in ESX-5 pathway this secretion Recently, in ESX-5 domain the PE of to activity (similar domain presence PPE the with a agreed of which extracellularly, secreted was LipY al. et Daleke ae)(dpe rmRef. a from (Adapted in a panel) surface in cell not the but on panel) LipY of localization immunogold Ref. i.2. Fig. ervdi hsht-ufrdsln PS o lns2ad4,teW strain WT nutrient- the were 4), cells and when 2 ef C); (lanes more controls, h 3; 6 far for and TAGs (PBS) 1 utilized saline (lanes were phosphate-buffered arrow) strains in by two (indicated deprived TAGs the accumulated in intracellularly of similar Levels TAGs. of zation idtp W)sri rLipY-de of or cultures strain hypoxic (WT) from type isolated wild lipids of chromatography layer Thin epne hshptei a encon been has hypothesis This response. h otimn ytmadmytigraspeci a trigger may and to accessible system be surface may immune it of that host hypothesis the observation the the to in the led from LipY LipY lipids Importantly, of of exposure of respectively. hydrolysis Both the action LBs, and machinery. the ILIs host ESX-5 within with TAGs via of agreement degradation domain in are N-terminal localizations PE/PPE cleavage the the LipY, the of of following of cytoplasm localization surface-associated cellular the cell dual or in a intracytoplasmic lipase suggest the results These of bacteria. retention which LipY, the with explain interactions may physical of capable proteins cellular outat-iYatbd epnei Bpatients TB in response antibody anti-LipY robust a oee,teatvt fLp nII a endif been has ILIs on LipY of activity the However, [16] hsooia ucinadda oaiaino iY (A) LipY. of localization dual and function Physiological .(B) ). nu sc LipY e

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lipY uatsri,poesdfrvisuali- for processed strain, mutant ) fi mdb h eeto of detection the by rmed yoatru marinum Mycobacterium fi .tuberculosis M. in uat(dpe from (Adapted mutant cient da h ytmfor system the as ed .marinum M. [35] [39] LipY eetwr by work Recent . identi sa is fi Tsri (left strain WT .tuberculosis M. A hydrolase TAG humoral c fi n the and ) dintra- ed fi i.2 Fig. utto cult [35] B). , . . Manuscrit d’auteur / Author manuscript Manuscrit d’auteuVersionr / Author postprint manuscript Manuscrit d’auteur / Author manuscript Journal homepage B Ve iochimie r si ptp apn taey muooiateioe were epitopes immunodominant 6 identi strategy, mapping epitope family Lip the of member al. et ufrcnann ,1o Msdu ardoyhlt NTC.NTCi iesl sdt ouiieislbelpd ern at cdwt ogcro hi.Teatvt on activity The chain. carbon long with acid fatty bearing lipids insoluble solubilize to used salt bile a is NaTDC (NaTDC). lengths taurodeoxycholate sodium carbon mM of 4 substrates or 1 0, containing buffer a rmRef. from elwl n h asl of the capsule both in the present and protein wall surface cell cell was a it is enzyme, LipC cytoplasmic that a demonstrated be to predicted Although protein. the host in roles important play TB active esterase surface cell immunogenic an LipC: FM. 3.2. the of in acquisition LBs the the in in present role TAGs key degrading a by plays lipids, starvation, host LipY TAG that nutrient long-chain postulate during may major hydrolysis one TAG the TAG in as in involved LipY emerges hydrolase of LipY role Because central the hydrolysis. validate results these Collectively, ciiyo eobnn iCwsmaue sn 8- using measured was LipC recombinant of activity and panel) (upper 3. Fig. puri was LipC recombinant relevance, biological its understand a o fetdi t rwhrt rviability or rate growth its in affected not was ersnsteol oolcrd iaeidenti lipase and teria monoglyceride respectively, only human, and the rat represents from lipases monoglyceride the host of degradation lipids in involved cell lipase monoglyceride host a Rv0183: the 3.3. of modulation the cell in novel, involved response. a immune possibly is and LipC nogenic that of demonstrate esterase surface-associated studies cells was epithelial these LipC pulmonary together, and addition, macrophages In in pro-in host. kines stimulating the from of substrates capable ester acyl fatty on hydrolyzes LipC chains whether investigate to tool useful a sents on ( activity esters esterase short-chain an revealing biochemically, characterized and on définitive du du m on définitive Mycobacterial lipolytic enzymes:Agold minefortuberculosis research. Biochimie, 95(1),66-73. , ayatgncpoen xrse uigteerysae of stages early the during expressed proteins antigenic Many v13sae 4 n 6 mn cdsqec dniywith identity sequence acid amino 36% and 34% shares Rv0183 Dedieu, L.,Serveau-Avesque, C.,Kremer, L.,Canaan, S.(Auteurde correspondance) (2013). oaiainadezmtcatvt fLp.()Mcbceilcl ufc oaiaino iC muolcrnmcocp futahnsectionsof ultrathin of microscopy Immunoelectron LipC. of localization surface cell Mycobacterial (A) LipC. of activity enzymatic and Localization [6] fi [32] d fwihmpe orsde xoe ttesraeof surface the at exposed residues to mapped which of 5 ed, [32]

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2wsudtcal.Tema auso speci of values mean The undetectable. was 12 95, p. [33] ) A B). D15 lwrpnl from panel) (lower CDC1551 sc .tuberculosis M. r eogn oteHLfml.Uigan Using family. HSL the to belonging .tuberculosis M. it ﬂ 66- .tuberculosis M. p maoyctknsadchemo- and cytokines ammatory ublié DOI :10.1016/j.biochi.2012.07.008 73, e Comment citer cedocument: ahgnitrcin.Shen interactions. pathogen DOI e e p dan ntohnl(N)etr ihcro hi egh agn rmC from ranging lengths chain carbon with esters (PNP) -nitrophenyl ( i.3 Fig. : : hti ihyimmu- highly is that 10.1016/j.biochi.2012.07.008 s s / Fi iCdltdstrain LipC-deleted nvitro in nvitro in ) na fotto effort an In A). ﬁ di mycobac- in ed l nal .Dde ta./Bohme9 21)66 (2013) 95 Biochimie / al. et Dedieu L.

n repre- and , utrs rbdwt abtat-iCatbde.()Lp a rfrnefrsotcanetr.Enzymatic esters. short-chain for preference has LipC (B) antibodies. anti-LipC rabbit with probed cultures, [32] ver Taken . si ﬁ ciiycrepnigt 1 to corresponding activity c on of theof on ma ﬁ ed os arpae a on oidc elaotssadto and apoptosis cell induce in to of expression found trigger was macrophages mouse patients rwho h SE_20dsutdmtn a impaired was that mutant indicated MSMEG_0220-disrupted (MO) the ( monoolein of in inactive with growth an performed or supplemented studies active growth medium an Moreover, either enzyme. encoding MSMEG_0220 gene a with mentation xii akddfeecswt epc otersbtaeand substrate their to respect homology. speci with enzymatic 66% differences and marked exhibit identity enzymes these that 52% showed characterization biochemical However, sharing related, appear (Rv3452) Cut4 structurally and (Rv1984c) Cfp21 enzymes, secreted two the in found genome been have family cuti- cutinase the genes if to seven related Even organisms, phytopathogenic phopholipids. in pro- found polyester and mainly a TAGs are is nases also which but cutin, leaves as plant such tecting substrates of panel large a cytotoxicity and activity broad-range Cutinases: 4. host. the of response immune the exogenous induce hydrolyzes strongly wall, may and cell lipids mycobacterial the of component ,TR6 TNF- TLR-6, 2, on ob oehmgnu ihls lmigo cells, of interactions clumping intercellular less the in with alteration ( homogenous an to more due presumably be to found osblt hti rvdsfe at cd omcbcei.To mycobacteria. to acids the fatty free raising the provides TAGs, address mono- it or for that diacylglycerols preference possibility to substrate compared strong likely a acylglycerols is exhibits it of point Rv0183 that enzymatic an hypothesis view, From lipids. the cell to host of leading hydrolysis in wall, involved cell the in Rv0183 i.4 (Fig. strain parental the to compared [40] in ortholog i.4 Fig. i.4 Fig. uiae r xrclua eieetrssal odegrade to able esterases serine extracellular are Cutinases hsmtn ipae nuepce ooymorphotype colony unexpected an displayed mutant This . nu ) oal,R08 ol edtce ntesrmo infected of serum the in detected be could Rv0183 Notably, B). G e sc [33,42] [31]

) ohpeoye eeprilyrsoe ycomple- by restored partially were phenotypes Both J). ri e .smegmatis M. pt publis 73 nvivo in m oevr eeooosepeso fR08 in Rv0183 of expression heterologous Moreover, . o N eesdprmnadprm rti,aepotd(Adapted plotted are protein, mg per and min per released PNP mol a [41] mn hs uaiectns-ieproteins, cutinase-like putative 7 these Among . ﬁ iis hl u4bhvssmlrt typical a to similar behaves Cut4 While cities. hs eut ml htR08 sastructural a is Rv0183 that imply results These . oeo hsezm,adsutdmtn fits of mutant disrupted a enzyme, this of role hed ﬂ 2 ammat oC to SE_20 a eeae ( generated was MSMEG_0220, , in 10 : ssbtae.TeePPetr eemxdin mixed were esters PNP These substrates. as

r atr uha L6 NF- IL-6, as such factors ory C e ) t iudcluewas culture liquid Its F). .tuberculosis M. .tuberculosis M. k i.4 Fig. ,TLR- B, H37Rv A) 69 Manuscrit d’auteur / Author manuscript Manuscrit d’auteuVersionr / Author postprint manuscript Manuscrit d’auteur / Author manuscript Journal homepage B Ve v13 oa rti (120 protein Total Rv0183. uiaefml,Ct R30)adishmlg in homologs the its and cutinases of (Rv3802) member Cut6 Another family, that cutinase lipids. host virulence/ degrading view the by in process participate the infection phospholipase and membranes its host support the of with observations interact consequence These a macro- as activity. induces presumably Cut4 lysis, that phage demonstrated macrophages on assays viability Corynebacterium h ooissoigruh(,E n )o moh()mrhlge.(G morphologies. (D) smooth or MSMEG F) of and Cultures broth. E, (C, rough showing colonies the .smegmatis M. aayial inactive, catalytically 10n) B oprsno rwho h bv tan of strains above the of growth of Comparison (B) ng). (100 nuaina 37 at incubation 70 ComMSMEG xiisalpltcatvt ntili n hwdapeeec for preference a showed and C triolein on activity lipolytic a exhibits speci by A phospholipase 4. Fig. .smegmatis M. iochimie 8 r si at clcan oose substrates monoester chains acyl fatty on définitive du du m on définitive Mycobacterial lipolytic enzymes:Agold minefortuberculosis research. Biochimie, 95(1),66-73. , oaiainadpyilgclfnto fmngyeielps,MMG02 of MSMEG_0220 lipase, monoglyceride of function physiological and Localization Dedieu, L.,Serveau-Avesque, C.,Kremer, L.,Canaan, S.(Auteurde correspondance) (2013). [43,44] D mc 20( n ) n ComMSMEG and I), and (E 0220

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95, p. al yrlzn hshlpd,Cfp21 phospholipids, hydrolyzing cally sc r (ComMSMEG it 66- p D 2010 FadJ.(C J). and (F 0220S110A ublié DOI :10.1016/j.biochi.2012.07.008 73, Comment citer cedocument: DOI e e D D 20sri opeetdwt orsodn ee(ComMSMEG gene corresponding with complemented strain 0220 2010;S1 orsod otectltcsrn fMMG02) ,rcmiatMMG02 puri MSMEG_0220 recombinant 6, MSMEG_0220); of serine catalytic the to corresponds S110 0220S110A; .smegmatis M. dan [8] .smegmatis M. : : . 10.1016/j.biochi.2012.07.008 ﬁ s s / Fi nvitro In drcmiatpoen 10n)wr odda olw:1 eobnn v13 2, Rv0183; recombinant 1, follows: as loaded were ng) (100 proteins recombinant ed .smegmatis M. .Dde ta./Bohme9 21)66 (2013) 95 Biochimie / al. et Dedieu L. e l nal n79aa upeetdwt ooli M) ooyfriguis(F)wr one fe 3 after counted were (CFU) units forming Colony (MO). monoolein with supplemented agar 7H9 on )Cmaio fcln opooisatr5dy ficbto t37 at incubation of days 5 after morphologies colony of Comparison F) e

tan fe aso nuaina 37 at incubation of days 5 after strains )Otclmcocp (magni microscopy Optical J) cytotoxic ver nvitro in si on of theof on ma or .smegmatis M. hoseaeatvt ugse hs nye a atcpt to participate biosynthesis may acid enzymes mycolic these suggested activity thioesterase ilvcieantigen vaccine tial with challenge a speci a .PopoiaeC Phospholipase 5. thsbe hw htmcbceilpopoiiscnmimic can phospholipids mycobacterial that shown been has vesicle. endosomal It early an of characteristics exhibits that vacuole a nifce macrophages, infected In ﬁ ﬁ nu n t ooo v13from Rv0183 homolog its and -ypoyersos n a bet rtc iefrom mice protect to able was and response T-lymphocyte c cation, sc e

73 ri pt publis 3 bevto of observation 63) n71 oi eim T( n ) MSMEG G), and (C WT medium: solid 7H11 on C n ec srgre sapoten- a as regarded is hence and tuberculosis , M. [36] . hed D 20;5 tancmlmne ihgn encoding gene with complemented strain 5, 0220); [45] .smegmatis M. in oevr u6wsal otrigger to able was Cut6 Moreover, . .tuberculosis M. dplcoa nioisaantrecombinant against antibodies polyclonal ed : .tuberculosis M.

n71 oi eim o iwof view Top medium. solid 7H11 on C el rw nTen8-re7H9 80-free Tween in grown cells A SE_20poue in produced MSMEG_0220 (A) . .smegmatis M. eie within resides D 20( n H), and (D 0220 mc ﬁ 2 5 T 3, WT; 155 dprotein ed e asof days 5 [40] ). Manuscrit d’auteur / Author manuscript Manuscrit d’auteuVersionr / Author postprint manuscript Manuscrit d’auteur / Author manuscript Journal homepage B Ve iochimie r si eecutrwihi ato h D eoi eeini this in deletion genomic RD5 the of gene, remaining part the and a strain, is which the lacks model cluster naturally TB, gene mouse bovine of agent a causative quadruple the bovis in , and M. infection triple of that shown phase has plc al. tuberculosis M. et Raynaud from Previous phospholipids. work membrane cell on to activity owing macrophages hydrolytic mouse their to cytotoxic are enzymes four all that recombinant pure four all of cluster otlpd n nii uledctcmtrto ftepathogen- the of maturation vacuole endocytic containing full inhibit and lipids host suooa euioaplcH aeruginosa Pseudomonas in detected [47] been have activities sphingomyelinase eoecristreajcnl oae hshlps (PLC) C phospholipase located ( adjacently genes three carries genome muesse a o eflypie gis antigens against primed fully the be where not patients infected may newly is system of problem immune diagnosis this the and TB, in of acute detection more precise the in tools diagnostic biomarkers diagnostic TB new as enzymes Lipolytic 6. of function exact pathogenesis. the in identify enzymes to these failed have PLCs mycobacterial the to lysing by possibly membrane phagolysosomal activity, cytolytic contact-dependent their insertion ( gene truncated fourth a and genes tgso cieifcin ossetwt t oaiaina the in at response antibody localization an induce its to early found with was the LipC Consistent surface, during bacterial infection. expressed active was of LipC stages that showed DNA genomic actively are bacilli when disease replicating. the of of also with phase levels active expression those the higher ELISA during to Cut4 related from be Cut4-based may TB an This active infection. The latent indicating a with assay. patients test, this discriminating serological allowed of Cut4 sensitivity for culture-positive excellent positive but TB smear-negative were active of 105 patients 87% and addition, individuals In patients. healthy 149 including performance population best speci individuals. the a presented healthy Cut4 with markers, BCG-vaccinated four in these in Among and background low volunteers a to healthy compared antigens, these against patients 5 in demonstrated as been such intra- species has in bacterial factor virulence phospholipase a of as role and survival The cellular pathogen. this of virulence the oeo h puri the of Some aibesniiiyadspeci and sensitivity variable u4 n vlae nErpa ainswt ihratv Bor TB active either with patients and laten European Cfp21, in Rv0183, humoral evaluated LipY, and anti-LipY to Cut4, extended strong recently area a endemic were an observations of in living induction patients TB in the response reporting study a TB. active of biomarkers cells host of yohsz htteepoen a lopriiaei the in participate also to may response investigators immune proteins the cell of prompted these modulation host the above that in involvement discussed hypothesize their and degradation surface, cell cell lipid the the in at enzymes or lipolytic wall mycobacterial the of localization The B) on définitive du du m on définitive Mycobacterial lipolytic enzymes:Agold minefortuberculosis research. Biochimie, 95(1),66-73. , eea eot aehglgtdtefiueo conventional of failure the highlighted have reports Several muocenn fa xrsinlbayof library expression an of Immunoscreening h s fLp sasrdansi akrwas marker serodiagnostic a as LipY of use The Dedieu, L.,Serveau-Avesque, C.,Kremer, L.,Canaan, S.(Auteurde correspondance) (2013). [31] . t .tuberculosis M. Bcmae onnifce ouaingop ( groups population non-infected to compared TB plcA [49] togIgG-speciﬁ Strong .

(201 [51] aaaN Bakala . , plcB ﬁ [5,8] L nye a otiuet Bifcinthrough infection TB to contribute may enzymes PLC . iyo 46 n estvt f9.%i study a in 90.5% of sensitivity a and 94.6% of city 3 ), ﬁ and hs nye a hrfr ecniee as considered be therefore may enzymes These .

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95, p. itramonocytogenes Listeria hshlps ,popoiaeD and D, phospholipase C, Phospholipase . htaesmlrt the to similar are that ) uoa epne eedtce nTB in detected were responses humoral c sc [52] .tuberculosis M. ﬁ r and it iyi eooia et o TB for tests serological in city plcD 66- p oee,svrlsuisdevoted studies several However, . [5] ublié plcN DOI :10.1016/j.biochi.2012.07.008 73, eei nciae ya IS6110 an by inactivated is gene eeal ooti ihyields high obtain to able were [35,36] Comment citer cedocument: plcD DOI ee,wihcnrbt to contribute which genes, e e [50] dan Lsaddemonstrated and PLCs ,sprtdfo this from separated ), : : nteohrhand, other the On . n/ri h invasion the in and/or 10.1016/j.biochi.2012.07.008 s s / Fi [48] [35] ﬁ .argns plc aeruginosa P. s ugse in suggested rst . .tuberculosis M. .tuberculosis M. l nal .tuberculosis M. .Dde ta./Bohme9 21)66 (2013) 95 Biochimie / al. et Dedieu L.

hs initial These . i.5 Fig. ver plcABC si and A on of theof on ma [53] [54] . . ilvciecniae sdmntae yWs tal. et West poten- by demonstrated as therapeutic as considered candidates such vaccine be in tial also consideration could enzymes into Lipolytic taken safety between approaches. be important interactions must raising that possible thus lipases, issues to human due and inhibitors occur these may effects sirable o uueppiebsdsrdansi tests serodiagnostic peptide-based future identi for were protein the outer of on surface exposed are individuals. that TB-uninfected LipC of from epitopes immunodominant patients Lastly, TB ( distinguish HIV with to not helped or co-infected TB active with patients losis signi a able induce were vaccines to DNA as delivered cutinases that showed authors etctresfrdvlpn uueat-Bdus ned speci Indeed, drugs. anti-TB future as developing also for targets perhaps peutic exit and to catalytic as ILIs pathogen entities. of throughout the structural involved of assimilation be to life-cycle seem for the and enzymes required carbon these of Thus, TAG be dormancy. sources intracellular as may Conversely, used stage. be and hydrolases persistent to mycobacteria the TAGs during intracellular of energy by form released up the The taken in lipids. stored then lipolytic host. cell survival are host the mycobacterial acids and of in fatty secreted hydrolysis virulence the persist process, catalyze the to enzymes infection in bacterium enzymes the role this of During by central family employed a this strategies play properties, to biological unusually appears their an on of observation studies in the lipases/phospholipases and of chronic on number a large Based establish to infection. and immunity persistent host evade to ability its prospects future and Summary 7. microscopy. smear sputum standard by simple missed detec- in cases inexpensive of application and tion sensitive, where rapid, allow countries, TB may better endemic immunoassays a in for and warranted particularly control, is This observations. initial nldn agrnmeso eaaenwrqie ocon to required now speci are sera and of numbers sensitivity larger including high TB with latent from infections active distinguishing allow would enzymes lipolytic in potential their for Therefore, evaluated diagnosis. TB. be TB should from PLCs suffering LipF, children with in together response cell B humoral to shown was enzyme phosphatidylcholine hydrolyze recombinant The activity. C phospholipase exhibiting LipF, protein, wall-associated cell additional an possesses I-netdidvdasi onre ihyaf smear-negative, highly in countries TB in of individuals diagnosis for HIV-infected exploited potent These be infections. as TB may appear latent background enzymes from and TB the active vaccination In distinguish BCG to override markers. candidates prior to diagnostic from arising allowed new signals markers as these considered lipolytic particular, patients, being in are seroreactivity of enzymes occurrence the and disease, lokona ritt nihbtro ietvslpssue to signi used to lipases found digestives of was inhibitor or obesity, tetrahydrolipstatin, an prevent example, treat Orlistat, to an as able As known be state. also may persistent mycobacteria the cell host infecting shorten of the consumption by the with lipids interfere would that inhibitors ttni o h nycniaeadohrdusaebigtested being are drugs other and candidate only right the not is including statin species, several on smegmatis M. ioyi nye a ec ecniee sptn thera- potent as considered be hence may enzymes Lipolytic of success the in feature key A h bv tde ugs htteesrnl immunogenic strongly these that suggest studies above The nve fteripratrl nteptoeei fthe of pathogenesis the in role important their of view In infection. nu now sc

naaei n nutillbrtr.Hwvr unde- However, laboratory. industrial and academic in ri e pt publis 73 [10] ﬁ atpoeto namuemdlof model mouse a in protection cant n xiisptn nimcbceilactivity anti-mycobacterial potent exhibits and hed in : .tuberculosis M.

nvitro in fi .tuberculosis M. fi dadpooe singredients as proposed and ed atyipc h rwhof growth the impact cantly [55] n nueasigni a induce and .tuberculosis M. fi iy ute studies, Further city. [32] [23] fl ce yHVand HIV by icted saptoe is pathogen a as . Tetrahydrolip- . .tuberculosis M. i.5 Fig. [42] .tubercu- M. fi ) which C), mthese rm n the and , Indeed, . fi cant fi 71 c Manuscrit d’auteur / Author manuscript Manuscrit d’auteuVersionr / Author postprint manuscript Manuscrit d’auteur / Author manuscript Journal homepage B Ve 72 0% h uiyo ahrcmiatezm a veri was enzyme recombinant each of purity The 100%. the using pce,te ersn huge a represent they species, TB adults. a in to that from responses different immune be their can as infection infants, in investigated should be diagnosis in also enzymes lipolytic of potential The infections. TB response, the of values mean the represent lines continuous Horizontal (B). de Cut4 values or cut-off (A) the Rv0183 represent enzymes, lines lipolytic dotted recombinant whereas against tested individuals healthy BCG-vaccinated 5. Fig. eobnn iCwt eafo Bptet (HIV patients TB from sera with LipC recombinant aanadLuetKee,b h IACro Institute Carnot LISA the by Kremer, n ANR Laurent (Convention and framework Canaan 2009 the MIEN in (ANR Recherche grant ANR-Foamy-TuB la the de of Nationale Agence the by funded Acknowledgments TB. combating thus and understanding are in enzymes research bio- for their lipolytic mine of gold mycobacterial diversity a the The about functions. learned far be logical so to studied been remains have much number and limited a only which of enzymes, cystic such disorders as for lung underlying by with include, patients in pathogen by caused emerging These caused infections diseases. ulcer pulmonary mycobacterial Buruli other instance, for also but iochimie r si eas hs nye r hrdb eea mycobacterial several by shared are enzymes these Because hswr n h otdcoa elwhpt u eiuwere Dedieu Luc to fellowship post-doctoral the and work This on définitive du du m on définitive Mycobacterial lipolytic enzymes:Agold minefortuberculosis research. Biochimie, 95(1),66-73. , muoeooia epnet eobnn ioyi nye nT ainsadhatyidvdas AadB,IGhmrlrsos nsrmsmlsfo Bptet or patients TB from samples serum in response humoral IgG B), and (A individuals. healthy and patients TB in enzymes lipolytic recombinant to response Immunoserological Dedieu, L.,Serveau-Avesque, C.,Kremer, L.,Canaan, S.(Auteurde correspondance) (2013). mg J” “Image fi brosis.

(201 otaeb eemnn h nest ftercmiatLp-Hsbn gis niLp oprdt htaantat-i nioyta a osdrdas considered was that antibody anti-His against that to compared anti-LipC against band LipC-6His recombinant the of intensity the determining by software 3 .tuberculosis M. ),

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95, p. sc fi r l fepoainntol o TB for only not exploration of eld it xrse ltoao lipolytic of plethora a expresses 66- p yoatru abscessus Mycobacterium ublié yoatru ulcerans Mycobacterium DOI :10.1016/j.biochi.2012.07.008 73, þ TB Comment citer cedocument: fi þ e ntesuy BCG study. the in ned DOI e e n HIV and fi db D-AE(dpe rmRefs. from (Adapted SDS-PAGE by ed dan e : : 00)t Stéphane to 00904) 10.1016/j.biochi.2012.07.008 s s / Fi TB þ .Dde ta./Bohme9 21)66 (2013) 95 Biochimie / al. et Dedieu L. each , l nal

n þ ¼ ver D C-acntdboddnr (n donors blood BCG-vaccinated BD, )adcnrlsbet (PPD subjects control and 6) ,an si and on of theof on ma References manuscript. help the for editing Alahari in Anuradha thank Authors Université. Aix-Marseille 8 .Shé .Mui,R hub ..N J.C. Dhouib, R. Maurin, D. Schué, Tailleux, M. L. [8] Fornes, P. Pietri-Rouxel, F. Hernandez-Pando, R. Neyrolles, O. [7] Canaan, S. Carriere, F. Caro, de A. Chahinian, H. Douchet, I. Dhouib, R. Côtes, K. [6] N Bakala J.C. [5] 4 .Gro,H hitne,D inkn .Aebl,M ae,Intracellular Barer, M. Adegbola, R. Minnikin, D. Christensen, H. Garton, N. [4] 3 .Nyols .Giht eetavne ndcpeigtecnrbto of contribution the deciphering in advances Recent Guilhot, C. Neyrolles, O. [3] 1 H,Goa ueclssCnrl2011, Control Tuberculosis Global WHO, [1] 2 ..Mnii,L rmr ..Dvr ..Bsa h ehlbace forti methyl-branched The Besra, G.S. Dover, L.G. Kremer, L. Minnikin, D.E. [2] [31,32] ..Pyn .Pvr,Y odt .Aulr ..Peot .Ptt .Gcul Is Gicquel, B. Petit, C. Prevost, e43. M.C. (2006) Aguilar, for place D. a Bordat, tissue Y. adipose Pivert, E. Payan, J.A. 417 (2007) 408 from J. Biochem. lipase monoglyceride exported tuberculosis an of Characterization .Crir,S aan w uiaelk rtisscee by secreted proteins cutinase-like Two Canaan, S. Carriere, F. 1305 (2010) 1801 Acta Biophys. Biochim. of macrophages, effects cytotoxic 148 Microbiology sputum, in 2951 and (2002) vitro in mycobacteria of inclusions lipophilic 21)187 (2011) tuberculosis Mycobacterium ain of cations global_report/en/index.html ). nu sc e PPD ,

73 yoatru tuberculosis Mycobacterium e ri osbyivle ntemtbls fhs elmmrn lipids, membrane cell host of metabolism the in involved possibly ’ e oa .She .Crir,A ero,S aan vdnefrthe for Evidence Canaan, S. Geerlof, A. Carriere, F. Schue, M. goma, 195. pt publis 2958. þ HIV , ¼ 0,T,atv Bptet (n patients TB active TB, 50), þ TB yoatru tuberculosis Mycobacterium each , yoatru tuberculosis Mycobacterium hed e . iist ahgnss ueclss(dn. 91 (Edinb.) Tuberculosis pathogenesis, to lipids 427. n in ¼ : ) eaieimnratvt a calculated was immunoreactivity Relative 6).

hm il 20)545 (2002) 9 Biol. Chem. , http://www.who.int/tb/publications/ ’ oa .Dlre .Lambeau, G. Delorme, V. Goma, ¼ 0) C muoeciiyof Immunoreactivity (C) 105). hshlps towards C phospholipase essec?Po N 1 ONE PLoS persistence? e 1313. Mycobacterium Mycobacterium e 553. ﬁ - Manuscrit d’auteur / Author manuscript Manuscrit d’auteuVersionr / Author postprint manuscript Manuscrit d’auteur / Author manuscript Journal homepage B Ve iochimie r si 1]M ätran .Senühl eta ii oisi rkroe:recent prokaryotes: in bodies lipid Tri- Neutral Wenk, Steinbüchel, M.R. A. Dick, Wältermann, T. M. Pethe, [14] K. Bendt, A.K. Shui, G. Bardou, Rao, P.S. F. Low, Botanch, K.L. C. [13] Levillain, F. Poquet, Y. Vaubourgeix, J. Peyron, P. [12] Watching Canaan, McKinney, S. J.D. Munoz-Elias, Dukan, E.J. S. [11] Carriere, F. Hubert, P. Ducret, A. Dhouib, R. [10] 3]G hn .Snh .Cada .SreuAeqe .Mui,S Canaan, S. Maurin, D. Serveau-Avesque, C. Chandra, D. Singh, K. Shen, G. [32] Lecou M. Brust, B. [31] (2011). Acta triacylglycerol Biophys. Biochim. Liver lipases, Lehner, R. Quiroga, A.D. [30] virulence: in enzymes hydrolizing Lipid Kaur, J. Jadeja, D. by Singh, Esters G. Acids Fatty [27] of Respir. Hydrolysis Rev. Desbordes, Am. J. Andrejew, mycobacteria, A. of activity [26] Lipase Steenken, W. Hawkins, Balor, J.E. S. [25] in activity Bifani, Lipase Mahadevan, P. P.R. Talati, Gibson, S. K.J. [24] Dobson, G. Chastellier, de C. Kremer, L. [23] 2]NM ars,JD ik ..Bsa,Mcaim fltnyin latency of Mechanisms in Bishai, W.R. biosynthesis Dick, lipid J.D. for Parrish, N.M. sources [21] carbon of Use Ratledge, C. Wheeler, P.R. [20] Chen, B. Miczak, A. Munoz-Elias, E.J. Bentrup, zu Honer K. McKinney, J.D. Kolattukudy, [17] P.E. Dubey, V.S. Abomoelak, B. Sirakova, T.D. Daniel, J. Deb, Morbidoni, C. H.R. [16] Abomoelak, B. Sirakova, T.D. Dubey, V.S. Deb, C. Daniel, J. [15] 1]KL o,G hi .Nte,WK e,SD olen .Dc,SP Rao, S.P. Dick, D H. T. Mattos, Kohlwein, K.A. S.D. [19] Yeo, W.K. Natter, K. Shui, G. Low, K.L. [18] 2]B ilig rcn h aeo itr at cd:mtblcsuiso post- of studies metabolic acids: fatty dietary enzyme of fate interfacial the extremophilic Tracing an Fielding, B. lipase: Gastric [29] Carriere, F. Aloulou, A. [28] 3]ST oe .Boc,J akil .Grir .Cuce,D ars ..Gordon, S.V. Harris, D. Churcher, C. Garnier, T. Parkhill, J. Brosch, R. Cole, S.T. [33] [22] 9 .Dne,H amr .Db ..Srkv,PE Kolattukudy, P.E. Sirakova, T.D. Deb, C. Maamar, H. Daniel, J. [9] on définitive du du m on définitive Mycobacterial lipolytic enzymes:Agold minefortuberculosis research. Biochimie, 95(1),66-73. , Dedieu, L.,Serveau-Avesque, C.,Kremer, L.,Canaan, S.(Auteurde correspondance) (2013). omnylk hntp nlpdlae arpae,Po ahg 7 Pathog. PLoS macrophages, lipid-loaded in e1002093. (2011) phenotype dormancy-like a tuberculosis tuberculosis clfnto,FSBJ 4(00 1893 (2010) 24 J. 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