The Mechanisms of Antibacterial Action of Some Nonionic Surfactants by Suzanne Louise Moore ...

261 Pages·2016·12.13 MB·English

TheM echanisomfsA ntibacterial
ActioofnS ome
NonionSiucr factants
by
SuzannLeo uisMeo oreB,S c(Hons),
GRSC
A thesis subimnpi atrttieadl
fulfilomfte hnetr equirements
oft heU niverosfiB tryi ghton
fotrh ed egreoefD octoorf
Philosophy
December
1997
TheU niverosfiB tryi ghtionn
collaborwaittiUhon ni lever
ResearcPho,rtS unlight
THEM ECHANISMOSF A NTIBACTERIAALC TOIN OFS OME
NONIONSIUCRF ACTANTS
SUZANNE MOORE
L.
ABSTRACT
Anbtaicrtieaaglne taser composoefad d viersgeo rupa ndm ansyu ch
agetnsh avee nteedrc ommouns agteho rugehxp eirnecew itlthilt e
ifnortmiaoonnt ehimre chanoifas ctmi oSnt.u odfty hmee chiasnomfa ction
ofa na nmtiiocbriaagle pnortv idaenis n gshtii ntroe sistmaecnhcaiens ms,
tocxoiliocgpaorlbl emasn dt hdee siagnndd evleopemnto fn ew agentosr
combtiinoans.
Thep riymt aarrogfemt os ta ntiimcroabgiean(lte csxl udainbntigtio icists)h e
cyptloasmmeimcba nrean da ssotceiedan yzmes. Maenmebc-rtaiavgee nts
canc ausae c hanigne ftlhuteyi a dn/idopre rmieilatobyft hceyt oplasmic
membarneS.uc hc hangceasnb ed ertmeinde byt hleae kagoefc elalru l
consteinttsusu cahsp ostsauimi onnsu,co litedsea ntdh iecron stnittasun ed
aminaoc i.d Tshe efoffae nca tnb taicrtieaagle notnt hceyt oplasmic
membarnec ana lsboe d etreminbeyde liudctaintgh ee ffeocftt h e
antcietbraiaagle notnt heac tiivty meomfb arneb-ounednz ymesa nd
subsattruept ake.
A ragneo fa nioannidnc o nico snufiractawneetr ssc erenefdop ro tieanlt
anbtaicrtieaaclt ibvyia t gyor wtihn htiibomineh todF.r otmh isee nsicnrg
porceash so molosgeoirsueo sfa lcoehtohlo txeywsle arceh osfeonsr t udy.
Thes eirecso annitedt hsea mhee adogurps iez (6E)b utd fieferdi nt he
numbeorfc arbaotno mistn h eiigrl r 'otfuarpo'1m 0t o 1 .6
A rnagoef p orbeosfc ytoplamsemmibacnr dea mawgeeer u setdoe luctei da
mechanoifs mta icionnf otrimfoaontr h siesr iTeest.rl aizusoaml wteseru sed
asa m akreorfd amatgoem embanre-bodueynhddor gesneae nyzme.Ts he
twoh ighmeerm bse rtohfh eo molosgeoirue(ssC1 4E6a ndC 16E6) ihna d,
genaelltr liteef foence tny zmea ctiitcvyo mpeadwr itthho et ehrt wmoe mbse r
oft hsee ires tahneedfr olrietm eiadn tcietbiraalo ppterires.e fTothrheeer
futrhesrt udcioensc aetnetodrnt hoeht etrw moe mbseo rft he ise(esCr O1E 6
anCd 12E6.)
BotCh1 O E6a ndC 12E6 aer capabolfde a maigntgh ec ytoapslimc
membarneh,ow evret,h eea rppetaor sbueb tdfliefeer necsi nt hmeo doef
actiooftn h tew sou fratcan.tC s1O E6ha vse yrl titelfef beecwlt o iMtIswG h list
C12E6e xteeridt esf feactst usb -lMveIesGl; b otshurf actaanrtpeesr s eanst
monomsae trt hecsoen caetnitornIists.p o rpostehtdat hedsfief ereinnc es
itnreaocntai rdeu teo t hree lastziievsoe ft hhey rodphotbaiicl tsh eosfe
compounThdest a.lgi oruopf C 12E6i sa popxrimtaitnhgtao fp hospholipid
andi sl eilktyoi nstie trsienltafon dfp lf-iloapco rsts hmee mbarneT.h e
shtorerc halientn hog fC 1O E6ic nerasteshh ey rodphiiictlyt hosifurs f actant
whiaccch umlutaeast t heext eroifto hrbe li ayuentria l s atuariton tpi osi n
recaheadn dgo rsmse mbanred amaogcce usr.T heu soef D SCa ndE SR
may opvricdoen saibdlieenrg shtii tnot hpiorsp osietnde rtaicon.
Contents
Page
LISOTF F GIURSE
LITS OFT ABLES
ACKNOWLEDEGMENTS
TEXTA BBREVITAIONS
CHATPERO NE Itnroudction
11. ThSet rucotftu hbreae tc ercielal..l. . . 1
.. . ...... ......................... .........
1. 1 T.h1ce e wlalll . 1
........... ................................. ............................
1.21 T.hpee riplsapsacmei c 6
.................. .....................................
1. .13 T hcey tloapsm . 7
........................................... ................... ......
11.4. Thoeu tmeerm brane 7
............................ ..............................
1. .15 T hcety oplamsemmbirca ne . 1 2
............................. ................
1. .16T he stero uft chctetyu orplamsemmiabcnr e 14
..................
12. Boichmeiacpla htwyas bacteria 17
in ........... ............... .....................
12..1E mdben-yMeerhof-oPrga lcryonlayspta itchw .ay1 7
.. ......
12..2P etnospeh ostpeph aathway 17
............................... ........... ..
12..3E nt-nDeoruodfpofar thway 19
................................ ................
12..4T ribcoayxrlaiccic cydl e . 20
.....................................................
12..5E lcetrtoarnn stpc ohraainnod x idative
phosryplhaotion 20
........................ ........ ......................... ........
13. AntcitbeaArgineatls 24
............................... ................ .......... ........ ... ....
1. 31. Mechiasnmosfa citoonfa ntibaacgteenrtisa l2 4
...............
1. 32 .Thee ffoefac ntt ibaacgteenortnits ah le
cyotplasmmeimcab nre 30
................. .....................................
1. 23..1G enaelmr embarne mpeebariitleyf fect3s1
.........
13...12C ytopliamcse mmabnre-ubneodn zyme3s 1
.......
Page
14. Surtfaancts 33
... .............................. ......................................................
1. .41D enftiiiocnlsa,s asnegdse naelpro rperties
ofs urfact.a..n.t...s . .. .. 33
.. ... . ......... ............. ............ ....................
14 2 Chemiccaolm posoifst ufiraocnt a.n.ts . 40
. . .... ... ........... .........
14..3S urfaacntatd sorpt.i.o..n.. . . ......4 2
....... ... ... ..... ....... ........ .... ... . ..
14..4T hein atcetriosnurf aocfta wnittmshe mabnre..s. ...50
. . .. .. .
14..5S urfactaansat nst imiacgreonbtisa l ....5 9
.............. ............ ..
14..6S tructurer-etfliuaonincspso thfsi urfoanc tants6 5
..............
1. 5 Aimasn Odb jecti.v..e..s. ............. .. 7 4
.... .. ............ ... ......... .... ..... ..... . .....
CHATPERT WO Anitmircoibaslc erengi nsutdieosf a
rnageo fn o-nionsiurcf actants
2.1ltn rodu.c.t.i.o.n. ...................................................................7....5.... ......
2.2C hemiucsaelds ... ..7 6
............................... .. .............. ........................ ...
2.12 S.urfactants . ... .. .76
................ ................... .............. ...... .... . ......
2.2C.u2lt uMreed ia .. 76
............................ ......................................
2.3Wa sihgno fG alssw.ar.e. ... ...........7.8
. ......... .. ................ ..... ........ ..... ...
2.4G rowtahn Mda itnenaonfMc cieor ogarnis.m.s .... .7 8
... ... ..... .. .. .. ....
24.1. PorducotfCi oeCnla lil baitroCnur ves 79
............. ...............
2.4P.or2d utcioofGn ro wtChu ervs. ..........................8...1... ...........
2.4R.e3s lut.s.. .....................................................................8...4... ...
2.5Ch oiocfBe ue ffr . .. ... .. . .87
.................... ...... . ..... .......... .......... ............... ..
2.15 P.erpaatriaonnsd t leairitsioonfM cllvaibnuef'f.se. 8r.7 . ..
.
2.6A ntimoibciprarlorp teioefsr n aag eo fs urfact.a..nts .8.7
.. ............. .
2.16 .DeteiranitmoonfM inimIunbmhi iyt or
Conctertnaio(nMs)I C........... . 87
... . .. .... ............. ... .............. .....
2.6R.e2s ults . .... . .8.8
........................... ... .. .. ................................... .....
Page
2.7 Antimicrpobrioartplie eosfa h omologosuesr ioesf
alcoheotlh oxylates. 91
..... ... ............. ...... ......... ..... ............ ................ ...
27..R1e sluts .. ..............9 .1. .....
...... ........... .. .............. ................... ... ....
2.8D sicussion 92
... ............ ......... ..... .... .............. .................................... .....
CHATPERT HREE Phsyiaclp roeprtioefas h omoglouos
serioefsa lcohehtoolyx lates
31. ntIrod.u..c.t..i.o.n .............................9..4.. ..............................................
3.2 DeterminaotfCiro int iMciacle lCloe ncterinaotn(CsM C). ..9.5. ....
3.3L opg d eetrminatoifto hnes uarfcta.n.t.s. ..............1..0 ...2... ........ ......
3.4 Cloupdo indte terminatoifto hnase l hcooelt holaxtesy. ...1.20. ......
3.5Ds icuss.i..o..n... ............... .........1.50. .
.. . . .. .............. ..... .... .............. .... . . ... .
CHATPERF OUR Thee ffeocfat cl ohoelt ohxyalteosn the
cellluamre atboilsmo f NCTC8 196
Escherichicao li
41. Oxygen UptakSet udies ..............1..0. .7.. ..
............ ......................... .... . .
41.1. Calaitbioroftn h Cel aorxky geelnte rco.d.e. .....10. 7.
. . . ... .... ..
41.. 2 Thera toefo xyegnu ptakeE. cboyNl CiT C8 196.. ....10. 8. .
4..13 T hera toefo xygueptna keE. cboyNl CiT C8 169
in ptrhees eonfc eC 1.. ...O...E...6... ................................1...0 . 9
41..4R esluts ................ 1 1 1
.... ............................. .. . . .... .................. .... ...
4.2T hue seo f2 .3.5-triphenyltcehtlroarzaiodslae i nu m
dehydroegnasae cttiyv i............ .131
indicator of ........ . . ... ...... ...... ..... .
4.12 D.evleopmtoe fen xpericmoenndtia.tl..i ..o...n..s.. ..1.41
. . .. .. . .
4..21.D1e teirntmaioofna pprotpewr aivlaenegth
atw hitcomh e sauerf ormazan 141
............. ............
Page
4.122.. Thee ffeocfta eratioonfn o rmazan
production. .. ... 151
...................................... ..... .. ... ......
4.123.. Thee ffeocftc elclo ncentraotni on
formazparno ducti.on 191
.... .................................. ...
4.2T.h2ee ffeocfta lcoheotlh oxylaotnfe osr mazan
product.i.on. ...... . 122
....... .... .......... ................................. ............
4.22..1Th ee ffeocfta lcoheotlh oxylaotne s
formazpanr odcutiouns inggl ucose
subs/ tr...a...t..e... .................................................... 1 22
4.2.T2h.ee 2f feocfta lcoheotlh oxylates
formazparno ductuisoinn sgu ccinate
substrate 12 3
...............................................................
\
4.2R.e3s ults 12 8
..................................................................................
4.3Bi olumicneenAstT PA ssa..y... ...............................:... ..... ....1..43.. .. ....
4.13 D.eterminaotfEi xotnr acCtoinodni ti..o...n...s... ............... 153
4.3V.a2li daotifto hnea ssay ....... 193
............................ .. . .. . . .............
4..32.Th1e e ffeocftc eldle broinst hea ssay 193
...............
4.3.D2e.te2r minaotfti hoenA TPc oncentration
rangoef t he ass.ay .. ....1 .93. .
.. ........ . ....................... ..
4.3.D2e.te3r minaotfti hoens tabiolfit thye
ligohutt put. . . 1 4 1
.... ............. .......................................
4.34. V2a.riatiboent weebna tchoefsm onitoring
reagent . .... .. . 1 4 1
........................... ... ..... ................... ....
4.3.T2h.ee 5ff eocfts urfactaonntl su ciferase 114
.............
4.3..6T2 hee ffeocfts urfactaonntt sh ee xtraction
ofA TP ....... 154
.............................. . ..... .........................
Page
4.3.A3T.Py 1i elfdr omE .c oNlCiT C8 196i nt he
absencoefs urfactants. .. 184
............. ...... ... ..............
4.3.A3T.Py 2i elfdr omE .c oN/CiT C8 196i nt he
presenocfes urfacta.nt.s .. .184
.. ....... ......... . ........... ...
4.3R4e sults 15 2
...................................................................................
4.4Gl ucouspet akset udie..s . . 15 3
... . .................. ................... .....................
4.14 P.henoslu lphuarciicad s say. .. . . .1 45
.. ........ .. ............... .... ........
4..4.11G lucosuep takbey E .c oNlCiT C8 196
usi/nt gh ep henol-sulapchiuadrsi sca ..y... ...... 145
44.1.2. Thee ffeocfti ntcreallumlaatre rioantl h e
phenoslu lphuarciicad s say .155
.......................... ..
4.4G.l2u coosxei daasses ay . 195
.......................... ...........................
4.41. A2s.savya lidation 195
............................................ .......
4.241... 1P reparatoifco anl ibractuirvoens . 195
...... ...........
4.41.2.2 S.talbiiotyfo xidisoe-dd ianisidine 126
........... ......
4.4.G2l.uc2o suep takbey E .c oNlCiT C8 196
usinag g lucoosxei daasses ay 126
.................. ......
4.4.G2l.uc3o suep takbey E .c olNiC TC8 196i n
thep resenocfea lcoheotlh oxylates 156
................
4.4.R2e.su4l. ts . 16 6
........................... ...... ...................... ..........
4.5Di scuss..i.on .... . . . .. .1 96
. . ............. ... ... .......... ....... .......... ........ . .........
Page
CHATPERF IEV Thee ffeocfat l ochole tohxlyateosn the
permaebiitloyf t hec yotpalsmicm embraen of
NCTC 8196
Escherichicao /i
51. Theef feocfta lcoehtoohyxlal teosnp otsasiuimno
laekgaef roE.m c o/NiC T8C 916. .............................................1.7. 1
51.1. Methdoedv eleon.pt.m 171
............ ...........................................
51..2M ethVoadla itdion ..............................1.37
............ . . . . . . ... .. . .... .. . .. ... .. . . .
5..1.21C alaiitboornft hPee kriEnl emrM 110
AtomAibcs orpStpieocnt roepthr.eo .t.o1.m37. ......
5..122 . Tehfef oeffcl ittt riaoonnt hmee auserd
potiasuismo cno ncaeitno.tn.r .........1..47.. .......... ....
5..12.T3h eef foefac lt coehtohlo xyolnta htee s
meauserdp ostsauimi ocno nctertnai.o.n. 1.47. ....
5..13.L1e akaogfpe o tiasuismo nfrso m
E. coNlCiT C8 16.9 ...............................1...57... ..... ...
5..13.D2e termionfa tttihroeanl c liaeurnl
potiasuismo n pE.o ocolN/ CioT fC8 196. .1.57. ..
5..13.L3e akaogfpe o stsauimi onfrso m
E. coN/CiT C8 196i tnh peer seen cof
alcoehtohlot xey.sl. .a.. ...... .. 167
.... . . .. ...... .. . . .. . .... ... ....
51.4. Resluts 167
..................................................................................
5.2Th ee ffoefac ltc oehhtooylxal teosn lekaagoef2 6n0 m
the
absormbatienrgfi raoE.lm c o/iNC T8C 196. .......1.18. ...................
5.2D.e1t enratmiiooftn h mee tbaolpiocoo lf2 6n0 m
absroibnmgat erfiraoElm.c olNiCT C8 196. ....1.18. ..........
5.22 M.ethVoadl idation . . 138
..................................................... ........
Page
5.2L.e3a kagoef2 6n0m absorbimnagt erifarlo m
E.c oN/iC CT 8196. .......................................................1..38.. ..
5.2L.e4a kagoef2 6n0m absorbimnagt erifarlo mE .c oli
NCCT 8196i nt hep resenocfea lcohol ethoxy1l48a tes
.....
5.2R.es5u lts .. . .. 148
...................................................................... ..... ....
5.3Di scussion. . . ..............................1..68.
...................................... . . .. . ...... ...
CHAPTERS IX DICSUSSION
6.G1r owtihn hibitteiso/..t. n.s. . ..............................................................1 88
6.2St ructure-fruenlcoatntisiohni ps. ...............:... .......1.9 .1. .
............. . . . . . . .
6.3Su rfactaadnsto rption . .... ... ..139
............................... .. . ....... ... . ............ ..
6.4Da maget ot hec ytoplasmmemibcr ane. ...................19 4
. . ..... .. . . . .. .. . .
6.5Fu rthweorr k . . ................2 .0.3.
. .................................................. .. . . .. .. . . ...
205
APPENDI.C...E...S.... .....................................................................................
.....................................'....... .....................................2..0..7
REFERENC. ES . . . . . . . . .
LiosftF igures on foolrwlo ing Page
Chpater1
Figeu .r1.1 . .D.i.aa.gm.or .ft. hb.ea. c tceelrila.l 2
.............. ..............................
Fige1u 2r.. ..T.h.se. t.r.ru.eco. tfa.u G arm-posbiatcitveer ia 3
.... ............. .....
Fiuger1 3. .T.h se trrueco tfau G ram-gnaetbiavcet eria .. 4
......... ................. ..
Fiuger1 .4 . Tshtert uuecor ft hpel asmmeamb rane . 1 6
.......... .......... ............. ..
Fige1u 5r. .Embdeny-eMrehof-oPrgay lrcnoalspy attihcw ay1 8
......... . .........
Fige1u 6r.. ....P..e.n.t.po.hs oes tpeph aathway .... ..1.8.
.................. ............... .. .
Fige1u 7r.. . Ent-nDeoruodfpofar th.w.ay . ...1.8
. .. ...... . . ............. .... ................... . ..
Fige1u 8r.. ..T.r.i.c.ay.rl.abic.ocic.x dy c l.e. . .. . .2.1
..... .... . .............. . ... ............ .... ..
Fige1u 9r.. ..T.h.ge. e.na.et.ri.oo.fnp oart omno tifvoer. .c.e .. .2.1
. .................
Fige1u 1r.0 Theel ecttrraonntsc phoaorifn c E.o /i. .. 21
......... ..... . ....................
Figeu .r111 Them aitna rgfeontros an nt-itbiico
.........
antibaacgteenrti.sa. l . 2 6
....... .. ..................................... ...........
Figeu1 1r.2 Thec onseencqeuosf a ntcietbiraal-aignedenudtc
.........
damage 27
.................................................................................
Fiuger1 1.3 Retliaonbsehtiwpeb eanc teriaonsdt atic
.........
bacrtieceifdfaelc t.s 2 8
........ ..................... .................. ..............
Figeu .r11 4.. Chnageisnc onctertnaidoenp enydo efan w cide
..... . .
vaertioyfp hsyicohemipcoraple rtaioreusn tdh e
critmicicealcllo en ctaertni.o.n .......39
.. .............. ............... ....
Figeu1 1r.5. .At ypiLcaanlug miard soripstoito.hn.e rm ...34
.... . .. . ............... ....
Fiuger1 1.6 A genaeilrseadd spoitroinst ohme srhowtihneg
.........
typoefss u fractaagngtr egeantcioouennd ot ne r
thseo lsiurfda cietn h vear iorueisgo ns.. :. .........44.
...... . .....
Figeu1 1r.7 Adsotripoofnn onisournficatca nCtx6sEo n
.........
Grahpon . 44
................. ..............................................................