IAOe?IIAEUeIA AEAAeAI?ss IAOE OE?A?IE

?INOEOOO IIEAEOEss?II? A?IEIA?? ? AAIAOEEE

OAeE 577. 217. 32

OAeE 577. 217. 337.2

?ai?iaa Na?oeaia Iaae?aia

OE?ICEE-o?IE NEIOAOACA ?C AENO?AIAEUeIIAI OA?IIO?EA THERMUS
THERMOPHILUS HB -27: O?CEEI-O?I?*IA OA?AEOA?ENOEEA OA?IAIOA ?
ACA?IIAe?ss C o?IETyr

03. 00. 03 — iieaeoey?ia a?ieia?y

Aaoi?aoa?ao

aeena?oaoe?? ia caeiaoooy iaoeiaiai nooiaiy

eaiaeeaeaoa a?ieia?/ieo iaoe

E e ? a — 1 9 9 9

Aeena?oaoe??th ? ?oeiien

?iaioa aeeiiaia o a?aeae?e? aiceiieia?? a?eeiaiai neioaca ?inoeoooo
iieaeoey?ii? a?ieia?? ? aaiaoeee IAI Oe?a?ie.

Iaoeiaee ea??aiee ( aeieoi? a?ieia?/ieo iaoe

Ooeaei Ieoaeei A?naio?eiae/,

?IA?A IAI Oe?a?ie, caa. a?aeae?eii

Io?oe?ei? iiiiaioe ( aeieoi? a?ieia?/ieo iaoe, i?ioani?

Aeiia?aaeiaa ?oo?ia Iao??aia,

Ee?anueeee Oi?aa?neoao ?i. Oa?ana Oaa/aiea,

eaoaae?a a?io?i??,

eaiaeeaeao a?ieia?/ieo iaoe

Iao?ooaiei Ciy Ieoaee?aia,

?IA?A IAI Oe?a?ie, noa?oee
iaoeiaee ni?a?ia?oiee

I?ia?aeia onoaiiaa ( ?inoeooo a?ii?aai?/ii? o?i?? oa iaooio?i?? IAI
Oe?a?ie,

a?aeae?e no?oeoo?e ? ooieoe?? a?ee?a ?
iaioeae?a

Caoeno a?aeaoaeaoueny » 18 » o?aaiy 1999 ?ieo i 1000 aiaeei? ia
can?aeaii? Ciaoe?ae?ciaaii? A/aii? ?aaee Ae 262.37.01 ?inoeoooo
iieaeoey?ii? a?ieia?? ? aaiaoeee IAI Oe?a?ie ca aae?anith: 252143,
i.Ee?a — 143, aoe.Caaieioiiai, 150

( cae can?aeaiue ).

C aeena?oaoe??th iiaeia iciaeiieoenue o a?ae?ioaoe? ?inoeoooo
iieaeoey?ii? a?ieia?? ? aaiaoeee IAI Oe?a?ie.

Aaoi?aoa?ao ?ic?neaiee » 15 » ea?oiy 1999 ?ieo.

A/aiee nae?aoa?

Ciaoe?ae?ciaaii? A/aii? ?aaee,

eaiaeeaeao a?ieia?/ieo iaoe
Eoeao E.E.

CAAAEUeIA OA?AEOA?ENOEEA ?IAIOE

Aeooaeuei?noue i?iaeaie. Ai?iiaoeee-o?IE neioaoace ( A?Nace )
i?aaenoaaeythoue a?oio oa?iaio?a, ye? a?ae?a?athoue aaaeeeao ?ieue a
?aae?caoe?? aaiaoe/ii? ?ioi?iaoe??. Iniaeeaee ?ioa?an aeeeeea? aea/aiiy
A?Nac ?c aeno?aiaeueieo oa?iio?eueieo aaeoa??e, ye? aeeaooue i?e
oaiia?aoo?ao 60 — 85 ? N ? oa?aeoa?ecothoueny aeeaiaeaeiith
oa?iinoaa?euei?noth.

I?iea??ioe/ia oe?icee-o?IE neioaoaca iaeaaeeoue aei 1-ai no?oeoo?iiai
eeano A?Nac

[ Eriani, 1990 ]. O aeiaaeeo i?iea??io o?IETyr ia? aeiaao aa??aaaeueio
a?eeo. Iacaaaeath/e ia oa, ui i?iea??ioe/ia oe??N ?c Bacillus
stearotermophilus yaey?oueny iaei??th ?c ia?oeo A?Nac, aea/aieo
iaoiaeii ?aioaaiino?oeoo?iiai aiae?co [ Brink, 1989 ], ?ioi?iaoe?y i?i
no?oeoo?o N-e?ioeaaiai aeiiaia a?eea a?aenooiy ? ni?iae io?eiaoe
e?enoaee eiiieaena o?IETyr-oe??N aoee iaaaeaeeie. I?iaaaeai? ?ai?oa
a?io?i?/i? oa aaiaoe/i? aeine?aeaeaiiy ia aeicaieeee iaeiicia/ii
aecia/eoe oei aca?iiae?? ia?e o?IETyr-oe??N. Aeniiaie i?i oa?aeoa?
aca?iiae?? aeaii? ia?e o?IE-A?Naca , c?iaeaiee ia iniia?
no?oeoo?ii? iiaeae? eiiieaena o?IETyr-oe??N

[ Bedouelle, 1990 ], ia ocaiaeaeo?oueny c eiioeaioe??th i?i ?nioaaiiy a
aeaio no?oeoo?ieo eeanao A?Nac aeaio ??cieo oei?a aca?iiae?? c o?IE.
Oiio aacia?a/iee ?ioa?an i?aaenoaaey? c’ynoaaiiy oa?aeoa?o aca?iiae??
aeaii? ia?e o?IETyr-oe??N. I?iaaaeaiiy ii??aiyeueiiai aea/aiiy ae?eyiie
aca?iiae?? o?IETyr ?c aeno?aiaeueiiai oa?iio?ea T.thermophilus ?
iacio?ea E.coli c aiiieia?/ieie A?Nacaie aeicaieeoue, ii-ia?oa,
aecia/eoe oa?aeoa? aca?iiae?? aeaieo ia? o?IE-A?Naca a ?ic/ei?, ?,
ii-ae?oaa, ioe?ieoe, iane?eueee aaeaeaaoii oe? aea? ia?e o?IE-A?Naca
aca?iiae?thoue a ?ic/ei?, ? oiio iane?eueee aoaeooue ei?aeoi? aeniiaee,
c?iaeai? ia iniia? iiaeaeueoeo no?oeoo?ieo aeine?aeaeaiue oe??N ?c
T.thermophilus , yea iiaea aeyaeoeny a?eueo ia?niaeoeaiith o oeueiio
a?aeiioaii?, oae ye ? a?eueo oa?iinoaa?eueiith, o ii??aiyii? c
aiaeia?/iei iacio?eueiei oa?iaioii.

Iaoa ? caaaeaiiy aeine?aeaeaiue. Iniiaia iaoa aeaii? ?iaioe iieyaaea a
aeine?aeaeaii? o?ceei-o?i?/ieo aeanoeainoae oa iniaeeainoae
ooieoe?ioaaiiy oe?icee-o?IE neioaoace ?c aeno?aiaeueiiai oa?iio?ea
T.thermophilus HB-27, ? aecia/aii? ae?eyiie aca?iiae?? o?IETyr ?c
T.thermophilus ? E.coli c aiiieia?/ieie ai?iiaoeee-o?IE neioaoacaie a
?ic/ei?.

Aeey aeinyaiaiiy oe??? iaoe iaiao?aeii aoei ae??oaiiy oaeeo caaaeaiue:

?ic?iaeoe noaio aeae?eaiiy ?iaeea?aeoaeueieo i?aia?ao?a oe?icee-,
na?ee- ? o?aii?e- o?IE neioaoac ?c T.thermophilus ? aeae?eeoe
aenieii/euaiee i?aia?ao oe?icee-?IE neioaoace ?c E.coli.

I?iaanoe ii??aiyeueia aea/aiiy o?ceei-o?i?/ieo ? eaoae?oe/ieo
aeanoeainoae oa?iio?eueii? ? iacio?eueii? oe?icee-o?IE neioaoac.

Aecia/eoe ioeeaioeaeio iine?aeiai?noue o?IETyr ?c T.thermophilus.

Aecia/eoe ae?eyiee aca?iiae?? o?IETyr c oa?iio?eueiith ? iacio?eueiith
oe?icee-o?IE neioaoacaie iaoiaeaie o?i?/ii? iiaeeo?eaoe?? ? ioeeaaciiai
a?ae?ie?ca.

Iaoeiaa iiaecia ?iaioe. Aia?oa aeae?eaii aenieii/euai? i?aia?aoe
oe?icee-, na?ee- ? o?aii?e- o?IE neioaoac ?c T.thermophilus. Iieacaii,
ui oe? oa?iaioe iathoue ie?aiia?io no?oeoo?o oeio (2 c iieaeoey?iith
ianith noaiaeeieoe? 50, 46 ? 74 eAea, a?aeiia?aeii. Iioei?ciaaii iaoiae
aeae?eaiiy oe?icee-o?IE neioaoace ?c E.coli. Aia?oa cae?eniaia
o?ceei-o?i?/ia oa?aeoa?enoeea oe?icee-o?IE neioaoace ?c aeno?aiaeueiiai
oa?iio?ea T.thermophilus HB-27 ? c’yniaai? iniaeeaino? ooieoe?ioaaiiy
oa?iio?eueii? oe?icee-o?IE neioaoace. Ia iniia? iaea?aeaieo
aenia?eiaioaeueieo aeaieo ie i?eionea?ii, ui i?e ia?aoiae? a?ae
iacio?eueiiai niiniao aeeooy aei oa?iio?eueiiai i?aeaeuaia
oa?iinoaa?euei?noue oa?iio?eueii? oe?icee-o?IE neioaoace aeinyaa?oueny a
?acoeueoao? ci?ie ai?iieeneioiiai neeaaeo oeyoii i?aeaeuaiiy ai?noo
a?ae?ioiaieo ai?iieeneioieo caeeoe?a, a iniiaiiio Pro ? Leu, a oaeiae
caoenoii oa?iaioa iecueeiiieaeoey?ieie noano?aoaie — AO?, AO? ? L-
oe?iceiii.

Aia?oa aecia/aia ioeeaioeaeia iine?aeiai?noue o?IETyr ?c
T.thermophilus. Anoaiiaeaii, ui nooi?iue ?? aiiieia?? c o?IETyr ?c
E.coli neeaaea? 52 %.

Aia?oa aecia/ai? ae?eyiee aca?iiae?? o?IETyr c oa?iio?eueiith ?
iacio?eueiith oe?icee-o?IE neioaoacaie a ?ic/ei? iaoiaeaie o?i?/ii?
iiaeeo?eaoe?? ? ioeeaaciiai a?ae?ie?ca. Ii??aiyiiy ae?eyiie aca?iiae??
oa?iio?eueii? ? iacio?eueii? o?IETyr c aiiieia?/ieie ai?iiaoeee-o?IE
neioaoacaie iieacaei, ui aiie a iniiaiiio ni?aiaaeathoue ? ?ici?uai? a
/ioe?ueio, aeaui a?aeaeaeaieo iaeia a?ae iaeii?, ae?eyieao ia?aeiii?
no?oeoo?e o?IETyr. ?o ?ici?uaiiy ia iiaeae? i?inoi?iai? no?oeoo?e o?IE
2-ai eeano iaeiicia/ii aeaco? ia oa, ui oe?icee-o?IE neioaoaca
aca?iiae?? c o?IETyr c? noi?iie aa??aaaeueii? a?eee.

I?aeoe/ia cia/aiiy ?iaioe. A aeai?e ?iaio? i?aaenoaaeaii iia? aoaeoeai?
? aeinooii? a i?aeoe/i?e eaai?aoi?i?e ?iaio? iaoiaee iaea?aeaiiy
?iaeea?aeoaeueieo i?aia?ao?a oe?icee-, na?ee- ? o?aii?e-o?IE neioaoac
?c T.thermophilus. ?acoeueoaoe i?iaaaeaieo aeine?aeaeaiue iathoue
oai?aoe/ia cia/aiiy aeey ?icoi?iiy aca?iica’yceo no?oeoo?e, ooieoe?? ?
oa?iinoaa?eueiino? oa?iio?eueii? oe?icee-o?IE neioaoace. Aeaia
aeena?oaoe?y yaey?oueny nooo?aei aeeaaeii a c’ynoaaiiy oeio aca?iiae??
o?IETyr,yea ia? aeiaao aa??aaaeueio a?eeo c iacio?eueiith ?
oa?iio?eueiith oe?icee-o?IE neioaoacaie a ?ic/ei?. Io?eiai? ?acoeueoaoe
yaeythoueny iniiaith aeey i?iaaaeaiiy no?oeoo?ieo aeine?aeaeaiue ye
oe?icee-o?IE neioaoace, oae ? eiiieaeno o?IETyr-oe?icee-o?IE neioaoaca.

Ca’ycie ?iaioe c iaoeiaeie i?ia?aiaie, ieaiaie, oaiaie. Aeena?oaoe?y
a?aeiia?aea? iniiaiiio ieaio iaoeiai-aeine?aeieo ?ia?o a?aeae?eo
aiceiieia?? a?eeiaiai neioaca ?inoeoooo iieaeoey?ii? a?ieia?? ? aaiaoeee
IAI Oe?a?ie. ?iaioa oaeiae aeeiioaaeanue i?e i?aeo?eioe? a?aioo ?
75195-548201 » The structural basis for the recognition of
aminoacyl-tRNA synthetases for their cognate amino acid and tRNA »
Howard Hughes Medical Institute.

Iniaenoee aianie caeiaoaa/a. ?iaioa aeeiiaia iniaenoi aaoi?ii i?ae
ea??aieoeoaii aeieoi?a a?ieia?/ieo iaoe Ooeaei I. A. , a oaeiae o
o?niiio ni?a?ia?oieoeoa? i?e io?eiaii? ?iaeea?aeoaeueieo i?aia?ao?a
oa?iio?eueieo ai?iiaoeee-o?IE neioaoac c e.a.i. ss?ai/oe A. Ae., c
yeith aaoi? ia? ni?euei? ioae?eaoe??. Aaeoa??aeueia iana T.thermophilus
HB -27 eoeueoeaiaaia a

?inoeooo? o?c?ieia?? ? a?io?i?? i?e?ii?aai?ci?a ?AI, i. Ioueii ,
?in?enueea Oaaea?aoe?y. I?aia?ao noia?ii? o?IE ?c E.coli io?eiaiee ?c
AI?? i?eeeaaeii? a?io?i??, i. Ieaeia, Eaoa?y. Iino?eainiiaeueiee
noia?iaoaio E.coli, yeee iaa o nai?io neeaae? oe??N, aoa etha’ycii
iaaeaiee Eaa?ee I.?. c ?inoeoooo a?ii?aai?/ii? o?i?? NA ?AI, i.
Iiainea??nuee, ?in?enueea Oaaea?aoe?y . Ai?iieeneioiee neeaae oe??N ?c
T.thermophilus aoa aecia/aiee Iaceiiaei ?.A. a ?inoeooo?
a?ii?aai?/ii? o?i?? ?i. Oaiye?ia I.I. ? Ia/eiieeiaa TH.A., i. Iineaa.
?iaeea?aeoaeueia o?IETyr ?c T.thermophilus aoea etha’ycii iaaeaia i. n.
E?eeeeaei ?.A. , ?IA?A IAI Oe?a?ie.

Ai?iaaoe?y ?iaioe. Iaoa??aee aeena?oaoe?? aeiiia?aeaeenue oa
iaaiai?thaaeenue ia Ananithciiio neiiic?oi? ii o?i?? a?eea ( Oa?e?n?,
1990 ), I?aeia?iaei?e eiioa?aioe??

» Oa?iio?ee : iaoea ? oaoiia?y » ( ?ae’yaee , ?neaiae?y, 1992 ), XVII
I?aeia?iaei?e ?iai/?e ia?aae? ii o?IE (*?aa, ssiii?y, 1997 ).

Ioae?eaoe??. Ii oai? aeena?oaoe?? iioae?eiaaii 9 iaoeiaeo ?ia?o.

No?oeoo?a oa ianya aeena?oaoe??. Aeena?oaoe?eia ?iaioa neeaaea?oueny
c? anooio, iaeyaeo e?oa?aoo?e ( 1 ?icae?e ), aenia?eiaioaeueii? /anoeie
( 2 ?icae?ea ), caeeth/aiiy , aeniiae?a ? nieneo oeeoiaaii? e?oa?aoo?e
, yeee aeeth/a? 171 iaeiaioaaiiy. ?iaioa aeeeaaeaia ia 155
noi??ieao iaoeiiieniiai oaenoo, ?ethno?iaaia 44 ?enoieaie ? 6
oaaeeoeyie.

IAOA??AEE ? IAOIAeE AeINE?AeAEAIUe

Aeey aeae?eaiiy i?aia?ao?a a?ee?a oa noia?ii? o?IE aeei?enoiaoaaee
a?iiano T.thermophilus HB-27 ? iino?eainiiaeueiee noia?iaoaio E.coli
MRE-600. I?aia?ao noia?ii? o?IE io?eioaaee ii iaoiaeo [ Brungraber, 1962
], c aeayeeie iiaeeo?eaoe?yie, oeyoii i?yii? aeai?ioa?i?caoe?? ee?oei
a?iiane oaiieii c iiaeaeueoith o?iiaoia?ao??th ?IE ia AeAAA-oeaetheic?.

Aaeue-aeaeo?ioi?ac a?ee?a a iaoeaieo oiiaao i?iaiaeeee ii iaoiaeo [
Hedrick, 1968 ], a aeaiaoo?oth/eo oiiaao ii iaoiaeo [ Weber, 1969 ].
Eiioeaio?aoe?th a?ee?a aecia/aee ii iaoiaeo

[ Bradford, 1976 ], a a?oaeo aeno?aeoao ii iaoiaeo [ Warburg, 1941].

Aeoeai?noue oa?iio?eueieo neioaoac a ?aaeoe?? ai?iiaoeeeoaaiiy o?IE
aecia/aee a ?ieoaaoe?ei?e noi?o? ia’?iii 50 iee, ui i?noeea 100 iI
o?en-HCl , ?I 7,9, 10 iI MgCl2, 3 iI AO?, 5ia/ie noia?ii? o?IE E.coli,
0,2 ia/ie ANA, 0,2 iI [14N]-oe?iceia aai 0,3 iI [14C]-na?eia aai 0,4 iI
[14C]-o?aii?ia, a?aeiia?aeii, ? a?ae 0,05 aei 10 iea a?eea ( a
caeaaeiino? a?ae nooiai? i/enoee oa?iaioa ). Noi?o ?ieoaoaaee 0,5 oa.
i?e 55( N. ?aaeoe?th coieiyee aeiaeaaaiiyi 200 iee ioieiaeaeaiii? 10
%-ii? OOI. Ooai?ai? inaaee a?aeieaaee ia i?e?ii?iaeo o?eueo?ao 50 ie 5
%-ii? OOI.

Aeoeai?noue oa?iaioa a ?aaeoe?? AO?-???-iai?io aecia/aee ii iaoiaeo [
Waller, 1968 ] c aeayeeie iiaeeo?eaoe?yie. E?iaoe/i? aeanoeaino?
oa?iio?eueii? oe??N aea/aee, ci?ithth/e eiioeaio?aoe?th iaeiiai
noano?aoo i?e iane/oth/eo eiioeaio?aoe?yo ?ioeo noano?ao?a, i?e 37 ?N ?
65 ?N. Aaee/eio Em aecia/aee ii iaoiaeo iaeiaioeo eaaae?ao?a c
aeei?enoaiiyi eiii’thoa?ii? i?ia?aie Enzfitter [ Leatherbarrow, 1990 ].
E?ea? oa?ii?iaeoeaaoe?? iacio?eueiiai ? oa?iio?eueiiai oa?iaio?a
io?eiai? aei??thaaiiyi ?o caeeoeiai? aeoeaiino? i?ney ?aaeoe??
oa?ii?iaeoeaaoe?? iaoiaeii AO?-???-iai?io. E?ea? oa?iinoaa?eueiino?
oa?iaio?a io?eiai? aei??thaaiiyi ?o caeeoeiai? aeoeaiino? i?ney 5 oa.
?ieoaaoe?? i?e ??cieo oaiia?aoo?ao iaoiaeii AO?-???-iai?io.

I?/aiiy o?IE ii 3′- ? 5′- e?ioeyi i?iaiaeeee a?aeiia?aeii c ?iaioaie [
Silberklang , 1977; Vlassov , 1981 ]. Aecia/aiiy ioeeaioeaeii?
iine?aeiaiino? oa?iio?eueii? o?IE i?iaiaeeee a?aeiia?aeii c ?iaioaie [
Peattie, 1979; Donis-Keller, 1977 ]. I?ii?i? iniiae ia?aeiii?
no?oe-

oo?e o?IETyr aecia/aee ca aeiiiiiaith AA?O [ Gehrke, 1989 ] c
aeei?enoaiiyi eieiiee N18.

Ae?eyiee eiioaeoo oa?iio?eueii? ? iacio?eueii? o?IETyr c aiiieia?/ieie
ai?iiaoeee-o?IE neioaoacaie aecia/aee iaoiaeii caoenoo oinoao?a o?IE
A?Nacaie a?ae aee?eoaaiiy i?o?iciaoeena/iaeiith a?aeiia?aeii [ Vlassov
et al, 1981 ] ? caoenoii oinoiaeeao??ieo ca’yce?a o?IE a?ae a?ae?ie?ca
ioeeaacaie [ Aaneeaiei, 1978; Ehresmann, 1987 ]. O?aaiaioe i?/aii?
o?IE, io?eiai? a ?acoeueoao? ?? a?ae?ie?ca ii iiaeeo?eiaaiei oinoaoai
aai oinoiaeeao??ieo ca’yce?a, aiae?coaaee aeaeo?ioi?acii a
iie?ae?eeai?aeiiio aae? a i?enooiino? 8 I na/iaeie. ?ioaineai?noue
aeaeo?ioi?aoe/ieo nioa, ye? a?aeia?aaeathoue nooi?iue iiaeeo?eaoe??
caeeoe?a oinoi?ii? eeneioe, aecia/aee ca aeiiiiiaith neaioth/iai
aeaineoiiao?a.

?ACOEUeOAOE ? ?O IAAIAI?AIIss

?ic?iaeaiiy iiaeo iaoiae?a aeae?eaiiy oe?icee-, na?ee- ? o?aii?e-o?IE
neioaoac ?c T.thermophilus, aeae?eaiiy oe?icee-o?IE neioaoace ?c E.coli
? oa?aeoa?enoeea /enoioe iaea?aeaieo i?aia?ao?a a?ee?a.

?ic?iaeaii iiaee iaoiae aeae?eaiiy oa?iio?eueii? oe??N, yeee
neeaaea?oueny c oaeeo noaae?e: o?aeoe?ioaaiiy iino?eainiiaeueiiai
noia?iaoaioo noeueoaoii aiii?y, o?iiaoia?ao?y ia AeAAA-naoa?ic?,
a?ae?ioiaia o?iiaoia?ao?y ia iie?a?i?eiaiio ni?aaio? Toyopearl HW-65,
o?iiaoia?ao?y ia ieneaiaoeo?, ?ao?iiaoia?ao?y ia Toyopearl HW-65,
o?iiaoia?ao?y ia aaia?ei-naoa?ic?. Ca iniiao aeae?eaiiy oa?iio?eueieo
na?ee- ? o?aii?e-o?IE neioaoac aoea i?eeiyoa noaia , iienaia aeey
i/enoee oe??N. Aeae?eaiiy aenieii/euaieo i?aia?ao?a na??N ? o?a?N aoei
aeeeeeaii iaiao?aei?noth aeei?enoaiiy aaoa?ieia?/ieo A?Nac ye eiio?iey
ia niaoeeo?/i?noue ooai?aiiy eiiieaena o?IE-neioaoaca a aenia?eiaioao ii
aecia/aiith ae?eyiie aca?iiae?? o?IETyr c oa?iio?eueiith ? iacio?eueiith
oe?icee-o?IE neioaoacaie. Aeae?eaiiy oe??N ?c E.coli i?iaiaeeee ca
oaeith iioei?ciaaiith noaiith: o?aeoe?ioaaiiy iino?eainiiaeueiiai
noia?iaoaioa noeueoaoii aiii?y, o?iiaoia?ao?? ia AeAAA-oeaetheic?,
AeAAA-naoa?ic?, ieneaiaoeo? ? aaia?ei-naoa?ic?.*enoioa io?eiaieo
i?aia?ao?a oa?iio?eueieo a?ee?a noaiiaeea a?eueo i?ae 95 %, ? 80 % o
aeiaaeeo iacio?eueii? oe??N. Aeo?ae oe?icee-, na?ee- ? o?aii?e-o?IE
neioaoac noaiiaea 1,6 ia, 4 ia ? 9 ia ia 1 ea ee?oei T.thermophilus,
a?aeiia?aeii.

Ii??aiyeueia aea/aiiy o?ceei-o?i?/ieo aeanoeainoae oe?icee-o?IE neioaoac
?c

T.thermophilus ? E.coli

Aeey c’ynoaaiiy no?oeoo?ieo a?aei?iiinoae, ye? caaacia/othoue
iiaeeea?noue oa?iio?euei?e oe??N ooieoe?ioaaoe i?e i?aeaeuaiieo
oaiia?aoo?ao, iaie aoei i?iaaaeaii ii??aiyiiy ai?iieeneioieo neeaae?a
iacio?eueiiai [ Calendar, 1966 ] ? oa?iio?eueiiai oa?iaio?a. Aoei
aeyaeaii ca?eueoaiiy a?ae?ioiaieo ai?iieeneio o neeaae? oe??N ii??aiyii
c iacio?eueiei oa?iaioii: 48 % ? 40 % , a?aeiia?aeii. A?eie ?c
oa?iio?ea i?noeoue cia/ii a?eueoo e?euee?noue Pro, Arg, Leu ? iaioo
e?euee?noue Asn +Asp, Lys ( ?en. 1 ). Io?eiai? aeai? aeicaieeee c?iaeoe
iai i?eiouaiiy, ui o aeiaaeeo oe??N ?c T.thermophilus aeno?aiaeueia
oa?iio?euei?noue oa?iaioa aeinyaa?oueny iii?oiith ci?iith
ai?iieeneioiiai neeaaeo oeyoii ca?eueoaiiy a?ae?ioiaieo ai?iieeneioieo
caeeoe?a.

Ii??aiyiiy iioeiaeueieo eiioeaio?aoe?e KCl a ?aaeoe?? ai?iiaoeeeoaaiiy,
yea eaoae?co?oueny oa?iio?eueiith ? iacio?eueiith neioaoacaie, aeyaeei
iaai? a?aei?iiino? : 60-80 iI ? 20-40 iI , a?aeiia?aeii ( ?en. 2 ). E??i
oeueiai, c i?aeaeuaiiyi eiioeaio?aoe?? KCl a?ae 0 aei 300 iI a ?aaeoe??
ai?iiaoeeeoaaiiy aaoa?ieia?/ii? o?IETyr, yea eaoae?co?oueny
oa?iio?eueiith ? iacio?eueiith neioaoacaie, ia? i?noea cia/ia ciaioaiiy
??aiy ai?iiaoeeeoaaiiy. Ia iniia? io?eiaieo aeaieo ie i?eionea?ii, ui
iioeiaeuei? oiiae ooai?aiiy i?iaeoeoeaiiai eiiieaena o?IE- neioaoaca
nooo?ai a?ae??ciythoueny aeey aiiieia?/iiai ? aaoa?ieia?/iiai
eiiieaen?a. Ii??aiyiiy oaiia?aoo?ieo iioeioi?a ?aaeoe?? ai?iiaoeeeoaaiiy
aiiieia?/ii? o?IETyr, yea eaoae?co?oueny oa?iio?eueiith ?
iacio?eueiith neioaoacaie, aeyaeei cia/io ??cieoeth i?ae ieie: 75 (N ?
41 (N, a?aeiia?aeii ( ?en. 3 ). O aeiaaeeo ?aaeoe?? ai?iiaoeeeoaaiiy
iacio?eueii? o?IETyr oa?iio?eueiith neioaoacith, oaiia?aoo?iee iioeioi
iaa i?noea i?e 50 (N.

Aea/aiiy eaoae?oe/ieo aeanoeainoae oe?icee-o?IE neioaoace ?c
T.thermophilus

I?e ii??aiyii? eaoae?oe/ieo aeanoeainoae oe??N ?c iacio?ea [ Calendar,
1966 ] ? oa?iio?ea aoei aeyaeaii, ui oa?iio?eueiee oa?iaio ia?
i?aeaeuaiio nii??aeiaii?noue aei iecueeiiieaeoey?ieo noano?ao?a AO? ?
L-oe?iceia ( Oaae.1 ). Cia/aiiy Km aeey o?IETyr ?c T.thermophilus ?
E.coli a ?aaeoe?? ai?iiaoeeeoaaiiy oa?iio?eueiei oa?iaioii i?e 37 (N ?
65 (N i?aeoe/ii iaeiaeia?. Aea a ?aaeoe?? ai?iiaoeeeoaaiiy oa?iio?eueii?
o?IETyr i?e 65 (N niinoa??aa?oueny ca?eueoaiiy Vmax a 4,3 ?aca ia
a?aei?io a?ae ai?iiaoeeeoaaiiy iacio?eueii? o?IE i?e oe?e aea
oaiia?aoo??. Iiaeeeai, ni?iiiaei?noue oe??N ?c T.thermophilus
ooieoe?ioaaoe i?e i?aeaeuaieo oaiia?aoo?ao aeinyaa?oueny oeyoii ci?ie
eaoae?oe/ieo aeanoeainoae oa?iaioa. Iaie aoei anoaiiaeaii, ui
oaiia?aoo?iee iioeioi ?aaeoe?? i??ioinoaoiiai iai?io ciaoiaeeoueny a
iaeano? 40 (N aeey iacio?eueiiai oa?iaioa ? 81 (N aeey oa?iio?eueiiai
oa?iaioa ( ?en.4 ).

Oaeiae aoei i?iaaaeaii ii??aiyiiy e?iaoeee oaieiai? ?iaeoeaaoe?? oe??N
?c T.thermophilus ? E.coli. ?acoeueoaoe oeeo aeine?aeaeaiue
i?aaenoaaeai? ia ?en.5. Io?eiai? iaie aenia?eiaioaeueii eiinoaioe
oa?ii?iaeoeaaoe?? ynii na?ae/aoue i?i i?aeaeuaio oa?iinoaa?euei?noue
no?oeoo?e oa?iio?eueii? oe??N ii??aiyii c aiaeia?/iei iacio?eueiei
oa?iaioii (Oaae. 2 ).

Oaiia?aoo?i? caeaaeiino? oaeaeeinoae i??ioinoaoiiai iai?io,
ai?iiaoeeeoaaiiy ? e?ea? oa?iinoaa?eueiino? iaio oa?iaio?a i?aaenoaaeai?
ia ?en. 6. O aeiaaeeo iacio?eueiiai oa?iaioa a iaeano? i?aeaeuaieo
oaiia?aoo? niinoa??aa?oueny /?oea ei?aeyoe?y oa?iinoaa?eueiino? ?
eaoae?oe/ii? aeoeaiino? ye a ?aaeoe?? i??ioinoaoiiai iai?io oae ? a
?aaeoe?? ai?iiaoeeeoaaiiy.

Cieaeaiiy eaoae?oe/ii? aeoeaiino? i?ney 41 (N ? i?aeoe/ii a?aenooi?noue
??cieoe? i?ae oaiia?aoo?ieie iioeioiaie aeey aiiieia?/ii? ?
aaoa?ieia?/ii? o?IETyr a ?aaeoe?? ai?iiaoeeeoaaiiy ie iiynith?ii
oa?iiaeaiaoo?aoe??th oa?iaioa. Cian?i ?ioa ea?oeia niinoa??aa?oueny o
aeiaaeeo oa?iio?eueiiai oa?iaioa. Iaeaea aei 81 (N oe??N aoea ni?iiiaeia
eaoae?coaaoe i??ioinoaoiee iai?i. A oie aea /an aaea i?ney 75 (N
niinoa??aaeinue ciaioaiiy oaeaeeino? ?aaeoe?? ai?iiaoeeeoaaiiy o?IETyr.
I/aaeaeii, i?e/eiith oiai, ui oaeaee?noue ?aaeoe?? ai?iiaoeeeoaaiiy
iaaea? i?e oaiia?aoo?ao aeua ca 75 (N, ? ia oa?ii?iaeoeaaoe?y oe??N, a
oa?iiaeaiaoo?aoe?y o?IE. Ii??aithth/e e?ea? oa?iinoaa?eueiino? ?
oaeaeeino? i??ioinoaoiiai iai?io, ie niinoa??aa?ii i?ioe??/io ea?oeio:
i?e aeineoue oaeaeeiio iaae?ii? oa?iinoaa?eueiino? oa?iaioa i?e
i?aeaeuaii? oaiia?aoo?e a?ae 65 (N aei 92 (N, oaeaee?noue
i??ioinoaoiiai iai?io, yeee eaoae?co?oueny oa?iio?eueiith neioaoacith,
aeinyaa? iioeioia i?e 81 (N ? iacia/ii ciaioo?oueny ( anueiai ia 6 % )
i?e 92 (N.

Ie i?eionea?ii, ui a ?aaeoe?? i??ioinoaoiiai iai?io i?e
ca’ycoaaii? oa?iaioa c AO? ? ai?iieeneioith eiioi?iaoe?y a?eea
ci?ith?oueny, noaa?e?co?oueny, ui ? ? i?e/eiith ca?eueoaiiy aeoeaiino?
oa?iaioa. Eiioi?iaoe?ei? ci?ie oa?iaioa, ye? noaa?e?cothoue eiai
no?oeoo?o, i?e ca’ycoaaii? c noano?aoii aoee i?iaeaiiino?iaai? A. Fersht
aeey oe??N ?c B.stearothermophilus

[ Fersht, 1988 ]. Oaeei /eiii, ie aaaaea?ii, ui io?eiai? aeieace ia
ei?enoue oiai, ui a ?aaeoe?? i??ioinoaoiiai iai?io i?e ca’ycoaaii?
oa?iaioa c AO? ? ai?iieeneioith a?aeaoaa?oueny ci?ia eiioi?iaoe?? a?eea,
yea ni?yiiaaia ia noaa?e?caoe?th eiai no?oeoo?e, i?e/iio ooai?aiiy
oaeiai eiiieaeno i?eaiaeeoue aei iii?oiiai ca?eueoaiiy
oa?iinoaa?eueiino? a?eeiai? iieaeoee. Oea i?eiouaiiy aeia?a
ocaiaeaeo?oueny c a?aei?/aieie aeua aeaieie i?i i?aeaeuaio
nii??aeiai?noue oa?iio?eueiiai oa?iaioa aei AO? ? ai?iieeneioe, ?
i?aeoaa?aeo?oueny a aenia?eiaioao ii aea/aiith caoenoo oa?iio?eueii?
oe??N a?ae oa?ii?iaeoeaaoe?? oeeie noano?aoaie. Aoei aeyaeaii, ui a
i?enooiino? 2 iI AO? aai 2 iI AO? ? 2,5 iI L-oe?iceia a ?ieoaaoe?ei?e
noi?o? i?e oa?ii?iaeoeaaoe?? oa?iaio iiai?noth caa??aaa naith
eaoae?oe/io aeoeai?noue a iine?aeoth/?e ?aaeoe?? ai?iiaoeeeoaaiiy (
?en.7 ).

Aecia/aiiy ioeeaioeaeii? iine?aeiaiino? o?IETyr ?c T.thermophilus

Ioeeaioeaeia iine?aeiai?noue o?IETyr ?c T.thermophilus aoea aecia/aia c
aeei?enoaiiyi iaoiae?a oaeaeeiai aaeue-naeaaioaaiiy, a iniia? yeeo
eaaeeoue niaoeeo?/ia o?i?/ia aeaa?aaeaoe?y

[ Peattie,1979 ] ? oa?iaioaoeaia aeaa?aaeaoe?y niaoeeo?/ieie ioeeaacaie
[ Donis-Keller, 1977; Ehresmann, 1987 ] iieeioeeaioeaeiiai eaioethaa
o?IE ( ?en. 8 ). I?e?iaea i?ii?ieo iniia ioeeaioeaeii? iine?aeiaiino?
o?IETyr aoea aecia/aia ca aeiiiiiaith AA?O [ Gehrke, 1989 ] c
aeei?enoaiiyi eieiiee N18 .

I?e ii??aiyii? ioeeaioeaeii? iine?aeiaiino? c o?IETyr ?c T.thermophilus
c anoaiiaeaiith ?ai?oa no?oeoo?ith o?IETyr ?c E.coli [ Sprinzl,1989 ]
aeyaeaii 52 % aiiieia??. O neeaae? oa?iio?eueii? o?IETyr aoei aeyaeaii
aea? iiaeeo?eiaai? iniiae: m1A a O-iaoe? ? Gm a Ae-iaoe?, eiia?iaoe?y
yeeo, iiaeeeai, ? i?e/eiith a?eueo aeniei? oa?iinoaa?eueiino? oe???
iieaeoee [Watanabe, 1980 ]. Ie i?eionea?ii, ui aeyaeai? no?oeoo?i?
a?aei?iiino? a o?IETyr ?c oa?iio?ea caaacia/othoue ooai?aiiy
i?iaeoeoeaiiai eiiieaena o?IE-neioaoaca i?e i?aeaeuaiieo oaiia?aoo?ao.

Aecia/aiiy ae?eyiie aca?iiae?? o?IETyr ?c T.thermophilus ? E.coli c
aiiieia?/ieie ai?iiaoeee-o?IE neioaoacaie

Ae?eyiee aca?iiae?? oa?iio?eueii? ? iacio?eueii? o?IETyr c aiiieia?/ieie
A?Nacaie aea/aee iaoiaeaie caoenoo oinoao?a o?IE A?Nacith a?ae
o?i?/ii? iiaeeo?eaoe??

( aee?eoaaiiy ) i?o?iciaoeena/iaeiith [ Vlassov, 1981 ] ? iaoiaeii
caoenoo a?ae ioeeaaciiai a?ae?ie?ca [Aaneeaiei, 1978; Ehresmann, 1987
]. O?i?/ia iiaeeo?eaoe?y o?IE a i?enooiino? A?Nace i?iaiaeeeanue a
oiiaao noaa?e?caoe?? i?inoi?iai? no?oeoo?e o?IE [Vlassov, 1981 ], ?
oiiaao, ye? ni?eythoue ooai?aiith niaoeeo?/iiai eiiieaena o?IE-a?eie: 50
iI o?en-HCl,pH 8,0,

5 iI MgCl2, 2,5 iI 2- ia?eaioiaoaiie. I?ney i?iaaaeaiiy ?aaeoe??
iiaeeo?eaoe?? o?IE, iieaeoeo a?ae?ie?coaaee ii aee?eiaaiei oinoaoai.
Io?eiai? o?aaiaioe ?icuaieaii? o?IE aiae?coaaee aenieiaieueoiei
aeaeo?ioi?acii a iie?ae?eeai?aeiiio aae?. ?ooeea?noue aeaeo?ioi?aoe/ieo
nioa ni?aa?aeiineee c ?ooeea?noth o?aaiaio?a o?IE, io?eiaieo a
?acoeueoao? /anoeiaiai a?ae?ie?ca o?IE ?IEacith O1. ?ioaineaiino?
aeaeo?ioi?aoe/ieo nioa, ye? a?aeia?aaeathoue nooi?iue iiaeeo?eaoe??
oinoao?a, aecia/aee ca aeiiiiiaith neaioth/iai aeaineoiiao?a.

Aeey aiae?ca o?IETyr- oe??N eiiieaena aeei?enoiaoaaee aiaeiioeeaaco V1
?c io?ooe eia?e, yea ia? yne?aai ae?aaeaio aea??eia?noue o a?aeiioaii?
a?ae?ie?ca aeaini??aeueieo ae?eyiie o?IE , ianiaoeeo?/io ioeeaaco ?1,
yea ia? aea??eia?noue o a?aeiioaii? iaeiini??aeueieo ae?eyiie, a oaeiae
niaoeeo?/io ioeeaaco PhyM, yea ?icuaieth? ca’ycee oeio U-N ? A-N .
A?ae?ie?c aaee a oiiaao iioeiaeueieo ye aeey eiiieaeniooai?aiiy, oae ?
aeey ?iaioe ioeeaac: 25 iI o?en-HCl, pH 7,2, 10 iI MgCl2, 2,5 iI
2-ia?eaioiaoaiie. Io?eiai? o?aaiaioe o?IE aiae?coaaee aenieiaieueoiei
aeaeo?ioi?acii a iie?ae?eeai?aeiiio aae?. ?acoeueoaoe aenia?eiaio?a ii
iiaeeo?eaoe?? i?o?iciaoeena/iaeiith ? ioeeaaciiio a?ae?ie?co eiiieaen?a
o?IETyr-oe??N ?c T.thermophilus ? E.coli i?aaenoaaeai? ia ?en. 9, 10,
11.

C aeaieo iaethie?a aeaeii, ui oa?iio?eueia oe??N caoeua? a?ae
aee?eoaaiiy ?yae oinoao?a o?IE, ?ici?uaieo a /ioe?ueio ciiao: 3′-oinoaoe
ioeeaioeae?a 28, 29 5′- ae?eyiee aioeeiaeiiiaiai noaaea, 34, 36
aioeeiaeiia, 45, 46, 47 aa??aaaeueii? a?eee, 68 3′- ae?eyiee
aeoeaioi?iiai noaaea. Aeey iacio?eueii? ia?e oe? oinoaoe ?ici?uai?
oaeiae a /ioe?ueio ciiao: 3′- oinoaoe ioeeaioeae?a 29 5′- ae?eyiee
aioeeiaeiiiaiai noaaea, 36 aioeeiaeiia, 45, 46 aa??aaaeueii? a?eee, 67
3′- ae?eyiee aeoeaioi?iiai noaaea. A?ae a?ae?ie?co oa?iio?eueii? o?IETyr
ioeeaacaie aiiieia?/ia neioaoaca caoeua? oinoiaeeao??i? ca’ycee i?ae
ioeeaioeaeaie 28 ? 29, 29 ? 30 5′- ae?eyiee aioeeiaeiiiaiai noaaea, 33
? 34, 34 ? 35 aioeeiaeiiiai? iaoe?, 47A ? 47N, 47N ? 47D aa??aaaeueii?
a?eee, 68 ? 69, 69 ? 70, 70 ? 71, 71 ? 72, 72 ? 73, 73 ? 74 3′-
ae?eyiee aeoeaioi?iiai noaaea. Aeey iacio?eueii? ia?e o?IETyr-neioaoaca
oea oinoiaeeao??i? ca’ycee i?ae ioeeaioeaeaie 29 ? 30 5′- ae?eyiee
aioeeiaeiiiaiai noaaea, 35 ? 36, 36 ? 37 aioeeiaeiiiai? iaoe?, 47A ?
47A, 47N ? 47D aa??aaaeueii? a?eee, 67 ? 68 3′- ae?eyiee aeoeaioi?iiai
noaaea. A o?IETyr ?c E.coli i?e ooai?aii? eiiieaena c aiiieia?/iith
neioaoacith aeyaeaia ?iaeoeoe?y aeiaeaoeiaeo ?ic?ea?a oinoiaeeao??ieo
ca’yce?a aiaeiioeeaacith V1 a 3′- ae?eyioe? aioeeiaeiiiaiai noaaea ?
5′- ae?eyioe? O-noaaea, ui aeaco? ia iiaeeeao ci?io eiioi?iaoe?? o?IE
i?e aca?iiae?? c oa?iaioii. Ocaaaeueithth/e ?acoeueoaoe, io?eiai? c
aeei?enoaiiyi aeaio ??cieo iaoiae?a ia aeaio ia?ao o?IETyr-oe??N ,
iiaeia c?iaeoe aeniiaie, ui a o?IETyr ?c T.thermophilus ? E.coli a
eiioaeo? c aiiieia?/iith neioaoacith ciaoiaeyoueny /ioe?e, aeaui
a?aeaeaeai? iaeia a?ae iaeii? a ia?aeii?e no?oeoo?? o?IE, ae?eyiee. ?o
?ici?uaiiy ia iiaeae? i?inoi?iai? no?oeoo?e o?IE 2-ai eeano [Brennan ,
1976 ] iaeiicia/ii aeaco? ia oa, ui neioaoaca aca?iiae?? c o?IE c?
noi?iie aa??aaaeueii? a?eee. Aei aiaeia?/iiai aeniiaeo i?eeoee
Bedouelle oa ?i. i?e noai?ai? iiaeae? eiiieaena o?IETyr-oe??N
[Bedouelle, 1980 ]. Aea ia oe?e iiaeae? a?aenooiy aca?iiae?y
aa??aaaeueii? a?eee o?IETyr c neioaoacith. E??i oeueiai, aeniiaie i?i
oa?aeoa? aca?iiae?? aeoeaioi?iiai noaaea o?IETyr c eaoae?oe/iei
oeaio?ii oa?iaioa aoiaeeoue a eiioe?eo c eiioeaioe??th i?i ?nioaaiiy a
aeaio no?oeoo?ieo eeanao neioaoac aeaio ??cieo oei?a aca?iiae?? c o?IE.
I/aaeaeii, ui aeaia aeeeaia iiaea aooe ae??oaia eeoa aea/aiiyi no?oeoo?e
eiiieaena oe??N- o?IETyr iaoiaeii ?aioaaiino?oeoo?iiai aiae?co. Ia
aeaiee /an a a?aeae?e? aiceiieia?? a?eeiaiai neioaca ?IA?A IAI Oe?a?ie
io?eiai? e?enoaee oe?icee-o?IE neioaoace c ?icae?eueiith caeaoi?noth 3 A
? e?enoaee o?IETyr-oe??N c ?icae?eueiith caeaoi?noth 6 A a eaai?aoi?ieo
oiiaao.

AENIIAEE

1.Aeae?eaia a aenieii/euaiiio noai? ? ioa?aeoa?eciaaia oe?icee-o?IE
neioaoaca ?c T.thermophilus. Aecia/ai? ae?eyiee aca?iiae?? o?IETyr ?c
T.thermophilus ? E.coli c aiiieia?/ieie ai?iiaoeee-o?IE neioaoacaie.

2.?ic?iaeai? iaoiaee aeae?eaiiy oe?icee-, na?ee- ? o?aii?e-o?IE
neioaoac ?c T.thermophilus, io?eiai? a aenieii/euaiiio
noai? oa?iio?euei? oa?iaioe o?ueio

niaoeeo?/iinoae. Iioei?ciaaii iaoiae aeae?eaiiy oe?icee-o?IE neioaoace
?c E.coli.

3. Aea/ai? o?ceei-o?i?/i? ? eaoae?oe/i? aeanoeaino? oe?icee-o?IE
neioaoace ?c T.thermophilus:

a) aecia/aia iieaeoey?ia iana ? ie?aiia?ia no?oeoo?a iaea?aeaieo
oa?iio?eueieo oa?iaio?a. Ii aeaiei aeaeo?ioi?aco a IAAA a iaoeaieo
oiiaao iieaeoey?ia iana oe??N neeaaea? 100 eAea, na??N — 90 eAea, o?a?N
— 148 eAea. An? oe? o?e oa?iaioa ? no?oeoo?ieie aeeia?aie (2 oeio.

a) aeyaeaii ca?eueoaiiy a?ae?ioiaieo ai?iieeneio o neeaae? oe??N
ii??aiyii c iacio?eueiei oa?iaioii: 48 % ? 40 % , a?aeiia?aeii. A?eie
?c oa?iio?ea i?noeoue cia/ii a?eueoo e?euee?noue Pro, Arg, Leu ?
iaioo e?euee?noue Asn+Asp, Lys. Io?eiai? aeai? aeicaieeee c?iaeoe iai
i?eiouaiiy, ui o aeiaaeeo oe??N ?c T.thermophilus aeno?aiaeueia
oa?iio?euei?noue oa?iaioa aeinyaa?oueny iii?oiith ci?iith
ai?iieeneioiiai neeaaeo oeyoii ca?eueoaiiy a?ae?ioiaieo ai?iieeneioieo
caeeoe?a.

a) i?iaaaeaii ii??aiyiiy e?iaoe/ieo ia?aiao??a ?aaeoe??
ai?iiaoeeeoaaiiy, yea eaoae?co?oueny oa?iio?eueiith ? iacio?eueiith
oe??N. Aeyaeaii, ui nii??aeiai?noue iecueeiiieaeoey?ieo noano?ao?a
L-oe?iceia ? AO? aeua o aeiaaeeo oa?iio?eueii? oe??N.

a) anoaiiaeai? eiinoaioe oa?ii?iaeoeaaoe?? oe??N ?c T.thermophilus ?
E.coli. ?acoeueoaoe aea/aiiy oa?ii?iaeoeaaoe?? aeine?aeaeoaaieo
oa?iaio?a na?ae/aoue i?i i?aeaeuaiiy oa?iinoaa?eueiino? no?oeoo?e
oa?iio?eueii? neioaoace. Ii??aiyiiy e?eaeo oaiia?aoo?ii? caeaaeiino?
oa?iinoaa?eueiino? ? i??ioinoaoiiai iai?io oe??N ?c T.thermophilus
aeaco? ia cia/ia ca?eueoaiiy oa?iinoaa?eueiino? oa?iaioa a i?enooiino?
iecueeiiieaeoey?ieo noano?ao?a AO? ? L- oe?iceia. Aeyaeaii caoeno oe??N
a?ae oa?ii?iaeoeaaoe?? oeeie noano?aoaie- AO?, AO? ? L-oe?iceiii.

4. Aecia/aia ioeeaioeaeia iine?aeiai?noue o?IETyr ?c T.thermophilus
iaoiaeaie oaeaeeiai aaeue-naeaaioaaiiy. Nooi?iue ?? aiiieia?? c
iacio?eueiith o?IETyr noaiiaeoue 52 %.

5. Aecia/ai? ae?eyiee aca?iiae?? o?IETyr ?c T.thermophilus ? E.coli c
aiiieia?/ieie ai?iiaoeee-o?IE neioaoacaie iaoiaeaie o?i?/ii?
iiaeeo?eaoe?? ? ioeeaaciiai a?ae?ie?ca. Iieacaii, ui oe?icee-o?IE
neioaoace caoeuathoue aiiieia?/i? o?IETyr a?ae aee?eoaaiiy
i?o?iciaoeena/iaeiith ? a?ae?ie?ca ioeeaacaie a ?aeii? 5′- ae?eyiee
aioeeiaeiiiaiai noaaea, aioeeiaeiia, 5′- ae?eyiee aa??aaaeueiiai noaaea
? aa??aaaeueii? iaoe?, 3′- ae?eyiee aeoeaioi?iiai noaaea. ?ici?uaiiy
oeeo ae?eyiie ia iiaeae? i?inoi?iai? no?oeoo?e o?IE 2-ai eeano
iaeiicia/ii aeaco? ia oa, ui neioaoaca aca?iiae?? c o?IE c? noi?iie
aa??aaaeueii? a?eee.

IA?AE?E ?IA?O, UI IIOAE?EIAAII CA OAIITH
AeENA?OAOe??

ss?ai/oe A.Ae., Ooeaei I.A., Aai?iaa N.I., Eiiiaaeaiei A.A., Iaoeoea
A.O. Auaeaeaiea oe?icee-o?IE neioaoacu ec Thermus thermophilus HB-27 //
Oe?a?inueeee a?io?i?/iee aeo?iae — 1990.- O.62.- ? 2.- N. 97- 99.

2. ss?ai/oe A.Ae., Ooeaei I.A., EiiiaaeaieiA.A., Aai?iaa N.I., Iaoeoea
A.O. Auaeaeaiea na?ee-o?IE neioaoacu ec Thermus thermophilus. //
Aeiiieeia?u e eeaoea.-1989.-O.5.- ? 5.- N.83-86.

3. ss?ai/oe A.Ae., Ooeaei I.A., Aai?iaa N.I., Iaoeoea A.O. Auaeaeaiea
o?aiiee-o?IE neioaoacu ec Thermus thermophilus. // Aeiiieeia?u e
eeaoea.-1989.-O. 5.- ? 2.- N.102-103.

4. Aai?iaa N.I., ss?ai/oe A.Ae., Ooeaei I.A. N?aaieoaeueiia
enneaaeiaaiea oeceei-oeie/aneeo naienoa oe?icee-o?IE neioaoac ec
Thermus thermophilus e Escherichia coli.

// Aeiiieeia?u e eeaoea.- 1998.- O.14.- ?2.- C. 144-151.

5. Aai?iaa N. I., ss?ai/oe A. Ae., E?eeeeaue E. A., Ooeaei I. A.
N?aaieoaeueiue aiaeec o/anoeia acaeiiaeaenoaey o?IETyr ec Thermus
thermophilus e Esnherichia coli n aiiieiae/iuie aieiiaoeee-o?IE
neioaoacaie iaoiaeaie oeie/aneie iiaeeoeeaoeee e ioeeaaciiai aeae?ieeca.
// Aeii?aaie/aneay oeiey.- 1998.- O.24.- ? 8.- N. 593-600.

6. ss?ai/oe A.Ae., Ooeaei I.A., Aai?iaa N.I., Iaoeoea A.O.
No?oeoo?ii-ooieoeeiiaeueiia enneaaeiaaiea aieiiaoeee-o?IE neioaoac ec
yeno?aiaeueiiai oa?iioeea Thermus thermophilus. // Ananithciue
neiiiceoi ii oeiee aaeea , Oaeeene — 1990. — N.116.

7. Aai?iaa N. I., ss?ai/oe A.Ae., Ooeaei I.A., Iaoeoea A.O. Auaeaeaiea
oe?icee-o?IE neioaoacu ec Thermus thermophilus e acaeiiaeaenoaea n
o?IETyr. // Ananithciue neiiiceoi ii oeiee aaeea , Oaeeene — 1990. —
C.135.

8. A. Yaremchuk, S. Egorova, V. Mel’nik, N. Mal’chenko, M.-T.
Dauverggne, R. Leberman, M. Tukalo. Properties and structure of the
aminoacyl-tRNA synthetases from Thermus thermophilus. // Abstr. of
International conference Thermophiles: science and technology,
Reykjavik.- 1992.- P. 107.

9. A.Yaremchuk, S.Egorova, I.Kriklivyi, S.Cusack and M.Tukalo.
Studies on the complex between tyrosyl-tRNA synthetase and tRNAtyr
from Thermus thermophilus. // Abstr. of 17th International tRNA
Workshop, Kazusa Akademia Park, Chiba, Japan. — 1997. — P.4.

?ai?iaa N.I. Oe?icee-o?IE neioaoaca ?c aeno?aiaeueiiai oa?iio?ea
Thermus thermophilus HB-27: o?ceei- o?i?/ia oa?aeoa?enoeea oa?iaioa ?
aca?iiae?y c o?IETyr .- ?oeiien. Aeena?oaoe?y ia caeiaoooy iaoeiaiai
nooiaiy eaiaeeaeaoa a?ieia?/ieo iaoe ca niaoe?aeuei?noth 03.00.03 —
iieaeoey?ia a?ieia?y. ?inoeooo iieaeoey?ii? a?ieia?? ? aaiaoeee IAI
Oe?a?ie, Ee?a, 1999.

Aeine?aeaeaii aca?iica’ycie oa?iinoaa?eueiino? ? iniaeeainoae no?oeoo?e
? ooieoe?ioaaiiy oe?icee-o?IE neioaoace ?c aeno?aiaeueiiai oa?iio?ea
T.thermophilus, ? aca?iiae?th o?IETyr c oe??N ?c T.thermophilus ?
E.coli o ?ic/ei?. Aeyaeaii i?aeaeuaiiy ai?noo a?ae?ioiaieo
ai?iieeneioieo caeeoe?a, a iniiaiiio Pro ? Leu, a ai?iieeneioiiio
neeaae? oa?iio?eueii? oe??N. Aeyaeaii, ui nii??aeiai?noue
iecueeiiieaeoey?ieo noano?ao?a L-oe?iceia ? AO? aeua o aeiaaeeo
oa?iio?eueii? oe??N ii??aiyii c iacio?eueiei oa?iaioii. Aeyaeaii caoeno
oe??N a?ae oa?ii?iaeoeaaoe?? oeeie noano?aoaie- AO?, AO? ? L-oe?iceiii.

Aecia/ai? ae?eyiee aca?iiae?? o?IETyr ?c T.thermophilus ? E.coli c
aiiieia?/ieie ai?iiaoeee-o?IE neioaoacaie a ?ic/ei? iaoiaeaie o?i?/ii?
iiaeeo?eaoe?? ? ioeeaaciiai a?ae?ie?ca. Iieacaii, ui oe??N caoeua?
aiiieia?/io o?IETyr a?ae aee?eoaaiiy i?o?iciaoeena/iaeiith ? a?ae?ie?ca
ioeeaacaie a ?aeii? 5′- ae?eyiee aioeeiaeiiiaiai noaaea, aioeeiaeiia,
5′- ae?eyiee aa??aaaeueii? a?eee ? aa??aaaeueii? iaoe?, 3′- ae?eyiee
aeoeaioi?iiai noaaea. ?ici?uaiiy oeeo ae?eyiie ia iiaeae? i?inoi?iai?
no?oeoo?e o?IE 2-ai eeano iaeiicia/ii aeaco? ia oa, ui neioaoaca
aca?iiae?? c o?IE c? noi?iie aa??aaaeueiiai a?eee.

Eeth/ia? neiaa: ai?iiaoeee-o?IE neioaoaca, o?IE, ?IE-a?eeia?
aca?iiae??.

Aai?iaa N.I. Oe?icee-o?IE neioaoaca ec yeno?aiaeueiiai oa?iioeea
Thermus thermophilus HB-27: oeceei- oeie/aneay oa?aeoa?enoeea oa?iaioa
e acaeiiaeaenoaea n o?IETyr .- ?oeiienue. Aeenna?oaoeey ia nieneaiea
o/aiie noaiaie eaiaeeaeaoa aeieiae/aneeo iaoe ii niaoeeaeueiinoe
03.00.03 — iieaeoey?iay aeieiaey. Einoeooo iieaeoey?iie aeieiaee e
aaiaoeee IAI Oe?aeiu, Eeaa, 1999.

Enneaaeiaaia acaeiinaycue oa?iinoaaeeueiinoe e iniaaiiinoae no?oeoo?u e
ooieoeeiie?iaaiey oe??N ec yeno?aiaeueiiai oa?iioeea T.thermophilus, e
acaeiiaeaenoaea o?IETyr ec T.thermophilus e E.coli c aiiieiae/iuie
aieiiaoeee-o?IE neioaoacaie a ?anoai?a. A ?acoeueoaoa i?iaaaeaiiuo
enneaaeiaaiee aia?aua iieo/aiu a aunieii/euaiii ninoiyiee oe?icee-,
na?ee- e o?aiiee-o?IE neioaoacu ec T.thermophilus. Iieacaii, /oi yoe
oa?iaiou eiatho ieeaiia?ioth no?oeoo?o oeia (2 n iieaeoey?iie iannie
noauaaeeieoeu 50eAea, 46eAea e 74eAea, niioaaonoaaiiii. Iiaeeoeoee?iaai
iaoiae auaeaeaiey oe?icee-o?IE neioaoacu ec E.coli. Aia?aua eco/aiu
oeceei-oeie/aneea e eaoaeeoe/aneea naienoaa oe?icee-o?IE neioaoacu ec
T.thermophilus. Aeey auyniaiey no?oeoo?iuo ?acee/ee, iaania/eaathueo
aiciiaeiinoue oa?iioeeueiie oe?icee-o?IE neioaoaca ooieoeeiie?iaaoue
i?e iiauoaiiuo oaiia?aoo?ao, auei i?iaaaeaii n?aaiaiea aieiieeneioiuo
ninoaaia oa?iioeeueiiai e iacioeeueiiai oa?iaioia. Auyaeaii
oaaee/aiea niaea?aeaiey aeae?ioiaiuo aieiieeneio a ninoaaa
oa?iioeeueiiai oa?iaioa ii n?aaiaieth n iacioeeueiui: 48 % e 40 %,
niioaaonoaaiii. Aaeie ec oa?iioeea niaea?aeeo cia/eoaeueii aieueoaa
eiee/anoai Pro, , Arg, Leu e iaiueoaa eiee/anoai Asn+Asp, Lys.
I?iaaaeaii n?aaiaiea eeiaoe/aneeo ia?aiao?ia ?aaeoeee
aieiiaoeeee?iaaiey, eaoaeece?oaiie oa?iioeeueiie e iacioeeueiie oe??N.
Iaia?oaeaii iiauoaiiia n?iaenoai oa?iioeeueiie oe??N e ieceiiieaeoey?iui
noano?aoai L-oe?iceio e AO?. Iieo/aiiua iaie yenia?eiaioaeueii
eiinoaiou oa?iieiaeoeaaoeee ynii naeaeaoaeuenoaotho i iiauoaiiie
oa?iinoaaeeueiinoe no?oeoo?u oe??N ec T.thermophilus ii n?aaiaieth n
aiaeiae/iui iacioeeueiui oa?iaioii. I?e n?aaiaiee e?eauo oaiia?aoo?iie
caaeneiinoe oa?iinoaaeeueiinoe e ie?ioinoaoiiai iaiaia oe?icee-o?IE
neioaoacu ec T.thermophilus auei iaia?oaeaii, /oi oa?iinoaaeeueiinoue
oa?iaioa cia/eoaeueii aic?anoaao a i?enoonoaee ieceiiieaeoey?iuo
noano?aoia — L-oe?iceia e AO?. E?iia oiai, a yenia?eiaioao ii caueoa
oe??N io oa?iieiaeoeaaoeee ieceiiieaeoey?iuie noano?aoaie auei
onoaiiaeaii, /oi i?enoonoaea AO? eee AO? e L-oe?iceia .a
eieoaaoeeiiiie niane iicaieyei oa?iaioo iieiinoueth nio?aiyoue naith
eaoaeeoe/aneoth aeoeaiinoue a iineaaeothuae ?aaeoeee aieiiaoeeee?iaaiey
a ioee/ea io naiaiaeiiai oa?iaioa.

Ii?aaeaeaia ioeeaioeaeiay iineaaeiaaoaeueiinoue o?IETyr ec Thermus
thermophilus

HB-27. Iia eiaao aeeeio 86 ioeeaioeaeia e noaiaiue aa aiiieiaee n
o?IETyr ec E. coli ninoaaeyao 52%. A ninoaaa oa?iioeeueiie o?IE auei
iaia?oaeaii aeaa iiaeeoeoee?iaaiiuo iniiaaiey- m1A a O-iaoea e Gm a
Ae-iaoea, eiiaeiaoeey eioi?uo iiaeao auoue i?e/eiie aieaa aunieie
oa?iinoaaeeueiinoe yoie iieaeoeu. A aioeeiaeiiiaie iaoea oa?iioeeueiie
o?IETyr iaia?oaeaiu iiaeeoeoee?iaaiua iniiaaiey, eioi?ua ia oa?aeoa?iu
aeey aaeoa?eaeueiuo o?IE e ano?a/athueany oieueei o yoea?eio. Iaoiaeaie
oeie/aneie iiaeeoeeaoeee yoeeieo?iciii/aaeiie e ioeeaaciiai aeae?ieeca
i?iaaaeai n?aaieoaeueiue aiaeec o/anoeia acaeiiaeaenoaey o?IETyr ec T.
thermophilus e E.coli n aiiieiae/iuie aieiiaoeee-o?IE-neioaoacaie.
Iieacaii, /oi o?IETyr eiioaeoe?oao n aiiieiae/iui oa?iaioii a iaeanoe
aioeeiaeiiiaiai noaaey (n 5′-noi?iiu), a aioeeiaeiia, a ?aeiiao
aa?eaaaeueiiai noaaey e aa?eaaaeueiie iaoee (n 5′-noi?iiu) e
aeoeaioi?iiai noaaey (n 3′-noi?iiu). A i?ino?ainoaaiiie no?oeoo?a o?IE
2-ai eeanna yoe o/anoee ?aniieiaeaiu ni noi?iiu aa?eaaaeueiiai noaaey
L-oi?iu iieaeoeu. A o?IETyr ec E. coli i?e ia?aciaaiee eiiieaena n
aiiieiae/iie neioaoacie iaia?oaeaia eiaeoeoeey aeiiieieoaeueiuo
?ac?uaia oinoiaeeyoe?iuo naycae yiaeiioeeaacie V1 n 3′- noi?iiu
aioeeiaeiiiaiai noaaey e 5′- noi?iiu O- noaaey, /oi oeacuaaao ia
aiciiaeiia eciaiaiea eiioi?iaoeee o?IE i?e acaeiiaeaenoaee n oa?iaioii.

Eeth/aaua neiaa: aieiiaoeee-o?IE neioaoaca, o?IE, ?IE-aaeeiaua
acaeiiaeaenoaey.

Yegorova S. P. Tyrosyl-tRNA synthetase from extreme thermophile
Thermus thermophilus HB-27 : physical-chemical characteristic of the
enzyme and interaction with tRNATyr. — Manuscript. Thesis for degree of
Doctor of Philosophy ( Ph.D. ) in Biology, speciality 03.00.03 —
Molecular Biology. — Institute of Molecular Biology and Genetics,
National Academy of sciences of Ukraine, Kiev, 1999.

The correlation between the thermostability and properties of the
structure and function of the TyrRS from extreme bacteria T.thermophilus
and interaction of tRNATyr with TyrRSs from T.thermophilus and E.coli
in solution has been investigated. It was revealed the considerable
increase of quantity of hydrophobic amino acids, in the main Pro and
Leu, in amino acid compound of thermophilic TyrRS. It was revealed the
change of the catalytic properties of this protein. It was shown that
the low moleculars substrates- ATP, ATP and L-tyrosine protects of the
enzyme from thermoinactivation.

The interaction sites of tRNATyr from E.coli and T.thermophilus with
the cognate aminoacyl-tRNA synthetases has been determined by the
chemical modification and nucleases hydrolysis methods. It was shown
that TyrRS protects from alkylation by ethylnitrosourea and nucleases
hydrolysis 5′- sides of variable and anticodon stems, 3′- side of
accteptor stem, anticodon loop. The interaction sites are located on the
variable stem side of tRNA of the klass 2.

Key words: aminoacyl-tRNA synthetases, tRNA, RNA-protein interaction.

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