(SANITIZED)ENGLISH LANGUAGE TRANSLATION OF A POLISH PAPER ENTITLED, "PROTEIN COMPOUNDS OF VITAMIN B 12 AND ITS ANALOGS,"1965(SANITIZED)

• Document Type: CREST
• Collection: General CIA Records
• Document Number (FOIA) /ESDN (CREST): CIA-RDP80T00246A074600230001-8
• Release Decision: RIPPUB
• Original Classification: C
• Document Page Count: 20
• Document Creation Date: December 27, 2016
• Document Release Date: May 23, 2014
• Sequence Number: 1
• Publication Date: April 30, 1964
• Content Type: REPORT

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ChairALtuLToc` gom.,z)College
of AciTieul:tre
Pot,- ,f.Po,anglen Misizovsicleka
1-1,0 ; .14,liove4tOwelita.6
K,T14:130.5:ow*kaaktiv 171,lowalzylf: r)r,b .(1 ttroir.
? . f.01F 4 00MPOITIOr:3Oi
Rtporb pexsidd.3
-02t.iiervi 1
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le
dgr to get et me Dicre infornAtio= tm. the nature
of the protein bound,Cor:ciltoids,experimilts ware
conducted *which the properties tY2 different.
prepanations hithekto imaatedt fran Propiontbaotezim
sherranii 1t v: omotred.
The at tieal studis ? the amino acid
oompositiono elootrophol,c3,sis and neasurenents oT the
Womerption or*mtra ofthvariolist Qom-pounds..
Th iotad protainboua4corrInciets Vaen subjected
to di ferent pul?eivation pnocedureso showeartarkod
litrations in the amlno acid-condans No vetine or
vsteino couli be evidenooa ohrozatograpitical
the orudg preputtion. rwri?rstAldior3oreculted
it the separation itto fraotiohov my inaictite the
heterogeplty of the investigated conple2os. Thus the
isolated fractions tere 4ydrolysed and subjected to.
ftmito acid, anal7sis.
The absorption spectr&. of the protein :bad corrinoids
differed mattmlly from tba ot.vitamin B120
Attempts vivre also t1,9:de to chracterie the protein
octpouadg of lisht scanitive oorritoids.
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?
riPrRINFOTA.I1
====2:4.4.""r=r2=011==
Material and methods
rtiopionibeeterium shermanii was usea ea source oZ
the oorrinoid protein compounds? The isolation and
purification procedures of these copylemus wore deaw,
eribed in the previous annual reportvAttezptc were
also made to isolate the protein compourds of the
light sensitive corrinoidso Studies intble direction
were performed according to Null/ and Sabsidtsexcevt
that a1 'the operations were tarried out in dark or
in the presence of red lisbto
Isolation of the tight sensitive compoundoll To 800 g
or tresh oell paste 3 1 ofet".--7.91 04301101 were addede
A fine suspension was obtained. by azin,S the natorial
in a tarring blenlor for 10 in The suspeision was
centritusede the superlIntarli reamed, whereas the rests
dual tateria1 was sabjected to three sinilar treattlents,
10 1 a seam, 1 were usea for all the extractions,
The o lear stipernatanto were combined and evitipomteq
under reducaq pressure at 30?G. To the residual aqueou0
solutiono watm was adage's to obtain a final
'trauma of 4 lityear, To this solution a portion of 1100g
of solid stmonites solphste was added& Men a clear so
-
lotion was,obtalned, a treefold extraction was perform
ma(151 by warv a4e 3017, TIII-ciortion$: of y-butanole
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l.)1\ -num
10
Subsequent40 the codbined butanol extracts we:6 p0,ssed
through a columm of previouslz; activated Al2d30
column was washed it with 1000 n1 of 9CYX acetones
and then with 10D0 ml of 805 acetone.
This vas followed 127 vasaine thyouel a suitable volume
of distilled waters rtsoulting it the elution of a
brick-coloured nmteriaL
The residual acetone war) remavea fm the aqueous eluate
by evaporation under reeuced preissure,, The clear soln-
bloat) thus obta.Sneds wt i aUbjected to several subsequent
=tractional, un g small portions of phenol? The con61-
i
ned. phenol extracts Imre carefu;ly washed with distii.
led waters followed by treatment with a required amount
of ethyl ethers to got a clear solutiont,
The coloured compowes were reevered pentitaiway
fa= this solution water extractiono Traces of phenol
were removed Vim tile aqueous extraest Tog washing it
with appreciable volts. ef ethyl others the !Atom beLlsg
remowet by owsporntlanE.
The *queen*? solution of the cerrltoid Ampounklesthus
obtaineds was muWoOted to seveml, phenol extractions.
The combined phenca extracts were =shed with distilled
water and subsequent:ty treated with nine volumes of
acetone. This solution was al1owe4 to stand for several
hours,' The precipitate of an amorthous brick,coloured
naterial was seperated br centrifugations wasaled with
ethanol and ethers ta.6d finally dried nvacuum over
B26040
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IV
All the hitherto isolated compounds were sUbjected to
further studinso Other moth es applied i* this research
are Oxert ln appropriate sections of thia text.
VSINITO OD DISCUSION 2
The following abbreviations are beinG used through out
this reporh m-a and M,2/or -qUifor preParationaf
isolated by the method of Muni and Schmid%
11-1 and 3-2 /or Bej2-W-for the compounds obtained by
the modified aedbore5 procedurep
B12-M.S'- for the 1ir satitlitiVQ corrinoid propara.'
tion.
f,:tu5tiorl_o4.91e0 amino note colvogtioa
of -M and B -B
12.
CMItlitt71,=.44ram,ItOMNSW,mwdm.s.lm.WWWPG0.64M
The q?uuntitative amino atad oempositions of the .B
and IS, preparations cs well as of the major' frac-
tions of those compoutds obtainod after electrophoretic
geporationo are presented' ;La Table 10
Eaah of the isolated compouna was hydrolysed by peons
of GN ECI in a waYr, described in the previous annual
report. The amino acid ana4sis of the-hydrollyzates
has boon performed using an automaticana ier /Phoenix..
Pkeeision I Model N.5000,..A/g,
,Tho amino acid composition of N-1 and M.2 was found to
be identioal. No significh variations could be also
stated between B=1 wad B-2, Data presented it Table I
are therefore mean values fmm the ficaros obtained for
both proparationav.resoective4. The results aro
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bux-i-HUN
40.? 11 cm
exoressed as percents of the amount of command anaZrzedo
The data illustrated in Table le bowevere indicate
Tv:r?ked differo7c?s in the amounts of individual animo
acid residues of the preparations* as ear es the no
of purification is bear; concernede Both compounds
41/42.41
and Bia.B/ contsineta sic:nifica craounts of ?arta=
mic acid an-d pralines and did not show the presence of
cystine.or.cysteine, All the protoin.bound forms of'
corrinoids0 hitherto isolated0 show. a 181 ratio / by
we1ght/0 of the protein moiety to the corrinold residuc
in the vitnole comn1e.x0
Studies an the amino aoid composition of the protein
compounds of corrinolds included Cleo the analysis og
fractions obtained after lonclhorasis of both 21241-
Cad B5k2-3 preparations, The ata of this researCh was to
fiat out which of -the olectrophoreticallyAelarated
teaCtion ShCV the hichest prate/Us oonttelec., Q.sonaltatiVo
nnino acid anaysia e the ingloated i4C,110,64vAtc
pertormode after acid 14,./e,.ro3;y3i0 iG 17M-0 22 !amp
in seeZed tdbes at 11094/0 on V:Ictran Xo4 Tiltqr
U w=butanol /acetic -acid/ water 481* WI? 48 brsi
and Pheno1 saturated with? p.al ;',2 phosphate buZfe?12
as solvent system, 1$3 cystinee, or c7cteino0 and tvroeilte -
could be feundt yeavayzatee of the major fractions*
obtained after electrophoreste of eit:lar ri143 or ri,T.24.
Ze absence of tyrosine differ$ re!rke4r- tba r oLa
woet a the aoproyriate prepara7;io:4 :)rior to* and
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after the electroohorotic; treatment, The 1%erteix4,.ng
'10=21" fraetlonso howetrerp vere found to contain the
?sine residua, The el,5,otrophoretite4y isolated
fractionsfi which enpeared to contain most of the rrotsin
wera sUbjected to further ouantitative studies on the
amino acid oonpositicno Results of this research are
presented iA Table 1b
jt-ak g, E-2
.T.NreDanationo
The absorption qpectwa of ti a isolated oomnlexes were
mearatred in aqueous solutions in the resion tg 200 .
700 mp using the Unlosm Ut.700 spectrophotometer.
0harattoristio msxlma wore observed at the following
wave lengths8
M . 1
- 35.7
zi220. 4676,
505 mp
X . 2
- 356i,
412g, 46?0
5o5 Tap
3
2
. 357T
4126 467t
526 up
.
2
- 356e
4229 476 and515 m.)10
The latk of a peak at 278 differs the absorption
spetra of the investigated preparations from that,)
typical for tho arystallibe vitamin. 222.0 shoving charao-
teristi maxima at 278D 361, 520 and 550 upp respeotive3
The remaining Desks were found also to be shifted some-
what into the region of altranioLet light,, The most
striking difforenoes appeared howeVero in the r gion.
between 370 . 470 1111 vglith might be due to the haterogc
nity of the preparations studied.
Fig. Eo 1 illustrates the adsorptl.on spectra of both the
111241 and B .B preparations
ro,
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bUAl -HLD
' eleprjaW
Taeotrophoresis of the 312-1 and )3/2,4 compounds
ii the resence of 0N-ions0
Mr..CAM2U IW?040..O4CP SIWOP*?...
FigAo 2 presents the Ioaation of dgferent fractions
obtatned during ionophoresim of the and 11/42,4
oomnounds in rresenee of ECE,'
The eloctrephoretil: studios worm illerror,ed according
to the iioldsworthe procedure, BletrOpheresis occurred
In 114 033000E .dif 207 to which 0,01,5 7";, mts added,
5a.iie oWhatman No filter paper wf.&x.i. subjecteato
a potent/elk gradient o 5 v per ea Tor 1:43v1rec
The B22441 complex separated ;.nto these mato. fractions?
numbered as 102 and 3n respetivey0 Mateo miditional
patterns were detected in trales*
Fraotiono contairiag most of the c01ou3 teria24
applied to the filter :covert appeared to 'be a neutral
one. 2he mobility of thio componete as well es its
colours, me found to be simile,: to that Of cristalline
vitemin.B The yiellow coloured fraot#n,Vo 2 shoved
12?
basic properties? The colour of this pattern would
indicate the absence of the eorr1;1 moiety,
rraction No 3 vb,s reddish and more 'hoz:10 as compared
with fraotion No 20
When Biza was eUbjected to elav.trophoretic studies,
similar patterns were demonstrated =opt the fraction
showing most basic properties, which was not evidenced,
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.7.
As illustrated on Figjo 2 both the position ana colour
of the major fraetions were fatar close to these?
obtained after elootraphoretio treatment of
Fraction. No I ahowed a 1w3elet7. acid chareater,,
The qualitative amino acid smayois of dIfferen
CTV.0151C4tv="6C.siC.1..Vrtaa r.7.=,.:510:42ittaftWoWAimnat ittat?Mr..-44.31,1=4Wr="4= CV CC, ~4:11^CV:62,2s..CM,
fractionsv eeperated electreohoreticany in tho
:presence of k.,45 aonzr,
46C7.4.er.roaceL,WV.,47.Noecamoramolfee.4.-=,=.041
Fractions go 1,2 ana 3t or.natinc frau both
and 312-B preparations5 roppectivolT, were hyaralmea
by means of GU ZO1 and IfUlljecteC t* ya.par chromatography -
as described 'else-them
Whereas fraction No 2 showed only tram; of the ninhy=
drinvositive comp&ds, the remainin6 ainjor fractto
No 2 and 5D contaiied. most of the amino acidu prtsemt
1:n the untreated salaplea of 7,312.2 and 5127b,,.
On the base of these fineinge A *onaluaion nould be ?
drawn. outv comerning the decorposition of those elo4p*--
=de wtthin tho course of elactrophoesis in ,lriasqnzi
or CivriOrevaltine: tn the now,xati.on of tht
constituent /mainly vitntlit L.12 or it am.). tho
torotei. molety4.
Electrophoreste of. the 444 &Ltd& A 00141141mm4
?.2
in the abseme of t;rionso
The omis4on of ar within the courue of elootrophorortio
led to a afferent separation of tf,o !tompourCe nnt -3od,
as compared,t1th the patterns obtalte Oaon. ?XI 'Ala'
added to the eIectrolvteoilig6
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.8.
The major fraotionsp teL from either BirM or
73- -43 numered as 2 and 50 had distinet basict propertio
12 v
Horsoverp the B".411 preraration separatea into three
tratefraction:46v, one of which moveid into .the position
of orystalline.vitamInBi2 and Showed a similar eolow,:o
The remaining two fraetions were foUnd. to be haslet:,
On the other? barely the Z12-B conDounds separatpd into
fractions Re 2 and 3, and.a weak:ij acid coimonent
frattion. NO I /in trancs/0
The red noloured fractions No .3 obtained after oleotro-
phoretie treatment of Bielp and Dlea as wen au ths
fraetion No 2 of the 17:1241 wore elutedt hydroZygoa and
adb600ted to the TAntitative amino acid anallysirto
The Crionse usulAyapplied 'IA the.olootrophoresis of
torrinoldag. were found to affect the ofabilit7 of 4a7le
investigated comolexes,t
TNe absorti,an ...r9ectra fvf frat'filem!if%
dd. .??????dr dir . ? ta 44. 44.,1.7 ????? ...r?????
4:IAA ...ad, .4. ?? A??? 4?Ne ?-? ? ? ?
. La '1 irc,Go GI 6 'foriz
? Ta? kAdq::41 ? .1...:,414 44 efe ? ?
Theso meavuremorte werG oonduttod tn'ordqv to oongirm
s date frou tb.fz altd..no ao Lei. ars ti f:t al.?1,$,W04.
to 4g3UM4 tbs.t deFraexItion 41,1: (i,.)r.11o4Av niaittoI
daz to the aotioA 01 Wiolls-
Dozmus4:4 af th,t; jdarfity of t.".'a
Ott/20r from B.., --1M or
4,2 '12
1):0(5Ta.roJA(J,i74E, 0.4s.a3u:re...c.71:o;
?
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9 -
of absorbency were United to one of tbe indicated ,
co1!?oun444 !Lonely B1241,
Decomposition of the px,eparetion use cleersr ervil1en04
/fig. No
fte absorption spectrtna of the most intensive fraction
No 1 wee found to be identical with tbat o crysW,line
vitamin 1332e whereas the absorption of traction n'O 2
could. not be ascribed to any Of the corrinolAs.
It seers very likely that traction So 3 was ,the only
one Attach con.0.1ned the watered ipoteixt 000plez of
corrincid. The absorbtion spectrum of tibia fraction
differed floes timt proparation-sparticularly in
the region 370 to 4-50 nu,
T.he a'bsorptia oPeetrit? of the riain tractions *WM;
'ilirademeoW4011C11411m1k041esoobw....W.Ilaroolariiiisralailwiro...
compoundt separated within the 'awares of elect:ea-
~OM. ilevirowittamilliMilletwookbilarnakikdwAyismbilawroks.oeligmmilmaishm101008011411101110.10
pharesis -the absent* of' 03rtions,
114rtorrowl^iibimortimpaudiviobilmneallhoefra...romititi60.4b6wOlmailmliMoolowsri
As 11.43.110. 5 illustre.tes the absorbtion bands of both 110 2
and3 show molt loom tutztiastit shottedtat shifted toward
the re#oa of short eaves. We phenomenon could be allots
observed tta the case of crude Sic preparatioa. The most
striking diteeranse between the crude covonn.4 and the
indicated be comonente appeared in the region of 370
to 450 mu; in which the spectrzi, t the.later compounds aro
may antler to that ot vitelain Due %UGC riniane
tbiAt the comowde Imre ,liberated from the law
terferring substanoe. A ditZerence between traotion No 2 -
andBo 3 could be noted in the regia lx5tween 450 and 550
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Table 1.
Amino acid contents of BieN and Ble preparations Alee.n
values from M..1 and 11,-20 34 an4 31-2e retpectively /and of
the major frictions? obtained duriqg electro oresis in the
absence of RCN
=ft. .. ..
. ?
4
4 Amino aoid
0
U.
M
=======?====
13 '41
2.474
Fraction, fractio4 Traction
of the whole com.st No 3 Po 2 0 No 5
pound
P
of the total of in acids
Lysine 08.00 103X I
fl Eitidine 0 2.65
Arginino 1029
1 . 1 . 1
Aspartic acid 165 4.12
Serino m...6 0m91 I 1.72
2.62
1 Threonine 0..? 2.24
Gletamic acid 840 1 9tL2
IPtolito .....
Glytine 4.... 1.34 1 1062
1303 1 7.34
0.71 1 1.83
Alanito .....
/
1'2 0$stine.. 0.
Vallno ttee,60 2.65 11..93
petlaionint .. 1.34 1.13
Iooloucine.4. 1,a3 i 2.83 246G
beutine ..... t 4.60 I 4.11
iI .
8.92
fe.,9,,,,,23 1 0.99
sine 61640
4.G5
I 1 P6 i 502$
Phonyloalaninei 0
A..,..t.-4,-r.-....clittt.41.-..r.....t...=*.r.,
-............ -..................................
3,3? ? 1,89 1 . t
1 4,29
! :
:::
2.40 2092 1 ,
2.43 1 367n 1 .
1 1
2 2
.84 i 'A , 90,21
1 4023 1109
,.........
8,5
1
4061 I $000 49
2r91 I 2,75 1 749
2L.2 119.2 20.7
1
254 ip07 14609
3.32 i 3.),86
Zs% ,40 1 54:3
' 14
9.1$' , 12,0i3) i 4.01
2.13 146 2619
I
i 5071
1 44"
t ..
4,05
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. 11
of 'L
omatitative amno girAd okmositian of ono
3'301. ?A0 f/vo41.omp Fl2-Al
o fa, *ai? ??.a4- t4
1a7?. prpa,ratxons in the absezze of ON ions.
<
? 4.? a 4.p...4. 4.. 4tiCSLIDSVIVT4.6.1?..
17 anAnomt residues wore determilatdi aetion Ivo 3
o.7!' eitMr tho "
/Table ".(A I/.
ilar4?No GI oxc,,,,,v4; sonple=es
. .
The nyn5ortaC kavino ao5A residlIcs was ovidenaed
in Z:caotitlaNt P of the B12-4 preparation t th* basir3
amino aoir:M Ilaine arginire a:se histidiao mre not
dotectd,? fractiaa has bow, provious27 deseribd
ae
E.,eatteo of tho-laok cf SA appreciable amoont'of the
enverlmeptal materiel the elootroplloretioal2w? seporated
fraotione aftcc sIutiono were not evapbrated to dryness
end wathto but hydrolyzed direet, Therefore the
ooncentratigan of IndIvidua/ amino aeid is being f3alou1aF-,
tea asspercmt og tht total amino acid contens dotormi?
vAdo mad assumed aa 1000
Tho some,ril,avan maft the determination of th* basie
enino aads tu Xraation NO 2 of the 73, gt,eppvatIon
ftriossiblec, o ponrere fradtion No 2 and Zio 3c both
originatea frov, .save oomooundo it has been assummod
that the contertn ofbasie amino acids .in fraation No 2
wou2d not Inclood the tiguro of 9rd,Th asSumption was
mado eacor4liers t* tho ,exportmexItall datA obtaina. for
the fraOtion BO T5 of i81,51V and for fr-eft=
:Go 3 of 13127,,00mpouna /9021W as .SallOWA in TalaXe I.
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oreier. tO, eCtipaZ8 ti.10 C.4;tt.,?ICiS a21,d coapositikIa tao
cy,:ads groparati.ol Vaith that o.t iM?,:bmotist.419,
ti-it.vt,oz c.i.orses1;7,1)..n.ditis titj tho rem,araticar? eacule
bq miltipli&4 by a faakTf 2.? SOGMB tO
beiumlst 1g1 bottmol. tbe ikirtall tioldAct$,
tAcidak att,d. Utielt thtl
(5ott,l,atiTif; tcy. t7,,mta,k frattion wc,
botkkndLut-g,Daralrlifto ead. act tfaem titP
eagttaYit 4ftims3).1e0 gm?eovoll`' at csatvtas
(71.0'016..i eiZt-6:17111L:S.O.,
wzal crugtxinelQat611, and. y,:,-krcbably
'TYAZ lowf,ti? hcsfe-4,7,v;t, tci; h1;.1:0.1111:geint 1,11 tb4,,
1W5atO glIZttPn.
:c 2iags.' Vac, B. -7 trkola-pr+.4,..-,14
51.47o.',U,?o -1401,1ine, and, al.esm Ixttrti...ene
cras `S..zotottorT, 51e 5 !D?E t324 44.1.tom*-vonqi.mez.i
*ortpawed tt UsTt tamdz la.'tparatLagl.6 rron, i41V.(1,4 talb
fx.atIvA las tiettontez ths ron0.1102.e4. AWizg
nald, rtprloa2roel ratebv.,wL rL bcrbh zubml.m.:4-
e6:)go Vats, ttMi&P P=id 00Myaatit= tractim Nc; a mna rq
are .Ettail,,,?,,r4 i.dth vxwptittistf., csaitilmo
4.10 ft:1;11y) 6,5 Irs;11 kik'; :17 .;1,14:"i'int>117.,* 1,:ct
'4" ? .# ,Th ..fr
-A, -? Oa: sera
contair, 20% OwtotrAe.*".,4A 13Ce' seiT,IJI
Etari. vo:!,e.1 ti2.0;
Fractl,g,trtis No 3 Oftu
illobses,h
?and,
ar,:.4..101;1.6 tra.41 1:1-Yuc.31.3'16,
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From the hitherto obtained data a soneral, Gonfilualco
could be dram. out* *commit* the hotezaoseni* of the
crude Sz241 preparationo This preparation seems to to/
a mixture of at least two comPonents aitzering iz sio
protein residue,
,
It awes very likely this property Could be taso
ascribed to the 33$0.410 commis/1G
4*" ?
Studies on the 1.1.ghtsnitive cimplams
The light sensitive compounds IS 444 isolated by the
procedure. described in the ,experimenta40 showed an
7? ,
ohracteriatic orangereddish co3.our*
1* .,....412420:2M1g*
This wao perforMede acoording to Boldawthoesoept
A the absence of light* In the presence of arions...
to preparation sei'Perated into two tractions., The neu.
trai one showed a,characterl.stie colo, specific to
vitamin B.R.1Al storasp the other contained moat ot the
? , .
orange-reddish coloured. tutorial ,M1, fileotrOphoreas
of th Bete.s coivound in the absence of oriOrievreltriacv
ted in a single fractione. whicb has not been, subjected
to further studiese within the caurse of research
,
covered by this report)
.? Samsk.rwaoom
ilustrates the fi,bsorption. spectra Which core
spend to the B -111-11 preparation as well as to the
preparations obtaine'd after eleotroph.oresiso The spectra
of /3 prepare.tien and of its oraxtge-reddish few.-
tione isolated elpotrophoraticallye are similar to
50X1 -HUM
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trnv 1_11 IRA
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that of 5p6.--dimothylhenzimidazole ib2 coenzyme reported
Baker iFed,irocto20 956 /1961/ The absorption
ectrint of fraction no I ma similar to that of crystal?
line vit.-1111in 11/42.
kormation of thith fraction within the course a electro?
,
phoresis V1Lprobably due to the action a crions the
t3.2 coenzyme Anothiar possibilityg, which might lead to
the elopeerance of thls fractiont would be the unsurZia.=
ant light protection during the isolation of the light
sensitive form of corrizoideo
309r,IAts.,jthu4....,,.....Ay..._ut? acid an sis.o.f.tizsai
atiUtious.
This was performed lay mesas of paper chroraatoaphare
Dutanol ifacetit, water aystom was used as solvente;
The proctedure for hydrolysis and chromatography was
identical with that desaribed elooWhere, Gine and
glutamia acid were the only amino acids found in eppreal=,
able emounteovihileert othox amino acids could be detected
only in tra01988
In, fraction Vo I which seems to be a vitamin Ike
corvound only Glutamic acid was, evidencedo On the other
handoglycine was the only one an3itse acid residue found
in the hydrolyzate of fraction Zo 2 tihrooenzyme.lite
colapwadC
The light?sensitive forme of corrinoids will be oubje?
ctod to further studiest, as indicated in the plan of
future research,.
50X1-HUM
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Chromatography on Sephadex A.25
COOF0,461WWWOMWOOMO.Pgalwel
An appropriate VOluDO of the M-2 water solution was
passed trought a column of Sephadex A.25 /medium
16 x 104 cm/0
This was followad by water elution of the coloured
fraction. No resolution could be observed. The recove.
red compound was hydrolyzed by GN BC1 /22 brae in
sealed glass tuber at 110?C, and after the acid has
been removed analyzed by means of paper chromatography
as mentioned elsewhere. .
All the amino aoidso commonly present in a protein
hydrolyzate were detectedo except cystine /or cysteine/
which could not be evidenced* This research will be
continued.
Microbiological assayst
amaibmatamenmemeAte=wwwwwwwW4010.4siborpaimao-ftwor
Assays?of the growht factor activity of the isolated
oomplexes for Leon were performed according to the
Burkholders tube procedure.0 slightly modified in this
laboratory. Leal 113-3 served as test o en0 The
activity of the isolated complexes /expressed as percent
of that of the crystalline vitamin B12/ for the E.coli
was as follows t
925210 AgItglE
m . 1 34
M 2 4o
- 1 2o
B IT 2 68
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HUM
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?
?
115
P4,1.3.,(mrring. tm.nr,s;gra..-.4,6o; fooltzte.01. rdtittz2. tho ow.a-sa
ZiAtd,7 vare carrx, .
, the -J.1**x; meglittyv skrrinold. prepapatita
ator 91T4atrvpht-tIo. tmRtmszt / DV Criftgl tddtdit
- twa.fraraiallA thep ,Tisht so.aitiftearrit.pta
wvaration,cmpated iobtmhtrotioa3l7it Vilt pTesexee
- the? patAT,Ioxi ?3f tl,v6 e:lootrcaettml fmotion
o(5.spontfgg t14 tl\st mgystaltlae vaAmmin 111.,
U,nt tth.bairti?:, traffic% nzvidiat* the poniti=
ot faetw
Tam absoxptiem apsqlitra ot theat oampments nay auzgoot
? thv cormonicin tho ,proteig-A cArroysid. ck light
zezzilAve zwotaltinri,i Vare: torre.44itui (It
tata 61, - C2rfez,Fial Trit:ZEtir. E120
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Declassified in Part - Sanitized Copy Approved for Release 2014/05/23: CIA-RDP80T00246A074600230001-8
bUAl -HUM
.L. of :.itzttave Yre.oevx