Document zQ65rOJGBJbO4RDRMEDdQkvJ6
, Monsanto
mM <NMI ft LM'
UftJItt RirilMNCi
TO :
W. B. Papageorge - General Offices, BIND
March 4, 1972* PCB BIODEGRADATION
Dr. E. S. Tucker T3P
"Wff S. Bergen - B2SL W. H. /Duffey - EISA J. TK Garrett - A2SA R./E. Keller - TIB W; R. Richard - T3A t. P. Wheeler - A2SA
Enclosed Is a copy of a recently Issued patent describing
a process for degrading polychlorinated biphenyls by
using specific strains of bacteria. I am interested in your evaluation of the proposed process.
/bt
Enclosure
W. B. Papageorge
HONS 028923
Arnrovi'lcx
latent: (t1'991i
(in 3,771',"-5 <> ('15! D-jv. 18, l'J7?
($41 MifKOaiAI. PKCUADATION OP t,OLVC:ii.CM!N.vrs;i> m.niKWi.s
1951 I nventor; Kdwnrd N. Azui-owjc/, Vienna, Va.
| 7.1J Af.mnre: I uvtiAv.Itje Alp\.mi!ib. \`;i.
nc., '
(22j l ilcJ:
Apr. 7,1972
(21) Appl. No.: 24.2,192
(52J ISM |5KJ
l\$. Cl............................................ 195/2. 195/5 II lit. cs............... ........................................ C12b wt> Meld of Search.................... 195/2. .2 If, I. 2S ft,
195/51 K. U\ 424/95; 210/2, 10, 11. P
1561
2,6l(i,20< 5.6)4.227
Ufcrn\rs Cited
UNITED STa'ILS PATENTS
10/1971 I.inn............................................. 195/2 1/1972 J'jllerson........................... 195/23 R
Primary fisaminer~~A. Louis Monnccl)
Assijtnnt k'\o/t::r.,r--ft. B. lYnhn.d Attorney-- ftwymond C. Slewin' vl id.
157) All.STMACT
A process leu the microbial degradation of polvchtorinutcd lip{;vn\!s (PCIis) which comprises treatlag the PCfU will* certain norvpati.o-vnic, hulnuarbon-utili/iny, s'.uiiu of Clyi.,op..fiuiu eladosporioi.l.s. Candida Upobdica, Noenu'in ylob.Muh, Nova,da* rubra aiuJ/or S;\cherorn)cos rcrevisbv until ti'5 I'Cb, have been substantially degraded. 'Mie process is ap plicable defrn .!:! PCDs ns they may in? presem as pollutants orconianinacus in water, in todvsirbd eft*'.'cnls, in vaiion; land areas such y; iiulu.iit:il sites at.d die like or in varied leboratcm or coin-u^rciul install.'lions. The process nay also Ik* u.ed to clean up degrade mixture* of rCBs am* \yrious hydrocarbon oils or petrochemicals whenever their presence consti tutes a deleterious pollution.
29 Claims, No Dr." wings
s HONS 02892*
1
3,779,866
' M,T!U>;i!/,l, DXUilAOATION W H>:.vos)i.<m;.natt:i>
very Severe and harmful ifV< n. from Pc/j |-oi<p.in>t `I'bcrc is some evidence that IT!', alter liver n-.ue, in
HACKfiUOUN!) Ol-' 'rill: INVENTION
hibit the pnv.vsb *f Ui)ti;n <l re.'.'x acut allwt ;> \ ;>n.l\ f-f enzyme r-sslvnts and the ;;emdio niatcrial m veil., hut
This iov-.-ntio.n icl.dev to a process (W the miuTnbtnl J in mo..: <.*;<>, the mevhi.nKn of anion is s:f?l unver-
degradation of pohcbl.uinnivd hipVoy N. More panic- tain. In .my ' '.erd, the p.wnve of TOfU in cite enwo.u.
titiirly, it relates to a method fti dc jr.-ding p.-ly-
menl h is been a front cmu-cr.i to i;vi .'n'.ist, end other
cMolimdvd biphenyl* by mews of in?vror.r;-urd>rii.< it. pe'vlc co;KCin.-vl with b.-jlih, safety ami volt >rr. atn*
nuar U' ttc*m up a.:J chru-tuic such organic pedal.:n.
tl.cra has l'.;va a .u'at i;e.d ft:-
il.-velupaieut of a
U*. they may occur in Xhc open r.ca. iid.utd t;oh. waters, 10 procedure f.n tley.icdtnp, t'.v^.- v.iV..,t;me.'>.
luf.il punk, h,minus and the like. in.lottrnl effluent
Aceoohnply, one of the o 'j. tt:. of the prevent inven
charges, su'VtijfC r.ud o'hcr pipeline systems, farm .cil tion is to piovidc a method ('< dv-y.ra,[tn.;
or other land areas ir.oludiiu: iivJustfud j-ilos, etc . The chlorinated tiiph-.-ov Is and sini'kir Imloeerutev! mole*
ihsci'.den is applicable :ot on.'y for cloa-aav.-. op
cuius, such :;S DIM*, wh-r.vet they may appeal as coo-
ctdoriiuud biphenyls i-s contamm. nts srd pollutant* 15 tamin.od ; or p.diutnnts.
In open environmental systems, but also in cto.cd syv
Another object ol the pu<eai iiv.entio.v is to provide
umaa.;, forcAUinpk .it. industrial. tnmrnerci.d or gov- .a process fist the mieiobial degradation of poly
etmuent plums ruul installations and in vrriou* labora chlorinated biphcnyU whoever Jesiicd, for exampte,
tory operations.
C5 <1 r.uar.s of ch-aainj; up cto-cd ; ii*l open wmerv, in-
liiHitotunvnt.d ch-nn-up j of modi concern to the dustriol cStb *nt di-charpcs, polkd-d umrsldamts, i-tu
country and to the. wot id today. Polychlorinated biphv- {tries, mnrioe m\irom.itnts, dispoial lagoons, undami-
.y!* (hoicinftticr referred to ws iCU>) :tfc int1a>tri^l n.ded imlustui'.! .nev.s and fiiiodeiMi.s. arid in otlvr -itur.-
cht'aiici'.U that ere widely used ns pl-ardicirejs, fire tolar- lions where t'Cds may become acctmtul.-sed,
<Iiut painl mjjrcuicnu. hydraulic fluids fmJ beat ex*
A further object of the invention 'v- to provide a pro-
clumj'.ti fluids. They have recently been detected m var cedurc for degrading* pulych!otir:Uvd biphcnyl.e lead-
ious vmlct sources, in human tissue ;nid in many species iiy, efficiemly nnd relatively ecunomically.
of biidr. nn*J fi>U. Heinj} relutivdy heavy organic mole
A still fuiihet object of the invention is to provide mi
cules, VCll* persist imioficholy under nnturrl condi tions. Moreover, they tend to accumulate ir. the food
croorganisms that arc capable of dci/rading poischlorinntml biphenyls, leaving u detoxified and beru/i-
clttiiu, smd oitu im;c .ted, they are stored in the tissues -ly ci.d cell mass, ihcr.y microorganisms being completely
of birds, mv.mnud-t and fish toed as human food. As with m.-ny other cMorina'ed hydrocarbons, such as
r.o-palhi>enic to muiinc fauna mtd flora, humans and animals.
l>t>T. even sma'I dosages uf I`CIU cun be toxic.
Yet another object of the invention h to provide a
rCUs are relatively inert compounds that have low method tor the degradation of poly-chlmin,th-d hiphrlolubiotics m water. The commercially produced TCiis -1* nyls v.livrciu theic is no need lor the hcrnHiiig, trans ! Ht e.etrVIy mixtures of many of the i'-orners which arc porting and rbK.jgf cif Itoavy bulky ev|uip;uem.
possible among the various combinations of chloii-
These and other objects and advantages of the pres
nuted products that can he obtained from the chlorina ent invention will become apparent to those skilled m
tion of biphenyl. The rnoti common industrial mixtures the art front a co.rdJcratiou of the following upccifrcHcontain between about -*0 and 60 percent by weight of ^ tion iind claims.
chlorine, but the Isometric composition thereof is rchv
i lively unknown. Mow-cvcr, the more bodily chlorinated
Pf.T.s appear to persist in the environment for the Ion-
, SVJMMAHY Ol-THU INVliNTlON In accordar.cr. with the jrescnt invention, the nl.mc
objectives are attained nnd tm advantageous procedure
PCits me widely used as dinlcclric nuk'* in capacitors 45 for themictobi.il d-ccradatlrui of prdychloriratcJ biphe
end trausfoiruTS. as hydraulic fluids. as beat transfer nyls ftfii been discovered cntplosing particular ttrams
figcrds and as plasticizers and solvents in various .'tribe- of rmctoottMtirm;. 'H;.` d;>5imn. tmunir* ;*t c.f
fives, fa-r-len'.;, pai"t and printing inks. M.u.; >f the re tiiC present invcniic-u i* that all of tin: materials uv.)
lease. of Pv.T-s into the cuvircu.nmnt probly occurs from me dciived orf;:t.'Ky from edible vjh-umces which are
jitdustiial ei.aimit discharges and Uiirnpiup. r.<] hmkage ^ t.Ot toxic. It h SipphciibU* equally to the degrad.Umii >f
of luhriv.-mti, hythnubc. fluids nnd I car t/isn.tfcr fluids industrial svjsivs i) p.-ner.d a.; well as to th.e deju i.la-
Into tb*.* vatcr'vriys and
frr.m materials containing tion of I'CU*. in <-p:i> and timed aqueous systems uo
PCHJs ris ( hi<tici/.:r into the atmosph.^.*. nml a a re sult of Ic.u'liiniT f.oni dumps and lurulfilk
Allhcaj'.li tlicrr is stnne evidenco that I'Crts cun be dcehlotiauted by ultraviolet Ivj'ut and, thy ;, lead to the formedun of byrlroxyl derivatives and other jud-ir com
1 pounds that rue more readily deg-ra.luMt, {`Cits me
veiy peo.ivtvm tin.l are e*oemi-)y di.iUidj to <hy.inde once they ;uc present in the unv-rooment. i'CU n-siducs have In'cn pytiicuhuly not'd in fish, bird; na.l mam
i mals, v.hieh lacltuie those used for human fio.).'. as well ns the s.ilJ .u.iiiv h, Ahbftigb the data nr.* i.icomplete t the ju-ew.nt lime, than- i> si^niruMP.I e-idviKo that Most huuu-ns have some residue:-. tf PCIIs in their listuc. Th.c cfieel on hum.ms of such reshh.vs it J.ko un . deni at the presom time, but name uvi.iear mdk.Ucv
matter how the sysU-.u was con!.'.minuted and polluted. In the present inwntioa, ndvimta.-c has been taken cf * judiciously chnosimj n;eioo:tvi;;iius which ere capa ble of degrathup 1'C`di mid u-dr:^ them ns h eaibon
smitL'O for jyrov'tlt. |ht: prerent im-.-utiou comprises v. partly biological
process in which certain Scl-.ctc.l tnicroorgu.i-ms * break down tt-.c l'Cb- and convirt the rontaiaia.nos
into m.ijscs oi cdb!., mm-toxi: cell;, iv, a pro:ehi nirxi. In ar> outdour j'pplicalivm, this cell m.i.s t aa be tharva.-'.'d into 'In' food ch;::;i t. feed h-g.her fi-imi of . fife nd, Ihu1., a very advantagcmi.s end ivtiit uachieved in iu'.ditiou to solving the psiddoni <? Pt'iJ conti*ruiiiiUii?M. 'I'bcrc i< no t*eed lor aucill.ity cle.ih-iip optta'ions when PC':.', arc degraded in accord.oct
028925
<A
I
3,779,866 34
with Ok* insertion ur.d, as pointed out above. there is also tio n.'i'J for l!;c handbag, tr.>i.sp..rii,;g and storing
large quantities of the lul'iucs. If necessary, sm anti foam agent can he cmplojnl therein, for 1 wimple, f)ow
of lic;t'>, buUy equipment.
Antifo.un A. or crude oil.
Thu Mlov.ing, n-.kinoig anisais. ..II completely novel
lienee, either a svr.fh* tie culture medium ornatural
and umdmous, sac utili/id in the picvcnt invention 5 nutrient r.criiim* is su'hri hr for the gross th of Pie micro
The>e i.mu'oi;.',! nis.sis **ru special species v.Inch have organism Strains employed in the pcen-nt invention as
been adapted l i uildcvc the objective* t.f the ir-vev lunp. as it cont-urn, the essential nutrients (<< t>iv- growth
tiun. They 1.;tvc h.mn il*.*|'o>ilci! with ibo American of the particular microi.ipnnirn thrum m str:drtv u;ed.
Typo Odtutc C*'M:clion in itockvillc. Maryland mtd Such nutrients me well known in the art ami include
have been given the dc-ijnuUd ATCC catalogue mnn- 10 substance* such as u eatiton source, a t.-ilrov.en sou.c..1,
be is:
inorganic compounds and the fil e which are Mi!; 'i d by
Cluiiosfnuium clndosporioidev (HI ?A02) ATCC
the microorganisms employed in appropriate amounts.
20251
The microorganisms used in the pi> ant ia-endon
Candida lipolytics (HI :-002> A1CC 2CC55
grow end rnif-ive in an ticp.eous mitriaiit meJium con-
Kocauli.J glclvtuin (HI 10>0) A'l CO 215*55
15 tuinirs;: a hydtoctirboa or a mixture of h>dioc: 'boMS :is
Nocmilia ruSr.t (l>! 1002) ATCC 2150?''
the nain carSoi source. Such hydrocaibans include
Sncchaiomyccv coio\iii'.c (Ul 2009) ATCC 20252
straight urfl branched sbaia pi'iaffin; (ttlhani-s) rang
The clwMiUTV-tios and properties M th-:-c raieroor-
ing from i:;?o*JS ulVun-.-s, such as methtme in-.l pro
gstniuns arc deisrihed In copcnding application Ser.
pane, liqtiid or seosi-rolid tdkanes, such r.s u-oeniunc.
No. 43,226. fried on June i<, 1970. entitled -'Microbial 20 n-octane, n-J cc?:c, r.-u'odecune, rt-hev.'.: -c.me, i-o-
Degradation of Petroleum". .
pentane, ioov*tane, ami including long-cl'.t.in
pat
Various media can to employed in Itamlling these
affins having high melting points, cyclopar;.fi'i:is -nch as
cultuic*. Alt of tlu microorganism* employed in the
cyclobc-x:.nc and ey ;loi.t t;RC, straight- end branched-
present invention will prow on media with I0D percent marine water, will* part imilne water and pari tjp wa
25
chain olefins such as par.tcnc-2, li'jACi n-l. oclvne-l, Oclcnc-2, etc., eyeloolciins such ui cyclohcscnc, arn-
ter, or in distilled water. Tlu following medium has been round to be quite satisfactory us u general use, all*
mntie hydrocarbons such as benven-a, o-xylcM, napltllialenc, phenamhrcne.s, unthiucenes, etc , and mix
purpose m-.ilium for maintaining stock cultures:
tures thereof, as well us mixed hydroc.ubmis such a-;
All-Purpose Medium
30 kerosene, ligltf oils, heavy oils, p/trafYiri oils, petroleum
Heart infusion broth (Difeo) -- 23.0 g.
crudes, jit fuel;, gasoline, etc. Oiltcr organic sub stances, such as tdcohols, aldehydes, kcto;.cr., organic
.Yens! extract (Difeo) -- 3.0 jj.
,
acids, phenolics and aromatic heterocyclic and carho*
Glycerol -- .*>.0 ml. Glucose -- 5.0 g. Aflat -- 1 .VO g.
cyclic compounds, are utilized by the microorganism-*
33 described in the present application cation. Small amounts of other carbon sources such as car
Water -- 1000 ml. The standard 1/UKhncll-llnas ttu-tl; has been found to
bohydrates, for example, gh.:cv>se, fru:tovc, maltose.
be ijuite suitable ns p. grow th mctliuni on the laboratory
RL'cro;c, stiuch. .%tatcb hydrolysate, r.udavscs, eta., or
settle. A typical medium comprises the following ingre
dients: Ycast-nilrojen base (Difeo)
1.0 g.
<0
an)' other suitable carbon source such as ghxrrcd, man nitol, sorbitol, organic acids, etc. may he ucd in the culture movluim ah.i.-p with the hydrocarSun. These
Yeast extract (Difeo) -- 1.0 g.
substances may be used either singly or in Mixtures of
MgSO - 0.2 g.
two or more.
CftCI, - 0.02 g. KM1PO4, Monobasic -- l.Og.
As a nitrogen source, various kind; of inorganic or
1 4$ organic sab* or compounds, such as urea or iminimmim
ir.UPO.. Dtiv.sic -- 1.0 ft.
. tolls such ns ammonium chloride, ammonium sulfate,
Nil,NO, - 1.0 R-
ammonium rdtr.vc, amtiKvmm* phosphate., etc., or :.-
FeCl# - 0.05 g-
or more than one amino acid or crude protein mi.\*d m
nto:n;h>r.u)l Hue -- 0.08 g.
combination, or r,dural Mibvtanccs c\ai. itiiuj* uii.i--
Wider - 1000 ml.
SO gen, such :u cornstc"p liquor, yeast erh.-.u, nict'.i C'-
The following medium has been found to be particu
tiact, Hs!. rr.e-i!, peptone, bouillon, ca-ein hyJrol>.ite*;,
larly advantageous for the large-scale ptouuciion of the
f*h soluSlcs. lice bran extract, etc., may he employed.
desired cultures: Skim milk (0.-* percent) -- 2.0 lbs, Cottonseed ntc;d (1.8 percent) - 9.0 ll>s.
These substances may :d>o be used either singly or in
combinations of two or more.
55 .
Inorganic compounds which may t>e added to the cul
Marine Salts (0.02 percent) -- 0.1 lb.
ture mcdUt.n include rnapnetium sulfiit-.', sadiian phos
(NJIt),!)l 04. /Abash: (0.1 percent) - 0.5 lb.
phate, potassium tlihydrog.cn pho-phate, po:usiuiii
Hydrocarbon (0 44 percent) -- 2.2 ibs.
ntonohyibugen pht sj.liat.-. non stdiate or other iron
,
Top Water -- 60.0 pal. Instead of the marine salts and tap water, native sea
60
salts such, ns ferric trichloride, niangan.'s,: chloride, cal cium chloride, Mxliiim chloride, autmotiium nitratr,
water can be used Iri the tbove medium. Aeration is etc*.
Usually pros hied to supply oxygen to the fermentor ves
The microorganism-; employed in the present inven
sel or tank. (i; r.cr. Ily, the mtcioorv.jni mi culls me har
tion are cultured usiJcr; erobic ron-.Sitiau-;, u:c h s-.s at ;-
vested after about lh/v: or four day, of cultivation. A large batch \ev:il or for mentor '.cdvd with a young
<A
otic shah ini' of the culture or with -.rirrii.g ar.i av ratio a of u nd*inc-r;:cd culture, at a toiv.p-. ialme of, hu c..im-
culture equivalent t* about 2 prreeM or more of the pic. abenit 5* to 35*C\. pr-;fe.dly ?h#- 3U ('., and at :
total capacity of the fcrmenlor is used for producing
plf of, for eraniple, about 5 10 N, prcJW.ddy 7-7.6. T'f.u
HONS 028926
3.779.SG6
56
vwk'ioory. m ans urc hnrveMed at nn appropriate time unJ arc U a-U i". ('.is-iu.i'.vvl b-dnw*.
ATCC 2 l.'U5, No;;;rili;< ruhru A'l CO ? I 50k and Slicehnfo.iiy.v.s cercvisi.ie ATCC* 203S2 ix in >.n'.U'.d intn
DHTAiM-.n di:su:ution ok the ekei nnitrr) I-MUOPiMLNlS
h 250 ml. l-rlonmejur !*:. *< containin'- ( 2 (1,-ueiit by weiglit <*f Pydrnjl 3 J2A in I Oil ml. of !:-.* Ovs.-rib:-.! me-
5 diuui, tt:-. |l.:sk ix >l:.tkvn for three days on : i ll ikcr .11
Tin* pn:v->s of l!i.: invention can bo used to raroewe 27 'C., l:n-J lljy resiiliinp evil iij.ijs i. renov-nl 1 rntu ibe
polycldorbiHcd !-i|d ,.-:tyU frem loiv.tior.s vhe<Wof its flask. An. ly: is tf tko n-minning tiiptid by i-., ,m , o|
piccncc umxrtiau-% a deleterious pollution. Thus. with chiOMa'.c/t-;!\y *hows that the I'CH cone-.-m.-aiinn u
lliis p.M;-,--**, it hacr-mc* posihle M clv.m up and do- lcs< than fi,3 p.p.ra..
ftack I'Ci ' m the npsn sen, in harbors. rivers and 10 The same rcvjhs arc uhtaim-d \vh,n tlu- procedure is
other inland w,tiers. on various kinds of bu.vktft ssr.J Carried mt i 'ii:^ ot.'ioi inixt-jivs of two or i;uiie of s.-.id
Soils, in industrial effluent Systems, in sewage dispoxd syatetnx, in various Lb-oratory systems, clc..
niicrtvrp.misfv.v Mcnc..-, rh.- th-scribed i.iivfo.:e;t(n*.'.s may b-b u.wd either s:ny!v or in various ce.-mL-ir*.uiocts of
The feSi'/.virn; examples arc given merely ;:s illusfra* two ci more to dcpi.u'o J'CUx in arcoid.iiicu wiih die
live of the prevent invention and arc not to h$ consul- 15 ohjccbvvs nf the inwiuion.
eted ns Ue.htinv.
Ciunerally. the dcgrail.'iio-i of PCfJ< on n Ijrgc scale
EXAMPLE I
h cordtic'cd mi a nmlti-tur.k installation. I c.%s often, degradation ix done in outdoor cuviroMfie! open
Py (Irani 3 1 2 A. a hydraulic HuiJ c.or.tnining a mixture whter upplicatirms, it is ipi-ic fca'ible to . ed or dis
of Pen# and an oll-sn'uhle dye, is pc-ured into five K,p,i- 20 pel sc the i.i-vroor^imuins employed by m mi'scif beats,
Kill' 250 ml. I:IiT.ntcytv flask* to give u concentration uitcuvlt ot other vehicles ;$ approprisic. Even Humph
f 0.2 percent by weight of the Pydr.iul 3I?A in tlie the PCPs tire heavier thnn water, dei'radaliori takes
flasks, which each contain 100 ml. of the following me plucc since the mittoo'tnnisnts become dispersed
dium: Ycjtst-nitrogco has? (Difeo) -- 0.1 fi.
dowim.ml v.iih time to th: benthic mud. '}he mixture 25 employed prtfxTubly inrlud-is n cellulose ub.,o;bcnt
Yeast extreti (Difeu) -- 0.1 g. Peanut oil (0.2*.-!) -- 2.0 in).
uch as, for example, Shaw, bagasse, pir.e b ;tk mulch, gftwduM c*r tdhur forest or n^rirulturol prod; etc. Aswli-
Ms$0 - 0 02 fi.
tivc nutrients for the mierr-organisms arc also mixed
CnU, -- 0.002 g, KH,PO, Morohriwe - 0.1 g.
50
with the absorbent, such a> cottonseed Inexpensive ay.iLuliutal h;.-producis
protein or other s-n-j inorganic
K,M1'0, Dibasic -- 0.1 g.
gaits of nitrogen and phosphorus. The proc-rrs is appli
NH4NO, * 0.! 1;.
cable from just above fr.-e/img tcnipciaturo (about
tvo, - 0.005
4*C.) to about 37'C.. The I'CH degradatk-n will bccia
UcoutUiyrnol blue -- C.008 p.. Distilled vuler -- to make 1000 ml.
. wpon gp:c.i(`-:n/ the mixture on tlio surface of the- water. 35 Complete degradation may take place as tally as three
The first flii'k is looinlutcd with CUtdosporium cl?.- days Jo onu v.-cek, but could lake kinder depending
dovpoiioid-.s ATCC 20251, the second Husk with Can upon the (YTl coueentrotion and thu K-rnj-eraMire com
dida lipolytica ATCC 20255, the third flask v.ltfi No- ddions. 0.f course, it is not necusiury to me yn absor
<hrdia glot-rrula ATCC 21505. the fourth flask with bent, and the iniCK<orjariis;nscan be vi'.-.-J ft-.im or Nouardiy rubra ATCC 2I50J and thu fifdi flask with tu in a slurry, powdered or pelletimi form with vvLIed nu
$arcluroM)ee3 ccrevisinc ATCC 20252. The flasks rto trients.
shake 11 at 170 i.p.ia. at 27'C. for three days. At the end
With an replication on dry or damp sivi', the mixture
Of ih.ec days, 11 visual examination of the Husks shov.-j of microorjar.bfTis ami n.-ui-.-nts can h<* uppbed n* the
|i dis.vpctu,im;c f the globules of hydraulic find, which svrf heavier than writer. The resulting eel* inn;<
45
roil ::< the nr.ture v.il! work a.itohlc.-.ily risd anatrobicaily la ccprada the contaminant ITlPi. This proce
U s;panttvU Horn each of the flasks, iuul the rcr.j;tini:v dure is espe::ahy eifectiv; on extremely wet Mjrf.res
liquid k analysed hy mean* of gm cbromrstoy.rai'liy. Thu gas chrotnr.fogr'.pMc analysis shows that the PC it
Mich as turn .h binds, fnim la i.ii or tridaurial land sites such ;*.s t.ucl.ir.g areas end ra lroad sidings.
conccntiation in etch ll.uk is not detectable one! is thus less than 0.3 p.p.r.v.
50.
The most )>r.ictiv".d upp'-icaVM) of the *.re'ent inven tion involves, the degradation of S'CHs in vvyr-*u taatcri*
EXAMPLE 2
ai from Jude..trial plruuvor other types .*f oo.nmcreinl Installations, where f'CHs arc spilled accident.,lly by
MONS 0 2 8 9 2 7
A 250 ml. IWenmeycr fl.uk eonl-iining 0.2 percent leakage fiom heat cxch.-f'-eri. In this epphec'.ioa, th.c
by weight of Pydtoyl 3I2.A in the same medium de- 55 eftlucnt rr.;-.:-.'ti;-.! to he degraded can be placed into
ictihed in Example I it inoculated with a mixture of large hol-Ji v. bulks, for cxauijd-e, ami thu r.iixsi.rv of tm-
CLil-.ixporium ck:di.-.a).>sii:idev ATCC 20251 and
C'uorf.'uiij'n*. and no'.rientx mMed thereto, wlk-ruby-
did.1 lipolytica ATCC 20255. The Had* is shaken at 170 dc/tridaMon will lake place i:-` dyveribed above, btiiriug
i.p.m. at 2VT. for tines days. A visw.il examination of nrul .'ler.itio*: of the mixture inside the lank is a Jvnrd.v
Hit Ua>k shows a d'.s.tppcaf.tme of the byc'i.udic Huai, fciuluf.cr mp.mittun of the resulting tell m.nss, jasci.rv-
60
p.omisly er:ij r *y 01) to provi-l: :: 1,1.1 re rapid r!c ;'.i .illation. A kitrjj'e tar.k >y*Jem may bo used, nr ft nml'i-imA sys
ntajogiaphic eaaly sis indicates that line I'CH concentra tem may he iv.->h>yed wlvt''m th? effbuint material is
tion is lesv llun O.3 p.p nv.
moved from t".ul; to tJ.uk at xpecil'H-J times. AdU.tionM
nticioorp.niii-m ruhuiev oiid mitflj.i's lor tlu* 1 ,i-.*:i,ot-
EXAMPLE 3
45 gnnir.ms may Iv added to the Mihief^ueirt lard.s ile-
III llic same man/ier n.s described in Example 1, a mi>.- sireJ, for example, U* obtain a ih-sired u; uvtexs.ir;/
lure of Clado.po.'iurii chidosputioidvs A'ICC 20251, level of I'CH co.-ecitlraiion. `I ha der.ra.lerl ei'ilm-.)!, fur
Candida lipolytic;- A ICC 20255, Nocardla glol'crul.t example, ui.ty than be )i>aiinrjU'd into a lake, stream.
3,779,f!66 7 fi
marsh,etc.. 01 the water may he recirculated within the ganism selected from the group cottds|in;< of non-
imlusiri,.! plant.
palhogenic, t'elbmavi producin'1., hydrocarbon-ntth/in*
Sinew the microorganisms of the present iuevntion strains of t'i;,.!ox;*urim-i ctndo*porir;ides, Candida 'jj,,'.
ore nlo t;:p..3vo of dogrudiny. pcttcleurn and various lytica, Noc.iu'i.i gloheiala. Nocardi.i ndu.i and .*' iceli>-
petrochemical substances, i" described in copcnJitig, toraye;\ cercsisinc. for sufficient time until the
Ujpli.M'i.,:t Scri.tl Number
the
present irvention may he. cmploycJ
jiroce-.x of the v.'i'!t diluents
t'
chlorinate biphenyl* have been substantial')' d-:?,- I. 2. Tile process of cl.iim 1, v.herein nni-l mirmoo-ari^
which rtofnin petroleum or petrochemical ror.tnmi- isni is mixed wilts a celV.osic nrakfie',.
ntmU if* vJi i:v I'CIts, f.x example. ctnukifi.d cutting
3. I lie process ol
!, wherein j. .id microorMti-
oi!< or rolling mill cotduntK. The degradation of one 10 ism is mixed with n ccllulodc r.udeiinl, a nitre'cm
tubidtincc i.c not affected by the degradation of other source and a phosphorus source.
luhstaai e\, and a complete clean-up operation of, for
*. Tlu- process of i 'aim 1, v. herein sdd microor.mu-
example, a total iiulu>ifiul waste problem, can l effec ir.m is ndded to an niveous solution rontamin th-
tuated in this manner.
polychlorinated hiphrnyls in slurry form in ;m amount
The nd.-roo; nanism oi mixture of micioorp-iRisrm is I? of approximately I pere.j.it to 5 peivont by volune.
{itlvur.lug'.sMtvIy added in slurry form in r.a amount of
5. The piotess of claim 2, wherein s.iid ccii'iUnk ma
uboul I { erven', to 5 percent, preferably 2 p-.ucent, by terial is selected fiom the. group consisting vl w.vduM,
volume to an aqueous r.ululion cuaicinit?}* about 10 pine built, wood flour, cuttua luiters. < ottonseed Imll.,. p.p.m. to about 15 percent by weight of l*Ck for opti straw, hjigasse, nsursh hay ami flired'h'iJ paper. mum degradation action. If the mixture to he degraded 20 0. Hie process of clniiw 3, v,hetein said nitrogen contains oil and other organic pollutants besides POtts, source is roUim>;ul piulcin,
t concentration of several percent (NV/V) of microor-
7. The process of claim 1, wherein said microorgan
gtiftism-nitncnt mixture is usually sufficient to be ef ism Is employed in a slurry form.
fective. About I percent to 5 percent by weight is desir
8. The process of claim I, wherein said microorgan
able.'
. 25 ism Iscmplojed in a pelluti/cd form.
It is to he noted that a balanced nutritional medium,
9. The process of claim 1, wherein said microorgan
including n carbon rourcc and additive nitrogen and ism Is employed in a powdered form.
phosphorous nutrients, is provided for the microorgan
10. The process of claim I, wherein said microorgan
isms employed.' Since the additives used arc of agricul ism is employed in the form cf tt foam. tural or iorrsl sources, they are safe and non-toxic. Ac 30 11. A process for the mieiohiul degradation of poly
cordingly, the entire procedure is safe and non-toxic chlorinated biphenyls which comprises treating swid
since the microorganisms themselves arc not toxic to polychlorinated biphenyl:; with a: least one microor
humans, animals or fish. Hie carbon source can be ganism selected from the y.roup condiiing of Clado-po-
l*CBs alo:*.c, or u mixture of I'CD* and other hydrocar bons at ccr.ccMtratiojis of trace tin to about 15 percent
35
rium cladoapoiioidrs A'KiC 20251, Candida lipolytic.* ATCC 20?55, Noeardia >;loScrid:s ATCC 21505, No-
by weight.
cardia rubra ATCC 2I50.C and Succhaioniycex ccrcvi-
It can thus be seen that the present invention pro siau A'ICC 20252 for a sufficient time until the poly
vides a desirable and advantageous process for degrad chlorinated biphenyls have been subi.tv.ntiaiiy de
ing iiiul cleaning tip polychlorinated biphenyls, os well us petroleum n; petrochemical pollutants and conlanii-
40
graded. 12. The
process
of claim
11,
wherein
said
microor
tuials, by means of microbial degradation, so as to re ganism is mixed v.ith a c.lhdo.'ic material.
store the polluted innterial to an eeolojjicnlly-cluan
13. The process of chum II, wherein said microor
condition. This procedure i* carrird out safely ami rela ganism is mixed with n cdlulusic material, a nitrogen
tively economically without any harm whatsoever to source and a phosphorus source.
human, animal or marine life.
' 45 14. The process of claim II, wherein the pnly-
It is to be understood that the present invention em braces. the use. pot c.nly of the above-dascrihed micro organism*, which tire given merely for illustrative pur
chlorinatcJ biphenyls nrc contained in t*n r .iacous soluliotr and said microorysrdsm is added thereto in slurry form in an nmoun: of about I percent to 5 per
poses, but it al ;o includes the use of mutants jv educed from the specifically enumerated niiciosrrcamrmis, pro
50
cent by volume. 1*5. The process of c t.ii.u
|d, wherein ;.b*ut 2 percent
viding that they perform the same function. It is to be further understood that the invention includes the use
by volume of the raien'or';.ir.ism slurry is added to the solution of palyclihoin.-V'J biphenyl.*.
of suhcidtim-s obtained by various standard microbio
16. The process of claim Id, wherein the concentra
HONS 0 2 8 9 2 8
logical t.;r1ii;i'|uci. Such mutants and/or subcultures may differ in certain respects from the abovc-i)e<cr;bcd
55
tion of polychlorinated biphenyl; in nut! aqueous s,*Intion is about 10 p.p.m. to about 15 percent by wei/ht.
new strains, but the) will work to detrude I'Clls in op-
17. The process of claim 12, wherein s:*.Ul tcl.ulnvv:
proximate);.' the s;:mc manner ax disclosed above.
ma'erial is tgledcd from the group consisting of s.iw-
The Invention being thus J. scribed, it will U: obvious dust, pine bark, wood flour, cotton Haters, cottonseed
that the wunc may be varied in many ways. iluch variafrom are nut to be regarded us a departure from the
<.0
hull/, rtraxv. bagasse, marsh hay end shredded paper. 1 The process of claim 13, v.hrrmn stud nitrogen
Spirit tui J n ope of the invention. and all such modifica iourec is cottonseed pro'ein.
tions as v.r'.iid he ohtmus to one skilled ht the art are
19. The process of claim II, wherein said microOt.
IntcnJe.l |r be inciuded herein.
gatiism is employetl in a lurry form.
I claim: f. A prvcvu for the microbial degrmhit;on of poly-
65
20. The proves* of claitn 11, \vh<:u-ia said mieioorgansm i; employed hi a pelletiv.ed form.
chlorinate.1 biplutiy'v which comprises treating said
21. 'I he procctx of claim 11, whert-ia said microor*
pulychhvi'.Med biphenyls with at lea d tr:a microor ganism is employed u; a powdued from.
3,7?<\66 9 10
*22. Vt: puvos o}' claim U* sshctcm still jnicrcor- ptmism is Nucailin filuhcri'lu A'f C'C ? 1505.
jinni'iit is i n:;4 u.l i Oic form of a foam.
.
27. rh*.* procc>-< of claim 11, wherein laid ruiciorw*
2.'. I In- 5>inc:.s of
It, v herein viid mierour*
CaaiMH is NVicault:, iaOr. A I VC 2l5U$.
fc;i\M is ;n!*Vil to Oic pi*!ycMoiivt*-.nl Hplv.nyls .it n
tvii.jKiatu.- of ajyro.simnM) my..
lo nj-j-ryscmtelyy
5
2S\ Ihe protvxs of claim 11. wheiem said mkroorf.ftni-m is Siu'charomyccs cerevi.iac A I CC 2025"'.
24.
of claim'd, whvicm sin) microor*
29, The piece-'-, of claim 11. wherein ike poly,
ptiiMii i> ("I;- .'o'.notium ct.n!o*}'rM.'kles ATCC 20251. chlorinated biphenyl. it.c contained in cm mpicotis so
25. 'I lc |'i oivss of claim 11, v. iicrcin sa'i! n;a;rootum- lution which .nWiitorully contains hydrocaibon oils or
hm is Cai.Jk'a lipoKticw A'lCC 2)2S:.
to Other )>ClfOC1crMic.d,v
`
2ft. Ike puKCsv of claim 11, wherein uiid tmcroor*
15
20
2$
70
75
40
45
50
55 \
60
HONS 028929 65