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Indian Journal of Experimental Biology

Vol. 41, September 2003, pp. 967-971


Biorecovery of gold
Ronald Eisler*
U.S. Geological Survey , Patuxent Wildlife Research Center, Laurel, Maryland 207084019, USA

Recovery of ionic and metallic gold (Au) from a wide variety of solutions by selected species of bacteria, yeasts. fungi.
algae. and higher plants is documented. Gold accumulations were up to 7.0 g/kg dry weight (OW) in various species of
bacteria, 25.0 g/kg OW in freshwater algae, 84.0 g/kg OW in peat. and 100.0 g/kg OW in dried fungus mixed with kerati-
nOlls material. Mechanisms of accumulation include oxidation, dissolution, reduction. leaching, and sorption. Uptake
patterns are significantly modified by the physicochemical milieu. Crab exoskeletons accumulate up to 4.9 g Au/kg OW;
however, gold accumulations in various tissues of living teleosts, decapod crustaceans, and bivalve molluscs are negligible.

Keywords : Biorecovery of gold, Gold uptake

Extraction of gold from solutions is under active rapidly-growing, arsenic-resistant bacterial strains of
investigation using a variety of physical, chemical, Thiobacillus ferrooxidans (Beijerinck), LeptospiriLlium
and biological processes. Recovery of ionic gold from ferrooxidans (Beijerinck), and Thiobacillus ferrooxidans
dilute solutions usually involves either precipitation (Beijerinck). These bacterial species obtain their
by zinc dust, carbon adsorption, solvent extraction, or energy through the oxidation of ferrous to ferric iron
ion exchange resins. All of these are of low selectivity (T. ferrooxidans, L. ferrooxidans) or through the
and comparatively expensive I.2. Chemical methods reduction of inorganic sulfur compounds to sulfate
for the recovery of gold from ores include cyanidation (Thiobacillus spp.). Monetary costs of biooxidation
and thiourea leaching, which present environmental are reported to be about 50% lower than roasting or
and health risks3.4. Biorecovery of dissolved gold pressure oxidation 13. Adding Thiobacillus
from solution presents fewer environmental risks than ferrooxidans into the thiourea leaching solution
chemical methods, and is documented for produces a 20% increase in the extraction of gold.
. .
mIcroorganisms 25-20
' ,a Igae 1421 . , water lernsl' 22 , peat-?3 , The reaction describing gold dissolution in an acidic
alfalfa , seaweeds2., fungi 2. 18.26.29, yeasts l4 , and solution of thiourea in the presence of ferric ion IS
crab exoskeletons 3o . This account briefly reviews the described by Kai et a/.f:> as :
potential of living and dead plants and animals to Au + Fe3+ + 2CS(NH 2h ~ Au[CS(NH 2hf+ + Fe2+
accumulate gold from solution, and some of the
The use of bacteria in pretreatment processes to
processes involved - including biooxidation,
degrade recalcitrant gold-bearing arsenopyrite ores
dissolution, bioreduction, bacterial leaching, and
and concentrates is well established7 19. Recalcitrant
ores are those in which the gold is enclosed in a
Gold uptake by microorganisms, fungi, and higher matrix of pyrite and arsenopyrite, and cannot be
plants solubilized by direct cyanidation. Bacterial
Biomining processes are llsed successfully on a decomposition of arsenopyrite assists in opening the
commercial scale for the recovery of gold and other molecular mineral structure, permitting access of the
metals, and are based on the activity of obligate gold to cyanide. However, greater quantities of
chemoautolithotropHic bacteria that use iron or sulfur cyanide are required to solubilize gold after bacterial
as their energy source and grow in highly acidic treatment when ores contain high quantities of gold. A
media 13. Biooxidation of difficult to treat gold-bearing possible cause of this excessive cyanide is the
arsenopyrite ores occurs in aerated, stirred tanks and presence of the enzyme rhodanese, produced by
Thiobacillus caldus (Beijerinck), a common species
*For correspondence:
of bacterium encountered in biooxidation facilities l 9 .
Fax : 301-497-5744 Optimum microbiological leaching by Thiobacillus
Phone : 301497-5724 spp. and Sulfolobus spp. of refractory sulfide ores for

recovery of go ld in tanks is poss ible under controlled chl oride soluti ons doc umented fo r at least seven
conditions of pH, di ssolved oxygen, carbon di ox ide, genera of fres hwater cyanobacteria3 1. So me
sul fur balance, redox potenti al, tox ic metal bacteriform go ld is bi ogeni c - the res ult of
concentrati ons, and rate of leachin g7. precIpitati on by bac teri a - and may be usefu l
Several species of Fe3+ -reducing bacteri a (Bacteria indicators of gold deposits and of processes of go ld
spp., Archaea spp.) can precipitate go ld by reducing acc umul ati on. PLectonema terebrans Born et, a spec ies
Au' + to Au' with hydrogen as the electron donor"o. of fil amentous marine cyanobacteria, accumul ates
Rate of bacteri al ox idati on by ThiobacilLus go ld in its sheath fro m an aqu eo us solu tion of AuCl 3 .
fe rrooxidans and Leptosp irillium fe rrooxidans of Sheaths are among th e few stru ctures likely to be
three South Afri can refractory go ld ores of varying preserv ed in so me form in mi crofossi ls of ancient
go ld-arsenopyrite co mposition was dependent mainl y bacteri a. In marine medi a, it is ex pec ted th at AuCI3
on crystal structure R These go ld ores were c1 ass ifi ed (2.0 g Au/l ) will fo rm AuCI ~ , Au' ; , and AuCi ;31.
as refractory due to th e presence of go ld inclusions in
Biosorption of Au3+, as AuCi ~ , by dri ed Pseudomonas
arsenopyrite and pyrite, and submi croscopi c gold
ma inl y in arsenopyrite. Refractory go ld occ urs at sites strains of bac teri a was inhibited by palladium, as Pd 2+,
whi ch are preferenti all y leached by th e bacteri a. The and possibl y other metal ions 10 .
rate of go ld liberati on fro m sul fides is enhan ced Go ld adsorpti on fro m cyanide solutions by dead
durin g th e earl y stages of bac teri al ox idati on. Defects bi omass of bacteri a (Bacillus subtilis), fun gus
in crys tal structure influence the rate of bioox idati on (Penicillium chrysogenum Thom), or seaweed
and is directl y related to th e crystal stru cture of th e (Sa rgassum flu itans Linnae us) at p H 2.0 were 1.8 g
sulfid e mineral, the crystall og rapHi c ori entati on of th e Au/kg OW fo r bacteri a, 1.4 g/kg OW for fun gus, and
exposed surfaces, and diffe rences In chemi cal 0.6 g Au/kg OW fo r seaweed. Ani oni c AuC ;
composIti on and mechani ca l dev iat ions in the adso rpti on was th e major mechani sm in go ld bioso r-
crys tals x. Pretreatment of refractory go ld concentrates ption fro m cya nide so luti ons, being mos t efficient at
with the bacterium Thiobacilllls fe rrooxidans lower p H va lues 2 . L-cysteine increased go ld-cya nide
ul timately results in sulpHur and sulpHide ox idati on bi oso rpti on of Bacillus, PenicilliulIl, and Sa rgassum l R
by fe rri c ion s from bacteri al ox idati on of ferrou s ions. At pH 2, the max imum go ld uptakes were 4.0 g Au/kg
The max imum co ncentrati on of attac hed Thiobacillus OW fo r bac teri a, 2.8 g/kg fo r fungus, and 0.9 g/kg fo r
increases with increasing co nce ntrati on of Fe 2+ and seaweed, or 150-250% greater than in the absence of
decreases with increasing size of the refrac tory go ld cysteine. The ani on ic gold cya nide species were
concentrate particles '6 . In Chile, whi ch produced adsorbed by ioni zable fun cti onal groups on cys teine-
30,000 kg of go ld in 1990, Thiobacillus ferrooxidans loaded biomass; depos ited gold coul d be eluted fro m
was used to recover go ld from a co mple x ore under go ld-loaded biomass at p H 5.0 18
laboratory conditi ons9 . The ore contained 8.2% Fe, Go ld-resistant strains of bac teri a th at also
0.78% Cu, 0.88 % As, and 3. 5 g Au/ton, with pyrite, acc umul ate go ld are doc umented, although the
hematite, arsenopyrite, and chalcopyrite as the mai n fundamenta l mechani sm of resistance to go ld in
metal- bearin g minerals. Initi al go ld recovery by mi croorgani sms is not known or und erstood 15. One
co nve nti onal cya nid ati on on a cru shed ore sampl e was strain of Burkholde ria (Pseudomonas) cepacia
54%; co ncentrati on by fl otati on improved reco very to Burkholder co ntained millimolar concentrati ons of
56%. Conce ntrated sampl es (17 .0 g Au/ton) were Au+ thi olates. Burkholde ria cells were large,
leached in reactors at p H 1.8. In th e presence of accumul ated polyhyroxybutyrate and gold, and
bacteri a, all di ssolved iron was present as ferri c ion; excreted thi orin , a low molec ul ar weight protein into
gold recovery by cyanidati on increased fro m 13% fo r the cultu re medium . Thi s effect was not observed with
th e initi al concentrate to 97 % after 10 days of th e Au3+ co mpl exes tes ted, whi ch were reduced to
bacteri al leachin g. To furth er increase go ld recovery, metalli c go ld in the medium. Gold-res istant strains of
fl otati on tailings were submitted to cyanid ati on9 . fun gi and heterotrophic bacteri a are also kn ow n ' 5 .
Some mi croorga nisms isolated from go ld-bea rin g Rapid recovery of go ld from go ld-thi ourea
depos its are capabl,e of di ssolving go ld ; di ssolution solutions was doc umented fo r waste biomass of yeasts
was aided by the prese nce of asparti c acid, histidin e, (Saccha romyces cerevisiae), cyanobacteri a (Sp iruLina
serine, alanine, glyc ine, and metal ox idants5 . Bacleri f- platensis [Nordst]), and bac teri a (Streptomyces
a nn go ld is well know n, with uptake of Au3+ from erythralus [Waks manl)' 4 The process is p H-

dependent for yeast and bacteri a, and pH-indepe nde nt Uptake of Au 3+ by Chlorella vulga ris Be ij erinc k, a
fo r Spirulina. O f all stra in s of mi croo rgani sms uni cellul ar green alga, from soluti o ns conta ining 10.0
examin ed, Spirulina platensis has the hi ghest affinit y o r 20.0 mg A u 3+/l is documented . Chlo rella
and capac ity for gold , eve n at lo w pH values. G o ld accumul ated up to 16.5 g Au/kg OW . In acti vating the
uptake by Spirulina was 7 .0 g Au/kg bi o mass OW in a lgal cell s by variou s treatments resulted in so me
1-2 hr at p H 2. 0, and abo ut 3.0 g A u/~ g OW in 15 min enh ancement in uptake capac ity o ver the pri stin e
14 cell s. In acti vati o n by heat treatment yie lded up to 18.8
atpH 2 throu gh 7 .
Metabolicall y acti ve fun gal cell s of Aspergillus g/kg DW ; fo r alk ali treatment, thi s was 20. 2 g/kg
f umigatus Fresen and A. niger Ti egh re mo ved gold OW ; fo r form aldehyde treatment, 25.5 g/k g OW ; and
from cyanide leach liquor of a Brazili an gold extracti on fo r acid treatment, 25 .4 g/kg OW . E le me nta l go ld
pl ant more effi cientl y than did dri ed fun gal bio mass or (A u") was measured by X-ray pho toe lectro n
other species of Aspergillus tested . These two species spectroscopy o n the cell s urface, indi catin g th at a
of fung i removed 35 to 37% of go ld from so lutio ns reducti o n had occurred . Studies with li vin g Chlor-
containing 2.8 mg A u/I in 84 hr26. Gold re moval fro m ella vulga ris suggest th at accumul ated Au 3+ is rap id ly
cyanide-containing solutio ns is documented for a strain reduced to A u+, fo ll o wed by a slo w reducti o n to A U .
of Aspergillus niger, a fun g us iso lated from the gold With dead a lgae, A u ini tiates a seedin g process whic h
26 28
extracti on pl ant at Nova Linda, BraziI - . The leach res ults in the formati o n of e leme ntal gold .
liquor contained, in mg/I , 18 1.0 cyanide, 1.3 go ld , 0 .4 Sequeste rin g meta l io ns usin g li ving o r dead pl ants
sil ver, 7. 1 copper, 5.2 iron, and 4 .5 zinc. After 60-72 hr is a pro posed eco no mi cal means o f re mov ing go ld
of incubati o n, A. niger removed from so luti o n, and oth er meta ls via intracellul ar acc umul ati o n o r
probabl y by adsorpti o n, 64% o f the go ld, I 00% of the s urface adsorpti o n. H owever, in the case of li ve
si lver, 59% of the copper, 80% of the iron, and 74% of pl ants, thi s is freq ue ntl y a re lati vely slow and time-
the zinc; all gold was re moved after 120 hr. Use of thi s consumin g process. No nli vin g pl ant materi a l fo r
fungus to develo p a bi oprocess to reduce metal and s urface adsorpti o n offers several ad va ntages over live
cyanide levels as well as recovery of va lu abl e metals pla nts, in cl udin g redu ced cost, greate r avail abil ity ,
shows promj se 26-28. Uptake pattern s of gold from Au +
eas ie r regenerati o n, and hi g he r me ta l spec ific ity4. In
soluti ons by dead fungal bi o mass fo llowed South Afri can minin g e fflu e nts, go ld us uall y ranges
mathemati cal uptake mode ls of Lang muir and Freu- betwee n I and 10 mg/I. In studi es of 180-m in
ndlich; bi o mass was prepared from the fruiting body of durati o n, dri ed red water fe rn s, Azalla .filiculoides
a mu shroo m co ll ected fro m the forests of Kerala, La marc k, re moved 86 to 100% of A u 3+ fro m ini tial
29 3
Indi a . Dri ed fungus, Cladosporium cladosporoides so luti o n of 2 to 10 mg Au +/l , in creas ing with
Fresen, mi xed w ith keratino us materi a l of natural increased initi a l concentrati o n o f A u 3+ (ref. 22). The
origin to form a bead, proved effecti ve in absorbing bio mass gave >95 % re moval e ffic iency at all bi o mass
gold fro m so lutio n . The bi osorpent beads adsorbed concentrati o ns measured . Optimum (99.9 %) removal
100.0 g Au/kg beads fro m a soluti o n containing 100 .0 of gold occUlTed within 20 min at pH 2, 42% remova l
mg Au/ I. Max imum bi osorpti o n of 80% occ urred at at pH 3 and 4, 63 % at p H 5, and 73% re moval at p H
acid pH ( 1-5) in less than 20 min . The bi osorpe nt beads 6; remova l effic iency see med indepe ndent of
degraded in so il in about 140 days . The beads also te mpe rature 22 . Simil ar res ults were o bserved w ith fo ur
removed 55 % of the go ld from e lectropl ating so luti o ns spec ies of gro und dri ed seaweeds (Sargassum sp.,
containing 46.0 mg Au/I , w ith observed go ld loading Gracila ria sp ., Eisenia sp ., and Ulva Sp. )25 Treated
capacity of 36.0 g/kg beads 12. Dri ed bi osorpents seawecds re moved 75-90% of the go ld within 60 min
encapsul ated in po lysul fo ne were prepared fro m at pH 2 fro m so luti o ns co ntaining 5.0 mg A u +/l . Go ld
microorgani sms iso lated fro m pri stine o r ac id mine (A u +) ca n be sequestered f ro m ac id so luti o ns by dead
drainage enviro nme nts " . Biosorpent materi a l ri ch in bi o mass of a brown alga, Sargassum na(ans
exopolysaccharides fro m the ac id mine drainage site (Linn aeus), and depos ited in its e leme ntal fo rm , A U 24 .
bo und Au 3+ three times mo re e ffec ti vely th an did other The ce ll wa ll of Sa rgassum was the maj o r loca le fo r
materi als, and removed 100% of the A u3 + from go ld depos iti o n, w ith carbo ny l g ro ups (C = 0 ) pl ay ing
soluti ons containing 1.0 mg Au/ I within 16 hr at 23C a maj o r ro le in bindin g, and N-conta ining groups a
andpH 3.0. lesser ro le. Lik e activ ated carbo n, th e biomass of
Algal ce ll s, ali ve o r dead , rapidl y accumul ate A u3+ Sa rgassum natans is ex tre me ly po rou s, reportedl y
and begin to reduce it to A u and A u+ w ithin 2 d ays 33 . more tha n most bi o materi als, and acco unts, in part ,

fo r its ab ility to accumulate go ld 24 . Dried ground crustaceans ranged from 0.3 to 38.0 J1. g Au/kg OW ;
shoots of alfalfa, M edicag o sa/i va Linnaeus, were for fish musc le, the mean concentrati ons were O. I
effective in removing go ld from solution 4 .The J1. g/k g OW and 2.6 J1.g/kg ash weight 35 . In studi es with
acc umulation process in vo lved th e reduction of Au 3+ th e eastern oyster, C rassos/rea virgill ica (Gmelin ),
to colloidal Au', and was most effici ent at elevated the blue crab, Callinectes sapidus Rat hbun , and th e
temperatures and ac id pH. In so lutions containing mummichog Fundulus hele roclitus (Linnaeus), an
60 mg Au )' /I, abou t 90% of the Au 3+ was bound to estu arine cyprinodon tiform fi sh, all species were
dri ed alfalfa shoots in abo ut 2 hI' at pH 2 and 55C. exposed in cages under field conditi ons to sediment-
The mechanism to acco unt for thi s phenomenon is sorbed, carrier-free, 198 Au + :14. The max imum level of
unknown , but may in vo lve red ucti on of Au 3+ to Au +, rad iogo ld in the caged organi sms was detected in
the latter being un stabl e in water to form Au' and oysters 17 hr after contact with 198A u-spiked sed iments.
Au 3+ (ref. 4). Dri ed peat from a Brazilian bog Indi ge nous orga ni sms collected 4 1 hr after contact
acc umulated up to 84.0 g Au/kg OW within 60 min with the 198 Au-labeled sed iments contai ned no detec table
from so luti ons conta ining 30.0 mg Au3+/1 (ref. 23). rad ioactivit/ . In a 25-day study with blue crab,
north ern quahog clam Mercenaria mercenaria
Gold uptake by aquatic macrofauna (Linnaeus), and the sheepshead minnow Cyprinodon
Except for crab exoskeleton s, go ld recovery from variegatus Lacepede, all species were maintained in a
the medium by various species of li ving molluscs, 1000-1 aquarium containing bentonite clay and seawater
cru staceans, and fi shes is neg li gible. spiked with carrier-free 199 Au (phys ical half-life of 3.2
Cert ain chitinous materials, such as exos keletons of days), as AuCi ); crabs accumul ated the most radi oacti vi-
the swa mp ghost crab, Ucides co rda /u s (Linn aeus), ty, followed by clams, clay, and fi sh, in that o rd er:l~ .
ca n remove and concentrate go ld from ani oni c go ld Bioconcentration factors (BCF's) for metal s and
cya nide so luti ons over a wide range of pH values:10 aq uatic orga ni sms deriv ed from carrier-free radio-
The max imum AuCN ; uptake occurred at pH 3.7, tracers in th e medium are probably artificiall y hi gh,
co rres pondin g to a fin al va lue of 4.9 g Au/k g OW ; and should be interpreted with ca uti on 35 .36. For metals
exoskeletons burnt in a non-oxid izing atmosphere it is a general observati on that high BCF's are
removed 90% of th e go ld at pH 10. phenolic gro ups associated with low co ncentrations in th e medium.
created durin g th e hea t treatment seemed to be the and th at BCF's are especiall y hi gh when they are
derived from carrier-free radioi sotopes. Typicall y,
main functional group respons ibl e for AueN ;
3o BCF's for metals and other chemicals studi ed reach a
bind ing by burnt ac id-washed crab shells . plateau before declining with increasing concent ra-
Bioconcentrati on factors (BCFs) we re reco rded for ti ons in solution35 .36 . The maximum co ncentration of
ca rrier-free 19X Au+ (ph ys ical half-life of 2.7 days) in stable go ld meas ured in ti ss ues of living marine
fres hwater orga ni sms after immersion for 2 1 days in a orga ni sms was 38.0 J1.g/k g FW35.
med ium co nt aining 25,000 pCiIl =675 ,700 Bq1l 32 . In
goldfi sh, Ca rassills a llra/us (Linnaeus), th e hi ghes t Conclusion
BCFs meas ured were < I in mu cle (i.e., less th an Gold recovery by selected species of bacteria, algae,
675.700 Bq/kg FW musc le), 10 in viscera, and 9 in fun gi, yeasts, and hi gher plants from dilute solutions
who le fish. In th e freshwater winged tloater clam, under contro lled phys icochemical conditi ons seems
Anodonw /lIm a/liana Lea, th e maximum BCF was 7 eco nomica ll y viable, with concentrat ion: up to 100 g
in soft parts; for crayfish (A slacus sp. ), BCFs were < I Au/kg OW documented . Dried crab exos keletons also
ill mu scle and 14 in viscera. For marin e organ isms show promise for commercial go ld recovery from
solut ion (4.9 g Au/kg OW); however. go ld uptake by
immersed for 26 days in sy ntheti c seawater cont ainin g
li ving species of aq uati c macrofauna see ms negligible.
33.000 pCi/1= 89 1,900 Bqi l. max imum BCFs
The mechani sms of accumul at ion wh ich include
meas ured we re tl in musc le and 16 in visce ra of th e
ox idation, reduction, dissoluti on, leac hin g and sorption
red crab . Ca llce r produ c/ll s Rand all , 11 in so ft parts
are not knovin with certainty and merit additiona l
of the buller ciJ m, Saxidol1l11s gigan /ells (Deshayes),
resea rch effo rt .
12 in soft parts of the co mmon mussel, MYlilus edlllis
Linnae us, and < I in muscle and I in a whole gobiid Acknowledgement
fish. th e longjaw mudsucker, C illiclilhys l1Iimbilis The author th ank s Drs. Peter H. Albers and Barnett
Coope1}2. Max imum stabl e go ld concentrati ons A. Rattner for th eir cons tru cti ve co mments on an
recorded in soft ti ssues of marine molluscs and ea rl y draft.

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