Bio-Extraction of Precious Metals

from Urban Solid Waste

Subhabrata Das, Gayathri Natarajan, Yen-Peng Ting

Department of Chemical & Biomolecular Engineering,

National University of Singapore

INTERNATIONAL PROCESS METALLURGY CONFERENCE ‘16 NOVEMBER 10-11, BANDUNG

 Overview

 Introduction

 Biomining and brief history

 Biomining techniques

 From biomining to bioleaching of solid waste

 Bioleaching microbes and mechanisms

 Case Study: Electronic scrap material

 Conclusion

2
 Biomining
Biomining
Biomining:
“Commercial application of microorganisms
for metal extraction from iron or sulfidic ores”. Bio-oxidation
Bioleaching
Can it be applied to urban solid waste? (Beneficiation)

Bioleaching:
Use of microorganisms to transform solid
compounds, resulting in soluble
and extractable elements which could be
recovered. (E.g. Cu, Zn, U)

Biooxidation: (bio-beneficiation)
Microbial oxidation of mineral, with the metal
values remaining (enriched) in the solid Biox reactors for biooxidation of refractory gold at Fairview gold
residues. (E.g. Au in arsenopyrite) mine.jpg
http://wiki.biomine.skelleftea.se/wiki/index.php/Image:Biox_reactors_for_bi
ooxidation_of_refractory_gold_at_Fairview_gold_mine.jpg

2 FeAsS[Au] + 7 O2 + 2 H2O + H2SO4 → Fe(SO4)3 + 2 H3AsO4 + [Au]

3
 Biomining

History of Biomining:

• Microbial metal solubilization from acidic mine waters initially

thought to be a chemical reaction mediated process
• Commercial application started around mid 1940’s
• Biomining of secondary ores of copper, gold, cobalt, nickel,
zinc and uranium

• Copper and gold biomining are the most successful

• Copper bioleaching; coper recovered by heap and dump
leaching
• Gold biooxidation; via stirred tank technology

4
 Biomining techniques

Dilute H2SO4

Commercial bioleaching techniques Laboratory scale techniques

• In-situ leaching Percolator leaching
• Dump leaching Column leaching
• Heap bioleaching Submerged leaching
• Stirred tank leaching
Schippers, Axel, et al. , Geobiotechnology. Springer Berlin Heidelberg, 2013. 1-47.
5
 Worldwide industrial heap bioleaching operations for
secondary copper ores and mixed oxide/sulfide ores

Gahan, CS, et al., .Research Journal of Recent Sciences __ ISSN 2277 (2012): 2502. 6
 Industrial aerated continuous stirred tank bioleach
operations for pretreatment of gold concentrates

Gahan, CS, et al., Research Journal of Recent Sciences __ ISSN 2277 (2012): 2502. 7
 Bioleaching Microorganisms

Bioleaching microorganisms

Chemolithoautotrophic Heterotrophic Heterotrophic

bacteria bacteria fungi

Thermophillic
microorganims: Bacillus, Aspergillus
Mesophillic
microorganisms: Sulfobacillus Pseudomonas Penicillium
Acidithiobacillus Chromobacterium
Acidianus

Metallosphaera

8
 Bioleaching Mechanisms
Direct (contact) mechanism
• Physical contact between microbes and solid surface
• Microbes form an extra-cellular polymeric substance (EPS) layer to which
the metal sulfide surface is attached
• Mineral dissolution (due to electrochemical processes) occurs at the
interface between the bacterial cell wall and the metal sulfide surface
• Oxidation of Fe2+ and S for energy.
• Attached microbes produce the leaching chemicals Fe3+ and H+.

Copper bioleaching

https://microbewiki.

9
 Bioleaching Mechanisms
Indirect (non contact) mechanism
• Carried out by cells in solution surrounding the metal sulfide, which
oxidize Fe2+ ions or elemental sulfur in solution.
• Resulting Fe3+ ions or H+ ions come into contact with the mineral surface,
where they are reduced during oxidation of the ore, and enter the cycle
again.
• Indirect leaching by heterotrophs involves acidolysis, complexolysis,
redoxolysis, alkalolysis, or combination of these mechanisms

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 Bio - urban mining of solid waste

Concept applied to urban solid wastes with metal values: (a) (b)

Spent catalysts (Co, Mo, Ni, Al);

MSW Incineration fly ash (Zn, Al, Cu);
Electronic Scraps (E-wastes: Cu, Ni, Au, Ag, Pt, Pd), etc

Current management of solid wastes

(i) Resource recovery & recycling (Hydrometallurgy / Pyrometallurgy)
(i) Regeneration of spent catalysts; MSW Incineration
Spent Batteries Filter Dust
(iii) Ultimate disposal Fly Ash

Advantages of biomining: Bio-urban Mining of

Environmentally friendly – no secondary Solid Wastes
pollutants
Loss cost and energy requirements Electronic Wastes
Spent Catalytic
Spent Catalysts Converters
Detoxification of hazardous wastes
Reclamation of metal values (resource recovery)
Slow; Metal toxicity!
11
Y.P.Ting
 Bioleaching microorganisms
Type Organism pH range Leaching agent Temperature (oC)

Acidithiobacillus thiooxidans 2.0-3.5 Sulfuric acid 10-37 (mesophile)

Acidithiobacillus 1.4-1.6 Ferric ion, Sulfuric acid 28-35 (mesophile)
ferrooxidans
Chemolitho- Leptospirillum ferrooxidans 2.5-3.0 Ferric ion 30 (mesophile)
trophic
Sulfobacillus extremely Ferric ion, Sulfuric acid 50 (moderate
autotrophic
thermosulfidooxidans acidophilic thermophile)
bacteria
Metallosphaera sedula acidophillic Ferric ion, Sulfuric acid extreme thermophile
Chemolitho- Acidianus brierleyi 1.5-3 Sulfuric acid 70 (extreme
trophic thermophile)
archaebacteria
Bacillus megaterium - Citrate, Amino acids mesophile

Pseudomonas putida Citrate , Gluconate mesophile

-
Heterotrophic Chromobacterium Cyanide ion 30
bacteria violaceum - mesophile
Pseudomonas fluorescens
Aspergillus Citrate, Gluconate, 30 (mesophile)
Fungi - Oxalate, Lactate
Penicillium simplicissium Citrate, Oxalate 22-30 (mesophile)
- Gluconate,

Alkalinophiles from MSW landfill: pH 10-12. Tolerates up to 20% fly ash pulp density.
Bosecker, Klaus. FEMS Microbiology reviews 20.3-4 (1997): 591-604.
 Bioleaching of solid waste
Waste Metals Organisms
Leached
Catalysts Al, V, Mo, Acidithiobacillus thiooxidans, Acidithiobacillus
Ni, Co, Li ferrooxidans
Aspergillus niger, Penicillium simplicissimum
Al. Cu, Ni, Pb, Acidithiobacillus ferrooxidans, Acidithiobacillus
Electronic scrap Sn, Zn, Au thiooxidans
Aspergillus niger, Penicillium simplicissimum
Chromobacterium violaceum, Sulfobacillus
thermosulfidooxidans
Municipal solid Al, Fe. Mn, Ni, Acidithiobacillus thiooxidans, Acidithiobacillus
waste Fly ash Cd, Cr, Cu, Ni, ferrooxidans Pseudomonas putida, Bacillus
Pb, Zn, megaterium,
Aspergillus niger, Acidianus brierleyi
Spent battery Li, Co Acidithiobacillus species
waste
Belt filter press Cu Acidithiobacillus ferrooxidans
solids
Sewage sludge Cu, Mn Zn, Ni, Acidithiobacillus thiooxidans
Cd, Cr, Pb
Tannery sludge Cr Acidithiobacillus thiooxidans
Jewelry waste Ag, Au Chromobacterium violaceum,
Pseudomonas fluorescens
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 Factors affecting Bioleaching

1. Physicochemical parameters of bioleaching environment

Temp, pH, redox potential, oxygen, carbon dioxide, nutrient
availability, …

2. Microbiological parameters
Microbial diversity, population, metal tolerance, adaptation, …

3. Properties of solids to be leached

Composition, particle size, surface area, porosity, hydrophobicity, …

4. Processing
Leaching mode (in-situ, heap, tank leaching), pulp density, stirring
rate, ..

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Y.P.Ting
Case study: Electronic waste
 What is E-waste?

E-waste: Waste containing mainly electrical and electronic components.

Includes precious metals (e.g. Au, Ag, Pt) & toxic heavy metals (Pb, Cd, Cu, Hg).

In US: E-waste constitutes 2% - 5% of the municipal solid waste stream and is

growing rapidly. In Europe: volume of E-waste is rising by 3% to 5% per year - almost
three times faster than the municipal waste stream.
In Singapore, approx. 6000 ton of E-waste generated annually.

Hazardous Waste found in a typical PC

 Lead - cathode ray tube and solder
 Arsenic - older cathode ray tubes
 Antimony trioxide -flame retardant
 Polybrominated flame retardants - plastic casings,
cables and circuit boards
 Selenium - circuit boards as power supply rectifier
 Cadmium - circuit boards and semiconductors
 Chromium - steel as corrosion protection
 Cobalt - steel for structure and magnetivity
 Mercury - switches and housing
Greenpeace International. The e-waste problem, http://www.greenpeace.org/ international/en/campaigns/toxics/electronics/the-e-waste-problem/ (2005).
Let's Cut Down on E-waste, https://www.nccs.gov.sg/climatechallenge/issue10/Lets-Cut-Down-on-E-Waste.html
ETBC (2012). Electronics Take Back Coalition- Facts and Figures on E-Waste and Recycling
 Why is E-waste recycling important?

 Reduce environmental impact

 Recover precious metals

A ton of used mobile phones contains about
3 kg Ag, 300 – 500 g Au, 140 g Pd, 130 kg of Cu 1
Au in ESM (10 to 1000 g gold/ ton) cf natural gold ores
(0.5g to 13 g gold/ ton) 2

 Need for an alternative resource for metals to delay the depletion

of (non-renewable) natural ores.

“Rock crunching” microbes (Acidthiobacillus and Leptospirillum

Spp) to “computer munching” microbes (Chromobacterium
violaceum)
1Facts and Figures on E‐Waste and Recycling-Electronics Take Back Coalition
2Korte, F., M. Spiteller, and F. Coulston, Ecotoxicology and Environmental Safety, 2000. 46(3): p. 241-245
 E-waste recycling

Gold Cyanidation

Cyanide-producing Chromobacterium
micro-organisms (including
violaceum, Pseudomonas fluorescens, Bacillus megatarium)

ELSNER EQUATION

Unlike chemical cyanidation of gold, detoxicification of excess

cyanide by β-cyanoalanine 1,2.

1Rodgers, P.B. and C.J. Knowles, J General Microbiology, 1978. 108(2): p. 261.
2Brysk, M.M., Corpe, W., and Hankes, L. (1969). Journal of bacteriology 97, 322-327.
 Case Study: Electronic Scrap Material

• Bioleaching of precious metal Au

and base metal Cu Bioleaching of ESM
• Heterotrophic bacteria
Chromobacterium violaceum
• Bacteria cyanide complex gold (Au)
particles into soluble gold cyanide Gold dissolution by
(AuCN) cyanogenic bacteria
• Effect of presence of Cu on Au
leaching: pre-treatment for Cu
extraction before AuCN leaching.
• Enhanced metabolite expression Biomineralization with
through gene insertion Delftia acidovorans
• Biomineralization

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 Case Study: Electronic Scrap Material

Strategies to enhance gold

dissolution rates

Pretreatment of Mutation to adapt to Metabolic Spent medium leaching

E-waste high pH environment engineering at adjusted pH

Au
Ag Pt
Pd
Au

Plastics Cu
Fe
Zn
Bioleaching
Ni
Sb Soluble metals
Insoluble solid metals complexes

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 Elemental composition of ESM
Metals Untreated ESM Pre-treated ESM % Removed by
Composition (mg/g) Composition(mg/g) Pretreatment
Cu 150.40 ± 4.0 30.40 ± 2.1 80
Al 47.20 ± 2.8 14.40 ± 1.7 69.5
Fe 31.40 ± 2.0 9.00 ± 0.9 71.3
Pb 28.00 ± 3.2 9.40 ± 1.5 66.4
Sn 17.60 ± 2.2 2.00 ± 0.3 88.6
Ni 16.00 ± 1.8 6.00 ± 0.8 62.5
Zn 11.60 ± 1.2 5.20 ± 0.6 55.2
Ag 0.56 ± 0.09 0.20 ± 0.03 64.3
Au 0.28 ± 0.03 0.28 ± 0.04 -
Total 303 76.8

 30% (by mass) – Metals; 70% - ceramics, refractory oxides and plastics
 Copper constitutes the bulk of untreated ESM
 Cu, Ni, Fe, Zn form stable complexes with cyanide
 Pretreatment removes 80% of Cu and other metals to enhance gold cyanidation

Natarajan, G, and Ting YP.,. Bioresource Technology 152 (2014): 80-85.

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 Approach 1: Pretreatment of E-waste

Challenge
 High concentration of copper in E-waste (500 x gold)

 Stable cyanide complex with copper

Strategy

Nitric acid pretreatment of ESM for maximum removal of copper

Chromobacterium sp.

E-waste Copper
Removal Gold recovery
Bioleaching

Nitric acid

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 Bioleaching of untreated and pretreated ESM

 Pretreated ESM yielded higher Au bio-recovery compared to untreated ESM

 Metal leaching yield decreases as pulp density increases
 Higher metal extractions of base metals Cu compared to Au

Natarajan, G and Ting YP. Bioresource Technology 152 (2014): 80-85.

23
 Approach 2: Mutation of C. violaceum

Challenge 𝐻𝐶𝑁 ↔ 𝐻+ + 𝐶𝑁−

 Hydrogen cyanide is volatile at physiological pH of C. violaceum (pH 7-8)
 pKa of HCN is 9.3; availability of cyanide lixiviant is favored at high pH
Strategy
 Mutation of bacteria by mutagen, N-Nitroso-N-ethyl urea (ENU)

 Select mutants growing at alkaline condition

Mutated Chromobacterium sp.

E-waste Copper
Removal Gold recovery
Two-step
Bioleaching
Nitric acid pretreatment

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 Bioleaching using mutated C. violaceum

25

Gold recovery (%)

20

15

10

Wild and mutated strains at different pH

 Au bioleached from pretreated ESM at 0.5% pulp density using wild and mutated
strains
 C. violaceum mutated at pH 9.5 gave the highest gold recovery (22.5%)
 Wild strain (unadapted) grown at pH 9, 9.5 and 10 achieved Au recovery of 14%,
16% and 0%. (Poor growth and CN production by wild strain at higher pH.)

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 Approach 3: Metabolic engineering of
C. violaceum
Challenge
 Low yield of cyanide lixiviant (~ 20 mg/l)
 Tight regulation of cyanide producing operon under quorum control
Strategy
 Construction of two metabolically-engineered strains of C. violaceum
 Insert an additional set of cyanide-producing operon in wild strain to enhance
cyanide yield Intergenic region

glmS CV0678 Wild type C. violaceum

pBAD

glmS hcnABC CV0678 pBAD C. violaceum strain

pTAC

glmS hcnABC CV0678 pTAC C. violaceum strain

Tay, SB, et al. Scientific Reports , 3 (2013).

26
 Bioleaching using engineered C. violaceum

 Engineered strains showed significantly higher Au and Cu recovery

 At 0.5% w/v pulp density: wild, pTAC and pBAD strains recovered 11.3%, 24.6%
and 29.6% respectively by two-step bioleaching process
 Cyanide production of 34.5 mg/l (pBAD) and 31 mg/l (pTAC) achieved by
engineered strains (compared with 20 mg/l by wild strain)

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 Approach 4: Spent medium leaching

Challenge
 Cyanide concentration in the solution is highly pH dependent
 pKa of HCN is 9.3; availability of cyanide lixiviant is favored at high pH
Strategy
 Spent media\um leaching - Decouple bacterial growth/cyanide production from
Au complexation

Operate bioleaching at high pH

Chromobacterium sp.

E-waste Copper
Removal Gold recovery
Spent media
leaching
Nitric acid pretreatment
Natarajan, G, and Ting YP. Chemosphere 136 (2015): 232-238.
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 Bioleaching using engineered C. violaceum

 Highest Au recovery (30%) and Cu recovery of (95.7%) was obtained with spent
medium leaching at pH 10 at 0.5% w/v pulp density
 Metal leaching yield decreases as pulp density increases

Tay, SB, et al., Scientific reports 3 (2013).

29
 Gold recovery at 0.5% w/v pulp density with
various strategies
39.6
36.5

30 30

22.5

18

11.25

7.14

Original ESM Pretreated Mutation Spent medium Spent medium Genetic Bioreactor - 2 Bioreactor -
ESM approach leaching leaching Engineering step (optimized Spent medium
(physiological (pH=10) approach conditions)
pH) (pBAD)

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 Gold Biomineralization

Conventional Methods
 zinc cementation  carbon-in-pulp
 Electrowinning  ion exchange
Disadvantage
 high operation cost
 low selectivity under multi-ion condition

Strategy
 Bacteria-mediated mineralization of Au ions from leachate

Gold
Gold precipitate
Bioleachate

Delftia acidovorans

Das, S, Dong,B, and Ting YP.," Advanced Materials Research. Vol. 1130. 2015.

31
 Gold Biomineralization

Biomineralization studies: Delftia acidovorans

 D. acidovorans secretes a secondary metabolite delftibactin; toxic effect of gold.

 Precipitates Au3+ as gold nanoparticles in extracellular medium
 Maximum Au recovery observed after 7 h of inoculation at pH 8

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 Gold Biomineralization
TEM/EDX Analysis
70

60

50

Weight (%)
40

30

20

10

0
C O N Au

 GNPs synthesized by D. acidovorans were mostly spherical in shape and

crystalline in nature
 Average size of 31±2 nm

33
 Conclusion
• Biomining of metals from ores can be applied to urban waste (e.g fly
ash, electronic scrap)
• Urban waste can serve as secondary raw materials
• Intrinsic capability of microorganisms can be exploited for
(i) detoxification of hazardous waste, and (ii) reclamation of metal
values. Delays depletion of non-renewable resources!
• Indigenous microorganisms can be isolated (in abandoned mines) and
adapted to toxic environment to improve metal extraction yield
• Optimization of physicochemical parameters and genetic manipulation
of existing strains can enhance biomining for metal recovery

• Slow process. Metal toxicity; hence use of spent medium. Need for
stringent government regulations and research policies that favor
“green technologies” for process development.

34
 Acknowledgement:

Ministry of Education (Singapore)

Cimelia Resource Recovery Pte Ltd
National Environmental Agency, Singapore

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 Biomining - Mining of the Future?

Terima Kasih