Trans Indian Inst Met (2019) 72(3):559–579
https://doi.org/10.1007/s12666-018-1516-4
REVIEW PAPER
Metals Extraction from Sulfide Ores with Microorganisms: The
Bioleaching Technology and Recent Developments
Wasim Sajjad1,2 • Guodong Zheng1 • Ghufranud Din3 • Xiangxian Ma1
Muhammad Rafiq3 • Wang Xu1,2
Received: 1 January 2018 / Accepted: 7 November 2018 / Published online: 2 January 2019
Ó The Indian Institute of Metals - IIM 2019
Abstract Nowadays, due to fast global industrial progress
and near diminution of high-grade ore reserves, there has
been massive call to cost-effectively process the resources
of low-grade ores and industrial effluents for metal
extraction. However, conventional approaches cannot be
used to process such resources due to high capital cost and
energy, also causing environmental pollution. Alterna-
tively, bioleaching is highly environmental friendly and
economic method to process such resources. Metal recov-
ery from metal sulfide ore is carried out by chemolitho-
trophic bacteria like Acidithiobacillus ferrooxidans and
Acidithiobacillus thiooxidans. The same is done by het-
erotrophic microorganisms in non-sulfide ores. Addition-
ally, for gold and copper extractions, bioleaching is used to
extract cobalt, zinc, nickel, and uranium from low-grade
ores and industrial effluents. In this review, the funda-
mental process of bioleaching from low-grade metal sulfide
ores are discussed with emphasis on mechanism, types,
pathways, techniques, and bioleaching development.
Keywords Acidithiobacillus ferrooxidans  bioleaching 
Iron-oxidizing bacteria  Pyrite  Acid mine drainage
& Wasim Sajjad
wasim.sajjad71@yahoo.com
1
Key Laboratory of Petroleum Resources, Gansu Province/
Key Laboratory of Petroleum Resources Research, Institute
of Geology and Geophysics, Chinese Academy of Sciences,
Lanzhou 730000, People’s Republic of China
2
University of Chinese Academy of Sciences, Beijing 100049,
People’s Republic of China
3
Department of Microbiology, Faculty of Biological Sciences,
Quaid-I-Azam University, Islamabad 45320, Pakistan
•
1 Introduction
In human civilization, metal extraction has been a funda-
mental activity since Bronze and Iron times. Global rise in
human population and development of several nations has
increased demand of all natural resources, together with
metals. Various mineral product consumption has been
growing rapidly with time as technologies develop, which
eventually causes the reduction of high-grade ore reserves.
Therefore, more attention has been made on metal
extraction from low-grade ores and industrial and mining
wastes. Hydrometallurgical and pyrometallurgical tech-
niques are beneficial for high-grade ores; however, for low-
grade ores, these techniques provide low metal extraction
due to high process and energy cost. Such kind of mining
techniques also cause severe water, air, and land pollution.
Water drainage from abandoned mines is acidic in nature
and have soluble metals, that causes damage of water
bodies and lands. In this scenario, mining industry focuses
on economical, advanced, less energy demanding, and
environmental friendly technology for these low-grade ore
processing [26, 94].
Biohydrometallurgy is potentially more sustainable
approach where microorganisms are used for metal
extraction from their iron and sulfide ores as well as from
minerals (Fig. 1). Unlike conventional methods, this
approach is less energy demanding and eco-friendly having
no discharge of injurious chemicals and gasses into the
environment [8, 26]. Both in mineral formation and
decomposition process in earth’s crust, microorganisms
have been active since beginning of life on earth. This
natural ability of mineral decomposition by several
microorganisms is an ancient practice that dates back to the
first century BC to Roman era and possibly before the
Phoenicians. This ability of microorganisms was used in
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Fig. 1 Flow sheet of bioleaching for low-grade sulfide ores
old days for copper leaching without having the knowledge
that the microorganisms were responsible. Rossi [117]
mentioned in his book (biohydrometallurgy) that microbial
copper recovery was done in 1752 on commercial scale in
Rio Tinto, Spain. The role of microorganisms in the acid
mine drainage (AMD) [35], and description of pioneer
iron- and sulfur-oxidizing bacterium A. ferrooxidans [143]
solved the mystery. Biohydrometallurgical approaches
were implemented commercially since 1950s, due to var-
ious factors like metals prices [25, 26], production cost
[52], and hurdles in ore exploration via conventional
practices [26].
Currently, bioleaching is a reputable biotechnology that
is pragmatic globally. Several factors such as construction
progress, designing, and operational management of the
process, well-defined microbial application and monitoring
of activity has caused spreading of this technology. Natu-
rally available ores of copper, nickel, and zinc mainly exist
in metal sulfide form. Under optimized conditions,
77.68 ± 3.55% of copper and 70.58 ± 3.77% of zinc are
dissolved by using indigenous iron-oxidizing bacterial
consortia [124]. With bioleaching technique, Falagán et al.
[45] achieved more than 90% of copper extraction by using
defined bacterial consortium. Significant copper extraction
(91%) was obtained from copper slag material with bac-
terial consortia under aerobic conditions [131]. More than
75% of zinc was leached from zinc plant leach residues
(ZLR) collected from Tres Marias Brazil, using
Acidithiobacillus thiooxidans [134]. Ghassa et al. [53]
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extracted 98.5% zinc and 98% of cadmium under optimum
conditions. During bench-scale bioleaching experiment,
ore was inoculated with Halothiobacillus halophilus and
supplemented with formate (0.3%) that enhanced the nickel
recovery to 70-fold [149]. Chaerun et al. [30] processed
low-grade nickel limonite and saprolite ores by using
mixotrophic bacterial consortia and achieved 30% and
2.5% of nickel from limonite and saprolite, respectively.
Today, only bioleaching is envisaged to process low-
grade ores, mostly when they encompass deleterious ele-
ments, which lead to huge penalties in pyrometallurgical
processing. This review presents the fundamental process
of bioleaching from low-grade metals sulfide ore with
emphasis on mechanism, types, pathways, and techniques.
Furthermore, metals sulfide-oxidizing microbes, factors
affecting bioleaching, bioleaching developments, life-cycle
assessment and OMICs technology role in upcoming
microbial aspects exploitation in industrial bioleaching
have been discussed in succinct.
2 Bioleaching
Bioleaching is the metal-extracting practice from sulfides
and/or iron-containing ores by means of microorganisms. It
is a specified biohydrometallurgical approach in which
either the metabolic activities or microbe’s products are
used to convert insoluble compounds into soluble form,
which can then be recovered later [6]. Biological product
Trans Indian Inst Met (2019) 72(3):559–579
synthesis by these microorganisms includes certain organic
acids [10], chelates, mineral acids and sometime ammonia
or amines [29]. More specifically, it utilizes the catalytic
effect of metabolic activities from sulfur- and iron-oxi-
dizing microorganisms that ultimately results in the
chemical degradation of the ores [83]. Bioleaching is
slightly slower procedure compared to chemical leaching.
However, it has additional benefits like wastage con-
sumption as substrate by microorganisms during oxidation.
By getting energy from this, microorganisms produce
organic and inorganic acids that extract metals. Bioleach-
ing dramatically reduces the capital costs and eco-pollution
because it is carried out under mild environments, without
supplementing toxic chemicals. Main mechanisms that
solubilize metals are acidolysis, complexolysis, and
redoxolysis [22].
Natural ores of several metals such as Ni, Zn, Fe, As,
and Cu generally exist in metal sulfide form. Unlike oxidic
minerals, metals sulfide is insoluble in normal and weak
acidic conditions. Role of ferrous- and/or sulfur-oxidizing
microorganisms have been studied widely in metal
extraction from metals sulfide [94]. Oxidation of ferrous
into ferric and sulfide into sulfate is the requirement of
bioleaching and is carried out by acidophilic microorgan-
isms. Most of these microorganisms are reported from
AMD and are characterized in such a way that they can
tolerate high acidity, sulfate, and metal concentration.
Significant characteristic features of some microorganisms
are given in Table 1.
3 Bacterial and Archaeal Diversity in Bioleaching
Metals extraction with microbes is not a novel approach; it
has been applied for centuries; however, mechanism stays
unknown. Isolation of A. ferrooxidans proves the role of
microorganism in leaching. Today, A. ferrooxidans is most
extensively characterized and commercially used for
bioleaching [21]. Since 1970s, the number of microor-
ganisms that is involved in metals sulfide oxidation has
increased [23]. Several researchers have explored and
characterized microorganisms from certain extreme envi-
ronments like volcanic regions, hot springs, and mining
operations where these microbes inhabit. Some advanced
molecular techniques such as randomly amplified poly-
morphic DNA (RAPD), terminal restriction fragment
length polymorphism (T-RFLP), amplified ribosomal DNA
restriction analysis (ARDRA), real-time PCR (RT-PCR),
fluorescence in situ hybridization (FISH) and next-gener-
ation sequencing of metagenomic analysis have been
employed for examination and quantification of bioleach-
ing microorganisms [86, 123, 130]. Several bacteria like A.
thiooxidans, Leptospirillum ferrooxidans, A. ferrooxidans,
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and archaea such as Sulpholobus brierleyi and S. thermo-
sulphidoxidans are vital in bioleaching. Several microor-
ganisms are listed in Table 1 that can grow in low pH.
These metals sulfide oxidizers have autotrophic, mixo-
trophic, or heterotrophic nature. Physiological and other
related details of all these acidophilic microorganisms can
be found elsewhere [21, 33, 39, 62, 115].
3.1 Nutritional-Based classification
3.1.1 Autotrophic (Chemolithotrophic)
Autotrophs involved in bioleaching are acidophilic
microbes, which can fix CO2 and gain energy by oxidizing
the ferrous iron or reduced sulfur. The metabolic end
product in this process includes ferric iron or H2SO4. These
end products solubilize metal sulfide and decrease the
surrounding pH, which further enhances metals solubi-
lization [10], since microbial leaching is best in low pH
(1.5–3.0) where most of the metals stay in solution.
Extreme acidophiles are widely reported within Domain
bacteria (Table 1), like members of phylum Proteobacteria
(alpha, beta, and gamma classes), Firmicutes, Nitrospirae,
Acidobacteria, and Actinobacteria. Autotrophic bacteria
involved in leaching comprise both iron and sulfur oxi-
dizers such as A. ferrooxidans, L. ferrooxidans, and A.
thiooxidans. In anaerobic conditions, A. thiooxidans is
capable of propagating by using reduced inorganic sulfide
compounds (RISCs) and ferric iron as an electron acceptor
[140]. Application of Thiobacillus and other autotrophs in
commercial-scale bioleaching of fossil fuels originating
from shales, schist, and ores, is well establishes [7, 81].
Current exploration establishes the metals recovery, such
as Al, Co, Cu, Zn, Mo, U, V, and radionuclides via che-
molithotrophs from acidic environments [7, 93]. Such
microbes are utilized for metals extraction from their
respective ores [4, 165].
3.1.2 Heterotrophic (Chemoorganotrophic)
Heterotrophic microbes need organic carbon source for
their survival and metal leaching. Metabolism of organic
carbon excretes certain organic acids such as lactic, glu-
conic, citric, and oxalic as metabolites, which solubilize
metals from their ores. This is indirect bioleaching mech-
anism, which is appropriate for minerals having lower
sulfur and sulfide sources for H2SO4 production by auto-
trophs [10]. Basically, the organic acid produced by het-
erotrophs makes the medium suitable for metal
solubilization in pH range 4–6, whereas ferric iron is pre-
cipitated. Some heterotrophs can synthesise certain non-
acidic metabolites by protein catabolic activity, which can
be utilized in alkaline leaching condition, e.g., ammonia
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Table 1 Microorganisms and their physiological properties capable of bioleaching (adopted from [94–96, 127] with modification)
Domain Microorganisms Nutritional type Chief leaching agent pH range Opt. pH Temp °C

Archaea Acidianus ambivalens Chemolithoautotrophic H2SO4 2–5 2.5 80

Acidianus brierleyi Chemolithoautotrophic H2SO4 1.5–3.0 1.5–2 45–75
Acidianus infernus Autotrophic H2SO4 2.0 *90
Acidianus sulfidivorans Chemolithoautotrophic 0.35–3.0 0.8–1.4 74
Acidiplasma cupricumulans Facultative autotroph 0–4.0 1.4–1.6 45
Ferroplasma acidarmanus Mixotrophic 1.0–1.7 35–42
Ferroplasma acidiphilum Autotrophic Fe? 1.3–2.2 1.7 15– 45
Metallosphaera hakonensis Facultative autotroph 3.0 70
Metallosphaera prunae Chemolithoautotrophic Fe?, H2SO4 2–3 *75
?
Metallosphaera sedula Chemolithoautotrophic Fe Acidophilic 2–3 75
Picrophilus oshimae Heterotrophic 0.7 60
Picrophilus torridus Heterotrophic 0.7 60
Sulfolobus acidocaldarius Chemolithoautotrophic Fe?, H2SO4 0.9–5.8 2.0–3.0 55–85
Sulfolobus ambivalens Autotrophic Fe?, H2SO4 Thermophilic
?
Sulfolobus brierleyi Chemolithoautotrophic Fe , H2SO4 1.5–2.0 *70
Sulfolobus hakonensis Chemolithoautotrophic 1–4 3.0 70
Sulfolobus metallicus autotrophic 2–3 65
Sulfolobus solfataricus Chemolithoautotrophic Fe?, H2SO4 3–4.5 85
Sulfolobus tokodaii Mixotrophic 2.5–3.0 80
Sulfolobus thermosulfidooxidans Autotrophic Fe?, H2SO4 Thermophilic
Sulfolobus yangmingensis Facultative autotroph 2.0–6.0 4 80
Sulfolobus yellowstonii Autotrophic Fe?, H2SO4 Thermophilic
Sulfurococcus mirabilis Mixotrophic Fe?, H2SO4 2–2.6 70–75
Sulfurococcus yellowstonii Mixotrophic H2SO4, Fe? 40–80
Thermoplasma acidophilum Heterotrophic 1–2 59
Thermoplasma volcanicum Heterotrophic 2 60
Bacteria Acetobacter methanolicus Heterotrophic Gluconate 3–6 4
Acidiferrobacter thiooxydans Autotroph 2.0 38
Acidimicrobium ferrooxidans Facultative autotroph *2 45–50
Acidiphilium angustum Heterotrophic Citric acid and Tween 80
Acidiphilium cryptum Heterotrophic Organic acids 2.0–6.0 35–41
Acidiphilium symbioticum Heterotrophic Organic acids 3.0 Mesophilic
Acidithiobacillus albertensis Autotrophic 3.5–4 25–30
Acidithiobacillus caldus Autotroph H2SO4 2.0–2.5 45
Acidithiobacillus ferrivorans Autotroph 2.5 27
Acidobacterium capsulatum Heterotrophic Organic acids 3.0–6.0 Mesophilic
Acidocella spp.
Acidomonas methanolica Heterotrophic
Alicyclobacillus GSM Facultative autotroph 1.8 47
Alicyclobacillus tolerans Facultative autotroph 2.5 37
Arthrobacter spp. Heterotrophic
Bacillus spp. Heterotrophic
Aureobacterium liquifaciens Heterotrophic
Bacillus coagulans Heterotrophic 5.4–6.0 22
Bacillus megaterium Heterotrophic Citrate 4–7 5.0 25
Bacillus licheniformis Heterotrophic 37
Micrococcus varians Mixotrophic 3–5 3.0 30
Bacillus subtilis Heterotrophic 3–5 3.0 35

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Table 1 continued
Domain Microorganisms Nutritional type Chief leaching agent pH range Opt. pH Temp °C

Bacillus pumilus Mixotrophic 6–8 7.0 25

Corynebacterium vitarumen Heterotrophic 4–6 5.0 30
Bacillus polymyxa Heterotrophic
Chromobacterium violaceum Heterotrophic Cyanide
Comamonas testosterone Heterotrophic
Crenothrix spp. Facultative Autotrophic Fe? 5.5–6.2 18–24
Enterobacter cloacae Heterotrophic 22
Enterobacter agglomerans Heterotrophic 5.4–6.0 22
Ferrimicrobium acidiphilum Heterotrophic 2 35
Ferrithrix thermotolerans Heterotrophic 1.8 43
Gallionella spp. Autotrophic Fe? 6.4–6.8 6–25
Kingella kingae
Lactobacillus acidophilus Heterotrophic 37
Leptospirillum ferriphilum Autotrophic 1.3–1.8 30–37
?
Leptospirillum ferrooxidans chemolithoautotrophic Fe 2.5–3.0 30
Leptospirillum thermoferrooxidans chemolithoautotrophic Fe? 1.7–1.9 45–50
Leptothrix discophora Facultative Autotrophic Fe?, H2SO4 5.8–7.8 5–40
Metallogenium spp. Heterotrophic Fe? 3.5–6.8 4.1
Ochrobacterium anthropic Heterotrophic
Pseudomonas putida Heterotrophic Citrate, Gluconate
Propionibacterium acnes Heterotrophic 37
Pseudomonas cepacia Heterotrophic 5.4–6.0 22
Psychrobacter glacincola Heterotrophic
Serratia ficaria Heterotrophic
Siderocapsa spp. Heterotrophic Fe?
Stenotrophomonas maltophilia Heterotrophic
Staphylococcus lactis Heterotrophic 37
Sulfobacillus thermosulfidooxidans Chemolithoautotrophic Fe?, H2SO4 Extr. 50
acidophil
Sulfobacillus thermotolerans Facultative autotroph 2.0–2.5 40
Thiobacillus acidophilus Mixotrophic H2SO4 1.5–6.0 3.0 25–30
Thermothrix thiopara Chemolithoautotrophic H2SO4 Neutral 60–75
Thiobacillus albertis Chemolithoautotrophic H2SO4 2.0–4.5 3.5–4.0 28–30
Thiobacillus concretivorus Chemolithoautotrophic H2SO4 0.5–6.0
Thiobacillus capsulatus Chemolithoautotrophic H2SO4 3–7
Thiobacillus delicates Mixotrophic H2SO4 5.0–7.0 25–30
Thiobacillus denitrificans Chemolithoautotrophic H2SO4 5.0–7.0 30
Thiobacillus ferrooxidans Chemolithoautotrophic Fe?, H2SO4 1.4–6.0 2.4 28–35
Thiobacillus intermedius Facultative Heterotrophic H2SO4 1.9–7.0 6.8 30
Thiobacillus kabobis Mixotrophic H2SO4 1.8–6.0 3.0 28
Thiobacillus neapolitanus Chemolithoautotrophic H2SO4 3.0–8.5 6.2–7.0 28
Thiobacillus novellus Chemolithoheterotrophic H2SO4 5.0–9.0 7.8–9.0 30
Thiobacillus perometabolis Chemolithoheterotrophic H2SO4 2.6–6.8 6.9 30
Thiobacillus organoparus Mixotrophic H2SO4 1.5–5.0 2.5–3.0 27–30
Thiobacillus prosperus Chemolithoheterotrophic H2SO4 1.0–4.5 23–41
Thiobacillus pumbophilus Chemolithoheterotrophic H2SO4 4.0–6 5 27
Thiobacillus rubellus Chemolithoheterotrophic H2SO4 5.0–7.0 25–30
Thiobacillus tepidarius Chemolithoautotrophic H2SO4 6–7 43–45

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Table 1 continued
Domain Microorganisms Nutritional type Chief leaching agent pH range Opt. pH Temp °C

Thiobacillus thiooxidans Chemolithoautotrophic H2SO4 0.5–6.0 2.0–3.5 10–37

Thiobacillus thioparus Chemolithoautotrophic H2SO4 4.5–10.0 6.6–7.2 11–25
Thiomonas cuprinus Facultative heterotrophic H2SO4 3.0–4.0 30–36
Thiobacillus versutus Chemolithoautotrophic H2SO4 8.0–9.0
Eukarya Actinomucor spp. Heterotrophic Succinate 27
Fungi Alternaria spp. Heterotrophic Citrate, Oxalate 32
Aspergillus fumigatus Heterotrophic
Aspergillus awamori Heterotrophic 28
Aspergillus niger Heterotrophic Oxalate, Citrate, Gluconate, 30
Malate,
Tartrate,
Succinate
Aspergillus ochraceus Heterotrophic Citrate 28
Cladosporium spp. Heterotrophic
Aspergillus spp. Heterotrophic Citrate, 30
Oxalate
Cladosporium resinae Heterotrophic 28
Coriolus versicolor Heterotrophic Oxalate
Fusarium spp. Heterotrophic Oxalate, Malate,
Pyruvate,
Oxalacetate
Mucor racemosus Heterotrophic Citrate, Succinate 27
Paecilomyces variotii Heterotrophic Citrate, Oxalate
Penicillium chrysogenum Heterotrophic 28
Penicillium spp. Heterotrophic 25
Penicillium funiculosum Heterotrophic Citrate
Penicillium notatum Heterotrophic 26
Penicillium simplicissimum Heterotrophic Citrate, Oxalate, Gluconate 22–30
Rhizopus japonicas Heterotrophic
Trichoderma lignorum Heterotrophic 24–26
Trichoderma viride Heterotrophic 32
Yeast Candida lipolytica Heterotrophic 30
Rhodotorula spp. Heterotrophic
Saccharomyces cerevisiae Heterotrophic 28
Torulopsis spp. Heterotrophic
Trichosporon Heterotrophic
Latest information in the table has been included following literature research

leaching [8, 50]. Heterotrophs play a vital role in biomining
along with autotrophs. Genera such as Acidocella, Acidis-
phaera, Acidobacterium, and Acidiphillum, have been
reported from acidic environments accompanied with A.
ferrooxidans that consume several organic substrates but
not iron. Heterotrophic bacteria have been isolated and
characterized from extreme low-pH environment of AMD
and black shale [80, 121, 122]. Such microbes oxidize acid-
soluble metal sulfide and improve the biomining ability of
autotrophs. During oxidation, autotrophs excrete some
inhibitory agents, which can be consumed by heterotrophs
and assist in providing energy substrate to iron oxidizers by
reducing ferric into ferrous iron [106, 130]. Several het-
erotrophs reported from leaching environments are repor-
ted in Table 1.
3.2 Temperature-Based Classification
3.2.1 Mesophilic
Mesophilic acidophiles were the pioneers in mining
industry, where these were utilized for commercial bio-

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oxidation of pyrite-bearing gold ores, arsenopyrite, in
stirred tank reactors. Later with the development of
research, these microorganisms have been utilized for low-
grade copper ore bioleaching [34]. Several mesophiles,
such as A. thiooxidans, A. ferrooxidans, L. ferriphilum, and
L. ferrooxidans, are metal-tolerant and solubilizing bacteria
that are frequently used in bioleaching [95]. Furthermore,
certain mesophilic microorganisms belong to genus Sul-
fobacillus, Acidimicrobium, Alicyclobacillus, and Ferro-
plasma have been utilized for biomining [127, 130].
Nowadays, mesophilic acidophiles are favored microor-
ganism for heap bioleaching from low-grade chalcopyrite
ores because it works optimally at normal temperature [99].
3.2.2 Thermophiles
Several moderate thermophiles like A. ferrooxidans, A.
caldus, S. acidophilus, S. thermosulfidooxidans, S. disulfi-
dooxidans, T. acidophilum, Alicyclobacillus acidocaldar-
ius, and F. acidarmanus are used for bioleaching [127].
Likewise, extreme thermophiles include Acidianus infer-
nus, A. brierleyi, Metallosphaera hakonensis, A. sulfidivo-
rans, M. sedula, Sulfurococcus mirabilis, S. metallicus, M.
prunae, S. yellowstonensis, and S. yangmingensis. Metal
sulfide oxidation rate by thermophilic acidophiles is higher
compared to mesophilic acidophiles [34, 169]. Since the
metal sulfide oxidation is exothermic, temperature rise
occurs in the heap-bioleaching core, which boosts the
thermophilic-acidophilic growth [106].
Nowadays, thermophilic biomining is used for gold and
copper [107, 162]. In pyrite-bearing refractory gold con-
centrates and arsenopyrite-oxidizing bioreactors, the dom-
inant sulfur oxidizer is A. caldus at intermediate
temperature, whereas S. metallicus and Metallosphaera
spp. are dominant above 60 °C [166]. Industrial application
of thermophilic biomining is still under exploration, and
factors like cell wall sensitivity to high pulp density and
low tolerance toward metals compared to mesophiles have
restricted its commercial application [95]. However, stirred
tank bioleaching is the sole effective industrial application
for the treatment of high-grade gold ores and concentrates
[52, 127]. Heap bioleaching with thermophiles can be an
alternate fast chalcopyrite-processing method, where
desired temperature is obtained for high microbial growth
and activity [106, 170].
4 Fungal Diversity in Bioleaching
Heterotrophic microbes capable of bioleaching also include
filamentous fungi. Fungi can produce certain organic acids,
amino acids, and other metabolites, which are involved in
leaching from ores (Table 1). These metabolites dissolve
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metals either by metal ion displacement from the ore via
hydrogen ion or by chelates creation and solubilize metals.
Leaching with fungus is rather slower compared to bacte-
ria; however, despite this, fungal leaching is best if min-
erals hold enough quantity of organic material, such as
black shale [8, 9]. Fungus such as Penicillium and Asper-
gillus spp. play a vital role in metal bioleaching because
they excrete high concentration of organic acids [110, 72].
They have been used for metal recovery or mineral bene-
ficiation such as refractory gold and silver ores, copper
oxides and carbonates, manganese oxides, spodumene,
cobalt and zinc ores, oxidic ores of nickel, zircon, removal
of unwanted minerals from bauxites, silicate ores con-
taining chromium, iron, titanium and quartz sands and
silicates [127]. Several fungal strains were used for metals
extraction from black shale [9, 8], incineration fly [157],
waste material [65], power plant residues [110], and min-
ing deposits [82]. Fungal strains were examined for toler-
ability towards these metals and it was discovered that
several metals are crucial for cellular activity [150]. Nor-
mally, fungus carry bioleaching at pH 2.0–8.0 and tem-
perature 20–40 °C and tolerate heavy metals and were
therefore used in past for mining waste and carbonaceous
low-grade ore processing. Le et al. [69] examined that high
percentage of metals (Cr, Co, Al, Mn, Fe, Cu, Mg, Zn and
Ni) deactivate the cellular metabolism responsible for
organic acid production and mineral leaching. Drawbacks
like supply of organic substrate that enhance the processing
cost, possible contamination with undesired microbes, and
pathogen occurrence, prevent industrial applications of
heterotrophic microbes for leaching till today.
5 Bioleaching Mechanisms
Bioleaching of metal sulfide is principally based on innate
uniqueness of microbes that get their vital energy from
oxidation of ferrous iron and RISCs in acidic environment.
Bio-oxidation of metal sulfide happens by transporting
electrons from sulfur or/and iron moiety of mineral to the
final electron acceptor oxygen [6]. There are three mech-
anisms of bioleaching: direct and indirect bioleaching, and
later the terms are replaced with contact and non-contact
bioleaching. The third one is co-operative bioleaching
[52, 127].
5.1 Direct Bioleaching
Direct bioleaching means direct electron transfer from
metal sulfide to bacterial cells adhering directly to metal
sulfide. In this mechanism, the cell attaches to metal sulfide
with the help of extracellular polymeric substance (EPS)
layers, resulting in sulfide mineral dissolution at the
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interface via electrochemical process. This approach is
easy to implement, but dissolved metals can negatively
affect microbial growth and metabolism, which in turns
limits productivity [70]. However, the microorganisms in
direct bioleaching have high resistance toward heavy
metals as described in oil contaminated land study [18]. In
this mechanism, pyrite oxidation occurs, and ferric sulfate
is produced [135]. Reactions are the followings:
4FeS2 þ 14O2 þ 4H2 O ) 4FeSO4 þ 4H2 SO4 ð1Þ
4FeSO4 þ O2 þ 2H2 SO4 ) 2Fe2 ðSO4 Þ3 þ 2H2 O ð2Þ
Direct oxidation of pyrite via microorganisms is
summarized in reaction:
4FeS2 þ 15O2 þ H2 O ) 2Fe2 ðSO4 Þ3 þ2H2 SO4 ð3Þ
Non-iron metal sulfide such as chalcocite, covellite, galena,
sphalerite, molybdenite, millerite, cobaltite, and stibnite
can also be oxidized by A. ferrooxidans via direct
bioleaching [146]. Therefore, direct bioleaching for entire
non-iron sulfide can be described in the following reaction:
MS þ 2O2 ) MSO4 ð4Þ
However, direct bioleaching does not seem real since direct
electron transfer between metal sulfide and adhered cells
has not been established till now [95, 127]. Now it is an
established fact that bioleaching is an integrally chemical
process and microbes produce the essential chemical
agents like ferric iron and protons for metal sulfide leach-
ing and provide reaction compartment of EPS layer
[95, 152].
5.2 Indirect Bioleaching
Indirect bioleaching occurs through ferric ion (oxidizing
agent), generated by ferrous-oxidizing bacteria either
attached or planktonic [152]. This ferric acts as a lixiviant
in acidic solution and solubilizes metal sulfide.
 pathway to adopt [126, 128]. The crystal structure like
MS þ Fe2 ðSO4 Þ3 ) MSO4 þ 2FeSO4 þ S ð5Þ
In this reaction, ferrous iron oxidized to ferric by bacteria
participates in leaching process. In indirect bioleaching, the
cells are not in direct contact with the mineral surface, but
they have catalytic activity to speed up the reoxidation of
ferrous iron, which is otherwise very slow in the absence of
microorganisms. At pH 2.0–3.0, ferrous oxidation by
bacteria is 105–106-fold faster than chemical oxidation.
The sulfur generates is simultaneously oxidized to H2SO4
by sulfur-oxidizing bacteria At. thiooxidans.
 attacked by protons, and such metal sulfides are acid non-
2S þ 3O2 þ 2H2 O ) 2H2 SO4 ð6Þ
Role of sulfur oxidizers is to provide satisfactory acidic
condition for optimum survival and function of iron-oxi-
dizing bacteria. Another example of indirect bioleaching is
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Trans Indian Inst Met (2019) 72(3):559–579
the uranium recovery; oxidation of insoluble tetravalent
uranium occurring in soluble hexavalent. The lixiviant,
produced by A. ferrooxidans during pyrite oxidation, is
associated with uranium ore.
UIV O2 þ Fe2 ðSO4 Þ3 ) UVI O2 SO4 þ 2FeSO4 ð7Þ
Indirect leaching is facilitated with organic and inorganic
acids produced by microorganisms. Such bioleaching
occurs in two stages: First, the cells are grown in a medium
having optimum cultural conditions to synthesise active
metabolites vital for bioleaching. Secondly, under suit-
able conditions, the culture-free medium is utilized as
mineral-leaching agent [10].
5.3 Co-operative Bioleaching
Term ‘‘Co-operative bioleaching’’ refers to sulfur inter-
mediates, sulfur colloids and other mineral dissolution by
planktonic cells [111]. This mechanism involves direct and
indirect mechanisms that synchronously affect metal dis-
solution. Certain chemicals released by the attached cells
fulfill the energy requirements of planktonic cells [100].
Terms like this are beneficial for physical status description
of the cells that perform bioleaching. However, they do not
reflect an idea about original chemical mechanism of
metals sulfide dissolution. In an orthodox way, bioleaching
is established by collaboration of chemical and biological
oxidation. Specific significance must be credited to ferrous
and ferric iron cycle.
6 Pathways of Bioleaching
With research progression, now it is understood that metal
sulfide dissolution is not similar, and therefore, two dif-
ferent pathways ‘‘thiosulfate’’ and ‘‘polysulfide’’ describe
this dissolution. The mineral species determine the type of
monosulfide and disulfide does not regulate the pathway,
but metal sulfide reaction with protons is the appropriate
principle. This property is controlled by the electronic
configuration of metals sulfide [128]. By nature, metal
sulfides are conductors, semiconductors, or insulators, and
both sulfur and metal atoms are bound to crystal lattice
[158]. Both molecular orbital and valence band theories
reveal that orbitals of atoms and molecules form electron
bands having diverse energy levels. The metal sulfide with
valence band derived only from metal atom orbital cannot
soluble (thiosulfate pathway). Such metal sulfides are
resistant to proton attack, and their bond can only be bro-
ken through oxidant like ferric iron by multistep electron
transfer. However, the valence bands resulting from both
Trans Indian Inst Met (2019) 72(3):559–579
orbitals of metal and sulfide will be acid soluble (poly-
sulfide pathway). In such metal sulfides, along with ferric,
protons can also remove valence band electrons and thus
causing metal sulfur bond cleavage [118, 126]. Therefore,
two classes of metal sulfides are present having two dif-
ferent oxidation mechanisms. Intermediate sulfur com-
pound formation in both pathways depends upon metal
sulfide mineralogy and geochemical conditions like exis-
tence of certain oxidant and pH [129]. Microbes play a
vital role in oxidation of these intermediates and convert
them into sulfate and protons (H2SO4) [132].
6.1 Thiosulfate Pathway
Metal sulfides such as pyrite, tungstenite, and molybdenite
holds pair of sulfur atoms, which form nonbonding orbitals
[151], while the valence bands derive only from metal
atoms, therefore, are entirely oxidized via ferric iron. Pyrite
oxidation is extensively studied, and detail can be found
somewhere else [41, 129]. Pyrites are exclusively oxidized
by ferric iron. When ferric initially attacks on pyrite, the
sulfur part gets oxidized into soluble sulfur intermediates.
In such metal sulfides, the bond between metal and sulfur
does not cleave until six successive one-electron oxidations
have occurred and thiosulfate is liberated. Therefore, this
pathway is called thiosulfate pathway. During this path-
way, the disulfide part of pyrite is oxidized by hydrated
ferric ions into sulfonic acid group by extraction of several
electrons. The bond between sulfur and iron is broken, and
thiosulfate and hydrated ferrous ions are produced. The
primary soluble sulfur intermediate that is formed is then
oxidized to tetrathionate [133]. Further, tetrathionate
degrade into several sulfur compounds, i.e., trithionate,
pentathionate, sulfite, and elemental sulfur [41, 129]. These
intermediates are ultimately oxidized to sulfate by bio-
logical and/or chemical reactions. The central role played
by microorganisms in this pathway is the ferric iron
regeneration from oxidation of released ferrous iron
(Eq. 8). Thiosulfate pathway can be described by the fol-
lowing Eqs. 9 and 10:
2Fe2þ þ 1=2O2 þ 2Hþ ) 2Fe3þ þ H2 O ð8Þ
FeS2 þ 6Feþ3 þ 3H2 O ) S2 O2
3 þ 7Fe
2þ
þ 6Hþ ð9Þ
S2 O2
3 þ 8Fe
3þ
þ 5H2 O ) 2SO2
4 þ 8Fe
2þ
þ 10Hþ
ð10Þ
Stoichiometry of this pathway has been established in
pyrite oxidation with A. ferrooxidans and oxygen and
sulfur stable isotopes are found as products [17]. From
electron extraction reaction, it has been proved that only
ferrous oxidizers are able to solubilize acid non-soluble
567
metal sulfide. Only such bacteria can redevelop ferric iron
after consumption, since chemically at low pH (2.0), fer-
rous iron is not oxidized significantly [137].
6.2 Polysulfide Pathways
Unlike pyrite oxidation, the bond between metal and sulfur
in acid-soluble metal sulfide can be broken prior to H2SO4
oxidation. Metal sulfides like orpiment, realgar, chalcopy-
rite, troilite, pyrrhotite, hauerite, galena, and sphalerite can
be dissolved by protons. Such metals sulfide is dissolved by
collective action of ferric and proton, i.e., the binding of
proton by the sulfide moiety through valence band elec-
trons. This proton attack cleaves the bond between metal
and sulfur moiety and after binding two protons, liberation
of hydrogen sulfide occurs. Though, in the presence of
ferric, sulfur oxidizes in a single-electron step with proton
attack. At acidic condition, the sulfur moiety of metal
sulfide is oxidized to elemental sulfur [78]. Several reac-
tions essentially describe the elemental sulfur formation
through polysulfide pathway [114, 128], which have been
identified during the dissolution of, chalcopyrite [58],
galena [138], and pyrrhotite [144]. The initial sulfur com-
pound produced is sulfide cation (H2S?) that can dimerize
impulsively to free disulfite (H2S2) and further oxidize to
elemental sulfur by polysulfides and polysulfide radicals
[139]. Therefore, acid-soluble metal sulfide oxidation has
been called polysulfide mechanism. The elemental sulfur
can inert chemically in natural environment, or it can
oxidize biologically to H2SO4 and conserve acidity of
medium. In polysulfide oxidation, sulfides ([ 90%) are
converted into elemental sulfur in the absence of sulfur-
oxidizing bacteria. Some products such as thiosulfate,
sulfate, and polythionates are formed [128, 139]. Oxidation
by ferric iron is not the only condition for polysulfide
pathway but can also be oxidized by the action of sulfur
oxidizers. In ferric iron deficiency, these bacteria carry
oxidation of free sulfide (H2S) resulting from the proton
attack on metals sulfide to H2SO4 and regenerate the
consumed protons. Polysulfide mechanism can be
explained by the following equations [128].
MS þ Fe3þ þ Hþ ) M2þ þ 0:5H2 Sn þ Fe2þ ð n  2Þ
ð11Þ
3þ 2þ þ
0:5H2 Sn þ Fe ) 0:125S8 þ Fe þH ð12Þ
þ
0:125S8 þ 1:5O2 þ H2 O ) SO2
4 þ 2H ð13Þ
This pathway agrees with outcomes of bioleaching inves-
tigation through A. ferrooxidans in which stable isotopes of
sulfur and oxygen are identified in the products of spha-
lerite and chalcopyrite oxidation [16].
123
568
7 Bioleaching Techniques
Mineral bioleaching is an effective and simple technology
and widely used on practical scale for metals extraction.
The economics and effectiveness of this approach highly
depends on microbial activity and mineralogical and
chemical composition of ores. This, process optimally
applied on one type of ores cannot be optimum for other
type. Prior to technical application, optimal leaching con-
ditions must elaborate for each type of ores. Leaching
techniques may be classified into three areas based on
working volume.
7.1 Laboratory Techniques
7.1.1 Submerged Leaching
Simple percolator leaching is not efficient enough due to
insufficient O2 supply and unfavorable surface ratio. It is
gentle and the experiments usually last for 100–300 days.
Consequently, percolating leaching has been replaced by
submerged leaching by using finely grinded ores material
having particle size of \ 100 lm, which are suspended in
leaching liquid all the time and kept in motion by shaking.
Adequate aeration, precise control, and monitoring of
several parameters enhance the growth and activity of
microorganisms. Thus, the reaction time is significantly
reduced, and metal extraction considerably rises. Sub-
merged leaching can be performed in Erlenmeyer flask, or
in more refined manner, in bioreactor. In addition of
mechanically agitated system, an air lift reactor has been
provided for industrial waste products, ore concentration,
and bio-desulfurization of coal treatments [145]. It is very
easy technique and has advantage that it can easily be
regulated and controlled [100]. Particle size \ 100 lm
considerably increases the surface area and leaching level,
which are tough to leach with percolation. But, it has
drawback of constantly fluctuating conditions like dis-
solved O2 and pH. This fluctuation makes it hard to study
the influence of certain factors since they can be intensified
or negated by continuous fluctuation [54]. Submerged
leaching is implemented to examine the material’s
amenability toward bioleaching and optimization of several
biological and physicochemical parameters. This labora-
tory-scale processing provides a strong base for scaling up
the process.
7.1.2 Percolator Column Leaching
Column is specifically designed container inside which the
low-grade ore or industrial effluents of specific size are
subjected to bioleaching. Column designing is according to
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Trans Indian Inst Met (2019) 72(3):559–579
specific measurement in diameter and height to accom-
modate sample weight. Top of the column acts as an inlet
for the lixiviant, O2, and CO2 into column. Principle of
column leaching is same with percolator leaching and used
as a model for dump or heap leaching. Column bioleaching
ensures bacterial lixiviant flow rate optimization for metal
dissolution. Moreover, this technique offers a clear picture
of acid consumption pattern [97, 98], which can assist to
cost estimation for scale-up trials. Based on size, column
can be made of lined concrete, plastic, glass, and steel.
Most column systems have fixed devices at various dis-
tances for sample taking and installation of special instru-
ments for O2, CO2, pH, temperature, and humidity
monitoring. Ore sample is flooded with lixiviant having
microorganisms, which percolates through the column
under gravity. The solution trickling through the column is
again pump backed to column surface for recirculation.
Concurrently, air stream fulfills the aeration requirements
of the system. One of the characteristic features of column
leaching is that it forms several zones within the ore, which
have discrete chemical and physical gradients. Such zones
may show redox potential difference, elemental sulfur
formation, and iron precipitation. These zones can show
similarity with those in large-scale heap and dump leaching
processes [108]. This technology has few disadvantages
like being inefficient and slow extraction rate, and series of
experiments lasting 100–300 days are usual because of
insufficient O2 supply and the unfavorable surface ratio
[100].
7.2 Pilot-Plant Technique
Constructions of reliable model for industrial-scale plants
and the laboratory-scale techniques are not considered
reliable; however, it provides valuable information in short
period of time. Industrial-scale leaching is performed under
different conditions, and optimum environmental condi-
tions in laboratory-scale techniques can not be appropriate
for it. Highly dependable industrial-scale plant can be
designed under optimum and controlled environmental
conditions after pilot plant practices, such as column
leaching and agitated tanks [75, 141].
7.3 Industrial Leaching Techniques
Presently, bioleaching is industrially used for metal
extraction from low-grade ores having metal concentra-
tion \ 0.5% (w/w). First industrial-scale bioleaching
technique was patented in early 1950s by the copper
industry, successively followed by gold mining operation
[91], uranium [64] and zinc [2] from low-grade ores. The
simplest way of bioleaching is to stack the materials in
heap and let the lixiviant to trickle through it and recollect
Trans Indian Inst Met (2019) 72(3):559–579
the leachate. Since the bacterial sulfide oxidation is slower
as compared to other biotechnical approaches, leachate
recirculates.
7.3.1 Dump Leaching
This is an oldest process. It involves pile up of uncrushed
rocks (25–400 m height, 100–300 m width, and
100–2000 m length) and then flooded with dilute H2SO4 to
enhance the microbial growth [117]. Generally, these
dumps comprise of about 0.1–0.5% copper, which is very
low to extract with conventional methods. The dump size
varies extensively, and ore amount can range from hun-
dreds to thousand tons. Dump is sprinkled constantly from
the top or flooded momentarily. Lixiviant can be simple
water, acidified water, or acidic ferric sulfate solution
obtained from other leaching operation having similar
mining properties, and it depends upon the ore [37]. No
microorganisms are inoculated in dump; they are ubiqui-
tous and proliferate when the conditions become appro-
priate. Lixiviant sprinkling introduces enough air that is
vital for biological and chemical oxidation. Prior to recir-
culation, the leachates can pass over an oxidation basin,
where the bacteria and ferric ions are renewed.
7.3.2 Heap Leaching
Materials are specifically designed to be arranged in dump
shape having dimension of height, width, length, and slant
over water and acid proof membrane. The heap design is
somewhat slanted to allow drainage of leachate out of
reservoir [99]. Heap and dump bioleaching procedure is
similar. Heap leaching deals with extra regulation of
chemical, engineering, and biological factors than dump
bioleaching. This method is used for fine-grinded ores,
which cannot be concentrated by floatation, to enhance the
lixiviant–mineral’s interaction and make the base imper-
meable to stop the lixiviant loss and water body’s pollution
[37, 75]. The heap’s height is the controlling dimension,
which ranges from 2–3 to 7–8 m when the larger part of the
ore sample is 60–150 mm or greater [117]. Such leaching
is performed in large basins holding up to 12,000 tons ores.
Sometimes in heap bioleaching, pipes are inserted in
planned sites, to provide adequate O2 into deeper portion.
Principally, both in dump and in heap bioleaching, the
lixiviant applications are carried out on the surface and the
recovered leachate that seeps to the bottom by gravity flow.
Dilute H2SO4 is sprinkled on the surface that penetrate
downwards and lowers the pH and enhances the microor-
ganism’s growth. This runoff is collected at the bottom and
is pumped to recovery stations where the metals are
recovered by cementation, solvent extraction or
electrowining.
569
7.3.3 Underground Leaching
Underground leaching is generally performed in abandoned
mines. Galleries are drowned or ores from unmined or
mine waste inside tunnels are trickled or washed away
under pressure. Lixiviant having microorganisms are
introduced into boreholes in the ruptured orebody. The
leachate is collected in deeper galleries, shafts and pumped
into processing plant. The well-known use of this technique
is at Stanrock uranium mine situated at Elliot Lake in
Ontario, Canada [77]. Ore deposits, which are unable to
process by conventional methods since they are of low
grade, can be leached in situ. The entire practice needs
suitable orebody permeability and impermeability of the
gangue rock so that to prevent the pregnant leaching
solution seepage.
7.3.4 Tank Leaching
Tank leaching construction and operation are more
expensive than heap and dump leaching, but metal
extraction rate is high. This approach is superbly used for
refractory gold ore bioleaching. With the construction of
pioneer industrial tank bioleaching plant at Fairview Gold
Mine in South Africa for gold processing, efforts have been
made to process other ores [1]. For high-yield metal
extraction by submerged leaching, the use of bioreactor
instead of shaking flask was experienced very early [117].
Tank leaching establishes the most effective approach for
bioleaching and [ 80% of Zn is leached from zinc sulfide
[147].
8 Factors Affecting Bioleaching
Bioleaching involve microorganisms, therefore, highly
effected by biological, environmental, and physicochemi-
cal factors, which in turn affect the metal extraction yield.
The bioleaching efficiency largely depends upon the
competence of microorganisms and mineralogical and
chemical composition of ores. High metal extractions can
only be obtained when the leaching conditions (Table 2)
are an optimum range of bacterial growth conditions.
8.1 pH
For microorganisms that carry out bioleaching, pH is the
most important factor. Changes in pH greatly influences the
colonization and metabolism of microorganisms [95]. The
bioleaching medium acidity level results from the proton
balance between net-consuming reactions and reaction of
H2SO4 production and iron hydrolysis. Optimum pH is
necessary for optimal microbial growth and metal
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570
Table 2 Factors affecting bioleaching and their effects
Factors Parameters that affect bioleaching
Physical and pH, temperature, nutrient, redox potential, CO2 and O2 content, Affect microbial composition, activity and leaching rate
chemical homogenous mass transfer,
parameters
Fe(III) concentration and inhibitors, etc.
Biological Microbial diversity
parameters
Inoculum density
Metal tolerance,
Adaptation abilities of microorganisms
Microbial activities
Spatial distribution and attachment of microorganisms to ore
particles
Ore characteristics Composition
Particle size
Surface area
Porosity
Mineral type and distribution
Acid consumption, hydrophobic galvanic interactions, Jarosite
formation and formation of secondary minerals
solubilization because most of the metals are soluble at low
pH. Optimum pH range for microbial ferrous and sulfide
oxidation is 2.0–2.5. Generally, bacteria like A. ferrooxi-
dans are unable to grow on ferrous iron at pH [ 3.0.
Optimal pH range varies among different system and
microorganisms [92]. From thermodynamic point of view,
higher the pH, quicker will be the acid production. On the
other hand, low pH close to 1.0 is injurious for microbes
and only the acid-tolerant species can withstand, making
the biological system delicate. To balance the pH during
process, a neutralizing agent like limestone can be added,
but it boosts the jarosite formation. pH range of 1.4–1.7 is
possibly a good compromise between the above-mentioned
risks and technical feasibility of pH controlling in big
stirred tank [100]. The influence of wide-range pH condi-
tions on microbial community shifts over DDGE analysis
exposing that At. caldus and S. thermosulfidooxidans
dominate at initial pH 1.0, whereas At. caldus, L. fer-
riphilum, and S. thermosulfidooxidans are always identified
at pH 2.0. The L. ferriphilum growth is repressed at pH 1.0
and 3.0 [162]. The bacterial growth usually starts at lower
pH and as the pH increases upto medium value, the growth
continues without affecting the cellular activity [11]. It has
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Trans Indian Inst Met (2019) 72(3):559–579
Effect
Low pH is needed to achieve high leaching rate and to
keep the ferric iron and metals in solution
In chemical and biological oxidation of metals sulfide an
electron acceptor (Ferric iron) is needed
Mixed culture tends to be more strong and efficient than
pure
High inoculum density tends to increase the leaching rate
High metal concentration may be toxic for
microorganisms
Leaching conditions are usually harsh, so microbes must
be able to adopt such conditions
Ferrous and ferric iron is due to microbial activity. It
must be an optimum range
The proper arrangement and attachment of
microorganism upon ore surface is essential for
leaching
It provides electron donor and trace elements for growth
Affects the available minerals/liquid contact area
Bioleaching is proportional to the increase mineral
surface area
Particles pores and cracks give rise into internal area and
facilitate solution penetration
Minerals having lowest potential is generally oxidized
first. Minerals distribution in ores also affect
bioleaching
All these generally influence the microbial potential of
bioleaching
been reported that the microbial activity increases many
folds by the bacterial strain adaptation to the growth
medium at optimum pH [43].
8.2 Temperature
Microorganisms are categorized based on optimally growth
temperature: mesophiles (30–40 °C), moderate ther-
mophiles (around 50 °C), and extreme thermophiles (above
65 °C). Cells become inactive below optimum temperature
and destroy above optimum temperature. Sulfide mineral’s
bio-oxidation is an exothermic process and, therefore,
generates significant heat in stirred tank reactor and heap.
Temperature control in heaps is a difficult task. Height of
heap is a significant factor for temperature rise, which rises
with square of heap height [106]. This is in association with
an aeration effect and irrigation rate [55, 99]. With tem-
perature alteration, the microbial diversity dynamics alters
[86]. Industrially, the heap inside temperature may reach
up to 50 °C on which both the chemical and biological
oxidation reactions cannot be structured, and the attain-
ment of an exceptionally high temperature will noticeably
prevent mesophilic bacterial activity. As temperature rises
Trans Indian Inst Met (2019) 72(3):559–579
to more than 40 °C, mesophiles are replaced by moderate
thermophilic sulfur and iron oxidizers. Moderate ther-
mophiles can be replaced by extreme thermophiles when
temperature rises above 60 °C [106]. Inside the heap,
temperature is determined by several factors, as described
by Petersen and Dixon [104]. Optimum temperature for A.
ferrooxidans to oxidize ferrous and sulfide is 28–30 °C. At
lower temperature, reduction in metal extraction occurs,
but at 4 °C, two competing factors affect the process: first
the reaction rate enhancement which are usually carried by
raising temperature, and second the reduction of microbial
activity with temperature rise and upper tolerance limit of
the microorganisms. This competition becomes greater if
mesophiles and psychrophiles are used. Therefore, appro-
priate temperature range should be adjusted for bioleaching
to keep conditions constant for microbial growth and
reactions.
8.3 Nutrients
Microorganisms need nutrients for their cellular metabo-
lism, new cells and metabolites biosynthesis. In bioleach-
ing, most of the microorganisms are chemolithotrophic and
needs inorganic compounds. Generally, the mineral nutri-
ents are acquired from the material to be leached. For
optimal microbial growth, sulfur and iron compounds must
be supplied into the media together with salts like ammo-
nium, magnesium, and phosphate. Furthermore, nutrient
supplement must be in optimal concentration; too low or
high concentration may affect the microbial growth. In
case of heterotrophs, sucrose has been found to be the best
carbon source for citric acid production by A. niger fol-
lowed by glucose and fructose [56]. Magnesium and
potassium are often available in the ore minerals. It has
been considered that jarosite precipitation depends upon
the ammonium concentration in the slurry, which can cause
the bacterial activity reduction by sulfide surface passiva-
tion [100].
8.4 Oxygen
Both aerobic and anaerobic microorganisms are capable of
bioleaching processes. For optimum growth and leaching
activity of bacteria, adequate supply of O2 is necessary.
Inadequate O2 supply will slow down the microbial growth
rate and eventually the leaching rate. Bacteria require O2
during iron and sulfur oxidation. O2 solubility in water at
35 °C is 8 g/m3; it decreases with increase in ionic con-
centration in the solution [63]. As per the stoichiometry,
bacterium iron oxidation reaction needs 0.07 g of O2 per
gram of ferrous oxidation, and this amount cannot be
supplied by the solution in view of O2 solubility. Thus, the
additional O2 supply must be from outside. More than
571
0.2 g/dm3 of dissolved O2 concentration is adequate for the
metabolic activates of bacterium [73]. In laboratory, it can
be achieved by doing aeration, shaking, or stirring. In
industrial stage, mainly in dump and heap leaching, ade-
quate O2 supply can cause hurdles. Air supply to heap may
be carried through pipe network installed at the base. Holes
of 50 mm diameter are bored at the bottom for air distri-
bution in pipes [24]. Blowers or low-pressure high-volume
fans are used to inject air into heaps.
8.5 Carbon Dioxide
Microorganisms that are used at commercial scale are
mostly autotrophic. They use CO2 as a carbon source.
Adequate CO2 supply is the subject of discussion as an
operating parameter in commercial bioleaching. However,
there is adequate CO2 in air to fulfill the requirements
through air sparging. At higher microbial growth rate,
carbon uptake rate reaches to the levels where CO2 supply
by the injected air is not further adequate and carbon
transfer become a rate limiting step for sulfide oxidation.
However, the fact that either the CO2 excess will amplify
or reduce the bacterial growth may vary among microor-
ganisms [100].
8.6 Inoculum Density
Inoculum density is one of the controlling factors in
leaching kinetics both in direct and indirect mechanisms.
Denser microbial population may increase the rate and
bioleaching efficiency. But several limiting factors like O2
and nutrient supply restrict the inoculum density. There-
fore, an optimum inoculum density should be supplied to
achieve maximum bioleaching efficiency. Maximum
microbial population concentration varies between 103 and
109 cells/mL in continuous stirred tank [63]. Therefore, to
enhance the kinetics, the microbial population must be
increased. The bacterial population in lixiviant can be
increased by harvesting the biomass through centrifugation
or membrane filtration and re-inoculate it in lesser volume
of solution. However, these practices are costly and need
complex instrumentation. These challenges can be over-
come by using a biofilm type of reactor known as bacterial
film oxidation [57]. Most of the acidophilic microorgan-
isms have distinctive attraction toward jarosite. Thus, if a
thin film of jarosite develops under optimum conditions,
these microorganisms will get essential site for extra
growth. Pesic and Kim [103] suggested jarosite formation
mechanism and growth of microbes. Other alternative to
increase microbial population is to apply electrical poten-
tial. Bacterial population depends on the amount of ferric
iron oxidized. In a rough estimate, for 1 g of biomass
production, it requires 100 g of ferrous iron oxidation.
123
572
Ideally, if iron oxidation and reduction rates are equivalent
by applied potential, it supports high bacterial population
and oxidation potential and enhances sulfide dissolution
kinetics [87].
8.7 Microbial Activity
Activity of microbes is measured in terms of sulfur and
iron oxidation rate. The oxidation rate varies from strain to
strain [20], due to several factors, like morphology varia-
tion [136], variation in lipopolysaccharides composition
[60], variation in nutrient metabolism [109], resistance
toward organic matter [155], and DNA base ratio [112].
Best microbial strains can be obtained from mines, UV
irradiation and by genetic engineering the activities can be
enhanced [145]. Bacterial activity can be enhanced by
adaptation techniques [15] as most of the metals are toxic
and adaptation increases the tolerance in alien condition
and curtail the lag period, hence increasing the leaching
kinetics [13].
8.8 Metal Resistance of Microorganisms
Metals concentration in leachate increases as the sulfide
mineral leaching progresses. Existence of certain heavy
metals such as arsenic, zinc, iron, and copper in high
concentration may inhibit the microbial growth [46, 47].
Toxicity of several metals on bacteria in 9 K medium was
studied by Escobar and Lazo [44]. High concentration of
metals demonstrates toxic activity due to four reasons: (1)
functional groups blockage of biologically important
molecules, (2) dislocating or replacement of essential
metals from biomolecules and functional cellular units, (3)
inducing conformational changes in polymers, and (4)
influencing the membrane integrity and transport practices
[49]. Sensitivity to heavy metals varies between different
genus of microorganisms and even strains of the same
species. Thus, microbes that have high tolerance towards
heavy metals in leach suspension should be selected.
8.9 Attachment of Microorganisms
Several studies have confirmed the bacterial attachment to
the ore’s surface and/or presence in the pregnant leach
solution in commercial procedures. A typical model for
bioleaching was proposed by Sand et al. [125], which is
principally proceeded by ferric ions complexation by the
secreted EPS substances. This model, initially considered
for A. ferrooxidans, and applied for L. ferrooxidans, since
both these iron-oxidizing bacteria share EPS quality that
enable the bacterial attachment to the ore surface [51, 116].
Ferric ion only did not bring any local pyrite corrosion,
which specifies that the reactions between bacterial
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Trans Indian Inst Met (2019) 72(3):559–579
attachment and original pyrite surface are responsible for
bioleaching that leads to corrosion.
8.10 Size, Chemistry, and Pulp Density of Substrate
Substrate size also affects the leaching rate. Smaller par-
ticle size increases the total particle surface area; therefore,
high metal yield can be obtained without changing the
particles’ total mass. Best metals solubilization rate have
been studied to occur in particle size of the order of few
tens of microns [117]. Higher the pulp density, higher will
be the leaching process because it increases the total par-
ticle surface area. On the other hand, higher pulp density
rises the toxic substances in the leaching environment,
which cause the microbial growth inhibition. Therefore, the
metal extraction process is often best at lower pulp density.
Any two ores are not similar, and each ore deposit displays
heterogeneity and holds different mineralogical composi-
tion and inclusion, acid demand characteristics, and ele-
ments concentration. Bioleaching profiles of each ore
sample are distinctive because of these differences. The
mineralogical composition of substrate is of primary
importance because it affects the bioleaching rate and
efficiency. Pyrite containing minerals are highly important.
Microbial growth may be inhibited by the presence of
highly toxic metals and reduction of the bioleaching rate
and efficiency occurs. Even in solid substrate, the chemical
state of the metal compound also disturbs the bioleaching
efficiency. The water and acid-soluble forms of metals
leach out easily compared to others. Porosity in ore
material greatly enhances the solution penetration into the
orebody [106].
8.11 Irrigation
Two types of irrigation exist, continuous, and discontinu-
ous, while it is largely believed that discontinuous irriga-
tion favor metal extraction. In this type, the solution is
irregularly sprayed onto the heap surface and is allowed to
penetrate before fresh solution supply. Through irrigation,
the liquid is drawn into the ore mass by capillary forces.
When irrigation finishes, the liquid with dissolved metals is
drained out of capillary and stays on the surface and fresh
irrigation carries with it and the practice starts again with
the supply of fresh liquid into the capillary. Thus, discon-
tinuous irrigation is highly effective for harsh ores than
continuous irrigation, since this alternative draining and
drying of the capillaries is significantly rapid than simple
ionic diffusion through a static capillary full of fluid. This
alternative draining and drying assist to leach coarse par-
ticles and washes soluble salts from the surface and
enhances O2 and CO2 diffusion to the ore surface [106].
Some studies reveal that daily irrigation is more effective
Trans Indian Inst Met (2019) 72(3):559–579
in bacterial presence compared to irrigation after 1 week
[12]. Industrially, the evaporation rate and metal concen-
tration in liquid phase determine the irrigation frequency
[28]. Though, the irrigation frequency is a vital factor to be
considered.
8.12 Jarosite Formation
Jarosite formation in bioleaching is problematic. After
formation, jarosite precipitates on mineral surfaces and
reduces the efficiency of reagents and mineral interactions,
and even metal oxidation inhibition mediated by microor-
ganisms. This inhibition of bioleaching especially in
chalcopyrite is due to the creation of an intermediate sul-
fide passivation layer. This layer is less reactive compared
to original chalcopyrite and can prevent the flow of elec-
trons and oxidants towards and from chalcopyrite [106].
But exact nature of this layer is complex and it is described
that it forms from one or both sources: (1) either Fe(OH)3
carry the jarosite [KFe3(SO4)2(OH)6] formation and coats
the unreacted material to produce a passivating layer and/
or, (2) the elemental sulfur formed during the process tends
to coat the surface. Ferric iron precipitation and jarosite
formation are pH dependent and jarosite precipitation is
more prominent within pH range of 1.9–2.2. But using
thermophilic bacteria rises the temperature to [ 60 °C
disrupting the passivating layer on chalcopyrite in a high-
temperature heap-bioleaching procedure [148].
9 Developments in Metals Sulfide Bioleaching
Due to depletion of high metal ore deposits or growing
depth of economically feasible deposits, all the conven-
tional processes such as smelting, roasting, floatation and
grinding will not be more applicable economically. With
conventional processes, the power demand is inversely
proportional to the metal present in the ore; that’s why with
low metal contents these conventional methods become
expensive. However, this gap can be compensated with
finding new high-grade ore deposits and/or employing new
bioleaching strategies or methods that can enhance pro-
duction. Followings are some strategies and methods that
can enhance extraction rate in bioleaching.
9.1 In Situ Bioleaching Consideration During Pre-
mine Planning Stage
Caving method, e.g., block caving method, during under-
ground mining from porphyry copper deposits is equivalent
both in production capacity and in expenditures to open-pit
mining. Hence, it can be employed for low-grade ores
especially when the practical and economic depth edge for
573
open-pit mining process is reached. During advanced block
caving methods, 25–30% of the orebody has left in situ
[153]. Nobody has yet considered employment of in situ
bioleaching for the abandoned ores at the time of block
caving during pre-mine planning periods to employ in situ
bioleaching for the abandon ores during block caving. The
worth of mining technology and resource efficiency can be
enhanced by considering in situ bioleaching processing
during planning stage of large-scale mining projects. The
new underground mine at Chuquicamata, Chile, which will
be commissioned in 2018, will employ block caving min-
ing to produce 350,000 t of copper annually [127].
Therefore, with an expected ore left over of 30% because
of caving mining, about 105,000 t per annum of copper will
remain in the abandon ores. Additionally, employing a
typical in situ post-bioleaching extraction for copper (50%)
from dump or stockpile bioleaching can contribute 50,000 t
of copper recovery annually. For concentrates that are
difficult to process by conventional methods due to pollu-
tant compounds such as bismuth and arsenic and their
unfavorable polymetallic structures or huge transportation
expenses, bioleaching is the best alternate way to exploit.
9.2 Use of Thermophilic Microorganisms
To date, bioleaching was generally limited to using
mesophilic bacteria during heap bioleaching. Temperature
rise inside heap generally restrict the proliferation of bac-
teria and subsequently minimize the efficiency of
bioleaching. However, recently the use of thermophilic
bacteria during bioleaching has been extensively studied
[53, 45, 90, 120] by amended modeling and monitoring of
thermophilic microbial communities, and high metal
extraction rate has been achieved. The material and setup
such as equipments and process control used for high-
temperature bioleaching already exists [42]; the only need
is to obtain competent microbial community.
9.3 Microbial Consortia
Further study has now been focused to use consortia
instead of single kind of microorganisms in bioleaching.
Encouraging results have been achieved by using microbial
consortia, because consortia has great potential to with-
stand against arsenic, copper, and chloride ion [67], prevent
iron contamination of leach liquor during manganese
extraction and desirable for downstream processing of
metal recovery [5], and hold barotolerant (higher pressure
resistant) ability that can contribute in anaerobic iron and
sulfur cycling in deepest ore deposits that are extremely
relevant for in situ bioleaching process [168].
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9.4 Use of Solar Energy
However, inside heap, the process is exothermic, but the
efficacy can be further enhanced with an additional heating
source suitable for thermophilic bacteria. The use of solar
energy can provide suitable conditions within heap and
enhance metal recovery rate and reduce cost in a sustain-
able way. Primarily the copper mines in Chile are sited in
the Atacama Deserts, which receives huge solar radiation.
These radiations can be unified into heap bioleaching to
rise the temperature inside heap and enhance the copper
extraction rate. Recently, Murray et al. [85] studied a
bioleaching process simulated over 1 year, including solar
thermal collector field and ponds using HeapSim (Heap
Bioleach Simulation tool) and TRNSYS (Simulation Stu-
dio to model the heat gain from the solar collector field).
Maximum copper extraction rate without solar energy was
67% with 7 kg/h m2 solution flow rate, while extraction
rate of 85% was achieved with collector field to heap area
ratio of 1:1 having 10 kg/h m2 solution flow rate. Capital
cost of the solar energy structure to the income generated
from extra copper leached was economically compared.
The NPV (net present value) and internal return rate were
positive for collector areas ranged from 10,000 to
150,000 m2 within 200,000 m2 of heap area. Highest NPV
happened at 50,000 m2 and 76% of extraction was
achieved by leaching over 1 year.
9.5 Use of Bio-surfactants
Bio-surfactants produced by bacteria are capable of alter-
ing the surface of several metals and aggregates on inter-
phases and enhancing metal extraction process. The use of
bio-surfactant promotes the interaction between ore and
bacteria. These natural bio-surfactants can reduce or even
replace chemical surfactants, and metals extraction can be
achieved in a more environmentally sustainable way. They
can enhance the metal recovery that is conventionally hard
to extract such as the molybdenum content. Bacteria such
as Bacillus subtilis, Brevibacillus parabrevis, Pseu-
domonas aeruginosa, and Bacilluspumilus has been
reported to produce bio-surfactants and are used in metal
extractions [66, 113]. These natural bio-surfactants have
qualities of emulsification, detergency, dispersion, and
foaming and show advantages over synthetic counterparts,
like biodegradability, low toxicity, and selectivity
[68, 105]. Diaz et al. [40] achieved 63% Fe and 70% Zn
extraction by using combined bioleaching and rhamnolipid
bio-surfactants; however, recovery rate was 50% (Zn) and
19% (Fe) with bioleaching alone. In recent studies, Yang
et al. [160] used the bio-surfactant (glycolipid) producing
Burkholderia sp. Z-90 for extraction of heavy metal con-
taminated soil and achieved significant results for Zn
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Trans Indian Inst Met (2019) 72(3):559–579
(44%), Pb (32.5%), Mn (52.2%), Cd (37.7%), Cu (24.1%)
and As (31.6%). Bio-surfactant can enhance the decon-
tamination of heavy metals from the sludge and signifi-
cantly influence the electro-kinetic process [142]. Bacteria
that need iron for their metabolism are already exploited in
the commercial scale for metal extraction. Nowadays, the
focus is on to study bio-floatation: a technique yet to be
fully explored where there will be no more need of living
microorganisms, but these active substances can alter the
mineral surface. These may be cell components, biomole-
cules or metabolic products. Once appropriate bacteria are
reported, and the bio-surfactants are isolated, further aim
will be to study the interaction of mineral surface and
individual molecule. In a long-term perspective, bio-
floatation can be integrated into conventional processes.
Furthermore, these bioactive substances can degrade nat-
urally within environment. How these substances behave
inside the process is the matter to explore further to make
them technologically and commercially viable.
9.6 Solvent Extraction and Electrowinning
Recently, bioleaching combined with solvent extraction
(SX) and electrowining (EW) technique has been suc-
cessfully implemented to extract copper. However, with
SX, several organic chemicals entrain in the form of
emulsion and get dissolved in aqueous raffinate. These
chemicals are usually entrained during raffinate recycling
in bioleaching and contaminate the system inevitably. The
impact of these organic chemicals on bioleaching ability of
A. ferrooxidans was studied by Liu et al. [74]. It was
reported that the organic chemicals present in raffinate are
directly proportional to LIX984 N concentration, and
concurrently decrease the bacterial metabolic activity.
Contamination with organic chemicals in SX affects the
cellular ultrastructure and morphology. The SX organics
adsorb the surface of minerals and hamper its contact with
bacteria that leads to lower extraction rate of metals [161].
Therefore, it is strongly recommended that the effect of SX
process must be considered during bioleaching process for
the safe recycling of raffinate, which is usually skipped.
Chen et al. [32], successfully investigated a combined
technique of bioleaching–solvent extraction–electrowin-
ning (L–SX–EW) for sludge-born copper extraction. The
copper solubilization ratio has been found to be more than
90% and extraction efficiency reaches more than 95% by
using 2% solvent extractant M5640 under optimum con-
ditions. Elemental copper powder of more than 90% is
achieved through electrolysis of acidic aqueous phase.
During this process, no waste water generation has been
reported, and therefore, L-SX-EW technique has potential
to exploit for metal recovery. Ruan et al. [119] discussed
the use of bioheap leaching–SX–EW technique used at
Trans Indian Inst Met (2019) 72(3):559–579
Zijinshan copper plant in China to treat secondary copper
sulfide ores. Megaw et al. [79] presented an alternate
process for copper sulfate production that includes crushing
and screening of the ores, heap-bioleaching, SX, and cop-
per sulfate EW at remote location for Central African
copper belt. Same technique has also been implemented in
South America with full success and targeted to copper
sulfate as a final product mainly for the animal feed addi-
tive market. Basically, the satellite copper sulfate produc-
tion is beneficial because the copper units provided to
refinery for cathode production need less secondary pro-
cessing compared to dense media separation (DMS) con-
centrate. The final product is favorable for copper cathode
processing as that have unused SX and EW capacity. When
copper sulfate is added to SX, the associated sulfate is
collected as sulfuric acid in the raffinate and fulfill the acid
requirement of bioleaching.
9.7 Combination of Electro-Kinetic Technology
and Bioleaching
Combination of electro-kinetic technology and bioleaching
not only dissolves or stabilize fraction of the heavy metals
but also improves recycling of heavy metals and eradicate
their hazard [102]. The bioleaching part is to extract metals
as ionic fraction that migrates toward cathode, precipitates
and is recycled. Also, bioleaching can be combined with
advanced oxidation process (AOPs) such as Fenton-like
reaction (Fe3þ/H2O2) that reduces shortages and enhance
the metal recovery [159]. Bioleaching is performed first by
providing acidic environment and catalysts for Fenton-like
reaction, which in turn shorten the bioleaching period
[171]. The iron-oxidizing bacteria convert ferrous to ferric
iron. The ferric iron not only oxidizes metal sulfide but also
enhances H2O2 conversion to OH, which oxidizes organic
components and destroys extracellular polymeric sub-
stances (EPS) and releases metals [61]. Conditions have
been successfully optimized for bioleaching combined with
Fenton-like reaction to enhance trace metal recovery and
dewaterability of dredged sediments [167]. Zhu et al. [171]
successfully recovered Cu (75.3%), Zn (72.6%), Pb
(34.5%), Cd (65.4%) by using this technique.
9.8 Pre-treatment of Ore Bodies
Microwave and ultrasonication treatment of ores can
improve bioleaching efficacy by causing grain boundary
fracture. This approach has potential advantage in down-
stream recovery stage such as energy conservation during
following comminution because of particle weakening and
high exposure or release of value grains. Such kind of
treatment enhances metal recovery, as kinetics and extent
of bioleaching depend upon grain accessibility to lixiviant.
575
Improved metal recovery has been reported by treating zinc
sulfide ores in microwave due to internal cracks created by
microwave [31].
9.9 Using Catalytic Additive Agents
Currently, several catalytic agents have been used during
bioleaching to enhance the metal extraction from metal
sulfide ores. Such agents can decrease the recalcitrant
behavior of sulfide ores for leaching the metals and improve
dissolution behavior of the ores. Several studies have
reported the use of silver as catalysts to improve dissolution
of low-grade ores in the presence of meso-acidophiles
[46, 47, 84, 163]. The use of sodium chloride to boost copper
dissolution rate has been proposed recently [19, 46, 47, 71].
However, these catalysts are uneconomical, sometime
related with deleterious effect on microorganisms, for
example silver is expensive and chloride ion create stress
conditions and inhibit growth of microorganisms for which
they try to combat these challenges with several adaptation
mechanisms [164]. The use of suitable catalytic agent is
favorable for bioleaching such as the waste newspaper used
by Panda et al. [94–96] is economical, eco-friendly, and
enhance the copper recovery (99%). Suitable catalytic
agents that can control oxido-reduction reaction during
bioleaching enhance the metal dissolution and are capable of
scaling up which will considerably contribute to enhancing
metal recovery from low-grade sulfide ores.
9.10 Controlled Slurry Potential
Metal recovery rate can be enhanced through controlled
slurry potential. The effect of redox potential on metal
dissolution has been extensively studied and it reveals that
redox potential plays crucial role in copper dissolution
[3, 59]. Recently, it has been reported that with electro-
chemical redox control in continuous bioleaching, higher
copper dissolution and faster leaching kinetics are achieved
[76]. The copper dissolution rate enhances significantly by
reducing the redox potential. The bioleaching and elec-
trochemical investigation of metals sulfide dissolution are
controlled by redox potential. Bioleaching of chalcocite
and pyrite is carried under constant redox potential
(\ 760 mV, vs SHE) and copper extraction of 90–98% is
achieved, which is tenfold time than iron recovery [156].
Under oxygen limited conditions, populations of iron oxi-
dizers are reduced; however, Acidithiobacillus having
sulfur reducing ability are an alternate controller of redox
potential. Therefore, maintenance of controlled redox
potential during bioleaching technique is one of the
effective approaches to balance iron and acid production.
Several other technologies are focused to optimizing the
commercial-scale metal extractions or passivation. These
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include ion exchange, filtration, chemical precipitation,
adsorption on activated carbon, electrochemical treatment,
membrane technology etc. [14, 154]. Although, great effort
has been made to improve bioleaching technology, yet
there are some limitations which delay the pervasive
application of technology.
10 Life-Cycle Assessment
High-grade ores of metals are falling worldwide as their
reserves are processed first and are depleted progressively.
Simultaneously, demand of metals extracted from these
ores has probably increased, even with high level of de-
materialization and reusing. Sustainability alarms have
emphasized the requirements to meet these demands, while
at the same time, reducing resource consumption and
environmental release. Several studies have been carried
out to undertake life-cycle assessment (LCA) methodology
to scrutinize several other processing ways for metal
extraction from low-grade ores. Norgate and Jahanshahi,
[89] used LCA methodology to inspect the impact of ore
grades (down to 25%) and grind sizes (down to 5 lm) based
on energy consumption and greenhouse gas emission of
nickel and copper extraction by conventional methods
(pyrometallurgical processing). The findings reveal that the
impact of declining ore grade on environment is significant
at grade below 1%, because extra energy is consumed, and
greenhouse gases are emitted during mineral processing to
move and treat additional gangue material. LCA of several
alternate routes of ore processing for metal extractions has
been studied by Norgate and Jahanshahi, [88] and they
reported the least embodied energy consumption and
greenhouse gas emission with different grade ores during
heap leaching. Compared to metal extraction through
conventional approaches at high temperature, the
microorganisms working in bioleaching are functional at
ambient temperature and produce very few direct con-
taminants and greenhouse gases. No external addition of
chemicals is required during bioleaching and the lixiviant
is processed under controlled environment and it prevents
water pollution.
11 Future Prospects
Future of bioleaching is quite encouraging. With fast
consumption of high-grade ores, it is essential to process
the low-grade ores and industrial effluents for metal
extraction. Today, many countries have a substantial
amount of low-grade ore assets, which on exploitation may
contribute to their economic development. Several devel-
oping countries such as Indonesia, Chile, Peru, Mexico and
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Trans Indian Inst Met (2019) 72(3):559–579
Zambia combinedly contribute 50% copper production
[106]. Currently, bioleaching is used for U, Au and Cu,
however, in future it will be vital for Co, Zn, Ni, and Mo
etc. recovery. In present scenario, demand and prices of
metals are increasing, thus, bioleaching application is also
increasing. Investment and processing cost of bioleaching
are much lower compared to conventional processes. Also,
the operational plant can be constructed in the close
vicinity of the ores to save the transport expenses. Besides
the metals extracted in the leachate, certain insoluble
metals are left behind in the residues like lead. Therefore,
the inferior lead sulfide concentrate can be converted into
high value concentrate by metal leaching, e.g., Cd, Zn and
Cu, which interfere in conventional extraction. Such pro-
cedures can also be applied for Ag and other precious
metals that are blocked by As, Cu, Fe and Zn sulfide.
Commercially, certain challenges are present in using
microbes for metals extraction from low-grade ores in
more engineered fashion. Mineralogical composition of
each orebody is different. Therefore, appropriate knowl-
edge of several process parameters like minerology, ore
characteristics and physical parameters is necessary for
effective metals recovery. For this purpose, detailed labo-
ratory and pilot scale testing is necessary to up-scale
commercially. Hence, modeling aspects in bioleaching
should be focused on dynamic aspects like actual predic-
tion of leaching behavior in freshly designed heap as well
as at different time intervals of leaching [38]. Also, the
microbiological aspects specifically the dynamics of active
communities is less-understood. Therefore, diverse simu-
lation in bioleaching on microbial community dynamics
will help to understand the interaction mechanisms, sur-
vival and diversity in such diverse and heterogeneous
systems.
Several environmental pollutants like industrial effluents
and AMD will be reduced by exploiting ultrasonically
enhanced bioleaching and bioremediation techniques.
Slowly, bioleaching is gaining importance in the waste-
treatment because microorganisms are the main agents
capable of metal dissolution. Mostly industrial waste
products such as fly ash and slag comprise high concen-
tration of valuable metals in oxide form rather than sul-
fides. It has been proved that these metals can be recovered
by the acid produced by microorganisms. Thiobacillus has
detoxification potential of soil and sediment polluted with
heavy metals and sewage sludge. Geomicrobiological
technologies intend to chase certifying commercialization
policy. Fresh strategic coalitions can help to improve the
full range of applications for new technologies and direct
authorization of the targeted mining companies will lead to
fast commercialization.
It is an established fact that the microbial consortium is
more effective in bioleaching compared to pure culture
Trans Indian Inst Met (2019) 72(3):559–579
[48]. Introduction and advancement of genetically engi-
neered microbes in consortia may enhance the bioleaching
and bioremediation [27]. No study has been conducted to
emphasize the novel consortium development that effi-
ciently dissolve metals. Recently, web server has been
projected based on bioinformatics applications, to predict
novel microorganisms to be used in bioleaching [101].
Such kind of bioinformatics prediction applications can
help to reshape the outlook of bioleaching technology. The
OMICS and bioinformatics approaches have been just
developing and such kind of more approaches can solve
many problems associated to microbial contribution. Fur-
thermore, these approaches will help in determining certain
new microbial characteristics that will arise from the genes,
proteins, macromolecules and environment interactions
[36]. The use of thermophiles in bioleaching will rise in the
future due to the quicker dissolution kinetics. Therefore,
further optimization studies can assist to recommend an
appropriate low-cost procedure for metal recovery in con-
trast to the currently established procedures.
12 Conclusions
This review provides basic materials on bioleaching. It is
generally an acceptable commercial technology for metal
recovery from low-grade ores, which are unable to exploit
by conventional techniques. So far, the bioleaching appli-
cation to recover metals from high-grade ores is very
limited due to slow kinetics. Some modified approaches,
such as bioleaching combined with other methods can
enhance the metal recovery rate in a more sustainable way.
Both prokaryotic and eukaryotic microorganisms and their
symbiotic association play role in metal dissolution. Dump,
heap and stirred tank are the key bioleaching processes. By
exploitation of this approach to enhance precious metal
recovery from refractory ores, great importance is now
being given to alter the kinetics. Further development is
crucial for both technical and biological aspects. In-depth
knowledge about reaction mechanisms, characterization
and microbial diversity and enzyme system is critical to
boost the kinetics by determining the rate limiting steps.
Notable progresses in OMICS and bioinformatics tech-
niques may enhance understanding about several microbial
aspects in bioleaching. It can be therefore be concluded
that soon more advanced, economical, and eco-friendly
bioleaching technology will be commercialized to process
low-grade ores.
Acknowledgement This work was sponsored by CAS-TWAS Pres-
ident’s Fellowship for international Ph.D. students to WS and par-
tially by the Natural Science Foundation of China (41572352).
577
Compliance with Ethical Standards
Conflict of interest There is no conflict of interest.
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