Subscriber access provided by University of Sussex Library
Article
Contact Behavior between Cells and Particles in the Bioleaching of Precious Metals from Waste Printed Circuit Boards Zhihui Yuan, Zhe Huang, Jujun Ruan, Yaying Li, Jian Hu, and Rong-Liang Qiu ACS Sustainable Chem. Eng., Just Accepted Manuscript • DOI: 10.1021/acssuschemeng.8b01742 • Publication Date (Web): 30 Jul 2018 Downloaded from http://pubs.acs.org on August 4, 2018
Just Accepted “Just Accepted” manuscripts have been peer-reviewed and accepted for publication. They are posted online prior to technical editing, formatting for publication and author proofing. The American Chemical Society provides “Just Accepted” as a service to the research community to expedite the dissemination of scientific material as soon as possible after acceptance. “Just Accepted” manuscripts appear in full in PDF format accompanied by an HTML abstract. “Just Accepted” manuscripts have been fully peer reviewed, but should not be considered the official version of record. They are citable by the Digital Object Identifier (DOI®). “Just Accepted” is an optional service offered to authors. Therefore, the “Just Accepted” Web site may not include all articles that will be published in the journal. After a manuscript is technically edited and formatted, it will be removed from the “Just Accepted” Web site and published as an ASAP article. Note that technical editing may introduce minor changes to the manuscript text and/or graphics which could affect content, and all legal disclaimers and ethical guidelines that apply to the journal pertain. ACS cannot be held responsible for errors or consequences arising from the use of information contained in these “Just Accepted” manuscripts.
is published by the American Chemical Society. 1155 Sixteenth Street N.W., Washington, DC 20036 Published by American Chemical Society. Copyright © American Chemical Society. However, no copyright claim is made to original U.S. Government works, or works produced by employees of any Commonwealth realm Crown government in the course of their duties.
Page 1 of 29 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
ACS Sustainable Chemistry & Engineering
1
Contact Behavior between Cells and Particles in the Bioleaching of
2
Precious Metals from Waste Printed Circuit Boards
3 4
Zhihui Yuan1, Zhe Huang1, Jujun Ruan1∗, Yaying Li1, Jian Hu2,
5
Rongliang Qiu1*
6
1. Guangdong Provincial Key Laboratory of Environmental Pollution Control and Remediation
7
Technology, School of Environmental Science and Engineering, Sun Yat-Sen University, 135
8
Xingang Xi Road, Guangzhou, 510275, People’s Republic of China
9
2. School of Environmental Science and Engineering, Yangzhou University
10
Corresponding author: Jujun, Ruan Tel: +86 20 84113620; Fax: +86 20 84113620; E-mail:
[email protected]; Rongliang, Qiu,
[email protected] 1
ACS Paragon Plus Environment
ACS Sustainable Chemistry & Engineering 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
11
Page 2 of 29
ABSTRACT
12
Bioleaching of precious metals from waste printed circuit boards (WPCBs) such
13
as using cyanogenic cultures for extraction of gold and silver has gained considerable
14
attention. It has the advantages of low cost and environment friendliness. However,
15
low efficiency hindered its industrialization. In this study, a novel strategy has been
16
proposed to improve bioleaching efficiency. We applied optical microscopy to
17
investigate the surface interactions between bacteria and silver particles in two-step
18
bioleaching. Results showed that bacterial adsorption and extracellular polymeric
19
substances (EPS) binding were the primary negative behavior that hindered the
20
leaching reaction. Functional group analysis by Fourier-transform infrared
21
spectroscopy (FTIR) indicated that carboxyl, hydroxyl, and amine groups were the
22
main chemical structures responsible for the negative influences. Bacteria–metal
23
interactions were reduced in the presence of polyvinylpyrrolidone (PVP) and the
24
silver recovery increased 1.8 times. Imaging analysis showed bacteria and EPS were
25
dispersed from silver surface in the case of PVP addition. Zeta potential analysis
26
indicated that PVP exhibited high affinity binding to silver particles and suppressed
27
the attachment of the microbial materials onto silver surface. Our results demonstrated
28
the utility of PVP addition for the bioleaching of precious metals.
29 30
Key
31
polyvinylpyrrolidone
32
word:
E-waste,
precious
metal,
bioleaching,
2
ACS Paragon Plus Environment
diffusion
control,
Page 3 of 29 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
ACS Sustainable Chemistry & Engineering
33
INTRODUCTION
34
E-wastes recycling is a worthwhile project not only from the perspective of
35
waste disposal but also from the context of resource recovery.1–3 Among recycled
36
materials, valuable metals in waste printed circuit boards (WPCBs) account for more
37
than 95% of the recovery value of e-wastes. Furthermore, precious metals contribute
38
to 80% of the monetary value of the recovered metals, and Au has the highest priority
39
to be recovered.4 Therefore, precious metal recovery has gained increased attention.
40
High concentrations of precious metals are found in electronic scrap samples.5
41
Physical methods, such as mechanical disruption, magnetic separation, and
42
electrostatic separation, are the preferred pre-treatment methods for WPCBs.6,7 These
43
methods have properties of low cost and low pollutant emission and can efficiently
44
obtain homogeneous materials. However, such methods present limitations in terms of
45
purification of various valuable metals from metal composites. Precious metals
46
exhibit high melting point and strong stability. Pyrometallurgical process requires
47
high energy consumption and creates pollution to the environment, and traditional
48
hydrometallurgical method produces large quantities of secondary pollutants.8
49
Therefore, alternative environment-friendly technologies must be developed for
50
reclamation of precious metals from e-wastes.
51
In precious metal bioleaching, hydrocyanic acid produced by some bacteria
52
serves as biogenic lixiviant. Precious metal dissolution can be achieved during cell
53
cultivation. Initially, metal composites obtained after physical separation were directly 3
ACS Paragon Plus Environment
ACS Sustainable Chemistry & Engineering 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
54
subjected to bioleaching. This method exhibited low efficiency due to interference
55
from exogenous metal. Pretreatment of metal components through biological
56
pre-oxidation to remove base metals in the leached concentrates leads to high precious
57
metal recovery.9,10 In addition, the researchers explored two-step leaching pattern and
58
spent medium leaching.11–13 In two step bioleaching, solid metal samples were added
59
when the lixiviant (cyanide) concentration reached maximum levels. In spent medium
60
bioleaching, cells were separated from the culture after it reached maximum cyanide
61
concentration, and only cell-free metabolites were used for bioleaching.13 However,
62
spent medium leaching is not easy to achieve for industrial upscaling by means of
63
centrifugation, filtration, and natural sedimentation. Bioleaching can also be enhanced
64
through other measures, included adding hydrogen peroxide, co-cultivation of
65
cyanogenic bacteria,14 changing leaching conditions,13 and using alkali tolerant
66
bacteria or metabolically engineered bacteria.9,15 Cyanide concentration produced by
67
bacteria is too low, though Au recovery has been nearly doubled using metabolically
68
engineered strain.15 Thus, increasing the availability of lixiviant (biogenic cyanide and
69
oxygen) will make important significance. However, investigations on mutual effect
70
between bacterial and metal particles are rarely conducted. Thus, potential
71
improvement strategies for high leaching efficiency are often overlooked. In our
72
previous study,3 the medium components used in cyanide production were optimized,
73
and a novel model was developed to predict cyanide production. However,
74
bioleaching kinetics are often determined by diffusion process.17 In this paper, 4
ACS Paragon Plus Environment
Page 4 of 29
Page 5 of 29 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
ACS Sustainable Chemistry & Engineering
75
two-step bioleaching was conducted and contact behavior between bacteria and metal
76
particles was studied. Effect of PVP on enhancing metal leaching was reported and
77
the potential mechanisms were discussed.
78 79 80
METHODS AND MATERIALS
81
Materials
82
In the present work, pure silver particles instead of crushed WPCBs were used to
83
simulate the bioleaching process. Ag powder (200 < mesh < 300, purity 99.99%) was
84
obtained from Changsha Tian Jiu Metal Material Co., Ltd. (Changsha, China). PVP
85
(average molecular weight 24,000 Da) was purchased from Aladdin Chemistry Co.,
86
Ltd. (Shanghai, China). Tryptone and yeast extract were obtained from Oxoid Ltd.
87
(Hampshire, UK). Other reagents used were all of GR grade. Distilled water was used
88
throughout the experiment.
89
Microorganism and culture conditions
90
Pseudomonas fluorescens P13,18 was used for cyanide production and silver
91
bioleaching. Cells were mixed in potassium phosphate buffer containing 50% glycerol
92
and stored at –20 °C. It was revived in 250 mL shaken flasks containing 60 mL of
93
Luria–Bertani (LB) medium (10 g/L tryptone, 5 g/L yeast extract, and 10 g/L NaCl).
94
Log phase bacteria (optical cell density at 600 nm wavelength ~0.6) was sampled and
95
inoculated in 250 mL flasks each containing 60 mL of the cyanogenic medium and 5
ACS Paragon Plus Environment
ACS Sustainable Chemistry & Engineering 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
96
with 5% (v/v) inoculum. The cyanogenic medium contains 6 g/L tryptone and 5 g/L
97
yeast extract.3 All the flasks used in the cell preparation and cyanide production were
98
shaken at 150 rpm and 25 °C. These conditions could produce the maximum
99
concentration of cyanide.
100
Leaching experiments
101
Two-step bioleaching was conducted. Bioleaching experiments of silver particles
102
was performed in flasks. 100 mg of silver particles were fed into 250 mL flasks. Then
103
the flasks were autoclaved at 121 °C for 15 min. Based on our previous study (ref. 3),
104
cyanide amount in the cultures reached the maximum (approximately 7 mg/L) after
105
15–18 hours of incubation (during early stationary phase). Thus, 50 mL of the cultures
106
cultivated for 15 hours were transferred into the Ag containing flasks. To investigate
107
the effect of PVP on silver leaching, PVP was immediately added into the flasks to a
108
final concentration of 240 mg/L. The flasks without PVP addition acted as control
109
group. The leaching condition was 25 °C and rotation speed of 200 rpm. After 96
110
hours, the biologically leached solutions were sampled and centrifuged at 10,000 rpm
111
10 min. The supernatant was passed through a membrane filter with pore size 0.45 µm.
112
The filtrate was digested in triplicates using nitric acid and perchloric acid. After acid
113
digestion, the metal composition was analyzed for Ag+ using Inductively Coupled
114
Plasma Optical Emission Spectrometer (ICP-OES, Perkin Elmer Optima 5300 DV).
115
Free cyanide and oxygen analysis
116
Free cyanide produced in the cultures was tested using 4-pyridinecarboxylic acid 6
ACS Paragon Plus Environment
Page 6 of 29
Page 7 of 29 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
ACS Sustainable Chemistry & Engineering
117
color comparison method (Pack Test, WAK-CN; Kyoritsu Chemical Check Lab. Corp.)
118
For details of cyanide measurement, please refer to ref. 3. Free oxygen of the leaching
119
cultures in the silver containing flasks was measured by oxygen meter (YSI 550A).
120
FTIR measurements
121
Infrared spectra of the cyanogenic bacteria were recorded on an FTIR instrument
122
(Nicolet, iS10) to identify the functional groups on the bacteria. Cultures at 3, 12, 48
123
hours of bioleaching were extracted and thoroughly dried. Sample was prepared using
124
the method described in ref. 19.
125
Contact angle measurements
126
The hydrophobicity of bacteria was determined by water contact angel
127
measurement on Dataphysics OCA40 Micro. Bacterial layers were obtained by
128
filtering the bacterial suspension on a 0.45 µm microporous membrane. Contact angle
129
was measured after air drying the bacterial sediments for about 10 min to remove
130
excess water. Three measurements were conducted, and the average value was
131
obtained.
132
Zeta potential measurements
133
The surface property of bacteria treated with PVP was investigated using zeta
134
potential measurements (Brookhaven Instruments Corporation, New York, NY, USA).
135
Bacteria at the end of exponential phase (15 hours of incubation) was harvested,
136
centrifuged at 5000 rpm for 5 min, and washed three times with saline. Then the
137
sedimented bacteria were resuspended in various concentrations of PVP solutions (30, 7
ACS Paragon Plus Environment
ACS Sustainable Chemistry & Engineering 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
138
60, 120, 240, 480 and 960 mg/L). The final optical cell densities of the suspension at
139
400 nm wavelength were approximately equal to 0.78, an appropriate bacterial
140
concentration in favor of zeta potential measurement. The pH of the suspension was
141
adjusted by 0.01 M NaOH or 0.01 M HNO3 solution to pH 7.9 (pH of bacterial
142
culture after 15 hours of incubation increased to 7.9). In addition, to investigate the
143
zeta potential change affected by silver addition, 100 mg of silver particles were
144
dispersed in 50 mL of the prepared suspension. All the samples were measured five
145
times and the average value was obtained.
146
Microscopic analysis
147
Optical microscopy imaging of silver particles was conducted using a
148
fluorescence microscope (Leica AF6000). Sampling was conducted at 3 min, 1 h, 2 h,
149
3 h, and 4 h after bioleaching started. Samples were extracted by the micropipet
150
technique and dropped on cleaned microscope slide. Coverslip was pressed on the
151
surface enough to disperse particles. Photomicrographs were taken immediately. The
152
transmitted light intensity was adjusted correspondingly to obtain optimum imaging
153
results.
154
SEM measurements
155
The collected bacterial cells (end log phase) were fixed with 2% glutaraldehyde at
156
room temperature. After eliminating the remaining glutaraldehyde, the dehydration
157
process was conducted with 20, 50, 80 and 100% of alcohol. The fixed cells was then
158
observed by scanning electron microscopy (SEM) (Quanta 400F). Silver particles 8
ACS Paragon Plus Environment
Page 8 of 29
Page 9 of 29 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
ACS Sustainable Chemistry & Engineering
159
were also determined by SEM.
160
To investigate the morphological properties of bacterial cells attached on silver
161
surface, 200 μL of leaching solutions (1-hour leaching) containing silver particles
162
were collected from the bottom of flasks. Then silver particles together with bacterial
163
solution were dropped on a tin foil and naturally air-dried. Images were obtained
164
using SEM.
165
Calculation of wet weight of bacteria
166
Force analysis of bacteria absorbed on silver surface was performed assuming
167
that bacteria does not move anymore when it starts to dehydrate. Thus bacteria density
168
was approximately equal to culture density. Wet weight of bacteria was estimated by
169
multiplying bacteria size by culture density. Bacteria size was estimated from
170
measurements of length and width at high magnification (20,000 fold enlargement,
171
Figure 1A). Culture density was obtained by determining the quality of a certain
172
volume of bacterial culture.
173 174
RESULTS AND DISCUSSION
175
A typical example of P. fluorescens in log phase was shown in Figure 1A. P.
176
fluorescens is rod shaped and has size of 0.5 µm (width) × 0.8–1.5 µm (length).
177
According to ref. 20, the size of P. fluorescens during the exponential phase was
178
0.7–0.8 µm × 2.3–2.8 µm, which decreased with age. In addition, growth of P.
179
fluorsescens generally experienced a lag phase (approximately 0-2 hours of 9
ACS Paragon Plus Environment
ACS Sustainable Chemistry & Engineering 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
180
incubation), a log phase (2-15 h), and a stationary phase (15-24 h).3 Figure 1B showed
181
the image of silver particles (round shaped, ~50 µm in diameter) used in this
182
investigation. For the density of silver is significantly greater than the density of
183
bacterial culture, silver particles would gravitate toward the bottom of the flasks. At
184
high shaking speed (150–200 rpm), silver particles tended to cluster at the center of
185
the flask’s bottom (Figure 2A). At lower agitation speed, silver particles would spread
186
out from the center and form a circle (Figure 2B). This behavior was obviously not
187
conducive to rapid leaching reaction. Thus, specially designed reactor such as an
188
end-to-end tubular allowing full contact of solid solution interface was recommended
189
in further investigation of bioleaching. Most importantly, in the current work, we
190
presented an alternative method for improving availability of cyanide lixiviant for the
191
settling silver particles, namely addition of PVP, as will be shown in the next
192
paragraphs.
193
Optical microscopy images of silver particles sounded by bacteria
194
Further observations on aggregate behavior of silver particles were performed
195
using optical microscopy. Figure 3 showed the microscopic view of the distributed
196
silver particles immersed in the bacterial bath. In its entirely, silver particles exhibited
197
high propensity to agglomerate due to the high interfacial energy. Figures 3A and 3B
198
showed there was little cellular material around the silver particles compared with
199
Figures 3D, and 3E, which were imaged after 1 and 2 h of bioleaching, respectively.
200
The dispersed silver particles in Figure 3B were likely caused by covering the 10
ACS Paragon Plus Environment
Page 10 of 29
Page 11 of 29 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
ACS Sustainable Chemistry & Engineering
201
coverslip. A magnified view of Figure 3B, namely (Figure 3C), showed silver
202
particles were continuously surround by bacteria and it implied there existed specific
203
adsorption of bacteria on silver particles. The brown and opaque bacterial fragments
204
in Figures 3D and 3E were thought to be aggregated form of biofilm. Biofilm is
205
commonly composed of structured micro-colonies and exopolysaccharide products. In
206
a biofilm, microbes are usually irreversibly bind to the biofilm surface and
207
encapsulated into a exopolysaccharide matrix.21 Closer observation of the stuck
208
particles (Figure 3F) showed the bacterial agglomerates consisted of loosely attached
209
EPS and bacteria. Acceleration of bacterial decline reinforced the biofilm formation
210
and exhibited tight adhesion to silver particles (Figure 3G). Figure 3H showed the
211
inner area of Figure 3G, double layer of silver particles formed as compared with that
212
showed in Figure 3A. In Figure 3H, the average spacing between particles in the
213
upper layer was smaller than that observed in Figure 3I. Increased EPS formation was
214
the primary cause of particle agglomeration. The production of EPS might be a
215
natural physiological response to the deterioration of the growth environment.
216
However, it caused undesirable effects on bioleaching.
217
SEM micrograph of bacteria attachment and force analysis
218
The definition, production, and function of EPS from microbes were extensively
219
studied. Here, only morphologies of EPS and bacteria adsorbed on the particles were
220
examined using SEM (Figure 4).Few materials existed on the top of silver particles
221
(Figures 4A and 4B). Emergence of surface roughening from the middle to the top 11
ACS Paragon Plus Environment
ACS Sustainable Chemistry & Engineering 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
222
half part of the particles might be attributed to the accretion of the downward
223
translocation of the attached bacterial material. Figures 4C and 4D showed that the
224
cells were monolayerly adsorbed on the upper hemispherical surface of silver particle.
225
The static friction that associated with bacteria attachment played a dominant role and
226
was analyzed in Figure 5. The calculated wet weight of a single bacterium ranged
227
from 0.159 × 10−15 to 0.299 × 10−15 kg, approximately 1.6 × 10−6 to 2.9 × 10−6 nN (g
228
= 9.8 N/kg). Magnitude of static friction force F1 = G = mg × cosθ. m refers to the wet
229
weight of bacteria. The static friction and gravity of the same bacteria were within the
230
same order of magnitude (0 < cosθ < 1). And it (about 10−6 nN) was far less than that
231
could be measured by atomic force microscopy (AFM). The forces measured by AFM
232
generally changed within ± 10 nN(+ represents the repulsion force, – represents the
233
attraction force) during the bacterial approach and retraction from the mineral (metal)
234
surface.22–24 Though the force interacted between bacteria and silver surface was very
235
little, it significantly influenced the adsorption, and reliable method could be found to
236
decrease the interaction. The approximate analysis might provide some useful
237
information in understanding the bacteria-material interaction.
238
Contact angle of bacteria
239
The aforementioned results demonstrated that the attachment greatly increased
240
due to the formation of biofilm during bioleaching. The contact angle of bacteria
241
before and after bioleaching decreased from 13.5 ± 2.2 to 10.7 ± 1.6 °.It was
242
demonstrated that bacterial hydrophilicity was slightly enhanced. Thus the force 12
ACS Paragon Plus Environment
Page 12 of 29
Page 13 of 29 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
ACS Sustainable Chemistry & Engineering
243
driving bacteria to silver surface might increase. In addition, silver dissolution might
244
give suitable sites for bacteria adhesion. These bio-chemical mechanisms would
245
enhance the surface passivity of silver during bioleaching.
246
Surface characteristic of P. fluorescens during bioleaching
247
The functional groups in P. fluorescens during bioleaching were investigated using
248
FTIR analysis to identify the substances that associated with adsorption (Figure 6).
249
All the spectrums showed similar absorption feature, which indicates the similar EPS
250
composition of the bacteria. The adsorption peak ranged from 3300 to 3500 cm−1
251
generally contained the characteristic peaks of the amine groups (−NH, from proteins)
252
and hydroxyl groups (−OH, from saccharides).19 But in this study it suffered a
253
significantly intensity loss revealing PVP had strongly interacted with bacterial
254
surface. The adsorption peaks at 2925 and 2960 cm−1, at 1456 cm−1 were assigned as
255
the −CH and –CH2 groups, respectively, and both are common components in the
256
phospholipid, peptidoglycan, and teichoic acid of bacteria.25 The peaks at 1654 cm−1
257
for the 3 h bacterial leaching, 1650 cm−1 for 12 h, 1650 cm−1 for 48 h were
258
characteristic of the amide groups (−CON−). The peaks around 1455 cm−1 were
259
regarded as amine groups (−NH−, −CN−, and peptidoglycan).19 The peaks around
260
1400 and 1312 cm−1 were assigned to the carboxylic acid groups (−COOH). In
261
addition, the unchanged peaks at 1238 and 1083 cm−1 represented the asymmetric and
262
symmetric phosphate (−PO2). These identified polar functional groups were all
263
available for biosorption,19 although it was possible that surface adsorption was not 13
ACS Paragon Plus Environment
ACS Sustainable Chemistry & Engineering 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
264
the main mechanism. In general, bacterial adsorption and EPS binding to solid
265
substrates were an important process in bioleaching, no matter this behavior was
266
positive or negative. However, in the bioleaching of precious metals, inward cyanide
267
and oxygen diffusion and outward metal-cyanide complexation might be impeded due
268
to the biological components strongly attached to the metal surfaces. Thus, precious
269
metal bioleaching efficiency decreased.
270
Effect of PVP as an additive on silver leaching
271
Many efforts have been devoted to increase recovery efficiency of precious metal
272
regardless of the extent of diffusion control on bioleaching. In this work, the addition
273
of PVP was shown to be effective in enhancing the recovery of silver (Figure 7). It
274
almost increased silver recovery by 1.8 times. Further experiments showed that PVP
275
had no effect on cyanide production in two step bioleaching process (data not
276
shown).Meanwhile PVP was unable to dissolve silver. According to equation 1, silver
277
could be dissolved by cyanide with the assistance of oxygen. Appropriate ratio of
278
cyanide/oxygen would benefit reaction. At the beginning of bioleaching, cyanide
279
concentration was approximately 7 mg/L, and the free oxygen concentration in the
280
leaching solutions ranged from 6.1 to 6.7 mg/L. Oxygen concentration was not a
281
limiting factor for leaching. In addition, PVP addition might cause damage to
282
bacterial cells, but the impact would be negligible because cell density and cyanide
283
concentration had reached maximum in two-step bioleaching. Thus, alleviation of
284
diffusion control was the most likely to occur. 14
ACS Paragon Plus Environment
Page 14 of 29
Page 15 of 29 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
ACS Sustainable Chemistry & Engineering
285
4Ag + 8CN − + O 2 + 2H 2 O → 4Ag(CN) 2− + 4OH −
286
Possible mechanisms in enhancing bioleaching by PVP
(1)
287
Potential beneficial changes occurred in the bacteria-silver surface could explain
288
the enhanced bioleaching performance upon the addition of PVP. (1) Binding
289
characteristic. The components of EPS, such as proteins, glycoproteins, lipoproteins,
290
and polysaccharides, contain both hydrophobic and hydrophilic sites that enable the
291
adsorption of cells onto the particles to form a biofilm. PVP is capable of suppressing
292
nonspecific binding, and can also disperse biological units, such as mycelia.27 Zeta
293
potential of the suspensions containing bacteria and PVP, with and without silver were
294
shown in Figure 8. PVP is positively charged in aqueous solutions and tend to interact
295
with the negatively charged bacteria surface.28 Thus a large increase in zeta potential
296
was observed when high concentrations of PVP was added. When silver particles
297
were added, the zeta potential of bacterial suspensions containing 120, 240, and 480
298
mg/L of PVP all decreased. This effect might be attributed to PVP’s affinity for silver
299
particles, which liberated many negative sites on the bacterial surface and lead to a
300
slight zeta potential reduction.
301
PVP has a strong anion binding property and the binding constants were 0.5, 3.3,
302
and 5.3 for Cl−, I−, and SCN−, respectively.29 Hence, PVP was expected to suppress
303
the negative adsorption of bacteria and EPS and liberate active surface areas on silver.
304
As shown in Figure 9, the biofilms aggregated and did not mix with silver particles.
305
PVP might have acted as a bridge and enhanced the binding interaction to promote 15
ACS Paragon Plus Environment
ACS Sustainable Chemistry & Engineering 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
306
Ag−CN− contact. (2) Dispersion ability. PVP is a nonionic dispersant that commonly
307
serves as capping agent in nanomaterial manufacturing.30,31 In the bioleaching process,
308
PVP might reduce the contact of silver particles to each other by specific silver
309
binding. Silver particles were mainly located at the center at the bottom of the flask
310
(Figure 2A) with a few dispersed particles spinning around the center. Particle
311
dispersion would enhance the conversion rate of the reactive components. (3)
312
Increased viscosity of the solution. At room temperature, PVP has a major impact on
313
the viscosity of aqueous solutions.32,33 In this study, 240 mg/L of PVP was added. This
314
concentration was effective in increasing the culture viscosity. Fluid shear stress is
315
proportional to fluid viscosity when it reached a steady state.34 A simplified formula
316
suitable for describing the shear force in shaken flasks is shown as γ = τt/µL, where γ
317
is the average shear rate, τt is the average shear stress, and µL is the fluid viscosity.
318
Thus, shear stress was speculated to increase when PVP was added. Meanwhile it
319
would offer a strong abrasion at the solid/liquid interface, resulting in the attenuation
320
of diffusion control. Recovering precious metals from WPCBs with bioleaching
321
technology occurred in the presence of oxygen (O2) and cyanide (CN−). The primary
322
site of metal particle dissolution was at the solid-liquid interface, and the interface
323
was immediately formed upon immersing the metal particle in the culture. Adsorption
324
of cell and EPS on the particle’s surface prevents direct collision and contact of active
325
components, which results in undesirable effects for the metal leaching. Though PVP
326
is demonstrated effective in increasing metal recovery, the solid-liquid interfaces 16
ACS Paragon Plus Environment
Page 16 of 29
Page 17 of 29 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
ACS Sustainable Chemistry & Engineering
327
deserves more attention.
328 329 330
CONCLUSIONS
331
In this study, we performed two-step bioleaching process for silver recovery. Contact
332
behavior between cells and silver particles were observed. After bioleaching, water
333
contact angle of the hydrophilic bacteria showed a minor decrease. This variation in
334
wettability indicated an enhanced bacterial affinity for polar materials. The
335
characterization analyses of functional groups on bacterial surface showed the
336
presence of carboxyl, hydroxyl, and amine groups, which were responsible for the
337
bacterial adsorption. Further observation found that bacteria formed aggregates on the
338
surface of silver. Biofilm formation caused negative effect on silver leaching reaction.
339
Subsequently, it was found that leaching of silver could be enhanced by the addition
340
of PVP. Considering that the positively charged PVP (in aqueous media) interacted
341
strongly with silver particles and bacteria, the overall mechanism can be summarized
342
as: (1) PVP promoted CN- migrate to silver. (2) PVP suppressed the bacterial
343
attachment. (3) PVP improved the interface exchange.
344 345
ACKNOWLEDGEMENTS
346
This work was supported by the 111 Project (B18060), the National Natural Science
347
Foundation of China (Grant No. 51308488, 51741409), the Science and Technology 17
ACS Paragon Plus Environment
ACS Sustainable Chemistry & Engineering 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
of
Guangdong
Province
(Grant
No.
2015B020237005
Page 18 of 29
348
Programs
and
349
2016A020221014), the Pearl River Star of Science and Technology (Grant No.
350
201710010032) and the Fundamental Research Funds for the Central Universities
351
(Grant No. 17lgzd22).
352 353
AUTHOR CONTRIBUTIONS
354
Z.Y. and J.R. designed the experiments; Z.Y. performed the experiments; Z.Y. and J.R.
355
wrote the manuscript; H.Z., Y.L. and R.Q. revised the manuscript.
356 357
COMPETING FINANCIAL INTERESTS
358
The authors declare that they have no conflict of interest.
359 360
REFERENCES
361
(1) Oguchi, M.; Murakami, S.; Sakanakura, H.; Kida, A.; Kameya, T. A preliminary categorization
362
of end-of-life electrical and electronic equipment as secondary metal resources. Waste Manage.
363
2011, 31, 2150–2160.
364 365
(2) Zhang, Z. Y.; Zhang, F. S.; Yao, T. An environmentally friendly ball milling process for recovery of valuable metals from e-waste scraps. Waste Manage. 2017, 68, 490–497.
366
(3) Yuan, Z. H.; Ruan, J. J.; Li, Y. Y.; Qiu, R. L. A new model for simulating microbial cyanide
367
production and optimizing the medium parameters for recovering precious metals from waste
368
printed circuit boards. J. Hazard. Mater. 2018, 353, 135–141.
18
ACS Paragon Plus Environment
Page 19 of 29 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
ACS Sustainable Chemistry & Engineering
369 370 371 372 373 374 375 376 377 378
(4) Wang, X.; Gaustad, G. Prioritizing material recovery for end-of-life printed circuit boards. Waste Manage. 2012, 32, 1903–1913. (5) Cui, J.; Zhang, L. Metallurgical recovery of metals from electronic waste: A review. J. Hazard. Mater. 2008, 158, 228–256. (6) Ruan, J. J.; Zhu, X. J.; Qian, Y. M.; Hu, J. A new strain for recovering precious metals from waste printed circuit boards. Waste Manage. 2014, 34, 901–907. (7) Zhang, L. G.; Xu, Z. M. A review of current progress of recycling technologies for metals from waste electrical and electronic equipment. J. Clean. Prod. 2016, 127, 19–36. (8) Zhuang, W. Q.; Fitts, J. P.; Ajo-Franklin, C. M.; Maes, S.; Alvarez-Cohen, L.; Hennebel, T. Recovery of critical metals using biometallurgy. Curr. Opin. Biotech. 2015, 33, 327–35.
379
(9) Işıldar, A.; van de Vossenberg, J.; Rene, E. R.; van Hullebusch, E. D.; Lens, P. N. L. Two-step
380
bioleaching of copper and gold from discarded printed circuit boards (PCB). Waste Manage. 2016,
381
57, 149–157.
382 383 384 385
(10) Natarajan, G.; Ting, Y. P. Pretreatment of e-waste and mutation of alkali-tolerant cyanogenic bacteria promote gold biorecovery. Bioresource Technol. 2014, 152, 80–85. (11) Brandl, H.; Bosshard, R.; Wegmann, M. Computer-munching microbes: metal leaching from electronic scrap by bacteria and fungi. Hydrometallurgy. 2001, 59, 319–326.
386
(12) Shin, D.; Park, J.; Jeong, J.; Kim, B.-s. A biological cyanide production and accumulation
387
system and the recovery of platinum-group metals from spent automotive catalysts by biogenic
388
cyanide. Hydrometallurgy. 2015, 158, 10–18.
389
(13) Natarajan, G.; Ting, Y. P. Gold biorecovery from e-waste: An improved strategy through
19
ACS Paragon Plus Environment
ACS Sustainable Chemistry & Engineering 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
390 391 392 393 394
spent medium leaching with pH modification. Chemosphere. 2015, 136, 232–238. (14) Pradhan, J. K.; Kumar, S. Metals bioleaching from electronic waste by Chromobacterium violaceum and Pseudomonads sp. Waste Manage. Res. 2012, 30, 1151–1159. (15) Tay, S. B.; et al. Enhancing gold recovery from electronic waste via lixiviant metabolic engineering in Chromobacterium violaceum. Sci. Rep. 2013, 3, 2236.
395
(16) Mishra, D.; Kim, D. J.; Ralph, D. E.; Ahn, J. G.; Rhee, Y. H. Bioleaching of spent
396
hydro-processing catalyst using acidophilic bacteria and its kinetics aspect. J. Hazard. Mater.
397
2008, 152, 1082–1091.
398
(17) Motaghed, M.; Mousavi, S. M.; Rastegar, S. O.; Shojaosadati, S. A. Platinum and rhenium
399
extraction from a spent refinery catalyst using Bacillus megaterium as a cyanogenic bacterium:
400
Statistical modeling and process optimization. Bioresource Technol. 2014, 171, 401–409.
401
(18) Yang, L.; et al. Promotion of plant growth and in situ degradation of phenol by an
402
engineered Pseudomonas fluorescens strain in different contaminated environments. Soil Biol.
403
Biochem. 2011, 43, 915–922.
404
(19) Cui, J.; Zhu, N.; Kang, N.; Ha, C.; Shi, C.; Wu, P. Biorecovery mechanism of palladium as
405
nanoparticles by Enterococcus faecalis: From biosorption to bioreduction. Chem. Eng. J. 2017,
406
328, 1051–1057.
407
(20) Karthikeyan, O. P.; Rajasekar, A.; Balasubramanian, R. Bio-oxidation and biocyanidation of
408
refractory mineral ores for gold extraction: A review. Crit. Rev. Env. Sci. Tec. 2014, 45,
409
1611–1643.
410
(21) Donlan, R. M. Biofilms: microbial life on surfaces. Emerg. Infect. Dis. 2002, 8, 881–890.
20
ACS Paragon Plus Environment
Page 20 of 29
Page 21 of 29 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
ACS Sustainable Chemistry & Engineering
411 412 413 414 415 416
(22) Zhu, J.; et al. Insights into the relation between adhesion force and chalcopyrite-bioleaching by Acidithiobacillus ferrooxidans. Colloid Surface B. 2015, 126, 351–357. (23) Lower, S. K.; Hochella, M. F.; Beveridge, T. J. Bacterial recognition of mineral surfaces: nanoscale interactions between Shewanella and α-FeOOH. Science. 2001, 292, 1360–1363. (24) Sheng, X.; Ting, Y. P.; Pehkonen, S. O. Force measurements of bacterial adhesion on metals using a cell probe atomic force microscope. J. Colloid Interf. Sci. 2007, 310, 661–669.
417
(25) Jiang, W.; Saxena, A.; Song, B.; Ward, B. B.; Beveridge, T. J.; Myneni, S. C. B. Elucidation
418
of functional groups on gram-positive and gram-negative bacterial surfaces using infrared
419
spectroscopy. Langmuir. 2004, 20, 11433–11442.
420 421
(26) Haycock, J. W. Polyvinylpyrrolidone as a blocking agent in immunochemical studies. Anal. Biochem. 1993, 208, 397–399.
422
(27) Archer, D. B.; MacKenzie, D. A.; Ridout, M. J. Heterologous protein secretion by
423
Aspergillus niger growing in submerged culture as dispersed or aggregated mycelia. Appl.
424
Microbiol. Bio. 1995, 44, 157–160.
425
(28) Zhai, L.; Lu, X.; Chen, W.; Hu, C.; Zheng, L. Interaction between spontaneously formed
426
SDBS/CTAB vesicles and polymer studied by fluorescence method. Colloid Surface A. 2004, 236,
427
1–5.
428 429
(29) Song, J. D.; Ryoo, R.; Jhon, M. S. Anion binding properties of poly(vinylpyrrolidone) in aqueous solution studied by halide NMR spectroscopy. Macromolecules. 1991, 24, 1727–1730.
430
(30) Aherne, D.; Ledwith, D. M.; Gara, M.; Kelly, J. M. Optical properties and growth aspects of
431
silver nanoprisms produced by a highly reproducible and rapid synthesis at toom temperature. Adv
21
ACS Paragon Plus Environment
ACS Sustainable Chemistry & Engineering 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
432
Funct. Mater. 2008, 18, 2005–2016.
433
(31) Huynh, K. A.; Chen, K. L. Aggregation kinetics of citrate and polyvinylpyrrolidone coated
434
silver nanoparticles in monovalent and divalent electrolyte solutions. Environ. Sci. Technol. 2011,
435
45, 5564–5571.
436 437
(32) Loftsson, T.; Frikdriksdóttir, H.; Sigurkdardóttir, A. M.; Ueda, H. The effect of water-soluble polymers on drug-cyclodextrin complexation. Int. J. Pharm. 1994, 110, 169–177.
438
(33) Mokkapati, V.; Koseoglu-Imer, D. Y.; Yilmaz-Deveci, N.; Mijakovic, I.; Koyuncu, I.
439
Membrane properties and anti-bacterial/anti-biofouling activity of polysulfone-graphene oxide
440
composite membranes phase inversed in graphene oxide non-solvent. RSC Adv. 2017, 7,
441
4378–4386.
442 443
(34) Koller, A.; Sun, D.; Kaley, G. Role of shear stress and endothelial prostaglandins in flowand viscosity-induced dilation of arterioles in vitro. Circ. Res. 1993, 72, 1276–1284.
444 445 446
22
ACS Paragon Plus Environment
Page 22 of 29
Page 23 of 29 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
ACS Sustainable Chemistry & Engineering
447
FIGURES
448 449
Figure 1. SEM images of P. fluorescens (A) and silver particles (B)
450 451
452 453
Figure 2. Distribution of silver particles in the flasks. To clearly observe the particle movement,
454
the bacteria were removed through centrifugation. The arrow points to the clarified solution
455 456 457
23
ACS Paragon Plus Environment
ACS Sustainable Chemistry & Engineering 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
458 459
Figure 3. Micrograph of bacterial and silver particle mixtures sampled at different time points
460 461
24
ACS Paragon Plus Environment
Page 24 of 29
Page 25 of 29 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
ACS Sustainable Chemistry & Engineering
462 463
Figure 4. SEM micrograph of naturally dried silver particles
464 465 466 467
25
ACS Paragon Plus Environment
ACS Sustainable Chemistry & Engineering 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
468 469
Figure 5. Simulation of static friction force of adsorbed bacteria
470 471
472 473
Figure 6. FTIR spectra of P. fluorescens
474 475
26
ACS Paragon Plus Environment
Page 26 of 29
Page 27 of 29 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
ACS Sustainable Chemistry & Engineering
476 477
Figure 7. Bioleaching of silver with and without the addition of PVP
478 479
480 481
Figure 8. Zeta potential of the bacterial solutions affected by the addition of PVP and silver
482
particles
483 484 485
27
ACS Paragon Plus Environment
ACS Sustainable Chemistry & Engineering 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
486 487
Figure 9. Effects of PVP on silver particle distribution
488 489
28
ACS Paragon Plus Environment
Page 28 of 29
Page 29 of 29 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
ACS Sustainable Chemistry & Engineering
490
TOC graphic
491 492
Recovery precious metals from WPCBs is a meaningful work in the area of
493
sustainable development of the world.
494
29
ACS Paragon Plus Environment