Mineral Bioflotation using the

Rhodococcus opacus bacteria

Antonio Gutiérrez Merma
Carlos Alberto Castañeda Olivera
Mauricio Leonardo Torem
Departament of Materials Engineering– PUC-Rio
Rio De Janeiro, Agosto 2014
 SYNOPSIS

1. Introduction
2. Objectives and Relevance
3. Materials and Methods
4. Results and Discussion
5. Conclusions
1. INTRODUCTION Mineral processing problems

Increasing demand for phosphate rocks and deposit

Increasing demand for environmental regulations depletion, processing:
• Wastewater and effluents • complex ores
• Chemical reagents • flotation tailings
• Fine mineral particles

Development of environmentally friendly

technologies

Biotechnologies

Effluents treatment Mineral Bioprocessing

Bioleaching Mineral biobeneficiation

Heavy metals uptake

Biofloculation Bioflotation
1. INTRODUCTION: Fundamentals
Water Chemistry

Speciation Diagrams
Water quality

Chemical dissolution and

precipitation Biochemical
Surface
Zeta potential Engineering
Chemistry Mineral
Biotechnology
Froth
Mineral Technology
Flotation Environmental
Bioprocessing
Thermodynamics, Biology
Adsorption fundamentals Biomass production

Ore and tailings

Structure of biomass
processing Transport and bioreagents
phenomena
interactions
Mineral and metal
extraction
1. INTRODUCTION

Bacteria of the genus Rhodococcus

 The Rhodococcus are aerobic actinobacteria, gram-positive, with high content of guanine
and cytosine contained in their DNA.

 They are widely distributed in the environment and live in various ecological niches such
as marine environments, Alpine soils, Arctic, Antarctic, in groundwater, guts of insects and
from healthy and diseased animals and plants.
1. INTRODUCTION

Gram positive and gram negative bacteria cell wall (Wiley et al., 2011)
1. INTRODUCTION

Rhodococcus opacus: contact angle = 70°

Equation of Dupré

 Δ 𝐺=𝛾 𝑆𝐺 −(𝛾 𝑆𝐿 +𝛾 𝐿𝐺 )

Equation of Young

𝛾  𝑆𝐺 =𝛾 𝑆𝐿 +𝛾 𝐿𝐺 . 𝑐𝑜𝑠 𝜃

Equation of Young-Dupré

 
1. INTRODUCTION

Bacterial adhesion

Attachment modes of bacteria on the surface of the

mineral (Li, yubiao et al., 2013).
 Bioflotation
Requisites of Biomass as a collector

 Adhesion to mineral surface:

 Hydrophobic behavior:
• Superficial characteristics;
• Presence of functional groups;
• Proteins and polysaccharides

Biomass
 Bacteria;
 Yeast;
 Seaweed;
 Fungus.

MEV images of (A) Hematite and (B) Quartz particles flotated with R.
opacus.

Fonte: Mesquita et al., 2003

 2. OBJECTIVES

General objective:
Study the mineral bioflotation of quartz and apatite the Rhodococcus opacus bacteria as bioreagent. This study includes the
effect of the cellular adaptation on the mineral flotability
3. MATERIALS AND METHODS
Preparation of the mineral samples
Mineral

Crushing

X-Ray
Homogenization Head
Fluorescence

Sample Collecting apatite

Crushing and milling Contac angle Purity 97.9

(5 x 5 x 10) mm
P2O5 39.96
Screening

CaO 53.14
Mineral Mineral Mineral Mineral
+ 104 µm, - 152 µm +75 µm, -104 µm +37 µm, -75 µm +20 µm, -37 µm CaO/ P2O5 1.285
Sample Collecting Quartz 1.5

Chemical and mineralogical Zeta potential apatite Talc 0.5

characterization experiments

X-Ray X-Ray Flotation

Fluorescence diffraction experiments

Schematic diagram of the mineral preparation

Growth medium
YMG

Component Solid (g/L) Liquido(g/L)

Glucose 10 10
Peptone 5 5
Malt extract 3 3
Yeast extract 3 3
Agar 20 -
pH 6.6 6.6
Microbial growth

This graph represents the four-phase pattern of growth of a microbial population

Preparation of the bacterial concentrate

R. Opacus strain

R.Opacus strain

Preparation and sterilization of the culture

Autoclave
medium.
Chapel– Filter Flux

Calibration curve
1º growth , liquid medium (24 2
h) y = 3,5978x 3 - 5,7393x 2 + 5,0952x - 0,0163

A b so r b a n c e
1,6 R² = 0,9995
Shaker – growth
1,2

0,8
2º growth, liquid medium (48 0,4
h) Calibration curve
0
0 0,1 0,2 0,3 0,4 0,5 0,6
Cellular Concentration (g/L)

Washing, concentration and

inactivation of the biomass
Centrifuge

Concentration determination
Cellular concentrate
Spectrophotometer UV -
1800
Experimental

Zeta potential determination of mineral

samples before and after bacterial
interaction Zetameter +4.0
1. Electrophoretic mobility

Zetameter - cell
pH solution and bacterial
3. Microflotation concentration effect;
Flotability evaluation

Identification of the bacteria

4. SEM adhesion

Microflotation – system, Hallimond tube

SEM, FEI Quanta 400
Zeta potential
measurements

Electrical double layer (Abd Karim Alias 2013)

The evaluation of the zeta potential profiles for the minerals was carried
out before and after interaction with the R.opacus biomass suspension,
and the pH was adjusted using HCl and NaOH concentrations.
4. RESULTS AND
DISCUSSION
Zeta Potential
Rhodococcus opacus Bacteria
Bacterial cells
10 Apatite-adapted
Quartz-adapted
Zeta Potential (mV)

-10
IEP bacterial cells: 2.8
-20 IEP apatite-adapted: 3.0

-30
IEP quartz-adapted: 3.4

-40
2 4 6 8 10 12
pH

Figure 1. Zeta potential of R. opacus. NaCl 10-3M as background

electrolyte. Bacterial concentration, 0.10 g/L.
Zeta Potential

R. opacus
Quartz before contact
10 Quartz after contact
Apatite before contact
Apatite after contact
0
Zeta Potential (mV)
-10

-20
IEP bacterial cells: 2.8
-30

-40

-50

2 4 6 8 10 12
pH
R. opacus strain obtained from CBMAI-UNICAMP, is a non-pathogenic, Gram-positive and chemoorganotrophic organism, with a high hydrophobicity (contact angle arou

Figure 2. Zeta potential of quartz and apatite before and after

interaction with R. opacus cells. NaCl 10-3M as background
electrolyte. Bacterial concentration, 0.10 g/L.
Microflotation
Effect of the particle size
20 Particle size range
(+38-75) micras
18 (+75-106) micras
(+106-150) micras
16
14
Flotability (%)

12
10
8
6
4
2
0
2 4 6 8 10 12
pH
Figure 3. Quartz microflotation as a function of pH and particle size; flotation
time: 2 min; Bacterial concentration: 0,15 g/L.
Microflotation
Effect of the particle size

Particle size range

70 (+38-75) micras
(+75-106) micras
(+106-150) micras
60
Flotability (%)

50

40

30

20

10

0
2 4 6 8 10 12
pH
Figure 4. Apatite microflotation as a function of pH and particle size; flotation
time: 2 min; Bacterial concentration: 0,15 g/L.
Microflotation:
Cellular adaptation
40 R. opacus cells
N/A
36 Adapted to apatite
Adapted to quartz
32
Flotability (%)

28
24
20
16
12
8
4
0
2 4 6 8 10 12
pH
Figure 5. Quartz microflotation as a function of pH using the bacterial cells
adapted to a mineral substrate; flotation time: 2 min; Bacterial concentration:
0,15 g/L.
Microflotation:
Cellular adaptation
70 R. opacus cells
N/A
60 Adapted to apatite
Adapted to quartz
50
Flotability (%)

40

30

20

10

0
2 4 6 8 10 12
pH
Figure 6. Apatite microflotation as a function of pH using the bacterial cells
adapted to a mineral substrate; flotation time: 2 min; Bacterial concentration:
0,15 g/L.
SEM: Scanning electron microscope studies

Figure 7. Scanning electron figures of quartz particles floated, showing the R.

opacus cells adsorbed onto the mineral surface.
SEM: Scanning electron microscope studies

Figure 8. Scanning electron figures of green apatite particles floated, showing

the R. opacus cells adsorbed onto the mineral surface.
 5. CONCLUSIONS
• The change observed on the zeta potential profile of the bacterial cells after cellular adaptation are an indicative of an
alteration of the functional groups present at the wall-cell. These studies also showed an adhesion of the bacterial cells on
the mineral surface.
• The flotability of both minerals was higher at a pH value of 5. The flotability of quartz increase as the particle size was
reduced, while the flotability of apatite present an opposing behavior.
• It was observed that the flotability of both minerals presented an increase when the mineral-adapted cells were used,
moreover, it was observed a kind of selectivity, thus, the use of the apatite-adapted cell increase the flotability of the
apatite and the use of the quartz-adapted cell increase the flotability of the quartz, this increase was higher at pH 3.
• This work demonstrated the potential of the Rhodococcus opacus bacteria on the mineral processing.
 Acknowledgements
 CNPq – Conselho Nacional de Desenvolvimento Cientifico e Tecnológico.
 CAPES – Coordenação de Aperfeiçoamento de Pessoal de Nível Superior.
 FAPERJ – Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro.
 Vale company trough to Edital CT-Mineral- VALE,

Many thanks !
Carlos Castañeda (caralcaso@gmail.com)
Antonio Gutiérrez (anguz21@hotmail.com)
Maurício Torem (torem@puc.rio-br)