Mineral Bioflotation using the

Rhodococcus opacus bacteria

Antonio Gutirrez Merma
Carlos Alberto Castaeda 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
Increasing demand for environmental
regulations
Wastewater and effluents
Chemical reagents

Mineral processing problems

Increasing demand for phosphate rocks

and deposit depletion, processing:
complex ores
flotation tailings
Fine mineral particles

Development of environmentally
friendly technologies

Biotechnologies
Effluents treatment

Heavy metals uptake

Mineral Bioprocessing

Bioleaching

Mineral biobeneficiation

Biofloculation

Bioflotation

1. INTRODUCTION: Fundamentals
Water Chemistry
Speciation Diagrams
Water quality

Chemical dissolution and

precipitation

Surface
Chemistry
Froth

Mineral Technology
Flotation

Zeta potential

Mineral
Biotechnology
Environmental
Bioprocessing
Thermodynamics,
Adsorption fundamentals

Ore and tailings

processing

Mineral and metal

extraction

Biochemical
Engineering

Transport
phenomena
interactions

Biology
Biomass production
Structure of biomass
and bioreagents

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 Dupr

Equation of Young

Equation of Young-Dupr

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

apatite

apatite

Schematic diagram of the mineral preparation

Purity

97.9

P2O5

39.96

CaO

53.14

CaO/ P2O5

1.285

Quartz

1.5

Talc

0.5

Growth medium
YMG

Solid

Liquido(g

(g/L)

/L)

Glucose

10

10

Peptone

Malt extract

Yeast extract

Agar

20

pH

6.6

6.6

Component

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 medium.

Autoclave
Chapel Filter Flux

1 growth , liquid
medium (24 h)
Shaker growth

2 growth, liquid
medium (48 h)

Washing, concentration
and inactivation of the
biomass

Calibration curve

Centrifuge

Concentration
determination

Spectrophotometer UV 1800

Cellular concentrate

Experimental
1. Electrophoretic
mobility

3. Microflotation

4. SEM

Zeta potential determination

of mineral samples before
and after bacterial
interaction

pH solution and
bacterial concentration
effect;
Flotability evaluation

Zetameter +4.0

Zetameter - cell

Identification of the
bacteria adhesion

Microflotation system, Hallimond tube

SEM, FEI Quanta 400

Zeta
potential
measureme
nts

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
Apatite-adapted
Quartz-adapted

Zeta Potential (mV)

10
0
-10

IEP bacterial cells: 2.8

-20

IEP apatite-adapted: 3.0

IEP quartz-adapted: 3.4

-30
-40

10

pH
Figure 1. Zeta potential of R. opacus. NaCl 10-3M as background
electrolyte. Bacterial concentration, 0.10 g/L.

12

Zeta Potential
R. opacus
Quartz before contact
Quartz after contact
Apatite before contact
Apatite after contact

10

Zeta Potential (mV)

0
-10
-20

IEP bacterial cells: 2.8

-30
-40
-50
2

10

pH

12

R. opacus strain obtained from CBMAI-UNICAMP, is a non-pathogenic, Gram-positive and chemoorganotrophic organism, with a high hydrophobicity (con

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
(+75-106) micras
(+106-150) micras

18

Flotability (%)

16
14
12
10
8
6
4
2
0

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
(+38-75) micras
(+75-106) micras
(+106-150) micras

70

Flotability (%)

60
50
40
30
20
10
0

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
Adapted to apatite
Adapted to quartz

36

Flotability (%)

32
28
24
20
16
12
8
4
0

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
Adapted to apatite
Adapted to quartz

Flotability (%)

60
50
40
30
20
10
0

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 mineraladapted 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 Tecnolgico.

CAPES Coordenao de Aperfeioamento de Pessoal de Nvel Superior.

FAPERJ Fundao de Amparo Pesquisa do Estado do Rio de Janeiro.

Vale company trough to Edital CT-Mineral- VALE,

Many thanks !
Carlos Castaeda (caralcaso@gmail.com)
Antonio Gutirrez (anguz21@hotmail.com)
Maurcio Torem (torem@puc.rio-br)