Arsenic removal by native

and chemically modified

lactic acid bacteria

By: disson Abdiel Rodrguez

 Abstract

Arsenic removal Aminated Lactobacillus

Native and chemically
capacity of three casei, remove As(V) but
modified forms
Lactobacillus not As(III) in water

Aminated methylated
pH
Study factors Concentration
Contact time
Tomado de: https://genome.jgi.doe.gov/lacca/lacca.home.html
Introduction
Why As?

China and India

Global health problem Rural areas of Finland
and USA

Tomado de: https://www.ecologiaverde.com/veinte-millones-chinos-

corren-riesgo-morir-envenenados-arsenico/

WHO < 10 g/L

 Introduction

Acidolactic bacteria (LAB)

Tomado de: http://aspergillusnigerhongo.blogspot.com.co/2010/10/bacterias-

utiles-en-la-elaboracion-de.html

LAB effectively remove cationic heavy metals,

Negatively charged arsenic
cadmium and lead. Cadmium and lead removal
in water may be more
was observed to occur rapidly to bacterial surface,
complicated
probably by an ion exchange mechanism

Tomado de: http://www.monografias.com/trabajos59/contaminacion-

arsenico/contaminacion-arsenico2.shtml
Material and methods
1) Bacterial strains and culture conditions 2) Chemical modification of
lyophilized bacterial cultures
All Methylated

methanol with 0,1 M HCl

L. casei L. crispatus L. acidophilus Incubated with agitation for 24 h at room temperature

culture conditions L. casei Aminated

Time (h) 24 and 40 crispatus

Temperature (C) 37
Lyophilized and
Stored (C) -20 0,5 M ethylenediamine and 0,2 M carbodiimide
MRS/ anaerobic
Medium conditions
5 g/l of bacteria was stirred (500 rpm) under N2 for 6 h
at room temperature.
The pH was adjusted to 4,75 with dilute HNO3
Material and methods
3) Characterization of the bacteria

Zeta potential () The efficiency of methylation

Potentiometric titration
Microelectrophoresis (Zetasizer 3000)
Comparison Native and methylated bacteria

Lyophilized bacteria were suspended in 1 mM KNO3
pH adjustment with dilute HNO3 or NaOH
Suspension was diluted to final concentration of about 108 cells/ml.
The pH range studied was from 2 to 12.
Lyophilized bacteria were suspended in 0,01 mM KNO3 to final
give a concentration of 5 g/L.
KNO3 had been purged with N2 for several hours in order to
remove solubilized CO2.
After pH adjustment to ,5 with dilute HNO3.
The suspension was titrated with 0.1 M KOH to pH 11 under N2.
 Material and methods
Effect of metal concentration
4) Binding assay
L. casei aminated
L. casei L. crispatus L. acidophilus

L. casei
0,1 to 5 mg/L pH =7 / time = 5 min

Assay conditions
Time (h) 24 1,5 mL was taken from the Desorption of bound As(V)
Temperature (C) 22 suspension. The sample was
pH 3,5,7 centrifuged (7000 rpm, 5 min), and Pellets of aminated L. casei
Lyophilized (g/L) 1 1,2 mL of the supernatant was
preserved with 100 L of ultra pure Ultra pure water, 1,5 mM HNO3 and 1,5 mM NaOH.
Metal concentration HNO3 (X 3)
10
(mg/L) Assay conditions pH =7 - time =5 min - As(V) =0,5 mg/L T=22C
Desorption conditions pH= 7 - time= 10 min - T= 22C
(11000 rpm x 10 min) 1,2 mL sample was taken
Material and methods

5) Metal analysis

As (V) was reduced to As(III) by

adding 1 mL 37 % and 0,6 mL 20%
Arsenic AAS (HG-AAS) (w/v) KI per 10 ml of sample and
concentrations
incubating the samples for 3 h at
22 C.
SpectrAA-300

VGA-76 Vapour
Generation Accessory

Tomado de: http://www.labwrench.com/?equipment.view/equipmentNo/14625/Varian/SpectrAA-300/

Results and discussion
1) Characterization of the bacteria

Change in PI > Methylation degree

Indicating that carboxylic groups are the most
abundant anionic groups on the surface
Results and discussion
2) As removal experiments

By aminated L. casei, up to about 6% of the initial

10 mg/L As(V) was removed in 1 h at pH 7
Native and methylated forms of all the studied strains were
also ineffective in removing As(V) from water at pH 7

The most efficient removal, 28,75,2%, was observed with

incubation time of 5 min at neutral pH.

1 mg/L Effect of metal concentration

Results and discussion

3) Desorption of bound As(V)

Recovery of all removed arsenic with HNO3

and NaOH washes proves that As(V) was
adsorbed to the bacterial surface rather than
accumulated inside the bacteria
Results and discussion
4) The maximum As(V) removal capacity
Maximum binding capacities of As(V) as high as 24,5 and 45,2 mg As(V)/ g
dry biomass have been reported for native biomasses of Penicillium
chrysogenum
(Loukidou et al., 2003)

qmax =31268 g/g

b = 0,0022 0.0013 L/g

Some other anionic groups, like phosphate groups of

(lipo)teichoic acids, may still have caused interference

Time = 5 min
pH = 7
Concentration range = 0,1 5 mg/L
Bacterial concentration = 1 g/L
Conclusions

Arsenic(III) removal is reported to occur by different mechanism than As(V) removal

mechanism was reported to be complex formation between arsenous acid and sulfhydryl
groups of Scytonema surface proteins.

Only aminated L. casei was observed to remove As(V) from wter.

The most efficient removal, 28,75,2%, was observed with incubation time of 5 min at
neutral pH.

Finally, this method may be uneconomical as modification of the biomass was required.
References

. vall-Jskelainen, S., Palva, A., 2005. Lactobacillus surface layers and their applications. FEMS Microbiology Reviews 29, 511529.
Ayotte, J.D., Montgomery, D.L., Flanagan, S.M., Robinson, K.W., 2003.
Arsenic in groundwater in eastern New England: occurrence, controls, and human health implications. Environmental Science and
Technology 372075372083.
Beveridge, T.J., Murray, R.G.E., 1980. Site of metal deposition in the cell wall of Bacillus subtilis. Journal of Bacteriology 141, 876887.
Boonaert, C.J.P., Rouxhet, P.G., 2000. Surface of lactic acid bacteria: relationships between chemical composition and physicochemical
properties.
Applied and Environmental Microbiology 66, 25482554. Cullen, W., Reimer, K., 1989. Arsenic speciation in the environment. Chemical
Reviews 89, 713764.
https://www.ecologiaverde.com/veinte-millones-chinos-corren-riesgo-morir-envenenados-arsenico/
https://embryology.med.unsw.edu.au/embryology/index.php?title=File:Bacteria_-_gram-stained_vaginal_smear_01.jpg&*
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC293699/pdf/jbacter00563-0446.pdf
http://www.monografias.com/trabajos59/contaminacion-arsenico/contaminacion-arsenico2.shtml