Nanoscale Biopolymers Nanoscale Biopolymers with Customizable Pro with Customizable Pro perties for Heavy Met perties for Heavy Met al Remediation al Remediation U Loi (Ann) Lao, Giri Prabhukuma U Loi (Ann) Lao, Giri Prabhukuma r, r, Jan Kostal, Mark Matsumoto, A Jan Kostal, Mark Matsumoto, A shok Mulchandani, and Wilfred Ch shok Mulchandani, and Wilfred Ch en en Chemical and Environmental Engineerin Chemical and Environmental Engineerin g g University of California, Riverside University of California, Riverside
30
Embed
Nanoscale Biopolymers with Customizable Properties for Heavy Metal Remediation U Loi (Ann) Lao, Giri Prabhukumar, Jan Kostal, Mark Matsumoto, Ashok Mulchandani,
This document is posted to help you gain knowledge. Please leave a comment to let me know what you think about it! Share it to your friends and learn new things together.
Transcript
Nanoscale Biopolymers Nanoscale Biopolymers with Customizable Propewith Customizable Properties for Heavy Metal Rerties for Heavy Metal Re
mediationmediationU Loi (Ann) Lao, Giri Prabhukumar,U Loi (Ann) Lao, Giri Prabhukumar, JaJan Kostal, Mark Matsumoto, Ashok Muln Kostal, Mark Matsumoto, Ashok Mul
chandani, and Wilfred Chenchandani, and Wilfred Chen
Chemical and Environmental EngineeringChemical and Environmental EngineeringUniversity of California, RiversideUniversity of California, Riverside
Heavy Metal Heavy Metal ContaminationContamination
2.4 million tons of metal wastes 2.4 million tons of metal wastes per year from industrial sources per year from industrial sources
2 million tons per year from 2 million tons per year from agriculture and domestic wasteagriculture and domestic waste
PbPb2+2+, Hg, Hg2+2+ and Cd and Cd2+2+ are ranked are ranked 2nd, 3rd and 7th, respectively, 2nd, 3rd and 7th, respectively, on the EPA’s priority list on the EPA’s priority list
Metal Chelating Metal Chelating PolymersPolymers
• Requires toxic solvents for synthesis
• Ultrafiltration is required
• Membrane clogging
Solution: Develop metal-binding materials that can be recovered by environmental stimuli
Metal Chelating Metal Chelating BiopolymersBiopolymers
Based on biological building blocksBased on biological building blocksNanoscale biopolymers that are specially Nanoscale biopolymers that are specially
pre-programmed within a DNA templatepre-programmed within a DNA templateEconomically produced by bacteria Economically produced by bacteria Environmentally friendlyEnvironmentally friendlyTunable properties based on changes Tunable properties based on changes in environmental conditions – pH or in environmental conditions – pH or temperaturetemperature
Elastin BiopolymerElastin Biopolymer Structurally similar to the repeating Structurally similar to the repeating
elastometric peptide sequence of the elastometric peptide sequence of the mammalian protein, elastin mammalian protein, elastin
VPGVG are the most frequently repeating unitsVPGVG are the most frequently repeating units Undergo a reversible phase transition from Undergo a reversible phase transition from
water soluble forms into aggregates as the water soluble forms into aggregates as the temperature increasestemperature increases
Low temperature: -turn
High temperature: twisted filament of -spirals
Genetic and Protein Genetic and Protein Engineering Engineering MethodologyMethodology
Synthetic gene
Recombinantplasmid
Plasmid
Enzyme
ELP biopolymers
Customizable Metal Binding BiCustomizable Metal Binding Biopolymersopolymers
Metal Binding Domain
Fine tune affinity with different binding sequences
Elastin Domain
Fine tune the transition temperature by controlling amino acid sequence and
number of repeating unit (VPGXG)n
Heavy Metal Removal by Heavy Metal Removal by Tunable BiopolymerTunable Biopolymer
+ CdT
Regeneration
soluble protein insoluble protein
Cd2+
Cd2+
Cd2+
Cd2+Cd2+
Cd2+
Elastin Biopolymers with MElastin Biopolymers with Metal-Binding Affinityetal-Binding Affinity
A B C D E
Biopolymer
A. ELP38H6B. ELP58H6C. ELP78H6D. ELP78E. ELP78H12
Kostal et al. Macromolecules, 34, 2257-2261, 2001.
Phase TransitionPhase Transition
Transition temperatures can be fine tuned from 20 to 40oC by controlling the chain length and salt concentration
Low TempHigh Temp
Cadmium RemovalCadmium Removal
Biopolymer
Cd2+ binding ( mol protein : mol Cd2+SD)
Ela38H6 1 : 1.04 0.04
Ela58H6 1 : 0.97 0.01
Ela78H6 1 : 0.95 0.01
Ela78 1 : 0.01 0.00
Ela78H12 1 : 1.50 0.02
SD - standard deviation
50 nmol of biopolymer were incubated with 100 nmol Cd2+
Repeated Cycles of Repeated Cycles of Metal BindingMetal Binding
1. Single Batch washing with distilled water 2. Two sequential batch washings with distilled water3. Single batch washing with 1.25 mg/ml (0.036 mM) ELP without the histidine tag
4. Single batch washing with 0.036 mM EDTA 5. Single batch washing with 1.25 mg/ml ELPH12 6. Single batch washing with 2.5 mg/ml ELPH12 7. Single batch washing with 5 mg/ml ELPH128. Two sequential batch washings with 1.25 mg/ml ELPH12
Less than 10% adsorption
•His6 or His12 tag can serve as a simple metal binding domain
•Both Tt and metal binding capacity can be easily regulated
•Biopolymers can be recycled
•Lack specificity and affinity
First generation biopolymers
Customizable Metal Binding BiCustomizable Metal Binding Biopolymersopolymers
Fine tune affinity with different binding sequences
Metal Binding Domain
Elastin Domain
Fine tune T by controlling amino acid sequence and number of repeating unit (VPGXG)n
Bacterial Bacterial mermer Systems Systemsm
er g
enes
on
E. c
oli
pla
smid
s or
tra
nsp
oson
s
MerRMerR
Mer
D
IM OM
MerT
Mer
P
MerA MerA
MerB
Hg2+
organomercury
Hg0
Hg2+
Hg2+
MerR can serve as a specific mercury binding domain
C123
C79
C114
C114
C79
C123
MerR-Hg complex
A B C kDaA, cell free extract
B, purified protein (80.1 kDa)
C, size marker
Production and purification of ELP153-
MerR
Binding of Mercury to ELP153-MerR
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.0 0.5 1.0 1.5 2.0 2.5 3.0
nmol Hg added
nmol
Hg
boun
d
Ela153-MerR
Added Mercury
Similar binding from pH 4 to 9
Selective Binding of Selective Binding of Mercury by Mercury by
Ela153-MerR BiopolymerEla153-MerR Biopolymer
0.00
0.10
0.20
0.30
0.40
0.50
0 25 50 75 100molar excess of Zn
met
al b
ound
(nm
ol)
Hg
Zn
0.00
0.10
0.20
0.30
0.40
0.50
0.60
0 25 50 75 100
molar excess of Cd
met
al b
ound
(nm
ol)
Hg
Cd
Acidic waste water (pH 4)
Kostal et al. ES&T, 37, 4457-4462, 2003.
Other Metal-binding Other Metal-binding DomainsDomains
Provide a novel, environmental frienProvide a novel, environmental friendly, and green engineering method fdly, and green engineering method for removal of heavy metal from wateor removal of heavy metal from water and soil.r and soil.