SCURC Presentation-RA 4.9.15-fix

A Toxic Metal “Sponge”

A Novel Technology For Water Remediation…

California State University, Long Beach

Phuc Nguyen

Acey Lab

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2015 ACS Southern California Undergraduate Research conference, UC San Diego

OUTLINE

The ProblemMetallothionein OverviewExperiments:•Purification•Metal Binding Properties•Metal Recovery technology

Conclusion

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THE PROBLEM

Wastewater Containing Toxic Metals

Lead | Mercury | Arsenic

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THE ANSWER: METALLOTHIONEIN (MT)

• Low Molecular Weight.• Cysteine rich.• Selectively Binds Toxic/Precious Metals

Hg, Pb, As, U Au, Ag, Pt, Pd

• High Affinity: 4 x 10-14 M-1 for Pb• Active At Extreme Conditions

Temperature - 100oC pH - 4 to 10

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EXPRESSION AND PURIFICATION OF RECOMBINANT MTMT Gene Cloned into pET 11a

Recombinant MT Expressed in BL-21 E.coli

Sonication

G-50 Sephadex Gel Filtration

Mono Q Anion Exchange

Superdex Gel Filtration

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SDS-PAGE ANALYSIS OF PURIFIED MT

MT6.5 KDa

Standards

26.6 KDa

16.9 KDa

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METAL BINDING ACTIVITY

Free-metal

MT-bound metal

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METAL BINDING ACTIVITY

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METAL BINDING IN ACETONE

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TECHNOLOGY FOR METAL REMOVAL

US Patent Number 7,273,962.

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METAL RECOVERY FROM SEA WATER

585 CPM 109Cd

4,612 CPM 109Cd

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CONCLUSION

•Recombinant MT is Expressed and Purified•MT binds a Number of Toxic/ Precious Metals •Active in 30% Acetone•Metal Binding Effective In High Salt

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Dr. Roger Acey

Dr. Katherine Kantardjieff

Paul Madera

Brent Wilkinson

Aaron Ong

Hieu Nguyen

ACKNOWLEDGEMENT