University of Arkansas, Fayetteville
ScholarWorks@UARK
Theses and Dissertations
8-2017
Peptide-directed Nanoparticle Synthesis With A
Denovo Pd-binding Sequence Fused To A
Reporter Protein
Rita Eloisa Tejada Vaprio
University of Arkansas, Fayetteville
Follow this and additional works at: http://scholarworks.uark.edu/etd
Part of the Nanoscience and Nanotechnology Commons, and the Other Chemical Engineering
Commons
Recommended Citation
Tejada Vaprio, Rita Eloisa, "Peptide-directed Nanoparticle Synthesis With A Denovo Pd-binding Sequence Fused To A Reporter
Protein" (2017). Theses and Dissertations. 2393.
http://scholarworks.uark.edu/etd/2393
This Thesis is brought to you for free and open access by ScholarWorks@UARK. It has been accepted for inclusion in Theses and Dissertations by an
authorized administrator of ScholarWorks@UARK. For more information, please contact , .
, Peptide-directed Nanoparticle Synthesis With A Denovo Pd-binding
Sequence Fused To A Reporter Protein
A thesis submitted in partial fulfillment
of the requirements for the degree of
Master of Science in Chemical Engineering
by
Rita Tejada Vaprio
University of Arkansas
Bachelor of Science in Chemical Engineering, 2015
August 2017
University of Arkansas
This thesis is approved for recommendation to the Graduate Council.
_______________________________
Dr. Robert Beitle
Thesis Director
_______________________________ _______________________________
Dr. Roger Koeppe Dr. Nicholas Bedford
Committee Member Committee Member
_______________________________
Dr. Lauren Greenlee
Committee Member
,Abstract
There is a need for low-cost nanoparticle materials in the context of new technologies for
catalyst development. The purpose of this work was to recombinantly produce a 45- amino acid
long metal binding peptide that is useful for nanoparticle synthesis. Using splicing by overlap
extension PCR, a synthetic gene containing the fusion of the metal binding peptide with Green
Fluorescent Protein (GFPUV) was constructed. The metal binding peptide, fused to reporter
protein GFPUV, was expressed using high cell density fermentation. Palladium nanoparticles of
an average diameter of 1.18 ± 0.45 nm were synthesized by using the crude cell extract
containing the fusion protein. Nanoparticle synthesis was also done using desalted samples
(removal of medium components) as well as enriched fractions from ion exchange
chromatography purification. In all cases, palladium, gold and palladium-gold nanoparticles
were successfully synthesized with good particle size for catalysis applications, control of
diameter, and lack of other metal precipitants, respectively.
This work illustrates that metallic nanoparticles can be synthesized using the soluble cell
extract containing the fusion protein without extensive purification or cleavage steps.
, Acknowledgements
First of all, I would like to thank my mom, Gioconda and my sisters, Sara and Carolina. Without
your never-ending love throughout my whole life, I would not be where I am today. I know it has
not been easy being away for almost 6 years, and I am incredibly grateful for the support you
have given me in every step of the way.
To the love of my life, Rudra, thank you for always encouraging me to be a better version of
myself and for your love and support (in and out of the lab) for the last 3 years. Thank you to Dr.
Beitle, the best advisor ever, for being so patient, optimistic and teaching me so much.
I would also like to thank Dr. Nicholas Bedford, for all the help provided with the nanoparticles,
and Dr. Lauren Greenlee and Dr. Roger Koeppe for being part of my committee. Thank you to
Dr. McKinzie Fruchtl for your help with the fed-batch fermentation.
ScholarWorks@UARK
Theses and Dissertations
8-2017
Peptide-directed Nanoparticle Synthesis With A
Denovo Pd-binding Sequence Fused To A
Reporter Protein
Rita Eloisa Tejada Vaprio
University of Arkansas, Fayetteville
Follow this and additional works at: http://scholarworks.uark.edu/etd
Part of the Nanoscience and Nanotechnology Commons, and the Other Chemical Engineering
Commons
Recommended Citation
Tejada Vaprio, Rita Eloisa, "Peptide-directed Nanoparticle Synthesis With A Denovo Pd-binding Sequence Fused To A Reporter
Protein" (2017). Theses and Dissertations. 2393.
http://scholarworks.uark.edu/etd/2393
This Thesis is brought to you for free and open access by ScholarWorks@UARK. It has been accepted for inclusion in Theses and Dissertations by an
authorized administrator of ScholarWorks@UARK. For more information, please contact , .
, Peptide-directed Nanoparticle Synthesis With A Denovo Pd-binding
Sequence Fused To A Reporter Protein
A thesis submitted in partial fulfillment
of the requirements for the degree of
Master of Science in Chemical Engineering
by
Rita Tejada Vaprio
University of Arkansas
Bachelor of Science in Chemical Engineering, 2015
August 2017
University of Arkansas
This thesis is approved for recommendation to the Graduate Council.
_______________________________
Dr. Robert Beitle
Thesis Director
_______________________________ _______________________________
Dr. Roger Koeppe Dr. Nicholas Bedford
Committee Member Committee Member
_______________________________
Dr. Lauren Greenlee
Committee Member
,Abstract
There is a need for low-cost nanoparticle materials in the context of new technologies for
catalyst development. The purpose of this work was to recombinantly produce a 45- amino acid
long metal binding peptide that is useful for nanoparticle synthesis. Using splicing by overlap
extension PCR, a synthetic gene containing the fusion of the metal binding peptide with Green
Fluorescent Protein (GFPUV) was constructed. The metal binding peptide, fused to reporter
protein GFPUV, was expressed using high cell density fermentation. Palladium nanoparticles of
an average diameter of 1.18 ± 0.45 nm were synthesized by using the crude cell extract
containing the fusion protein. Nanoparticle synthesis was also done using desalted samples
(removal of medium components) as well as enriched fractions from ion exchange
chromatography purification. In all cases, palladium, gold and palladium-gold nanoparticles
were successfully synthesized with good particle size for catalysis applications, control of
diameter, and lack of other metal precipitants, respectively.
This work illustrates that metallic nanoparticles can be synthesized using the soluble cell
extract containing the fusion protein without extensive purification or cleavage steps.
, Acknowledgements
First of all, I would like to thank my mom, Gioconda and my sisters, Sara and Carolina. Without
your never-ending love throughout my whole life, I would not be where I am today. I know it has
not been easy being away for almost 6 years, and I am incredibly grateful for the support you
have given me in every step of the way.
To the love of my life, Rudra, thank you for always encouraging me to be a better version of
myself and for your love and support (in and out of the lab) for the last 3 years. Thank you to Dr.
Beitle, the best advisor ever, for being so patient, optimistic and teaching me so much.
I would also like to thank Dr. Nicholas Bedford, for all the help provided with the nanoparticles,
and Dr. Lauren Greenlee and Dr. Roger Koeppe for being part of my committee. Thank you to
Dr. McKinzie Fruchtl for your help with the fed-batch fermentation.
