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Genetic engineering of novel protein polymers and their biosynthetic production in Escherichia coli and Pichia pastoris.

Record Type:	Electronic resources : Monograph/item
Title/Author:	Genetic engineering of novel protein polymers and their biosynthetic production in Escherichia coli and Pichia pastoris.
Author:	Wood, Nichole Lea.
Description:	178 p.
Notes:	Chairs: Jay D. Keasling; Susan J. Muller.
Notes:	Source: Dissertation Abstracts International, Volume: 64-09, Section: B, page: 4499.
Contained By:	Dissertation Abstracts International64-09B.
Subject:	Engineering, Chemical.
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3105407
ISBN:	0496529749

Genetic engineering of novel protein polymers and their biosynthetic production in Escherichia coli and Pichia pastoris.
Wood, Nichole Lea.

Genetic engineering of novel protein polymers and their biosynthetic production in Escherichia coli and Pichia pastoris. [electronic resource] - 178 p.

Chairs: Jay D. Keasling; Susan J. Muller.

Thesis (Ph.D.)--University of California, Berkeley, 2003.

A second class of protein-based materials, consisting of sequence disordered copolymers of leucine (L) and either serine (S), glutamate (E), or lysine (K) residues, were constructed and studied for expression in Escherichia coli. Statistically patterned protein polymers are of particular interest in the study of adsorption to multi-functionalized surfaces. Monte Carlo simulations and theoretical work by A. K. Chakraborty and co-workers suggest that disordered copolymers may preferentially adsorb to certain patterned surfaces when the statistics of the polymer sequence and the distribution of surface sites are related in a specific way. To probe such phenomena, the sequence statistics of disordered protein polymers could be studied for their effect on adsorption to similarly patterned surfaces, such as mixed SAMS consisting of functionalized alkanethiolates. Future development of synthetic systems that can mimic such recognition would have significant impact on applications ranging from high throughput nano-scale separation processes to viral inhibition and the development of sensors.

ISBN: 0496529749Subjects--Topical Terms:

226989
Engineering, Chemical.

Genetic engineering of novel protein polymers and their biosynthetic production in Escherichia coli and Pichia pastoris.
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100 0 $a Wood, Nichole Lea. $3 227339
245 1 0 $a Genetic engineering of novel protein polymers and their biosynthetic production in Escherichia coli and Pichia pastoris. $h [electronic resource]
300 $a 178 p.
500 $a Chairs: Jay D. Keasling; Susan J. Muller.
500 $a Source: Dissertation Abstracts International, Volume: 64-09, Section: B, page: 4499.
502 $a Thesis (Ph.D.)--University of California, Berkeley, 2003.
520 # $a A second class of protein-based materials, consisting of sequence disordered copolymers of leucine (L) and either serine (S), glutamate (E), or lysine (K) residues, were constructed and studied for expression in Escherichia coli. Statistically patterned protein polymers are of particular interest in the study of adsorption to multi-functionalized surfaces. Monte Carlo simulations and theoretical work by A. K. Chakraborty and co-workers suggest that disordered copolymers may preferentially adsorb to certain patterned surfaces when the statistics of the polymer sequence and the distribution of surface sites are related in a specific way. To probe such phenomena, the sequence statistics of disordered protein polymers could be studied for their effect on adsorption to similarly patterned surfaces, such as mixed SAMS consisting of functionalized alkanethiolates. Future development of synthetic systems that can mimic such recognition would have significant impact on applications ranging from high throughput nano-scale separation processes to viral inhibition and the development of sensors.
520 # $a As de novo designed protein polymers diverge from natural protein sequences, heterologous gene expression can drop significantly. Several artificial proteins, including disordered poly-EL and poly-KL polymers, displayed poor production levels in Escherichia coli. Possible reasons for this are presented and improved production from a yeast expression system using Pichia pastoris is reported. Finally, future directions and applications of artificial protein polymers will be discussed. (Abstract shortened by UMI.)
520 # $a With an interest in developing environmentally triggered protein-based materials, we have studied the microbial production of a self-assembling beta-sheet polymer, poly-EAK9, composed of the alternating amino acid repeat sequence, AEAEAKAK. Protein sequences of alternating hydrophilic (E = Glutamate, K = Lysine) and hydrophobic (A = Alanine) residues are known to adopt beta-sheet structures in aqueous solutions, and previous work by Zhang and co-workers on ionic self-complementary peptides have demonstrated their ability to self-assemble into macroscopic membranes, fibrils, and gels upon the addition of monovalent metal ions. Through the use of recombinant DNA techniques, we have constructed a high molecular weight analogue of the EAK sequence to probe its self-assembly and mechanical properties for potential use in biomaterial applications, such as local drug delivery, tissue engineering, and biosensors.
590 $a School code: 0028.
650 # 0 $a Engineering, Chemical. $3 226989
650 # 0 $a Engineering, Biomedical. $3 227004
710 0 # $a University of California, Berkeley. $3 212474
773 0 # $g 64-09B. $t Dissertation Abstracts International
790 $a 0028
790 1 0 $a Keasling, Jay D., $e advisor
790 1 0 $a Muller, Susan J., $e advisor
791 $a Ph.D.
792 $a 2003
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