Overexpression of pyruvate kinases and phosphoenolpyruvate synthase in Escherichia coli : physiological characterization and application /

Escherichia coli (E. coli) is perhaps the most widely studied

Bibliographic Details
Main Author: Patnaik, Ranjan, 1969-
Format: Thesis Book
Language:English
Published: [Place of publication not identified] : [publisher not identified] ; 1994.
Subjects:
Online Access:http://proxy.library.tamu.edu/login?url=http://proquest.umi.com/pqdweb?did=741965761&sid=1&Fmt=2&clientId=2945&RQT=309&VName=PQD

MARC

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099 |a 1994  |a Dissertation  |a P311 
100 1 |a Patnaik, Ranjan,  |d 1969- 
245 1 0 |a Overexpression of pyruvate kinases and phosphoenolpyruvate synthase in Escherichia coli :  |b physiological characterization and application /  |c by Ranjan Patnaik. 
264 1 |a [Place of publication not identified] :  |b [publisher not identified] ;  |c 1994. 
300 |a xiv, 91 leaves :  |b illustrations ;  |c 28 cm. 
336 |a text  |b txt  |2 rdacontent 
337 |a unmediated  |b n  |2 rdamedia 
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504 |a Includes bibliographical references. 
500 |a Vita. 
502 |b Ph. D.  |c Texas A&M University  |d 1994. 
500 |a "Major Subject: Chemical Engineering". 
530 |a Issued also on microfiche from University Microfilms Inc. 
520 |a Escherichia coli (E. coli) is perhaps the most widely studied  
520 |a prokaryote by both scientists and engineers in the area of  
520 |a biological sciences. On one hand scientists are trying to  
520 |a unravel and understand the biochemical and genetic regulation  
520 |a of various metabolic pathways as has evolved in the bacterial  
520 |a cell, while on the other hand engineers are constantly on the  
520 |a look out for constructing and designing new strains by  
520 |a altering existing pathways for improving production of  
520 |a chemicals. In most cases these engineered strains are quite  
520 |a different from the wild type. Engineers use the insights  
520 |a from the paradigms developed by the scientists to narrow down  
520 |a their choices for manipulating strains for a specific  
520 |a purpose, and the characterization of this engineered strain  
520 |a serves as feedback in the understanding of that paradigm,  
520 |a either strengthening it or invalidating it. In this study a  
520 |a combined attempt is made in which alterations in the central  
520 |a metabolism of E. coli are characterized from a fundamental  
520 |a perspective and then a new central metabolism is created that  
520 |a improves production of 3-deoxy-D-arabino-heptulosonate7- 
520 |a phosphate (DAHP) over an existing industrial strain. The  
520 |a central metabolism is altered by overexpression of a  
520 |a gluconeogenic enzyme, phosphoenolpyruvate synthetase (Pps)  
520 |a under glycolytic conditions and also by overexpression of  
520 |a pyruvate kinases (PykI, Pykll). These three enzymes along  
520 |a with the glucose phosphotransferase system (PTS) contribute  
520 |a significantly in the modulation of the intracellular levels  
520 |a of two key metabolites of the central metabolism,  
520 |a phosphoenolpyruvate (PEP) and pyruvate. Insights obtained  
520 |a from this study were successfully used in directing more  
520 |a carbon flux from PEP towards aromatics production. The  
520 |a following results were obtained: (1) Overexpression of PykH  
520 |a observable effect on the specific growth rate and the glucose  
520 |a consumption rate in E. coli. However, overexpression of Pps  
520 |a increased glucose uptake rate, oxygen consumption rate,  
520 |a fermentation product excretion, and showed activity dependent  
520 |a growth inhibition. (2) Overexpression of Pps in combination  
520 |a with DAHP synthase (AroG) and transketolase (TktA) increases  
520 |a the yield of DAHP production from glucose by almost twofold  
520 |a over the control strains to near theoretical levels. 
650 4 |a Major chemical engineering. 
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