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Title: Physiology and genetics of metabolic flux control in Zymomonas mobilis. Progress report

Abstract

This work seeks to understand the role of gene expression in regulating glycolytic enzyme synthesis in a balance that allows proper glycoltic flux control. The seven genes targeted for study in this laboratory have been cloned and sequenced, and molecular details of regulation have been investigated. Clear that glycolytic enzyme synthesis is coordinated to prevent the build up of toxic metabolic intermediates. The genetic mechanisms responsible for regulating balanced expression of the EntnerDoudoroff and glycolytic genes in Z. mobilis are beginning to be understood. Several layers of genetic control, perhaps in a hierarchal arrangement act in concert to determine the relative abundance of the glycolytic enzymes. These genetic controls involve differential translational efficiency, highly conserved promoter sequences, transcription factors, differential mRNA stabilities, and nucleolytic mRNA processing. The serendipitous cloning of the glucose facilitator, glf, as a result of linkage to several other genes of interest will have a significant impact on the study of Z. mobilis metabolism. The glucose facilitator is being characterized in a genetically reconstituted system in E. coli. Molecular genetic studies indicate that the ratio of glf expression to that of glk, zmf, and edd is carefully regulated, and suggests a critical role in metabolic control. Regulationmore » of glycolytic gene expression is now sufficiently well understood to allow use of the glycolytic genes as tools to manipulate specified enzyme levels for the purpose of analyzing metabolic flux control. The critical genes have been subcloned for stable expression in Z. mobilis and placed under control of a regulated promoter system involving the tac promoter, the lacI repressor, and gene induction in by IPTG. HPLC methods have been developed that allow quantitation of virtually all of the metabolic intermediates in the cell pool.« less

Authors:
Publication Date:
Research Org.:
Nebraska Univ., Lincoln, NE (United States). School of Biological Sciences
Sponsoring Org.:
USDOE, Washington, DC (United States)
OSTI Identifier:
10168249
Report Number(s):
DOE/ER/20002-T1
ON: DE92018480
DOE Contract Number:  
FG02-90ER20002
Resource Type:
Technical Report
Resource Relation:
Other Information: PBD: [1992]
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; GLYCOLYSIS; BIOCHEMICAL REACTION KINETICS; GENE REGULATION; GENE OPERONS; DNA SEQUENCING; PORINS; DNA-CLONING; PROGRESS REPORT; BIOLOGICAL PATHWAYS; TWO-DIMENSIONAL ELECTROPHORESIS; POST-TRANSLATION MODIFICATION; LIQUID COLUMN CHROMATOGRAPHY; MESSENGER-RNA; BACTERIA; 550500; METABOLISM

Citation Formats

Conway, T. Physiology and genetics of metabolic flux control in Zymomonas mobilis. Progress report. United States: N. p., 1992. Web. doi:10.2172/10168249.
Conway, T. Physiology and genetics of metabolic flux control in Zymomonas mobilis. Progress report. United States. https://doi.org/10.2172/10168249
Conway, T. 1992. "Physiology and genetics of metabolic flux control in Zymomonas mobilis. Progress report". United States. https://doi.org/10.2172/10168249. https://www.osti.gov/servlets/purl/10168249.
@article{osti_10168249,
title = {Physiology and genetics of metabolic flux control in Zymomonas mobilis. Progress report},
author = {Conway, T},
abstractNote = {This work seeks to understand the role of gene expression in regulating glycolytic enzyme synthesis in a balance that allows proper glycoltic flux control. The seven genes targeted for study in this laboratory have been cloned and sequenced, and molecular details of regulation have been investigated. Clear that glycolytic enzyme synthesis is coordinated to prevent the build up of toxic metabolic intermediates. The genetic mechanisms responsible for regulating balanced expression of the EntnerDoudoroff and glycolytic genes in Z. mobilis are beginning to be understood. Several layers of genetic control, perhaps in a hierarchal arrangement act in concert to determine the relative abundance of the glycolytic enzymes. These genetic controls involve differential translational efficiency, highly conserved promoter sequences, transcription factors, differential mRNA stabilities, and nucleolytic mRNA processing. The serendipitous cloning of the glucose facilitator, glf, as a result of linkage to several other genes of interest will have a significant impact on the study of Z. mobilis metabolism. The glucose facilitator is being characterized in a genetically reconstituted system in E. coli. Molecular genetic studies indicate that the ratio of glf expression to that of glk, zmf, and edd is carefully regulated, and suggests a critical role in metabolic control. Regulation of glycolytic gene expression is now sufficiently well understood to allow use of the glycolytic genes as tools to manipulate specified enzyme levels for the purpose of analyzing metabolic flux control. The critical genes have been subcloned for stable expression in Z. mobilis and placed under control of a regulated promoter system involving the tac promoter, the lacI repressor, and gene induction in by IPTG. HPLC methods have been developed that allow quantitation of virtually all of the metabolic intermediates in the cell pool.},
doi = {10.2172/10168249},
url = {https://www.osti.gov/biblio/10168249}, journal = {},
number = ,
volume = ,
place = {United States},
year = {Sat Aug 01 00:00:00 EDT 1992},
month = {Sat Aug 01 00:00:00 EDT 1992}
}