scholarly journals Clustering enzymes using E.coli inner cell membrane as scaffold in metabolic pathway

2017 ◽  
Author(s):  
You Wang ◽  
Yuqi Wu ◽  
Yang Suo ◽  
Huaqing Guo ◽  
Yineng Yu ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Nucleic Acid ◽  
Cell Membrane ◽  
Acid Synthesis ◽  
Construction Process ◽  
Metabolism Pathway ◽  
Synthetic Protein ◽  
Potential Applications ◽  
Membrane Target ◽  
Natural Cell

AbstractClustering enzymes in the same metabolism pathway is a natural strategy to enhance the productivity. Several systems have been designed to artificially cluster desired enzymes in the cell, such as synthetic protein scaffold and nucleic acid scaffold. However, these scaffolds require complicated construction process and have limited slots for target enzymes. Following this direction, we designed a scaffold system based on natural cell membrane. Target enzymes (FabZ, FabG, FabI and TesA’ in fatty acid synthesis II pathway) are anchored on the E.coli inner membrane, showing the enhanced metabolism flux without the requirement of the further artificial interactions to force the clustering. Furthermore, anchoring the enzymes on the membrane enhances the products exportation, which further increases the productivity. Together, the proposed system has potential applications in producing valuable biomaterials.

scholarly journals Fatty Acid Cosubstrates Provide β-Oxidation Precursors for Rhamnolipid Biosynthesis in Pseudomonas aeruginosa, as Evidenced by Isotope Tracing and Gene Expression Assays

2012 ◽  
Vol 78 (24) ◽  
pp. 8611-8622 ◽  
Author(s):  
Lin Zhang ◽  
Tracey A. Veres-Schalnat ◽  
Arpad Somogyi ◽  
Jeanne E. Pemberton ◽  
Raina M. Maier
Keyword(s):  
Gene Expression ◽  
Fatty Acid ◽  
Fatty Acid Synthesis ◽  
De Novo ◽  
Fold Increase ◽  
Acid Synthesis ◽  
Octadecanoic Acid ◽  
Isotope Tracing ◽  
Pathway Gene ◽  
Potential Applications

ABSTRACTRhamnolipids have multiple potential applications as “green” surfactants for industry, remediation, and medicine. As a result, they have been intensively investigated to add to our understanding of their biosynthesis and improve yields. Several studies have noted that the addition of a fatty acid cosubstrate increases rhamnolipid yields, but a metabolic explanation has not been offered, partly because biosynthesis studies to date have used sugar or sugar derivatives as the carbon source. The objective of this study was to investigate the role of fatty acid cosubstrates in improving rhamnolipid biosynthesis. A combination of stable isotope tracing and gene expression assays was used to identify lipid precursors and potential lipid metabolic pathways used in rhamnolipid synthesis when fatty acid cosubstrates are present. To this end, we compared the rhamnolipids produced and their yields using either glucose alone or glucose and octadecanoic acid-d35as cosubstrates. Using a combination of sugar and fatty acids, the rhamnolipid yield was significantly higher (i.e., doubled) than when glucose was used alone. Two patterns of deuterium incorporation (either 1 or 15 deuterium atoms) in a single Rha-C10lipid chain were observed for octadecanoic acid-d35treatment, indicating that in the presence of a fatty acid cosubstrate, bothde novofatty acid synthesis and β-oxidation are used to provide lipid precursors for rhamnolipids. Gene expression assays showed a 200- to 600-fold increase in the expression ofrhlAandrhlBrhamnolipid biosynthesis genes and a more modest increase of 3- to 4-fold of thefadAβ-oxidation pathway gene when octadecanoic acid was present. Taken together, these results suggest that the simultaneous use ofde novofatty acid synthesis and β-oxidation pathways allows for higher production of lipid precursors, resulting in increased rhamnolipid yields.

scholarly journals Gleevec (STI571) Influences Metabolic Enzyme Activities and Glucose Carbon Flow toward Nucleic Acid and Fatty Acid Synthesis in Myeloid Tumor Cells

2001 ◽  
Vol 276 (41) ◽  
pp. 37747-37753 ◽  
Author(s):  
Joan Boren ◽  
Marta Cascante ◽  
Silvia Marin ◽  
Begoña Comı́n-Anduix ◽  
Josep J. Centelles ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Nucleic Acid ◽  
Tumor Cells ◽  
Fatty Acid Synthesis ◽  
Enzyme Activities ◽  
Acid Synthesis ◽  
Carbon Flow ◽  
Metabolic Enzyme ◽  
Glucose Carbon

scholarly journals The specificity and metabolic implications of the inhibition of pyruvate transport in isolated mitochondria and intact tissue preparations by α-Cyano-4-hydroxycinnamate and related compounds

1975 ◽  
Vol 148 (1) ◽  
pp. 97-106 ◽  
Author(s):  
A P Halestrap ◽  
R M Denton
Keyword(s):  
Fatty Acid ◽  
Cell Membrane ◽  
Fatty Acid Synthesis ◽  
Pyruvate Dehydrogenase ◽  
Rat Heart ◽  
Pyruvate Carboxylase ◽  
Acid Synthesis ◽  
Pyruvate Dehydrogenase Kinase ◽  
Kidney Cortex ◽  
Rat Liver Mitochondria

1. Effects of α-cyano-4-hydroxycinnamate and α-cyanocinnamate on a number of enzymes involved in pyruvate metabolism have been investigated. Little or no inhibition was observed of any enzyme at concentrations that inhibit completely mitochondrial pyruvate transport. At much higher concentrations (1 mM) some inhibition of pyruvate carboxylase was apparent. 2. α-Cyano-4-hydroxycinnamate (1-100 muM) specifically inhibited pyruvate oxidation by mitochondria isolated from rat heart, brain, kidney and from blowfly flight muscle; oxidation of other substrates in the presence or absence of ADP was not affected. Similar concentrations of the compound also inhibited the carboxylation of pyruvate by rat liver mitochondria and the activation by pyruvate of pyruvate dehydrogenase in fat-cell mitochondria. These findings imply that pyruvate dehydrogenase, pyruvate dehydrogenase kinase and pyruvate carboxylase are exposed to mitochondrial matrix concentrations of pyruvate rather than to cytoplasmic concentrations. 3. Studies with whole-cell preparations incubated in vitro indicate that α-cyano-4-hydroxycinnamate or α-cyanocinnamate (at concentrations below 200 muM) can be used to specifically inhibit mitochondrial pyruvate transport within cells and thus alter the metabolic emphasis of the preparation. In epididymal fat-pads, fatty acid synthesis from glucose and fructose, but not from acetate, was markedly inhibited. No changes in tissue ATP concentrations were observed. The effects on fatty acid synthesis were reversible. In kidney-cortex slices, gluconeogenesis from pyruvate and lactate but not from succinate was inhibited. In the rat heart perfused with medium containing glucose and insulin, addition of α-cyanocinnamate (200 muM) greatly increased the output and tissue concentrations of lactate plus pyruvate but decreased the lactate/pyruvate ratio. 4. The inhibition by cyanocinnamate derivatives of pyruvate transport across the cell membrane of human erythrocytes requires much higher concentrations of the derivatives than the inhibition of transport across the mitochondrial membrane. α-Cyano-4-hydroxycinnamate appears to enter erythrocytes on the cell-membrane pyruvate carrier. Entry is not observed in the presence of albumin, which may explain the small effects when these compounds are injected into whole animals.

In silico Mapping and Structure Analysis of Key Enzyme Genes in Fatty Acid Synthesis of Soybean

2009 ◽  
Vol 35 (10) ◽  
pp. 1942-1947
Author(s):  
Wan-Kun SONG ◽  
Ming-Xi ZHU ◽  
Yang-Lin ZHAO ◽  
Jing WANG ◽  
Wen-Fu LI ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Structure Analysis ◽  
Fatty Acid Synthesis ◽  
In Silico ◽  
Acid Synthesis ◽  
In Silico Mapping ◽  
Key Enzyme
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