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Metabolic network analysis method for streptomyces roseosporus as daptomycin producing thalli

A technology of Streptomyces roseospora and daptomycin, applied in the field of metabolic engineering

Active Publication Date: 2011-11-16
NANTONG YIKAI MEDICAL DEVICES CO LTD
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Problems solved by technology

For the daptomycin-producing strain Streptomyces roseospora, there is no report on its metabolic network analysis. Therefore, constructing a metabolic network model of Streptomyces roseospora and calculating its intracellular metabolic flux distribution can better understand Metabolic properties of the strain

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  • Metabolic network analysis method for streptomyces roseosporus as daptomycin producing thalli
  • Metabolic network analysis method for streptomyces roseosporus as daptomycin producing thalli
  • Metabolic network analysis method for streptomyces roseosporus as daptomycin producing thalli

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Embodiment 1

[0016] According to the knowledge about metabolic reactions in biochemistry, construct the central carbon metabolic network, including glycolysis pathway, pentose phosphate pathway, citric acid cycle, anaplerosis reaction, cell synthesis reaction and daptomycin synthesis reaction; according to Daptomyces The amino acid composition of prime and the precursor substance of synthesizing these amino acids, express daptomycin as being synthesized by precursor substance, the precursor substance of amino acid synthesis refers to " metabolic engineering-principle and method " written by Stephanopoulos et al. (Zhao Xueming et al. Translated, Chemical Industry Press, 1998); the composition of precursor substances of cells refers to "Combining pathway analysis with fluxbalance analysis for the comprehensive study of metabolic systems", the obtained metabolic network includes 15 intracellular metabolites and 18 reactions.

[0017] Table 1 List of metabolic network reactions

[0018]

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Abstract

The present invention relates to a metabolic network analysis method for streptomyces roseosporus as adaptomycin producing thalli, which comprises the following steps of: (1) constructing a central metabolic network pattern according to a glucolysis pathway, a pentose phosphate pathway, a citric acid cycle, an anaplerotic reaction, and a daptomycin synthesis reaction belonging to a cell synthesis reaction; (2) converting the metabolic network pattern into a matrix S (m*n), collecting a fermented liquid, measuring the cell dry weight, the glucose consumption rate, the daptomycin growth rate and the cell growth rate, and calculating an unknown flux according to know fluxes. In the present invention, the metabolic network analysis method is applied to the metabolic flux analysis of streptomyces roseosporus,the metabolic fluxes of intracellular reactions are calculated by measuring the metabolic flux of extracellular substances, and the distribution condition of glucose utilized by thalli to the glucolysis pathway, the pentose phosphate pathway, the citric acid cycle and the anaplerotic reaction is reflected. The present invention provides a new method for quantitatively researching the intracellular metabolic properties of streptomyces roseosporus.

Description

technical field [0001] The invention belongs to the field of metabolic engineering, in particular to a metabolic network analysis method of the daptomycin-producing bacterium Streptomyces roseospora. Background technique [0002] "Streptomyces roseosporus" (Latin name: "Streptomyces roseosporus") is an actinomycete of the genus Streptomyces, and its synthetic daptomycin (English name "daptomycin") is N -Decanoyl-L-Tryptophanyl-L-Asparaginyl-L-Aspartyl-L-Threonyl-Glycyl-L-Ornithyl-L-Aspartyl-D-Propanyl Aminoacyl-L-aspartyl-glycyl-D-seryl-L-threo-3-methyl-glutamyl-L-kynurenyl ε1-lactone is composed of a ten-carbon The alkane side chain is connected with tryptophan at the N-terminal of a cyclic β-amino acid peptide chain, and it is a new type of lipopeptide antibiotics. [0003] Daptomycin is a calcium ion-dependent antibiotic. In the presence of calcium ions, daptomycin binds to cell membrane proteins in the form of non-covalent bonds. The daptomycin-binding protein on the c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G06F19/12
Inventor 闻建平王国英贾晓强黄笛
Owner NANTONG YIKAI MEDICAL DEVICES CO LTD
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