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An internal reaction and reaction technology, applied in the field of biological metabolism, can solve problems such as reproducibility, inability to replicate, and no substantial promotion
Inactive Publication Date: 2016-05-11
EAST CHINA UNIV OF SCI & TECH
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Problems solved by technology
Scientists can clearly see almost all in vivo reactions, and omics data has accumulated a lot, but no one has been able to reproduce an in vivo reaction system in vitro
Humans have been able to (at least) partially assemble genome systems, but have not been able to replicate metabolic systems that have been chemically unde
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Embodiment 1
[0116] figure 1 Flowchart for the system for reproducing in vivo responses in vitro.
[0117] Such as figure 1 Shown, a kind of in vitro reproduction system of in vivo reaction of the present invention comprises the following steps:
[0118] Step 1, turn on the computer, retrieve the relevant enzyme reaction kinetic parameters stored in the database, and then realize the metabolic network reaction in the computer according to the reaction mechanism in the body;
[0119] Step 2: For the minimal metabolic network system containing only cofactor metabolic intermediates CI, enzymes, substrates and products, perform intracellular and extracellular comparative experiments, and draw the cofactor metabolic intermediates CI, substrates and products. The curve of the corresponding concentration over time shows that the cofactor metabolic intermediate CI plays a global role in the metabolic reaction in vivo, and the molar content of the cofactor metabolic intermediate CI in the cell is...
Embodiment 2
[0128] figure 1 is a flow diagram of the system for reproducing in vivo responses in vitro.
[0129] Such as figure 1 As shown, the system for reproducing the in vivo response in vitro includes the following steps:
[0130] Step 1, turn on the computer, retrieve the relevant enzyme reaction kinetic parameters stored in the database, and then realize the metabolic network reaction in the computer according to the reaction mechanism in the body;
[0131] It is known that in the metabolic network in vivo, the ratio of double-substrate reactions is 70-80%, and the first and second substrates are both in excess, then the affinity of the two substrates to the enzyme can be obtained in the reaction of the metabolic network in vivo The constant Km, the catalytic constant kcat of an enzyme and the maximum reaction velocity Vmax;
[0132] Vmax=e 1,0 ×kcat;
[0133] The irreversible reaction formula of double substrate is: S 1 +B→P 1 +Q, the elementary reactions contained in this ...
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Abstract
The invention belongs to the technical field of biological metabolism, and provides a system for reproducing an in vivo reaction in vitro. The system comprises the following steps: opening a computer, and dispatching kinetic parameters of related in vivo metabolism stored in a database; carrying out an intracellular and extracellular comparative dry experiment on a minimal metabolic network system only containing a cofactor metabolic intermediate CI, an enzyme, a substrate and a product; carrying out an intracellular and extracellular comparative experiment on an unrestraint metabolic network system containing a non-cofactor metabolic intermediate NCI, the cofactor metabolic intermediate CI, the enzyme, the substrate and the product; obtaining the influence of the concentration level of CI on the steady state level of NCI at high, middle and low inhibition levels; and simulating a gene knock-in and knock-out experiment by adjusting the enzyme content in the reaction process to obtain the result that the enzyme content has influence on the NCI and the reaction rate and that the inhibition mechanism, the cofactor metabolic intermediate CI, the enzyme content and the non-cofactor metabolic intermediate NCI have influence on intracellular reactions, and then an extracellular metabolic reaction system is obtained. The system provided by the invention makes up the blank of the industry.
Description
technical field [0001] The invention belongs to the technical field of biological metabolism, and in particular relates to a system for reproducing in vivo reactions in vitro and its application in the fields of biology and the like. Background technique [0002] The metabolic process in the living body is the only way to express the characteristics of life. From gene expression, protein synthesis, to individual growth, all the characteristics of life are manifested through cell metabolism. Therefore, it is very necessary to understand the inner mechanism of metabolic reactions to reveal the whole process of life. [0003] On the other hand, the difference between in vivo (invivo) response and in vitro (invitro) response is an almost insurmountable gap in front of human beings for hundreds of years. Scientists can clearly see almost all in vivo reactions, and omics data has accumulated a lot, but no one has been able to reproduce an in vivo reaction system in vitro. Humans...
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