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Method for establishing and analyzing scale metabolism network model of actinoplanetes genomes

A swimming actinomycete and genome technology, applied in the field of systems biology, can solve the problems of large workload, long time, blind strain selection and fermentation optimization, etc., and achieve the effect of strong reliability and high accuracy

Inactive Publication Date: 2015-07-01
JIANGNAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

As the number of diabetic patients continues to increase, the demand for acarbose is also increasing, but there are some problems in traditional fermentation optimization: (1) Traditional strain selection and fermentation optimization are blind, time-consuming and seem to have reached the limit (2) Some metabolic mechanisms during the production of acarbose by the strain Actinoplanes are unclear, and the metabolic bottleneck is unknown
The conventional genome-scale metabolic network model construction is to extract genes with metabolic functions based on the published genome sequencing results, and then integrate biochemical databases and literature information to form a reaction list representing the known biochemical reactions in the target microbial cells, but this method works Large amount and time-consuming

Method used

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  • Method for establishing and analyzing scale metabolism network model of actinoplanetes genomes
  • Method for establishing and analyzing scale metabolism network model of actinoplanetes genomes
  • Method for establishing and analyzing scale metabolism network model of actinoplanetes genomes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 The effect of nicotinic acid addition on the fermentation of acarbose

[0042] The effects of niacin concentration (2 mg / L, 5 mg / L, 10 mg / L) on cell growth and acarbose synthesis are shown in Table 1. With the increase of niacin concentration (0-5mg / L), cell growth and acarbose production gradually increased, and the cell dry weight and acarbose production reached the maximum at 5mg / L (10.62g / L and 0.697g / L), increased by 13.1% and 53.5% respectively compared with the control group, and then with the increase of niacin concentration (≥5mg / L), cell growth and acarbose concentration gradually decreased.

[0043] Table 1 Effect of Niacin Addition on Acarbose Fermentation

[0044]

Embodiment 2

[0045] Example 2 Effect of Amino Acid Addition on Acarbose Fermentation

[0046] Table 2 shows the effects of adding 0mmol / L-2.0mmol / L arginine, histidine and glutamine on the cell growth in shake flasks respectively. It was found that when histidine was added, the dry weight of cells increased by 9.4%, while the addition of arginine and glutamine had no obvious promoting effect on cell growth.

[0047] The impact of adding amino acids on the production of acarbose in the total synthetic medium is shown in Table 2. Adding 2.0mmol / L glutamic acid, cysteine, lysine, glutamine and asparagine to the total synthetic medium increased the production of acarbose by 29.6%, 26.5%, 26.3%, 11.8%, respectively. % and 9.2%. Further studies have found that adding more than 0mmol / L to .2mmol / L of the above five amino acids alone or adding two or more amino acids in combination can increase the production of acarbose. In addition, the combined addition of 2mg / L-10mg / L niacin can increase th...

Embodiment 3

[0050] Example 3 Effect of pH on Acarbose Fermentation

[0051] The effects of different pH conditions (pH no control, pH 7.0, pH 5.5) on cell growth and acarbose production were compared. The results showed that the dry cell weight and acarbose production reached the highest value (9.53g / L and 0.875g / L) at pH7.0, which were increased by 21% and 7% (no pH control), 31% and 15% respectively. % (pH 5.5). This result shows that cell growth can tolerate certain acidic conditions (pH 5.5), but neutral conditions (pH 7.0) are more conducive to the growth of cells; for acarbose synthesis, neutral conditions (pH 7.0) are more favorable for the benefit.

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Abstract

The invention discloses a method for establishing and analyzing a scale metabolism network model of actinoplanetes genomes, and belongs to the field of systems biology. Cell growth simulation, essential growth essential genes and reaction verification are carried on the scale metabolism network model of actinoplanetes genomes established based on the method on a system level, a strategy is provided for simulating and improving acarbose yield, and an important platform is provided for comprehensively understanding the metabolic characteristics of actinoplanetes and improving the acarbose fermentation level. The invention provides multiple methods for improving the acarbose yield, the acarbose yield can be increased by 53.5% by adding nicotinic acid, and the acarbose yield is respectively increased by 29.6%, 26.5%, 26.3%, 11.8% and 9.2% by adding glutamic acid, cysteine, lysine, glutamine and asparagines.

Description

technical field [0001] The invention relates to a method for constructing a genome-scale metabolic network model of actinomycete mobilis and an analysis method, belonging to the field of systems biology. Background technique [0002] Type II diabetes is a worldwide disease. There are currently 3.2 million type II diabetes patients in the world, and the number of patients continues to increase. Clinically commonly used anti-type II diabetes drugs mainly include biguanides, sulfonylureas, α-glucosidase inhibitors, thiazolidinediketones and anisic acid. Acarbose is mild and persistent and non-toxic The characteristics of the function are widely used. [0003] As a common strain for the fermentation and production of acarbose, Actinomycetes mobilis is widely used. In 1977, German Bayer sieved Actinoplanes sp. SE50 (ATCC 31042) from the soil for the industrial production of acarbose. Subsequently, industrially produced strains have been continuously improved through strain sel...

Claims

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Application Information

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IPC IPC(8): C12P19/26G06F19/12A61K31/7036A61P3/10C12R1/045
Inventor 刘立明王雅丽
Owner JIANGNAN UNIV
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