Metabolic pathway for bio-converted methyl alcohol

A methanol metabolism pathway and biotransformation technology, applied in the field of biotransformation synthesis, can solve problems such as inability to transfer, poor portability, and complex metabolic processes

Active Publication Date: 2017-12-15
FUJIAN AGRI & FORESTRY UNIV
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Problems solved by technology

In nature, methylotrophic bacteria mainly oxidize methanol through the RuMP pathway and the Serine cycle. The metabolic process of this pathway is complex and requires the parti

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  • Metabolic pathway for bio-converted methyl alcohol
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Embodiment Construction

[0025] In order to make the content of the present invention easier to understand, the technical solutions of the present invention will be further described below in conjunction with specific embodiments, but the present invention is not limited thereto.

[0026] 1. Training method

[0027] 1.1 Medium

[0028] Seed medium (g / L): peptone 10, yeast powder 5, sodium chloride 10 (pH=7.0).

[0029] Fermentation medium (g / L): NaHPO 4 .12H2O 17.1, KH 2 PO 4 3. NaCl 0.5, NH4Cl 1

[0030] MgSO 4 0.12, CaCl 2 0.01, Methanol 3.2 (pH=7.0).

[0031] 1.2 Sterilization method

[0032] The seed medium and fermentation medium were sterilized at 121°C for 20 min, respectively, and the pH of the medium was adjusted to 7.0 with 4M NaOH before inoculation. MgSO 4 and CaCl 2 Sterilization needs to be configured separately; methanol cannot be sterilized by autoclaving, and the method of filter sterilization is used.

[0033] 1.3 Culture method

[0034] Take the glycerin tube of eng

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Abstract

The invention provides a metabolic pathway for bio-converted methyl alcohol. The metabolic pathway for methyl alcohol is artificially constructed in escherichia coli. The metabolic pathway is simple and can enter into the central metabolic pathway of the escherichia coli within two steps. The key enzyme, namely, formaldehyde lyase (formolase) on the pathway is subjected to orthogenesis, the beneficial mutant clone is acquired in the manner of screening through whole-cell catalysis, the enzyme activity of the formaldehyde lyase is increased, the related gene on the metabolic pathway is assembled for acquiring methyl alcohol, the engineering bacteria is utilized to culture in a culture medium taking methyl alcohol as a unique carbon source and the efficient utilization of the methyl alcohol is realized in the manners of optimizing culture medium and knocking out gene.

Description

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Claims

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Application Information

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Owner FUJIAN AGRI & FORESTRY UNIV
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