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Method for generating lactobionic acid through increasing microbial conversion

A technology of microbial transformation and lactobionic acid, applied in the biological field, can solve the problems of improving the difficulty of separation and purification, increasing the cost, and long fermentation time, and achieving the effect of improving the fermentation efficiency.

Active Publication Date: 2020-05-19
HANGZHOU BROTHER BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chinese patent technology "CN101988046A" discloses a method of utilizing Pseudomonas fluorescens to produce lactobionic acid. In this method, lactose is used as a conversion substrate, and nitrogen source and inorganic salt are added as a fermentation medium. Under the most optimal conditions, the conversion The yield can reach 90%, but the fermentation process takes a long time and lasts for 120 hours. In addition, other carbon sources need to be added, which increases the cost and increases the difficulty of subsequent separation and purification.

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  • Method for generating lactobionic acid through increasing microbial conversion
  • Method for generating lactobionic acid through increasing microbial conversion
  • Method for generating lactobionic acid through increasing microbial conversion

Examples

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Embodiment 1

[0033] A method for improving microbial conversion to generate lactobionic acid, comprising the steps of:

[0034] 1) Primary fermentation: Pseudomonas fluorescens ATCC17400 was used as the test strain, activated, then inoculated on the seed medium for cultivation, at 30°C, with an air flow of 0.3vvm, and cultivated to a concentration of 10 9 Pseudomonas fluorescens seed solution of cfu / ml, the Pseudomonas fluorescens seed solution is inoculated in the fermenter that contains fermentation medium according to 10% inoculum amount to carry out fermentation culture, 30 ℃, ventilation is 0.4vvm, with Cultivate at a stirring speed of 100rpm for 24 hours, then add promotion medium, the ratio of promotion medium to fermentation medium is: 3g:1L; continue to ferment for 30h, stop fermentation, and obtain fermentation liquid; throughout the fermentation process, control the pH by adding ammonia water is 6.0.

[0035] Seed culture medium: by weight percentage, 5% lactose, 2% glucose, 1%...

Embodiment 2

[0041] A method for improving microbial conversion to generate lactobionic acid, comprising the steps of:

[0042] 1) Primary fermentation: Pseudomonas fluorescens ATCC17400 was used as the test strain, activated, then inoculated on the seed medium for cultivation, at 30°C, with an air flow of 0.3vvm, and cultivated to a concentration of 109 Pseudomonas fluorescens seed solution of cfu / ml, the Pseudomonas fluorescens seed solution is inoculated in the fermenter that contains fermentation medium according to 5% inoculum amount to carry out fermentation culture, 30 ℃, ventilation is 0.4vvm, with Cultivate at a stirring speed of 100rpm for 24 hours, then add promotion medium, the ratio of promotion medium to fermentation medium is: 2g:1L; continue fermentation for 28h, stop fermentation, and obtain fermentation liquid; throughout the fermentation process, control the pH by adding ammonia water is 6.1.

[0043] Seed culture medium: by weight percentage, 5% lactose, 2% glucose, 1% ...

Embodiment 3

[0049] A method for improving microbial conversion to generate lactobionic acid, comprising the steps of:

[0050] 1) Primary fermentation: Pseudomonas fluorescens ATCC17386 was used as the test strain, activated, then inoculated on the seed medium for cultivation, at 30°C, with an air flow of 0.3vvm, and cultivated to a concentration of 10 9 The Pseudomonas fluorescens seed solution of cfu / ml, the Pseudomonas fluorescens seed solution is inoculated in the fermenter that contains fermentation medium according to 8% inoculum amount to carry out fermentation culture, 30 ℃, ventilation is 0.4vvm, with Cultivate at a stirring speed of 100rpm for 24 hours, then add promotion medium, the ratio of promotion medium to fermentation medium is: 2.5g: 1L; continue fermentation for 28h, stop fermentation, and obtain fermentation liquid; throughout the fermentation process, control by adding ammonia water The pH is 6.2.

[0051] Seed culture medium: by weight percentage, 5% lactose, 2% glu...

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Abstract

The invention belongs to the technical field of biology, and discloses a method for generating lactobionic acid through increasing microbial conversion. The method comprises the following steps of step (1) performing primary fermentation: performing inoculation on a fermentation tank containing fermentation culture mediums with a fluorescent pseudomonas seed solution, performing fermentation culture for 20-24h, then adding a promoting culture medium, continuing fermentation for 24-36h, and stopping fermentation to obtain fermentation liquor; and (2) performing secondary fermentation: performing filtration on fermentation liquor after primary fermentation with a ceramic membrane, collecting filtrate and wet thalli, adding the wet thalli back to the fermentation tank, then adding secondary fermentation culture mediums, continuing performing fermentation culture for 10-14h, stopping fermentation, performing filtration on secondary fermentation liquor, collecting the filtrate, and mergingthe filtrate obtained through filtration twice for subsequent separation and extraction of the lactobionic acid. According to the method disclosed by the invention, secondary utilization is performedon abandoned wet thalli, waste is turned into wealth, and the yield of the lactobionic acid is increased.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for improving the transformation of microorganisms into lactobionic acid. Background technique [0002] Lactobionic acid is an important pharmaceutical intermediate, which has been widely used in the field of pharmaceutical preparation technology. At present, the methods for synthesizing lactobionic acid mainly include chemical methods, enzymatic methods and biological methods. Lactobionic acid used in the market is mainly produced by chemical methods, which are energy-intensive and require expensive metal catalysts. The chemical reagents used in the chemical method are some toxic and expensive reagents; there will be by-products in the reaction process; the metal catalyst is used to synthesize lactobionic acid, and the metal catalyst is easily passivated; the chemical method requires stable pH and temperature It is relatively strict, and it is easy to produce ...

Claims

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Application Information

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IPC IPC(8): C12P7/58C12N1/20C12R1/39
CPCC12P7/58C12N1/20
Inventor 刘加林霍萧勇刘加海方朝杰张渊源
Owner HANGZHOU BROTHER BIO TECH CO LTD