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Fermentation method for preparing beta-carotin by using Blakeslea trispora

A technology of B. trispora and B. boulardii is applied in the field of fermentation for preparing beta-carotene, can solve the problems such as lack of comprehensive consideration of the fermentation process, unfavorable full utilization of fermenters, influence on the yield of target products and the like, and achieves low cost , Conducive to bacterial growth and product synthesis, the effect of reducing the amount of feed

Active Publication Date: 2016-04-20
INNER MONGOLIA KINGDOMWAY PHARMA LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the growth characteristics of the filamentous fungus B. trispora itself, its fermentation medium mostly uses viscous substances (such as starch, soybean powder, etc.) as carbon and nitrogen sources to facilitate the extension of filamentous fungal hyphae. Mycelium balls are finally formed, but the fermented liquid with high viscosity is a non-Newtonian fluid most of the time, which has a great influence on the transfer of oxygen; and in the early stage of fermentation, the rapid growth of bacteria makes it difficult to absorb dissolved oxygen and nutrients. The demand is relatively high. As the concentration of bacteria increases, the viscosity of the fermentation broth is further increased, which leads to dissolved oxygen becoming a key factor that limits the growth of bacteria and product synthesis, and even causes metabolic abnormalities, affecting the output of target products.
[0005] Therefore, in the fermentation process of producing β-carotene with B. trispora as the strain, researchers often screen excellent strains through mutagenesis or change the stirring form of fermentation and the way of dissolved oxygen supply, and add oxygen carrier during the fermentation process. Or add a surfactant that can increase the permeability of the bacterial cells to increase the oxygen concentration and oxygen utilization in the fermentation broth. These studies have a certain effect on the improvement of the fermentation process, but there are also shortcomings of one kind or another, or The final output is low, or the cost is high due to the complexity of the equipment, or it is only for one aspect without comprehensive consideration of the fermentation process
Chinese patent CN1331343A discloses a process for changing the form of the stirring blades. The upper and middle layers are wide blade propulsion type and the lower layer is a turbine type combined stirring blades, which improves the mass transfer performance of the fermented liquid, but changes After the stirring form can only be suitable for the production characteristics of the strain, it is not conducive to the comprehensive utilization of the fermenter; in the patent CN102787158A, the oxygen concentration entrained in the fermentation liquid is improved by performing colloid mill high-speed shear emulsification on the medium before inoculation. The requirements are relatively high, the cost investment is large, and the fermentation process has not been explored too much, and the subsequent extraction process has been studied in depth; the patent CN1193048A uses an air-lift fermenter to ferment B. trispora to prepare β -Carotene, this method can better avoid shearing force, is conducive to the growth of mycelia to obtain larger biomass, but the final yield is low, and it is not easy to carry out large-scale production

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1: a kind of fermentation method utilizing B. trispora to prepare β-carotene, it comprises the following steps: (1) strain activation; (2) primary seed cultivation; (3) secondary seed cultivation; ( 4) fermentation culture; wherein,

[0026] (1) Strain activation: under aseptic conditions, inoculate the positive and negative bacterial strains of B. trispora on the slanted PDA medium (potato dextrose agar medium, suitable for fungal growth), and place them in the incubator. Cultivate at 26°C for 4 days, after the positive and negative strains of B. trispora grow spores respectively, scrape and wash the spores in the slant medium with sterile saline under aseptic conditions, and prepare a uniform spore suspension. The concentrations of positive and negative bacteria spore suspensions are respectively: positive bacteria 10 4 spores / mL, negative bacteria 10 5 spores / mL; wherein, Blakesleatrispora positive and negative strains were purchased from ATCC, numbered ...

Embodiment 2

[0033] Embodiment 2: a kind of fermentation method utilizing B. trispora to prepare β-carotene, which comprises the following steps: (1) strain activation; (2) primary seed cultivation; (3) secondary seed cultivation; ( 4) fermentation culture; wherein,

[0034] (1) Strain activation: under aseptic conditions, inoculate the positive and negative bacterial strains of B. trispora on the slanted PDA medium (potato dextrose agar medium, suitable for fungal growth), and place them in the incubator. Cultivate at 30°C for 5 days, after the positive and negative strains of B. trispora grow spores respectively, scrape and wash the spores in the slant medium with sterile saline under aseptic conditions, and prepare a uniform spore suspension. The concentrations of positive and negative bacteria spore suspensions are respectively: positive bacteria 10 4 spores / mL, negative bacteria 10 5 spores / mL; wherein, Blakesleatrispora positive and negative strains were purchased from ATCC, number...

Embodiment 3

[0041] Embodiment 3: a kind of fermentation method utilizing B. trispora to prepare β-carotene, it comprises the following steps: (1) strain activation; (2) primary seed cultivation; (3) secondary seed cultivation; ( 4) fermentation culture; wherein,

[0042] (1) Strain activation: under aseptic conditions, inoculate the positive and negative bacterial strains of B. trispora on the slanted PDA medium (potato dextrose agar medium, suitable for fungal growth), and place them in the incubator. Cultivate at 28°C for 4 days, after the positive and negative strains of B. trispora grow spores respectively, scrape and wash the spores in the slant medium with sterile saline under aseptic conditions, and prepare a uniform spore suspension. The concentrations of positive and negative bacteria spore suspensions are respectively: positive bacteria 10 4spores / mL, negative bacteria 10 5 spores / mL; wherein, Blakesleatrispora positive and negative strains were purchased from ATCC, numbered A...

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Abstract

The invention discloses a fermentation method for preparing beta-carotin by using Blakeslea trispora, which comprises the following steps: (1) strain activation; (2) primary seed culture; (3) secondary seed culture; and (4) fermentation culture. The method has the advantages of low facility request, low cost input, wide raw material sources and low cost, obviously enhances the beta-carotin yield as compared with the unimproved technique, and thus, can implement large-scale production.

Description

technical field [0001] The invention relates to a fermentation method for preparing beta-carotene, in particular to a fermentation method for preparing beta-carotene by using B. trispora. Background technique [0002] β-carotene is a fat-soluble unsaturated aliphatic hydrocarbon, a typical representative of the carotenoid family, and the most common and stable natural pigment in nature. It is widely found in green and yellow vegetables and fruits. Its molecular formula is C 40 h 56 , the molecular weight is 536.88, the appearance is dark red to dark red, unstable to oxygen, heat and light, prone to redox reactions, not easily soluble in water, acid and alkali, easily soluble in chloroform, benzene and vegetable oil, slightly soluble in ethanol and ether. In addition to being commonly used as a coloring agent, β-carotene also has a variety of biological activities and is a medicinal active ingredient closely related to human health. Studies have found that because the mol...

Claims

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Application Information

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IPC IPC(8): C12P23/00C12R1/645
CPCC12P23/00
Inventor 钟锦潮蒋四富陈必钦文昌严必能赖荣乾戴昌华
Owner INNER MONGOLIA KINGDOMWAY PHARMA LTD
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