Method and fermentation device for increasing yield of riboflavin by using oxygen carrier

A fermentation device and riboflavin technology, applied in specific-purpose bioreactors/fermenters, microbial-based methods, fermentation, etc., can solve the complex reaction of pure chemical synthesis, affect the prospects of industrial production, and riboflavin production reduce problems such as improving the substrate conversion rate, shortening the fermentation cycle, and increasing the oxygen-dissolving capacity

Active Publication Date: 2020-01-17
HEBEI SHENGXUE DACHENG PHARMA
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, the large-scale synthesis of riboflavin is mainly divided into chemical synthesis and microbial fermentation. However, the reaction of pure chemical synthesis is too complicated, the cost is high and the yield is low, which greatly affects the prospect of industrial production.
[0004] The technology of riboflavin fermented by Bacillus subtilis has become relatively mature, replacing the previous complicated and inefficient chemical synthesis methods, but there are still some problems in the fermentation process. High, especially in the later stage of fermentation, the oxygen consumption of fermentation is high, once the dissolved oxygen is lower than a certain level, the riboflavin production will be greatly reduced
[0005] For a long time, most of the research on riboflavin production has been the transformation of genetically engineered bacteria, which is costly, time-consuming, and has a low success rate. The impact on production and cost caused by the oxygen consumption of the bacteria itself has been ignored. Therefore, how to meet the requirements during the fermentation process? The oxygen demand of Bacillus subtilis, increasing the production of riboflavin and reducing the cost are difficult problems that need to be solved at present. Therefore, we propose a method of using oxygen carrier to increase the production of riboflavin and its fermentation device to solve the above problems

Method used

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  • Method and fermentation device for increasing yield of riboflavin by using oxygen carrier
  • Method and fermentation device for increasing yield of riboflavin by using oxygen carrier
  • Method and fermentation device for increasing yield of riboflavin by using oxygen carrier

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Embodiment 1

[0038] This embodiment includes the following steps:

[0039] Seed culture: Inoculate the riboflavin-producing bacteria Bacillus subtilis into the seed medium, and culture it with shaking at 35°C and 200-240rpm on a fermentation device for 24 hours to obtain the seed medium;

[0040] Fermentation culture: Inoculate the obtained seed culture liquid into the fermentation medium, add sterile oxygen carrier n-dodecane at 0 hours after the fermentation starts, and the amount of n-dodecane added is 1%, and finally obtain the fermentation liquid containing riboflavin , the fermentation condition is: 500mL baffle Erlenmeyer flask, liquid volume is 30 milliliters, at 30-38 ℃, 200-240rpm fermenting equipment shaking culture for 120 hours.

[0041] Yield detection: the method is the same as that of Comparative Example, and the content of riboflavin in the fermented liquid of Example 1 is 12632 μg / mL, which is 10.6% higher than that of Comparative Example.

Embodiment 2

[0043] This embodiment includes the following steps:

[0044] Seed culture: Inoculate the riboflavin-producing bacteria Bacillus subtilis into the seed medium, and culture it with shaking at 35°C and 200-240rpm on a fermentation device for 24 hours to obtain the seed medium;

[0045] Fermentation culture: Inoculate the obtained seed culture liquid into the fermentation medium, add sterile oxygen carrier n-dodecane to the fermentation medium for 60 hours, and the addition amount of n-dodecane is 4%, finally obtain the fermentation liquid containing riboflavin, The fermentation conditions are: 500mL baffled Erlenmeyer flask with a liquid volume of 30ml, shake culture at 30-38°C, 200-240rpm fermentation equipment for 90 hours.

[0046] Yield detection: the method is the same as that of the comparative example. The riboflavin content in the fermented liquid of Example 2 is 13034 μg / mL, which is 12.4% higher than that of the comparative example, and the fermentation period is short...

Embodiment 3

[0048] This embodiment includes the following steps:

[0049] Seed culture: Inoculate the riboflavin-producing bacteria Bacillus subtilis into the seed medium, and culture it with shaking at 35°C and 200-240rpm on a fermentation device for 24 hours to obtain the seed medium;

[0050] Fermentation culture: inoculate the obtained seed culture solution into the fermentation medium, add sterile oxygen carrier n-dodecane to the fermentation medium for 50 hours, the addition of n-dodecane is 3%, and finally obtain the fermentation solution containing riboflavin, The fermentation conditions are: 500mL baffled Erlenmeyer flask with a liquid volume of 30ml, shake culture at 30-38°C, 200-240rpm on a fermentation device for 105 hours.

[0051] Yield detection: the method is the same as that of the comparative example, and the riboflavin content in the fermented liquid of Example 3 is 15820 μg / mL, which is 38.5% higher than that of the comparative example, and the fermentation period is s...

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Abstract

The invention discloses a method and a fermentation device for increasing yield of riboflavin by using an oxygen carrier. The method comprises the steps as follows: S1, seed culture: inoculating a seed culture medium with a riboflavin producing bacillus subtilis strain for culture to obtain a seed culture solution; and S2, fermentation culture: inoculating a fermentation culture medium with the obtained seed culture solution, then, adding the sterile oxygen carrier, and performing fermentation to obtain the riboflavin. The proper oxygen carrier is added to the culture medium, culture is performed in the fermentation device, the dissolved oxygen concentration in the culture medium is increased by chemical and physical methods, thallus metabolism of the riboflavin producing bacillus subtilisis promoted, the substrate conversion rate is increased, the synthetic amount of riboflavin is increased from 11424 [mu]g / mL to 15820 [mu]g / mL, besides, the fermentation cycle is shortened, and the production cost is reduced.

Description

technical field [0001] The invention relates to the technical field of microbial fermentation production, in particular to a method for increasing riboflavin production by using an oxygen carrier and a fermentation device thereof. Background technique [0002] Riboflavin is a kind of natural pigment soluble in water. It acts as a coenzyme to promote metabolism and exists in the form of flavin mononucleotide and flavin adenine dinucleotide in organisms. Riboflavin can be used to treat riboflavin deficiency, conjunctivitis, nutritional ulcers, and systemic nutritional disorders. Riboflavin lesions are mostly manifested as inflammation of the mouth, eyes and external genitalia, such as angular cheilitis, cheilitis, glossitis, conjunctivitis and scrotitis, etc. Therefore, riboflavin has a wide range of applications in food, feed, and pharmaceutical industries. application prospects. [0003] At present, the large-scale synthesis of riboflavin is mainly divided into chemical sy...

Claims

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Application Information

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IPC IPC(8): C12P25/00C12M1/38C12M1/36C12M1/34C12R1/125
CPCC12P25/00C12M23/48C12M41/12C12M41/34C12M23/46C12M27/16C12M29/00
Inventor 周景王绘砖魏伟林青曹建全
Owner HEBEI SHENGXUE DACHENG PHARMA
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