Method of preparing lycopene from trispore Bruce mould

A technology of Blakeslea and Blakeslea trispora, applied in the fields of chemical industry, medicine, and food, can solve the problems of limited oxygen supply, increased operating costs, and high viscosity of the medium, achieving fast oxygen transfer speed and less bubble generation , the effect of increasing biomass

Inactive Publication Date: 2007-02-28
BEIJING UNIV OF CHEM TECH
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AI Technical Summary

Problems solved by technology

[0005] In the method of using Blakeslea trispora to produce lycopene, since Blakeslea trispora is a highly aerobic and viscophilic microorganism, the medium used has a high viscosity and a high solid content, and also contains 5% vegetable oil (providing double for metabolism). key, resulting in a low dissolved oxygen rate in the fermentation system); during the growth process, it is easy to tangle into agglomerates, resulting in a far insufficient amount of dissolved oxygen in the logarithmic growth period, so the problem of oxygen supply is to restrict the biological reaction in the fermentation process of B. trispora and important factors of life process (Jeong JC.Lee IY.Kim SW.Park YH.1999.Biotechnol.Lett.21:683-686; Kim SW.SeoWT.Park YH.1997.J.Ferment.Bioeng.84:330 -332)
[0006] For most fermentations, the lack of oxygen will cause metabolic abnormalities, biomass and target metabolite yield reduction; and because the solubility of oxygen in aqueous solution is very low (its saturation is less than 7mg / L), the oxygen supply in fermentation is very low It is easy to become the main contradiction, so how to ensure the supply of oxygen in fermentation to meet the oxygen demand of production strains is one of the keys to improve fermentation production capacity and reduce costs
At present, the traditional mechanical oxygen supply methods such as stirring ventilation and shaking table oscillation have the following problems: when the operating conditions of the equipment are reduced, the oxygen supply is severely limited, and the formation of target metabolites is blocked; when the operating conditions are increased, the high mechanical strength will be enhanced The damage to the cells caused by shearing will cause the bacteria to be damaged and cannot metabolize normally, and the operating cost will also increase greatly; while improving the design of bioreactors, stirring paddles and air distributors, and using oxygen-enriched air will reduce production costs. greatly improved, so it is necessary to find a new way to strengthen oxygen supply

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] (1) Seed cultivation:

[0022] Inoculate the seed medium with B. trispora ATCC 14271 (+), ATCC 14272 (-) strains, the components and contents of the seed medium are: starch 40g / L, corn steep liquor 50g / L, KH 2 PO 4 1g / L, MgSO 4 0.1g / L, VB 1 0.01g / L, pH6.5. 28°C, 180rpm shaker culture for 40h, (+) (-) bacterial culture culture solution obtained by mixing 1:2 to obtain a seed solution.

[0023] (2) Fermentation culture:

[0024] Inoculate the seed liquid into the fermentation medium according to the inoculation amount of 10%. The components and contents of the fermentation medium are: starch 40g / L, soybean meal 20g / L, corn steep liquor 25g / L, KH 2 PO 4 1g / L, MgSO 4 0.1g / L, VB 1 0.01g / L, 20ml / L filter-sterilized n-hexane, pH6.5. A 500ml Erlenmeyer flask was used for fermentation culture, with a liquid volume of 100ml, and the cells were harvested after 4 days of culture on a shaker at 26°C and 220rpm. Add 1.5 g of pyridine or a tertiary amine compound blocke...

Embodiment 2

[0028] (1) Seed cultivation:

[0029] Inoculate the seed medium with B. trispora ATCC 14271 (+), ATCC 14272 (-) strains, the components and content of the seed medium are: starch 500g / L, corn steep liquor 40g / L, KH 2 PO 4 1.5g / L, MgSO 4 0.2g / L, VB 1 0.02g / L, pH6.5. 26°C, 200rpm shaker culture for 38h, (+) (-) bacteria culture culture solution obtained by mixing 1:2 to obtain seed solution.

[0030] (2) Fermentation culture:

[0031] The components and content of the fermentation medium are: starch 40g / L, soybean meal 20g / L, corn steep liquor 25g / L, KH 2 PO 4 1g / L, MgSO 4 0.1g / L, VB 1 0.01g / L, 15ml / L filter-sterilized n-dodecane, cultured on a shaker at 30°C and 230rpm for 6 days, and harvested the bacteria. Ferment to 50 hours and add 1.5g pyridine or tertiary amine compound blocking agent, other are the same as embodiment 1.

[0032] (3) Extraction

[0033] The fermented liquid was filtered with gauze to obtain wet thallus, and the wet thallus was placed in a ...

Embodiment 3

[0035] (1) Seed cultivation:

[0036] Seed culture process is with embodiment 1.

[0037] (2) Fermentation culture:

[0038] Inoculate the seed liquid into the fermentation medium according to the inoculation amount of 10%. The components and contents of the fermentation medium are: starch 50g / L, soybean meal 25g / L, corn steep liquor 20g / L, KH 2 PO 4 1.5g / L, MgSO 4 0.05g / L, VB 1 0.01g / L, tween-80 1g / L, 10ml / L filter-sterilized n-hexane, pH6.5. Harvest the cells after culturing on a shaker at 28° C. for 5 days at 250 rpm. Add 1.5g of pyridine or tertiary amine compound blocking agent after fermenting to 48 hours, and others are the same as in Example 1.

[0039] (3) Extraction:

[0040] The fermented liquid was filtered with gauze to obtain wet thallus, and after the wet thalline was placed in a vacuum drying oven at 50° C. for 48 hours, 5.08 g of dry bacterium powder was obtained. Dried bacterial powder is crushed, extracted with petroleum ether (take 5 μ l extract,...

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PUM

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Abstract

The invention relates the method for fermenting trisporic Bang's fungus to produce lycopersicin. The carrier of oxygen is added into fermentation culture medium to provide a carrier of oxygen fermentation system, the invention improves the oxygen concentration without consuming excess energy source, and it improves cell biomass and lycopersicin content, and it improves the productivity of lycopersicin.

Description

Technical field: [0001] The invention relates to a method for preparing lycopene by fermenting the three spora boulardii. It belongs to the fields of food, medicine and chemical industry. Background technique: [0002] Lycopene is a fat-soluble carotenoid and is an intermediate in the biosynthesis of many carotenoids. Compared with β-carotene, although lycopene is not the precursor of vitamin A, its ability to eliminate singlet oxygen and free radicals and its antioxidant effect are superior to β-carotene. A series of important physiological functions such as cardiovascular disease. [0003] Lycopene is a straight-chain hydrocarbon composed of 11 conjugated and 2 non-conjugated carbon-carbon double bonds. It is easy to oxidize and isomerize, so it is difficult to extract. At present, the commonly used extraction methods are: organic solvent extraction, supercritical extraction, saponification, and its detection methods mainly include spectrophotometry, paper chromatograph...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P5/02C12R1/645
Inventor 袁其朋徐芳朱艳
Owner BEIJING UNIV OF CHEM TECH
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