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Sedimentation-reverse perfusion-flow coupling method for preparing DHA-rich oil and its special cell sedimentation tank

A settling tank, cell technology, applied in microorganism-based methods, biochemical equipment and methods, specific-purpose bioreactors/fermenters, etc., can solve the problem of unstable DHA yield, reduced DHA yield, and reduced oil quality and other problems to achieve the effect of avoiding cell loss, avoiding stasis and senescence, maintaining activity and accumulation of fatty acids

Active Publication Date: 2018-02-16
RUNKE BIOENG FUJIAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

With the advancement of technology, the yield of DHA produced by intermittent fed-feed fermentation microalgae has gradually increased, but the proportion of DHA produced by these patents is less than 40% of the total fatty acids, and the proportion of saturated fatty acids is relatively high, which reduces the quality of oil; and The yield of DHA in the fed-batch fermentation process is unstable, and the improvement of the yield is largely limited by the concentration of the product and the fermentation time.
The invention patent with application number 94106621.5 adopts continuous culture method to cultivate ATCC30021 algae, the cell concentration in the fermenter reaches 15g / L, and the DHA production rate is 2.88g / L / d. Inhibitory effect, but due to the loss of cells, the concentration of cells in the tank is lower, reducing the yield of DHA

Method used

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  • Sedimentation-reverse perfusion-flow coupling method for preparing DHA-rich oil and its special cell sedimentation tank
  • Sedimentation-reverse perfusion-flow coupling method for preparing DHA-rich oil and its special cell sedimentation tank
  • Sedimentation-reverse perfusion-flow coupling method for preparing DHA-rich oil and its special cell sedimentation tank

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Preparation of oil rich in DHA by conventional cell settling tank sedimentation-reverse perfusion-discharge coupling

[0036] Preservation and cultivation of microalgae: Schizochytrium sp. CGMCC No. 7693 in this example was submitted to the General Microorganism Center of China Committee for the Collection of Microorganisms (CGMCC for short) for preservation on June 9, 2013. The composition of the solid preservation, expansion and fermentation medium of algae species is: glucose 10g / L, tryptone 1g / L, yeast extract 3g / L, sodium chloride 15g / L, magnesium sulfate 2g / L, potassium dihydrogen phosphate 0.5g / L, potassium chloride 1g / L, calcium chloride 0.5g / L, ammonium sulfate 0.2g / L, sodium bicarbonate 0.1g / L, boric acid 34.2mg / L, edetate disodium 40mg / L L, manganese chloride 2.5mg / L, ferric chloride 2.5mg / L, cyanocobalamin 0.1mg / L, zinc chloride 2mg / L, cobalt chloride 0.5mg / L, copper sulfate 0.02mg / L, pH = 6, add agar 15g / L to the solid medium. After the medium is prepared...

Embodiment 2

[0039] Preparation of DHA-enriched oil in a 10-liter fermenter using sedimentation-backflow-flow coupling

[0040] In the present embodiment, carry out the activation and expansion culture of bacterial classification by the method identical with embodiment 1, prepare 9L fermentation culture and be based on in 10L fermenter, the composition of culture medium is: glucose 20g / L, tryptone 1g / L, Yeast extract 5g / L, sodium chloride 25g / L, magnesium sulfate 1g / L, potassium dihydrogen phosphate 5g / L, potassium chloride 0.1g / L, calcium chloride 0.3g / L, ammonium sulfate 1g / L, carbonic acid Sodium Hydrogen 0.5g / L, Boric Acid 25mg / L, Disodium EDTA 50mg / L, Manganese Chloride 0.5mg / L, Ferric Chloride 5mg / L, Cyanocobalamin 0.2mg / L Zinc Chloride 1.5mg / L, cobalt chloride 0.05mg / L, copper sulfate 0.01mg / L, pH=6.5, add agar 15g / L to the solid medium. Cool after sterilization, inoculate into a fermenter according to 5% inoculation amount, the fermentation temperature is 20-30 DEG C, the pH is co...

Embodiment 3

[0042] Preparation of oil rich in DHA by coupling cell sedimentation-reverse perfusion-discharge in a special cell sedimentation tank

[0043] This embodiment provides an improved settling device for intercepting easily-settled microbial cells, so as to more effectively complete the cell concentration reflux in the process of settling-counterperfusion microalgae cell culture. What needs to be solved is the problem that the cells are not easy to settle due to the disturbance of the fermentation liquid in the ordinary settling tank, and the problems that the hollow fiber membrane is easy to be blocked, difficult to clean, and has a short life.

[0044] The settling tank specifically includes (as image 3 ): special settling tank, including two first-level settling tank 1 and second-level settling tank 2 arranged at intervals and conical in shape, and a cover 5 arranged on the first-level settling tank 1 and second-level settling tank 2, The opening 4 opened on the cover 5 is op...

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Abstract

The invention relates to a method for preparing grease rich in DHA and special equipment therefor. The method comprises the following steps: in an aeration fermentation tank containing a fermentation culture medium, cultivating schizochytrium CGMCCNo.7693 in a mechanical stirring manner; and extracting and recovering the grease rich in DHA from schizochytrium cells, wherein the culture process comprises continuous fed-batch cultivation of a medium, sedimentation-adverse perfusion of cells, and periodical discharge of fermentation liquor; the special cell sedimentation tank comprises a primary sedimentation tank, a secondary sedimentation tank, seal covers, openings and outlets; the primary sedimentation tank and the secondary sedimentation tank are arranged at an interval and are of conical shapes in appearance; the seal covers are arranged on the primary sedimentation tank and the secondary sedimentation tank; the openings are formed in the seal covers; the outlets are formed in the bottoms of the primary sedimentation tank and the secondary sedimentation tank; a liquid inlet pipe is arranged on the primary sedimentation tank; a supernate discharge pipe is arranged at the upper part of the secondary sedimentation tank; the primary sedimentation tank and the secondary sedimentation tank are connected through a communicating pipe; and the outlets are connected with a fermentation tank through concentrated cell reflux pumps and pipes. According to the method and the special cell sedimentation tank, the problems in a micro algae continuous cultivation technique that cells are difficultly intercepted and cell loss is serious are solved, the special cell sedimentation tank has the advantages of reduction of manual operation, easiness in automatic production, short production cycle and the like.

Description

technical field [0001] The invention relates to a preparation method and special equipment for oil rich in DHA. Background technique [0002] Known as "brain gold", DHA is an essential nutrient for the human body and has important physiological functions for the human body. Insufficient intake will cause discomfort in body functions. There have been many reports on the physiological function and practical application of DHA. DHA is an essential nutrient element for the development of nerves and vision. It has the function of regulating the central nervous system, and also has the effects of lowering blood pressure, regulating blood lipid balance in the human body, and preventing cardiovascular diseases. [0003] At present, DHA is mainly obtained from the fermentation culture of heterotrophic microalgae. Microalgae fermentation can adopt the following culture methods: batch, fed-batch and continuous culture. Batch culture is an operation mode in which the substrate matrix...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P7/64C12M1/00C12R1/89
Inventor 姜悦陈璇柳泽深
Owner RUNKE BIOENG FUJIAN
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