CN115369133A - Method for producing DHA grease by fermenting schizovibrio based on linseed oil - Google Patents

Method for producing DHA grease by fermenting schizovibrio based on linseed oil Download PDF

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CN115369133A
CN115369133A CN202210973369.5A CN202210973369A CN115369133A CN 115369133 A CN115369133 A CN 115369133A CN 202210973369 A CN202210973369 A CN 202210973369A CN 115369133 A CN115369133 A CN 115369133A
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fermentation
linseed oil
vibrio
dha
oil
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龚韧
金虎
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Hubei Ruishi Biotechnology Co ltd
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
    • C12P7/6436—Fatty acid esters
    • C12P7/6445—Glycerides
    • C12P7/6463—Glycerides obtained from glyceride producing microorganisms, e.g. single cell oil
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
    • C12P7/6409—Fatty acids
    • C12P7/6427—Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
    • C12P7/6434—Docosahexenoic acids [DHA]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
    • C12P7/6436—Fatty acid esters
    • C12P7/6445—Glycerides
    • C12P7/6472—Glycerides containing polyunsaturated fatty acid [PUFA] residues, i.e. having two or more double bonds in their backbone
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/89—Algae ; Processes using algae

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  • Biotechnology (AREA)
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  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

The invention provides a method for producing DHA grease by fermenting Vibrio fission based on linseed oil, which relates to the application of linseed oil as an inducer and a defoaming agent in the production of DHA by fermenting Vibrio fission, namely, a fermentation culture medium added with linseed oil accounting for 0.005-1% of the total volume of the fermentation culture medium is used for fermenting Vibrio fission; linseed oil was added to the medium as an antifoaming agent when foam was generated during fermentation. Experiments show that the added linseed oil is utilized in the method for producing DHA grease by fermenting schizovibrio based on linseed oil, so that the linseed oil not only has the function of a defoaming agent, but also can be used as an inducer to enrich grease in cells and DHA in the grease in the process of inducing the fermentation of the schizovibrio. The dry cell weight of the vibrio schizonepetae can be effectively improved, and the oil content in the cells and the DHA content in the oil can be improved after the fermentation of the vibrio schizonepetae is finished.

Description

Method for producing DHA grease by fermenting schizovibrio based on linseed oil
Technical Field
The invention relates to the technical field of marine microbial fermentation, in particular to a method for producing DHA grease by fermenting schizovibrio based on linseed oil.
Background
DHA is commonly called as brain gold and is a polyunsaturated fatty acid which is very important to human bodies. At present, the technology of directly obtaining DHA from marine fishes by utilizing marine microorganisms and replacing the marine microorganisms with a biological fermentation technology is becoming mature. However, it is common that the content of fats and oils in microbial cells and the DHA content in fats and oils are low. There are numerous application problems.
In the fermentation process of the schizovibrio, the oil content in cells is low, so that the production cost is high, and the DHA content in the oil is low, so that the DHA oil consumption in application is increased. Since microbial oils have a certain fishy smell, the taste of the microbial oils is impaired by an excessive amount of the microbial oils, and further, since microbial oils contain many other polyunsaturated fatty acids in addition to DHA. The grease is easily oxidized, forms instability of the product and influences the shelf life of the product.
Disclosure of Invention
Aiming at the defects in the prior art, the invention aims to provide a method for producing DHA grease by fermenting schizo vibrio based on linseed oil, so as to solve the problems in the background art, improve the dry cell weight of the schizo vibrio, and increase the grease content in cells and the DHA content in the grease.
In order to achieve the purpose, the invention is realized by the following technical scheme: a method for producing DHA grease by fermenting Vibrio fission based on linseed oil comprises the adding time and the adding amount of the linseed oil in the fermentation process of the Vibrio fission, wherein the linseed oil is used as a defoaming agent in the fermentation process of the Vibrio fission, and the defoaming effect is carried out on foams generated in the fermentation process in the production processes of batch fermentation (fermentation culture medium A) and semi-continuous fermentation (fermentation culture media B and C) of the Vibrio fission for fermenting the DHA. And added according to the following steps:
(1) A defoaming agent tank, wherein the linseed oil is put into the defoaming agent tank according to the proportion of 0.05-3% of the total volume of the fermentation liquor;
(2) Carrying out autoclaving on the linseed oil in the defoamer tank at the temperature of 121 ℃ for 20-30 minutes;
(3) And when foams are generated in the fermentation process of the schizo-vibrio, dropwise adding the linseed oil in the defoamer tank into the fermentation liquor until the foams are eliminated.
Furthermore, the linseed oil is used as an inducer in the fermentation process of DHA by the vibrio schizonepetae, and 0.005-1% of the linseed oil in the total volume of the fermentation medium is added into the fermentation medium (the fermentation medium A, B, C) for the batch fermentation and semi-continuous fermentation of DHA by the vibrio schizonepetae so as to induce the enrichment of the grease and DHA content of the vibrio schizonepetae.
Further, the fermentation medium added with linseed oil accounting for 0.005-1% of the total volume of the fermentation medium is autoclaved at 121 ℃ for 20-30 minutes.
Further, when foams are generated in the process of fermenting DHA by using the vibrio schizolyticus, the linseed oil is added dropwise.
Furthermore, except that the linseed oil is dripped when foams are generated in the process of fermenting DHA by the vibrio schizonepetae, at the later stage of 70-80 hours of fermentation, even if foams are not generated in the fermentation liquor, the linseed oil is dripped continuously for 2-5 times, and the dripping amount of each time is not more than 0.001-0.5 percent of the total volume of the fermentation liquor.
Further, the total addition amount of the linseed oil does not exceed 0.05-3% of the total volume of the fermentation liquor.
Further, the fermentation is batch fermentation or semi-continuous fermentation.
Further, in the batch fermentation, a fermentation medium is placed in a fermentation tank for sterilization, a defoamer tank, an acid-base tank (for adjusting the pH value) and the like are additionally arranged, the schizovibrio seed culture solution after the expanded culture is inoculated into the fermentation tank for closed culture, the pH value, the oxygen content, the defoaming and the like are adjusted as required in the culture process, the fermentation tank is adopted in the semi-continuous fermentation mode, the tank is larger in volume, and in general, in order to prolong the fermentation time and generate more microbial cells, a part of the fermentation liquid after the fermentation in the first fermentation tank is inoculated into the next fermentation tank for continuous fermentation culture.
Further, the components of the culture medium used for the semi-continuous fermentation are basically the same as those of the batch fermentation, and the culture conditions are basically the same.
The invention has the beneficial effects that:
1. according to the method for producing DHA grease by fermenting the vibrio schizolyticus based on linseed oil, the linseed oil is provided as an inducer and a defoaming agent in the DHA grease production by fermenting the vibrio schizolyticus, the linseed oil not only has the function of the defoaming agent, but also can be used as an inducer, and grease in cells and the DHA in the grease are enriched in the process of inducing the fermentation of the vibrio schizolyticus. Can effectively improve the dry cell weight of the schizovibrio.
2. According to the method for producing DHA grease by fermenting the vibrio schizolyticus based on the linseed oil, the linseed oil is added according to the amount of 0.005-1% in the fermentation process according to the condition of foam, and after the fermentation of the vibrio schizolyticus is finished, the grease content in cells and the DHA content in the grease can be improved.
Detailed Description
In order to make the technical means, the creation characteristics, the achievement purposes and the effects of the invention easy to understand, the invention is further explained by combining the specific embodiments.
Example 1: and (3) carrying out batch fermentation on the schizovibrio to produce DHA grease. The fermentation process of the 100L fermentation tank comprises the following steps: bevel seeds → shake flask seeds → 100L fermenter. Based on the batch fermentation medium A, 0.005%, i.e., 5g of linseed oil, was added thereto. Linseed oil is added in an amount of 0.005-1% during the fermentation process, depending on the foam. And (5) after the fermentation is finished for 120h, putting into a tank. In a 100L fermenter, the results of the experiments with and without linseed oil are shown in the following table:
Figure RE-GDA0003901613500000041
example 2: and (3) carrying out batch fermentation on the schizovibrio to produce DHA grease. The fermentation process of the 80T fermentation tank comprises the following steps: bevel seed → shake flask seed → primary seed tank → secondary seed tank → tertiary seed tank → 80 ton fermenter. Taking batch fermentation culture medium A as basic formula, adding 0.005% of 3Kg of linseed oil, and performing in-tank autoclaving at 121 deg.C for 20-30 min. The preparation method comprises the steps of preparing high-pressure sterilized linseed oil in a defoaming agent tank, and preparing 30Kg of the total amount of the linseed oil in the defoaming agent tank according to the proportion of 0.05 of the total volume of fermentation liquor. Linseed oil is added in an amount of 0.005-1% during the fermentation process, depending on the foam. During the fermentation process, glucose is fed according to the concentration of the glucose so as to maintain a carbon source required by the growth of the thalli. And (5) after the fermentation is finished for 120h, putting into a tank. In the 80T fermenter, the results of the experiments with the addition of linseed oil compared with the results of the experiments without the addition of linseed oil are shown in the following table:
Figure RE-GDA0003901613500000051
example 3: and (3) producing DHA grease by semi-continuous fermentation of the schizovibrio. Preparing the three-level seeds required by fermentation according to a batch fermentation process, inoculating the three-level seeds into an 80T fermentation tank, adding 0.005 percent of linseed oil (3 Kg) into the 80T fermentation tank by taking a culture medium B as a basic formula, fermenting for 48 hours, transferring 30 percent of fermentation liquor serving as seeds into another 80T fermentation tank, simultaneously supplementing 30 percent of culture medium with the formula C, continuously fermenting for 72 hours, putting the fermentation tank into the other 80T fermentation tank, and repeating the process in the other 80T fermentation tank. Linseed oil is added in an amount of 0.005-1% during the fermentation process, depending on the foam. During the fermentation process, glucose is fed according to the concentration of the glucose so as to maintain a carbon source required by the growth of the thalli. In the 80T fermenter, the results of the semi-continuous fermentation experiment with the addition of linseed oil are compared with the results of the batch fermentation experiment with the addition of linseed oil as follows:
Figure RE-GDA0003901613500000052
while there have been shown and described what are at present considered the fundamental principles and essential features of the invention and its advantages, it will be apparent to those skilled in the art that the invention is not limited to the details of the foregoing exemplary embodiments, but is capable of other specific forms without departing from the spirit or essential characteristics thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.
Furthermore, it should be understood that although the present description refers to embodiments, not every embodiment may contain only a single embodiment, and such description is for clarity only, and those skilled in the art should integrate the description, and the embodiments may be combined as appropriate to form other embodiments understood by those skilled in the art.

Claims (9)

1. A method for producing DHA grease by fermenting schizovibrio based on linseed oil is characterized by comprising the following steps: the method comprises the adding time and the adding amount of linseed oil in the fermentation process of the vibrio schizovici, wherein the linseed oil is used as a defoaming agent in the fermentation process of the vibrio schizovici, and the defoaming effect is performed on foams generated in the fermentation process in the production processes of batch fermentation (fermentation culture medium A) and semi-continuous fermentation (fermentation culture media B and C) of DHA (docosahexaenoic acid) fermentation of the vibrio schizovici. And added according to the following steps:
(1) A defoaming agent tank, wherein linseed oil is put into the defoaming agent tank according to the proportion of 0.05-3% of the total volume of the fermentation liquor;
(2) Sterilizing linseed oil in a defoaming agent tank at 121 ℃ for 20-30 minutes under high pressure;
(3) And when foams are generated in the fermentation process of the schizovibrio, dropwise adding the linseed oil in the defoaming agent tank into the fermentation liquor until the foams are eliminated.
2. The method for producing DHA oil through fermentation of Linseed oil-based Vibrio schizo fission according to claim 1, wherein: the linseed oil is used as an inducer in the fermentation process of DHA (docosahexaenoic acid) of the vibrio schizovici, and 0.005-1% of the linseed oil in the total volume of a fermentation medium is added into the fermentation medium (the fermentation medium A, B, C) for the batch fermentation and semi-continuous fermentation of DHA of the vibrio schizovici so as to induce the enrichment of the grease and DHA content of the vibrio schizonepetai.
3. The method for producing DHA oil through fermentation of Linseed oil-based Vibrio schizo fission according to claim 2, wherein: the fermentation medium added with linseed oil accounting for 0.005-1% of the total volume of the fermentation medium is autoclaved at 121 ℃ for 20-30 minutes.
4. The method for producing DHA oil by fermenting Linseed oil-based Schizovibrio according to claim 3, wherein: when foams are generated in the process of fermenting DHA by the vibrio schizonepetae, the linseed oil is dripped.
5. The method for producing DHA oil by fermenting Linseed oil-based Vibrio fission according to claim 1, wherein: except that the linseed oil is dripped when foams are generated in the process of fermenting DHA by the vibrio schizonepetae, the linseed oil is dripped continuously for 2-5 times in the later period of 70-80 hours of fermentation even if no foams are generated in the fermentation liquor, and the dripping amount of each time does not exceed 0.001-0.5 percent of the total volume of the fermentation liquor.
6. The method for producing DHA oil by fermenting Linseed oil-based Vibrio fission according to claim 1, wherein: the total addition amount of the linseed oil does not exceed 0.05-3% of the total volume of the fermentation liquor.
7. The method for producing DHA oil by fermenting Linseed oil-based Vibrio fission according to claim 5, wherein: the fermentation is batch fermentation or semi-continuous fermentation.
8. The method for producing DHA oil by fermentation of Linseed oil-based Vibrio fission according to claim 7, wherein: the fermentation culture medium is placed into a fermentation tank for sterilization in batch fermentation, a defoamer tank, an acid-base tank (for adjusting the pH value) and the like are additionally arranged, the schizovibrio seed culture solution after expanded culture is inoculated into the fermentation tank for closed culture, the pH value, the oxygen content, the defoaming and the like are adjusted as required in the culture process, the semi-continuous fermentation mode adopts fermentation tank fermentation, only the fermentation tank is larger in volume, and in general, in order to prolong the fermentation time and generate more microbial cells, a part of the fermentation liquid after the fermentation in the first fermentation tank is inoculated into the next fermentation tank for continuous fermentation culture.
9. The method for producing DHA oil by fermenting Linseed oil-based Vibrio fission according to claim 8, wherein: the components of the culture medium used for the semi-continuous fermentation are basically the same as those of batch fermentation, and the culture conditions are basically the same.
CN202210973369.5A 2022-08-15 2022-08-15 Method for producing DHA grease by fermenting schizovibrio based on linseed oil Withdrawn CN115369133A (en)

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