CN111235033A - Probiotic oil preservation process - Google Patents
Probiotic oil preservation process Download PDFInfo
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- CN111235033A CN111235033A CN202010068565.9A CN202010068565A CN111235033A CN 111235033 A CN111235033 A CN 111235033A CN 202010068565 A CN202010068565 A CN 202010068565A CN 111235033 A CN111235033 A CN 111235033A
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/04—Preserving or maintaining viable microorganisms
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Abstract
The invention discloses a probiotic oil preservation process, which specifically comprises the following steps: s1, thallus collection: the invention relates to the technical field of microorganisms, and discloses a method for culturing probiotics in logarithmic growth phase in a culture medium. The probiotic oil preservation process comprises the following steps of S1: placing the strain suspension cultured to the probiotics in the logarithmic growth phase in a centrifuge for centrifugation, collecting the strains, wherein the centrifugation temperature is 30-38 ℃, the centrifugation speed is 500-2000 r/min, the centrifugation time is 30-50 min, and S2 is added with substances: and (4) placing the thalli collected in the step S1 in a glass conical flask, dehydrating the compound oil by adopting a vacuum technology, adding the prepared compound oil into the thalli, and utilizing the strong oxidation resistance of the compound oil to play a more effective protection effect on the thalli so as to improve the survival rate of the probiotics after the preservation time is longer.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to a probiotic oil preservation process.
Background
The probiotics is an active microorganism which is beneficial to a host and is formed by changing the flora at a certain part of the host through colonization in a human body, and a single microorganism or a mixed microorganism with definite composition which is beneficial to the health of the host is generated by regulating the immune function of the host mucous membrane and the system or regulating the flora balance in the intestinal tract to promote the nutrition absorption and keep the intestinal tract healthy, and the probiotics exerts a microorganism additive which is beneficial to the host by improving the intestinal tract microorganism balance, and researches show that the probiotics has great physiological effects in the aspects of regulating the intestinal tract flora structure, enhancing the immunity of the organism, reducing cholesterol, eliminating carcinogenic factors and the like, has important significance for preventing and treating diabetes, hypertension, hyperlipidemia and other diseases, and has hidden troubles brought to the health of people by the environmental problems, the life style and the food sanitation at present, the probiotics have great potential in the aspects of maintaining health, reducing the risk of diseases and the like, so that the probiotics are more and more widely applied to various fields such as food, biological medicine products and the like.
The preservation process of probiotics in the prior art is still imperfect, although the traditional low-temperature strain preservation method is generally applicable to most strains, corresponding chemical protective agents such as glycerol, dimethyl sulfoxide and the like need to be added, the strain activity is easily affected, the probability of mixed bacteria pollution is high, and the survival rate of the probiotics after long-time preservation is low, so that a novel probiotic oil preservation process is provided, the probiotics can be preserved for a longer time, and the survival rate of the probiotics is high.
Disclosure of Invention
Technical problem to be solved
Aiming at the defects of the prior art, the invention provides a probiotic oil preservation process, which solves the problems that although the traditional low-temperature strain preservation method is generally applicable to most strains, corresponding chemical protective agents such as glycerol, dimethyl sulfoxide and the like need to be added, the strain activity is easily influenced, the probability of mixed bacteria pollution is high, and the survival rate of probiotics after long-time preservation is low.
(II) technical scheme
In order to achieve the purpose, the invention is realized by the following technical scheme: a probiotic oil preservation process specifically comprises the following steps:
s1, thallus collection: placing the strain suspension cultured to the probiotics in logarithmic phase in a centrifuge for centrifugal treatment, collecting the strains, wherein the centrifugal temperature is 30-38 ℃, the centrifugal rotating speed is 500-2000 r/min, and the centrifugal time is 30-50 min;
s2, addition of substances: placing the thalli collected in the step S1 in a glass conical flask, dehydrating the compound oil by adopting a vacuum technology, placing the dehydrated compound oil in a storage container, injecting the thalli in the glass conical flask into the storage container, wrapping the thalli by using the compound oil in the storage container, adsorbing the compound oil with the walls of the thalli, isolating the thalli from the outside, and finally sealing the bottle opening of the glass conical flask by using paraffin;
s3, low-temperature preservation: storing the glass conical flask at 3-5 ℃ for 6-10 hours, storing the glass conical flask at-2-4 ℃ for 6-10 hours, storing the glass conical flask at-8-10 ℃ for 6-10 hours, storing the glass conical flask at-14-16 ℃ for 6-10 hours, and finally storing the glass conical flask at-20-25 ℃.
Preferably, the composite oil raw materials comprise the following components in parts by weight: 80-90 parts of medium-chain triglyceride, 1-3 parts of tea polyphenol and 5-7 parts of organic solvent.
Preferably, the composite oil raw materials comprise the following components in parts by weight: 80 parts of medium-chain triglyceride, 1 part of tea polyphenol and 5 parts of organic solvent.
Preferably, the composite oil raw materials comprise the following components in parts by weight: 90 parts of medium-chain triglyceride, 3 parts of tea polyphenol and 7 parts of organic solvent.
Preferably, the composite oil raw materials comprise the following components in parts by weight: 85 parts of medium-chain triglyceride, 2 parts of tea polyphenol and 6 parts of organic solvent.
Preferably, the organic solvent is one or more of methanol, ethanol and acetone, wherein ethanol is a preferred embodiment of the organic solvent.
Preferably, the tea polyphenol is mixed with an organic solvent, and the tea polyphenol and the organic solvent are uniformly mixed by using an ultrasonic dispersion machine.
Preferably, the mixed solution of tea polyphenol and organic solvent is added into medium-chain triglyceride, and the mixed solution is stirred by a glass stirring rod at the stirring speed of 200r/min for 3 minutes.
(III) advantageous effects
The invention provides a process for preserving probiotic oil. Compared with the prior art, the method has the following beneficial effects:
(1) the probiotic oil preservation process comprises the following steps of S1: placing the strain suspension cultured to the probiotics in the logarithmic growth phase in a centrifuge for centrifugation, collecting the strains, wherein the centrifugation temperature is 30-38 ℃, the centrifugation speed is 500-2000 r/min, the centrifugation time is 30-50 min, and S2 is added with substances: placing the thalli collected in the step S1 in a glass conical flask, dehydrating the compound oil by adopting a vacuum technology, placing the dehydrated compound oil into a storage container, injecting the thalli in the glass conical flask into the storage container, wrapping the thalli by using the compound oil in the storage container, adsorbing the compound oil with the walls of the thalli, isolating the thalli from the outside, and finally sealing the bottleneck of the glass conical flask by using paraffin, wherein the step S3 is characterized in that: the method comprises the steps of storing the glass conical flask at 3-5 ℃ for 6-10 hours, storing the glass conical flask at-2-4 ℃ for 6-10 hours, storing the glass conical flask at-8-10 ℃ for 6-10 hours, storing the glass conical flask at-14-16 ℃ for 6-10 hours, storing the glass conical flask at-20-25 ℃, adding the prepared compound oil into thalli, and using strong oxidation resistance of the compound oil to achieve a more effective protection effect on the thalli and improve the survival rate of probiotics after the preservation time is longer.
(2) The probiotic oil preservation process comprises the following steps of S1: placing the strain suspension cultured to the probiotics in the logarithmic growth phase in a centrifuge for centrifugation, collecting the strains, wherein the centrifugation temperature is 30-38 ℃, the centrifugation speed is 500-2000 r/min, the centrifugation time is 30-50 min, and S2 is added with substances: placing the thalli collected in the step S1 in a glass conical flask, dehydrating the compound oil by adopting a vacuum technology, placing the dehydrated compound oil into a storage container, injecting the thalli in the glass conical flask into the storage container, wrapping the thalli by using the compound oil in the storage container, adsorbing the compound oil with the walls of the thalli, isolating the thalli from the outside, and finally sealing the bottleneck of the glass conical flask by using paraffin, wherein the step S3 is characterized in that: the method comprises the following steps of storing the glass conical flask at 3-5 ℃ for 6-10 hours, storing the glass conical flask at-2-4 ℃ for 6-10 hours, storing the glass conical flask at-8-10 ℃ for 6-10 hours, storing the glass conical flask at-14-16 ℃ for 6-10 hours, and finally storing the glass conical flask at-20-25 ℃, wherein the survival rate of probiotics during storage can be effectively improved by utilizing the descending temperature for storage, and the vitality of strains is not easily influenced by low temperature.
(3) The probiotic oil preservation process comprises the following steps of by weight: 80-90 parts of medium-chain triglyceride, 1-3 parts of tea polyphenol and 5-7 parts of organic solvent, wherein the organic solvent is one or a combination of methanol, ethanol and acetone, the ethanol is the preferred scheme of the organic solvent, the tea polyphenol and the organic solvent are mixed, the tea polyphenol and the organic solvent are uniformly mixed by using an ultrasonic dispersion machine, the mixed solution of the tea polyphenol and the organic solvent is added into the medium-chain triglyceride, a glass stirring rod is used for stirring the mixed solution, the stirring speed is 200r/min, the stirring time is 3 minutes, and the tea polyphenol is added into the medium-chain triglyceride, so that the oxidation resistance of the compound oil is greatly improved, and the probiotic protection effect of the compound oil is ensured.
Drawings
FIG. 1 is a flow chart of a process for preserving probiotic oils of the present invention;
FIG. 2 is a statistical table of comparative experimental data according to the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Referring to fig. 1-2, the embodiment of the present invention provides three technical solutions: a probiotic oil preservation process specifically comprises the following embodiments:
example 1
S1, thallus collection: placing the strain suspension of the probiotics cultured to logarithmic phase in a centrifuge for centrifugation, collecting the strains, wherein the centrifugation temperature is 30 ℃, the centrifugation speed is 500 rpm, and the centrifugation time is 30 minutes;
s2, addition of substances: placing the thalli collected in the step S1 in a glass conical flask, dehydrating the compound oil by adopting a vacuum technology, placing the dehydrated compound oil into a storage container, injecting the thalli in the glass conical flask into the storage container, wrapping the thalli by using the compound oil in the storage container, adsorbing the compound oil with the walls of the thalli, isolating the thalli from the outside, and finally sealing the bottleneck of the glass conical flask by using paraffin, wherein the compound oil comprises the following raw materials in parts by weight: 80 parts of medium-chain triglyceride, 1 part of tea polyphenol and 5 parts of organic solvent, wherein 1 part of tea polyphenol is mixed with 5 parts of organic solvent, 1 part of tea polyphenol and 5 parts of organic solvent are uniformly mixed by using an ultrasonic dispersion machine, a mixed solution of 1 part of tea polyphenol and 5 parts of organic solvent is added into 80 parts of medium-chain triglyceride, and the mixed solution is stirred by using a glass stirring rod at the stirring speed of 200r/min for 3 minutes;
s3, low-temperature preservation: the glass conical flask is stored for 6 hours at 3 ℃, stored for 6 hours at-2 ℃, stored for 6 hours at-8 ℃, stored for 6 hours at-14 ℃ and stored at-20 ℃.
Example 2
S1, thallus collection: placing the strain suspension of the probiotics cultured to logarithmic phase in a centrifuge for centrifugation, collecting the strains, wherein the centrifugation temperature is 38 ℃, the centrifugation speed is 2000 rpm, and the centrifugation time is 50 minutes;
s2, addition of substances: placing the thalli collected in the step S1 in a glass conical flask, dehydrating the compound oil by adopting a vacuum technology, placing the dehydrated compound oil into a storage container, injecting the thalli in the glass conical flask into the storage container, wrapping the thalli by using the compound oil in the storage container, adsorbing the compound oil with the walls of the thalli, isolating the thalli from the outside, and finally sealing the bottleneck of the glass conical flask by using paraffin, wherein the compound oil comprises the following raw materials in parts by weight: 90 parts of medium-chain triglyceride, 3 parts of tea polyphenol and 7 parts of organic solvent, wherein 3 parts of tea polyphenol and 7 parts of organic solvent are mixed, 3 parts of tea polyphenol and 7 parts of organic solvent are uniformly mixed by using an ultrasonic dispersion machine, the mixed solution of 3 parts of tea polyphenol and 7 parts of organic solvent is added into 90 parts of medium-chain triglyceride, and a glass stirring rod is used for stirring the mixed solution at the stirring speed of 200r/min for 3 minutes;
s3, low-temperature preservation: the glass conical flask is stored for 10 hours at 5 ℃, for 10 hours at-4 ℃, for 10 hours at-10 ℃, for 10 hours at-16 ℃ and for 10 hours at-25 ℃.
Example 3
S1, thallus collection: placing the strain suspension of the probiotics cultured to logarithmic phase in a centrifuge for centrifugation, collecting the strains, wherein the centrifugation temperature is 34 ℃, the centrifugation rotation speed is 1250 revolutions per minute, and the centrifugation time is 40 minutes;
s2, addition of substances: placing the thalli collected in the step S1 in a glass conical flask, dehydrating the compound oil by adopting a vacuum technology, placing the dehydrated compound oil into a storage container, injecting the thalli in the glass conical flask into the storage container, wrapping the thalli by using the compound oil in the storage container, adsorbing the compound oil with the walls of the thalli, isolating the thalli from the outside, and finally sealing the bottleneck of the glass conical flask by using paraffin, wherein the compound oil comprises the following raw materials in parts by weight: 85 parts of medium-chain triglyceride, 2 parts of tea polyphenol and 6 parts of organic solvent, wherein 2 parts of tea polyphenol and 6 parts of organic solvent are mixed, 2 parts of tea polyphenol and 6 parts of organic solvent are uniformly mixed by using an ultrasonic dispersion machine, a mixed solution of 2 parts of tea polyphenol and 6 parts of organic solvent is added into 85 parts of medium-chain triglyceride, a glass stirring rod is used for stirring the mixed solution, the stirring speed is 200r/min, and the stirring time is 3 minutes;
s3, low-temperature preservation: the glass conical flask is stored for 8 hours at 4 ℃, stored for 8 hours at-3 ℃, stored for 8 hours at-9 ℃, stored for 8 hours at-15 ℃ and stored at-23 ℃.
Comparative experiment
In a certain probiotic culture research laboratory, probiotics stored in examples 1 to 3 are respectively selected to perform a comparative experiment on activity and survival rate, as can be seen from fig. 1, the activity of the probiotics stored in example 1 is 6.5, the survival rate is 86%, the activity of the probiotics stored in example 2 is 6.9, the survival rate is 89%, the activity of the probiotics stored in example 3 is 7.7, and the survival rate is 93%, both the activity and the survival rate of the probiotics stored in example 3 are higher than those of the probiotics stored in examples 1 and 2, and the method is a preferred scheme, and by using the probiotic storage process in example 3, the activity and the survival rate of the probiotics after long-term storage can be greatly improved.
It is noted that, herein, relational terms such as first and second, and the like may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (8)
1. A probiotic oil preservation process is characterized by comprising the following steps: the method specifically comprises the following steps:
s1, thallus collection: placing the strain suspension cultured to the probiotics in logarithmic phase in a centrifuge for centrifugal treatment, collecting the strains, wherein the centrifugal temperature is 30-38 ℃, the centrifugal rotating speed is 500-2000 r/min, and the centrifugal time is 30-50 min;
s2, addition of substances: placing the thalli collected in the step S1 in a glass conical flask, dehydrating the compound oil by adopting a vacuum technology, placing the dehydrated compound oil in a storage container, injecting the thalli in the glass conical flask into the storage container, wrapping the thalli by using the compound oil in the storage container, adsorbing the compound oil with the walls of the thalli, isolating the thalli from the outside, and finally sealing the bottle opening of the glass conical flask by using paraffin;
s3, low-temperature preservation: storing the glass conical flask at 3-5 ℃ for 6-10 hours, storing the glass conical flask at-2-4 ℃ for 6-10 hours, storing the glass conical flask at-8-10 ℃ for 6-10 hours, storing the glass conical flask at-14-16 ℃ for 6-10 hours, and finally storing the glass conical flask at-20-25 ℃.
2. The process for preserving probiotic oils according to claim 1, characterized in that: the composite oil comprises the following raw materials in parts by weight: 80-90 parts of medium-chain triglyceride, 1-3 parts of tea polyphenol and 5-7 parts of organic solvent.
3. The process for preserving probiotic oils according to claim 2, characterized in that: the composite oil comprises the following raw materials in parts by weight: 80 parts of medium-chain triglyceride, 1 part of tea polyphenol and 5 parts of organic solvent.
4. The process for preserving probiotic oils according to claim 2, characterized in that: the composite oil comprises the following raw materials in parts by weight: 90 parts of medium-chain triglyceride, 3 parts of tea polyphenol and 7 parts of organic solvent.
5. The process for preserving probiotic oils according to claim 2, characterized in that: the composite oil comprises the following raw materials in parts by weight: 85 parts of medium-chain triglyceride, 2 parts of tea polyphenol and 6 parts of organic solvent.
6. The process for preserving probiotic oils according to any one of claims 2 to 5, characterized in that: the organic solvent is one or a combination of more of methanol, ethanol and acetone, wherein ethanol is a preferable scheme of the organic solvent.
7. The process for preserving probiotic oils according to any one of claims 2 to 5, characterized in that: the tea polyphenol and the organic solvent are mixed, and the tea polyphenol and the organic solvent are uniformly mixed by using an ultrasonic dispersion machine.
8. The process for preserving probiotic oils according to any one of claims 2 to 5, characterized in that: the mixed solution of the tea polyphenol and the organic solvent is added into the medium-chain triglyceride, and the mixed solution is stirred by a glass stirring rod at the stirring speed of 200r/min for 3 minutes.
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Cited By (1)
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| CN115088844A (en) * | 2022-07-14 | 2022-09-23 | 河北益存生物科技有限公司 | Lactic acid bacteria oil dropping liquid and preparation method thereof |
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