KR20150012452A - Microorganism additives compositions using glutinous rice paste as a cryoprotectant for fermentation of foods with enhanced survival rate of the microorganisms and method of preparing the same - Google Patents
Microorganism additives compositions using glutinous rice paste as a cryoprotectant for fermentation of foods with enhanced survival rate of the microorganisms and method of preparing the same Download PDFInfo
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- KR20150012452A KR20150012452A KR1020130087981A KR20130087981A KR20150012452A KR 20150012452 A KR20150012452 A KR 20150012452A KR 1020130087981 A KR1020130087981 A KR 1020130087981A KR 20130087981 A KR20130087981 A KR 20130087981A KR 20150012452 A KR20150012452 A KR 20150012452A
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- lactic acid
- glutinous rice
- cryoprotectant
- food
- additive composition
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/40—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by drying or kilning; Subsequent reconstitution
- A23L3/44—Freeze-drying
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- C—CHEMISTRY; METALLURGY
- 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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- C—CHEMISTRY; METALLURGY
- 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
- C12N1/20—Bacteria; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- 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
- C12N1/38—Chemical stimulation of growth or activity by addition of chemical compounds which are not essential growth factors; Stimulation of growth by removal of a chemical compound
Abstract
The present invention provides a microbial additive composition for food fermentation comprising a glutinous rice paste as a lactic acid bacterium and a cryoprotectant, and a method for producing the same.
The glutinous rice paste used as a novel cryoprotectant in the present invention is superior to the conventional cryoprotective agent in the production of a microbial additive composition for food fermentation and provides stability as a food grade material. Accordingly, the microbial additive composition for food fermentation according to the present invention maximizes the survival rate of microorganisms and enables high-quality food production, and enables lactic acid bacteria to be supplied at low unit cost as seeds for uniformizing quality of food and maintaining quality.
Description
The present invention relates to a microbial additive composition for food fermentation and a method for producing the same, in which survival rate is improved by using a glutinous rice paste as a cryoprotective agent.
Fermentation of food involves various lactic acid bacteria. Studies on microbial additives for food fermentation, which can be added during the production of food by separating and selecting excellent lactic acid bacterial strains in order to uniformize the quality of food and maintain the quality of fermented food, are under development. In order to supply microbial additives for food fermentation, formulation for preservation of microorganisms is required. Freezing / freeze drying is a method for effectively preserving most microorganisms for a long time. Freezing / freeze drying can be advantageous in prevention of pollution, storage, transportation, and economic efficiency. However, since the activity and survival rate of the microorganism are drastically reduced during the freezing / lyophilizing process, a method for maximizing the survival rate of microbial cells is required do.
Conventional processes for the preparation of lactic acid bacteria lyophilized powder include the fermentation of lactic acid bacteria in an anaerobic fermentation apparatus using an aqueous medium, centrifugation and microbial recovery using ultrafiltration, followed by rapid freezing and freeze drying. This process damages the cell membrane structure due to the formation of ice particles during freezing, and lactic acid bacteria powder is sensitive to air, moisture and temperature conditions, making it difficult to secure storage and distribution stability and processing stability. To improve this, a saccharide and a cryoprotectant such as amino acid, skim milk, gelatin, and citric acid are added singly or in combination as a means by which lactic acid bacteria can be maintained without being killed. The cryoprotectant exhibits a higher survival rate than the simple lyophilized lactic acid bacteria powder by imparting physicochemical stability to the lactic acid bacterial powder.
However, microbial formulation after lyophilization has been carried out for the main lactic acid bacterium. However, since the survival rate after the actual lyophilization has remained at 60-70%, the production cost has increased.
The present invention aims to provide a microbial additive composition for food fermentation which improves the survival rate of microorganisms after freezing or freeze-drying, and a method for producing the same.
The present invention provides a microbial additive composition for food fermentation comprising a glutinous rice paste as a lactic acid bacterium and a cryoprotectant, and a method for producing the same.
The microbial additive composition for food fermentation according to the present invention includes lactic acid bacteria known to be included in microbial additives for food fermentation in the past and includes glutinous rice paste as a cryoprotective agent capable of improving the survival rate thereof.
As can be seen in the following examples, when the glutinous rice paste was used as a cryoprotective agent in comparison with the conventional skim milk or garlic as a cryoprotectant for lactic acid bacteria, the survival rate of the lactic acid bacteria after lyophilization was significantly improved.
The glutinous rice paste used as a cryoprotective agent in the present invention can be directly manufactured or can be used by purchasing a commercially available product. The glutinous rice paste is a food grade material and is a stable substance which is used in various foods, and thus can be usefully used in the production of the microbial additive composition according to the present invention. In particular, when used as a microorganism additive in the form of a frozen form for short-term preservation and distribution, the production cost can be greatly reduced as compared with a microbial additive in the form of a freeze-dried powder.
The microbial additive composition for fermenting food according to the present invention may further contain ingredients known as a cryoprotective agent in addition to glutinous rice paste as a cryoprotectant.
In one embodiment of the invention the composition comprises one or more additional cryoprotectants selected from saccharides, amino acids, peptides, gelatin, glycerol, sugar alcohols, whey, alginic acid, ascorbic acid, yeast extract, skimmed milk, trehalose, . It was confirmed from the following examples that the use of the additional cryoprotectant in combination with the glutinous rice paste did not improve the survival rate of the microorganism. However, the use of the microbial additive composition according to the present invention, It is possible to use an appropriate combination of additional cryoprotectants in consideration of changes in flavor, texture, production cost of the product, and the like.
Meanwhile, the type of lactic acid bacteria that can be included in the microbial food additive composition for food fermentation according to the present invention is not particularly limited, and specific types of lactic acid bacteria are conventionally studied to be suitable for the fermentation of the food according to the type of the fermented food Anything that has come or been used can be used. In the present invention, the lactic acid bacteria may be used singly or in combination.
In one embodiment of the present invention, the lactic acid bacterium is a lactic acid bacterium derived from kimchi, and the food may be kimchi. In the following examples of the present invention, a microbial additive composition used for kimchi fermentation was prepared and the freeze-protection effect of the glutinous rice paste-derived lactic acid bacteria was evaluated.
In one embodiment of the invention, the lactic acid bacteria are selected from the group consisting of Weissella spp., Lactobacillus spp. Genus Streptococcus , Enterococcus genus, Bifidobacterium genus , Lactococcus genus and Pediococcus genus And at least one lactic acid bacterium selected from the group consisting of genus Escherichia. In a preferred embodiment of the present invention, the lactic acid bacterium is selected from the group consisting of Weissella cibaria , Lactobacillus plantarum, and Pediococcus pentosaceus .
The microbial additive composition for food fermentation according to the present invention can be formulated in the form of a frozen form or a lyophilized powder. It is advantageous to improve the quality of the food and the stability of the quality by preparing and supplying the microorganism additive in the form of a frozen form or a lyophilized powder rather than cultivating the liquid seed bacterium for food production. In particular, when the glutinous rice paste is used as a freezing protective agent and is prepared as a microorganism additive in a frozen form, the production cost can be significantly reduced as compared with the microbial additive in the form of a freeze-dried powder.
The present invention also provides a method for producing the microbial additive composition for food fermentation, which comprises mixing a glutinous rice paste as a lactic acid bacterium and a cryoprotectant and freezing or lyophilizing the mixture.
For example, lactic acid bacteria can be obtained by pure separation from foods such as kimchi, and they can be cultured by using a commercially available medium suitable for culturing lactic acid bacteria or a vegetable medium prepared from food materials such as Chinese cabbage. The thus cultured Lactobacillus was mixed with glutinous rice flour or an additional cryoprotectant at an appropriate concentration and incubated at -30 to -10 Freezing at a low temperature of -50 to -90 Deg.] C for 54 to 90 hours to prepare the microbial additive composition for food fermentation according to the present invention.
The concentration of the lactic acid bacteria can be appropriately adjusted according to the kind of the food to be produced, the kind of the lactic acid bacteria used, the degree of fermentation, etc., and the concentration of the glutinous rice paste can be adjusted depending on the presence or absence of additional cryoprotectant used together.
In one embodiment of the present invention, the mixture is prepared by mixing glutinous rice and sterilized water at a ratio of 1: 6 to 1:10 to lactic acid bacteria suspended in sterilized water at a concentration of 1 x 10 9 to 1 x 10 11 CFU / ml (V / v), for example 180 to 220% (v / v), based on the volume of the suspension. When the glutinous rice paste is used as a cryoprotectant within the above range, it exhibits superior microbial survival rate even in the case of long-term cryopreservation compared with skim milk or garlic.
The present invention also provides a method for freeze-protecting a lactic acid bacterium comprising adding glutinous rice paste as a cryoprotective agent to freeze-dried or freeze-dried lactic acid bacteria suspended in sterilized water in the preparation of a lactic acid bacterial preparation or a freeze- Namely, the present invention provides the use of glutinous rice paste for freeze-protection of lactic acid bacteria in the production of a freeze-dried lactic acid bacteria preparation or a freeze-dried lactic acid bacteria powder preparation. The configuration for implementing the above application is as described above.
The glutinous rice paste used as a novel cryoprotectant in the present invention is superior to the conventional cryoprotective agent in the production of a microbial additive composition for food fermentation and provides stability as a food grade material. Accordingly, the microbial additive composition for food fermentation according to the present invention maximizes the survival rate of microorganisms and enables high-quality food production, and enables the lactic acid bacteria to be supplied at low unit cost as a seed for uniformizing the quality of food and maintaining quality.
1 and 2 are graphs comparing the survival rates of lactic acid bacteria according to the type of cryoprotectant and the cryopreservation time.
Advantages and features of the present invention and methods of achieving them will become apparent with reference to the embodiments described in detail below. The present invention may, however, be embodied in many different forms and should not be construed as being limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art. Is provided to fully convey the scope of the invention to those skilled in the art, and the invention is only defined by the scope of the claims.
[ Example ]
In the following examples, a microbial additive composition used for kimchi fermentation was prepared and the freeze protection effect of the glutinous rice paste-derived lactic acid bacteria was evaluated.
Example 1. Isolation and Preservation of Lactic Acid Bacteria Derived from Kimchi
(1) Isolation of Lactic Acid Bacteria Derived from Kimchi
Lactic acid bacteria were isolated from Kimchi, which was directly manufactured by Kimchi Research Institute of Gwangju Metropolitan City, and weissella cibaria , Lactobacillus plantarum , Pediococcus pentosaceus .
(2) Cultivation and Preservation of Lactic Acid Bacteria Derived from Kimchi
Lactic acid bacteria isolated pure from kimchi were inoculated on MRS broth (Table 1) suitable for their cultivation and incubated at 30 ° C for 24 hours to grow the lactic acid bacteria well, followed by subculture at 4 weeks intervals. The microbial suspension was prepared at a final concentration of glycerol of 20% and stored at -75 ° C.
Example 2. Survival rate of lactic acid bacteria derived from kimchi according to cryoprotectant
(1) Survival rate of lactic acid bacteria derived from Kimchi according to the type of cryoprotectant
The three types of kimchi-derived lactic acid bacteria were inoculated into MRS broth and cultured at 30 ° C for 24 hours to obtain a concentration of 10 9 to 10 10 CFU / ml of kimchi microorganism. After centrifuging at 8,000 rpm for 10 minutes, kimchi-derived lactic acid bacteria were collected and washed three times with 0.9% physiological saline. The concentration of the lactic acid bacteria derived from kimchi was fixed to 1 × 10 10 CFU / ml, and a freezing protection agent (skim milk, garlic crush) was added at a final concentration of 10% (w / v) based on the volume of the lactic acid bacterium suspension. Lt; / RTI > When glutinous rice paste was used as a cryoprotectant, glutinous rice paste was prepared with 8 times the weight of glutinous rice and 200% (v / v) of the volume of kimchi microbial suspension fixed at a concentration of 1 × 10 10 CFU / Lt; / RTI > Thereafter, it was frozen and preserved in a -20 ° C low-temperature apparatus. After freezing and thawing, the kimchi microorganism preparation was smeared on MRS solid medium and the viable cell count was investigated.
The survival rate was calculated by calculating the percentage of viable cell count after freezing to the number of viable cells before freezing, and the survival rate according to the type of cryoprotectant and the cryopreservation time was shown in the graph of FIG. 1 and FIG. Weissella cibaria (WC), Lactobacillus plantarum (LP), and Pediococcus pentosaceus (PP) showed the highest survival rate after 14 days of cryopreservation when glutinous rice paste was used as a cryoprotectant. In particular, P. pentosaceus showed a high survival rate of 54% at 7 days after storage at 20 ° C, and survival rate of 43% or more after 28 days. When 10% of garlic was used as a cryoprotectant, the survival rate after 7 days was the best with 60% or more, but it showed 22% after 28 days. In the case of the control without cryoprotectant, most Kimchi microorganisms died within the storage period of 14-28 days and the survival rate was less than 10%. Weissella Cibaria also showed the highest survival rate (49%) when garlic was added as a preservative after 7 days of cryopreservation, but the survival rate decreased rapidly over the period of storage and reached 24% after 28 days And showed lower efficiency than glutinous rice paste. Lb. Plantarum also showed the highest survival rate when glutinous rice paste was used as a cryoprotectant. No synergistic effect on kimchi microbial survival rate was observed when garlic and skimmed milk were used together as cryoprotectants in all three strains.
Claims (9)
Wherein the composition further comprises at least one further cryoprotectant selected from sugars, amino acids, peptides, gelatin, glycerol, sugar alcohols, whey, alginic acid, ascorbic acid, yeast extract, skim milk, trehalose and garlic Composition.
Wherein the lactic acid bacterium is a lactic acid bacterium derived from kimchi, and the food is kimchi.
The lactic acid bacteria belong to the genus Weissella , Lactobacillus Genus Streptococcus , Enterococcus , Bifidobacterium In, Lactococcus and Pediococcus genus Wherein the microbial additive composition is at least one kind of lactic acid bacterium selected from the group consisting of soybean oil and soybean oil.
The lactic acid bacteria were Weissella cibaria , Lactobacillus plantarum and Pediococcus pentosaceus , wherein the microbial additive composition for food fermentation is at least one lactic acid bacterium.
Wherein the composition is in the form of a frozen or lyophilized powder.
The above mixture was added to sterilized water at a concentration of 1 x 10 < 9 > to 1 x 10 < 11 > CFU / ml, and the glutinous rice paste prepared by mixing the glutinous rice with sterilized water at a ratio of 1: To 250% (v / v) of the microbial additive composition.
A method for cryoprotecting a lactic acid bacterium comprising adding glutinous rice paste as a cryoprotectant to a suspected lactic acid bacterium suspended in sterilized water, followed by freezing or lyophilization.
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