KR102028076B1 - Method for the functional fermented-carrot juice using Lactococcus lactis - Google Patents

Abstract

The present invention relates to a method for producing a functional fermented carrot juice using Lactococcus lactis lactic acid bacteria, a method for producing a lactic acid bacteria fermented carrot juice of the present invention, Lactococcus Lactobacillus) inoculated to retain the antimicrobial effect, but by killing the lactococcus lactis safe lactic acid bacteria fermented carrot juice can be prepared.

Description

Method for preparing functional fermented carrot juice fermented with Lactococcus lactis lactobacillus {Method for the functional fermented-carrot juice using Lactococcus lactis}

The present invention relates to a method for preparing carrot juice, and more particularly, Lactococcus lactis ) relates to a method for producing functional fermented carrot juice using lactic acid bacteria.

 Carrots, which are one of the common vegetables in Korea, are added to foods or processed into juices and consumed. Carrots contain carotene as well as water-soluble vitamins such as vitamin B and vitamin C. Carotene is converted to vitamin A when absorbed into the body, making it a type of fat-soluble vitamin that is difficult to consume in diet. In addition, the carrot is rich in dietary fiber and iron and has a high sugar content has the advantage that can be used properly in processed foods.

On the other hand, many kinds of intestinal microorganisms live in human intestines. Their proper ratio keeps intestinal function healthy and helps to fight invasion and infection of foreign substances. However, due to alcohol intake, stress, meat intake, antibiotics, etc., the intestinal beneficial bacteria are killed or the balance of the bacterial layer is compromised. As a preventive measure of such a problem is lactic acid bacteria.

Lactic acid bacteria, also called lactic acid bacteria, is a generic term for bacteria that ferment sugars to gain energy and produce large amounts of lactic acid. Lactobacillus maintains a constant ratio of various intestinal microorganisms in the intestine, and contributes greatly to boosting immunity by itself.

The name 'lactic acid bacteria' is an idiomatic name and does not indicate a taxonomic position. It is appropriate to define the lactic acid bacteria are Lactobacillus bacteria (Lactobacillus), Lactococcus (Lactococcus), flow Stock Kono (Leuconostoc), Peddie Oh Caucus (Pediococcus), Bifidobacterium (Bifidobacterium). Both develop well in an environment with low oxygen to produce lactic acid from various sugars. It is much resistant to acid, and nutritional composition is very complicated and requires many kinds of amino acids and vitamins in addition to sugars.

Many efforts have been made to combine lactic acid bacteria and natural materials in order to obtain synergistic effects by substituting such advantages of lactic acid bacteria into natural materials. However, there has been a problem such as the destruction of nutrients of natural materials during the lactic acid bacteria fermentation process. However, studies to solve these problems are still insufficient. Therefore, research is required to maintain the function of lactic acid bacteria without destroying the nutrients of natural products.

Korean Patent Laid-Open No. 10-2010-0096287 (2010.09.02.) Describes a content of a method for producing dairy products containing carrots. Korean Patent Laid-Open No. 10-2013-0000519 (2013.01.03.) Describes a method for preparing a functional beverage using a lactic acid bacteria fermented vegetable juice and a citrus juice and a composition thereof.

The present invention is to develop and provide a lactic acid bacteria fermented carrot juice having an antimicrobial effect using a natural carrot and lactic acid bacteria.

The present invention comprises the steps of (a) to juice the carrot to obtain carrot juice; Centrifuging the carrot juice obtained above at 6,000 to 10,000 × g for 10 to 20 minutes to obtain a supernatant (b); (C) autoclaving the supernatant obtained above for 8-12 minutes at a temperature of 100-120 ° C .; And after the high-pressure sterilization, the step of inoculating the lactic acid bacteria and fermentation (d); provides a method for producing a fermented carrot juice comprising a.

The method of preparing the fermented carrot juice of the present invention, preferably, after the lactic acid bacteria fermentation of step (d), adding a bacteriophage (e); may further include.

In the method for producing fermented carrot juice of the present invention, the lactic acid bacteria is preferably added by 0.01 to 0.04% (v / v) relative to the volume of carrot juice.

In the method of producing fermented carrot juice of the present invention, the lactic acid bacteria, preferably may be Lactococcus lactis ( Lactococcus lactis ).

In the production method of the fermented carrot juice of the present invention, the fermented carrot juice may preferably have an antimicrobial activity.

According to the present invention, lactic acid bacteria fermented carrot juice can be prepared with a change in quality when retained for a long time while retaining the antimicrobial effect.

1 is a schematic diagram of a method for preparing fermented carrot juice of the present invention.
Figure 2 is a graph showing the number of lactic acid bacteria over time when carrot juice fermentation.
Figure 3 is a table showing the indicator strain used in the present invention.

The present invention comprises the steps of (a) to juice the carrot to obtain carrot juice; Centrifuging the carrot juice obtained above at 6,000 to 10,000 × g for 10 to 20 minutes to obtain a supernatant (b); (C) autoclaving the supernatant obtained above for 8-12 minutes at a temperature of 100-120 ° C .; And after the high-pressure sterilization, the step of inoculating the lactic acid bacteria and fermentation (d); provides a method for producing a fermented carrot juice comprising a.

Hereinafter, the present invention will be described by subdividing each step.

<Step (a): Get carrot juice>

This step is a process of obtaining carrot juice by juice carrots.

Carrots are preferably washed with water and peeled. In addition, it is good to slice and juice to 1 ~ 3cm in diameter.

<Step (b): supernatant obtained after centrifugation>

This step is a process of obtaining the supernatant by centrifuging the carrot juice obtained in the above at 6,000 ~ 10,000 × g for 10 to 20 minutes.

In the present invention, through the centrifugation, the bacteria remaining in the carrot juice can be removed first. In this process, centrifugation is recommended for 10 to 20 minutes at 6,000 to 10,000 × g. If the centrifugation is performed at less than 6,000 rpm or less than 10 minutes, there may be a problem in that residual bacteria cannot be effectively removed. If it proceeds in excess or for more than 20 minutes, there is a problem that other useful ingredients present in the carrot are removed, which is not preferable.

<Step (c): Sterilization of Supernatant>

This step is a process of autoclaving the obtained supernatant for 8 to 12 minutes at a temperature of 100 ~ 120 ℃.

The high pressure sterilization process of this step can remove the bacteria present in the carrot juice secondary. In this process, autoclaving is preferably performed at 100 to 120 ° C. for 8 to 12 minutes, since the remaining bacteria are effectively removed under these sterilization conditions, and the destruction of useful components present in the carrot is small.

<Step (d): Lactic acid bacteria fermentation>

This step is a process of inoculating and fermenting by lactic acid bacteria after the high-pressure sterilization.

In this process, fermentation is preferably performed for 12 to 24 hours, because fermentation for 12 to 24 hours can produce the largest number of lactic acid bacteria.

In addition, the lactic acid bacteria used in this step is preferably added by 0.01 to 0.04% (v / v) relative to the volume of carrot juice, more preferably 0.02% (v / v) by volume relative to the volume of carrot juice. .

In addition, the lactic acid bacteria used in the present invention may be preferably Lactococcus lactis ( Lactococcus lactis ), it is known that lactococcus lactis plays a role in boosting immunity and preventing the accumulation of cholesterol in the body.

<Step (e): Bacteriophage addition>

This step is a process of adding the bacteriophage to the lactic acid bacteria fermented carrot juice obtained in step (d).

The bacteriophage added in this step is preferably added by 5 to 10 log PFU / ml. If the bacteriophage is added and then kept at room temperature for 4 to 5 hours, the bacteriophage kills Lactococcus lactis, so that the lactic acid bacteria are no longer present in the fermented carrot juice. This prevents the deterioration of products due to over fermentation during distribution.

On the other hand, not only live lactic acid bacteria but also Lactobacillus microorganisms have been reported to have physiological functions, particularly immune control functions, as well as live lactic acid bacteria. Can be.

On the other hand, traditional methods of sterilization, such as heat treatment and sonication, are susceptible to damage to active ingredients having excellent physiological functions present in the mycelium and can destroy various useful substances from carrots present in carrot juice. However, the bacteriophage used in the present invention is a virus that targets only a host strain, and when the cell wall is lysed due to the endorisin produced by the bacteriophage, the bacteriophage does not cause enormous damage to the cell wall of the bacterium. It can prevent the destruction of various useful materials.

Hereinafter, the configuration of the present invention will be described in detail through the following examples and experimental examples. However, the scope of the present invention is not limited to the following Examples and Experimental Examples, but includes modifications of equivalent technical ideas.

< Example  1: Preparation of Fermented Carrot Juice of the Present Invention>

(1) of carrot juice Eradication  And sterilization

In order to prepare the fermented carrot juice of the present invention, after removing the carrot washed and peeled, the carrots were sliced into 2 × 2 cm size to make the juice smoothly. Then, the juice was extracted using a juicer, immediately stored in a sterilized bottle after extraction. Extracted carrot juice was isolated from the bacteria in the carrot by centrifugation (8000 g, 15 min), centrifugation supernatant was adjusted to pH 7.0, and short-time autoclaving (110˚C, 10 min) was performed. This process killed most of the bacteria and minimized the loss of nutrients. Carrot juice to be used in the final through the above process was prepared.

(2) Preparation of fermented carrot juice by adding lactic acid bacteria

Carrot juice prepared from the Lactococcus lactis (Lactococcus lactis ) was inoculated with 20 μl per 1 ml of carrot juice. Thereafter, the fermentation was performed for 12 hours in an incubator set at 30 ° C., and 9 log PFU / ml of bacteriophage solution was added to the fermented carrot juice. Then, after maintaining at room temperature for 4.5 hours, immediately refrigerated (FIG. 1). 1 is a schematic diagram of a fermented carrot juice production method of the present invention.

Experimental Example 1: Searching for Optimal Lactic Acid Bacteria Fermentation Time in Carrot Juice

When lactic acid bacteria were added to carrot juice, this experiment was conducted to set the optimum fermentation time. Fermented carrot juice was prepared by the method of Example 1, and the number of lactic acid bacteria was measured for 6, 12, 24, and 48 hours after inoculation (FIG. 2). Figure 2 is a graph showing the number of lactic acid bacteria over time when carrot juice fermentation. After inoculating Lactococcus lactis into carrot juice and fermenting for 12 to 24 hours, the largest number of Lactococcus lactis was identified.

< Experimental Example  2: for pathogenic strains Lactococcus Lactis  Confirmation of Antibacterial Activity of Lactic Acid Bacteria>

In order to confirm the antimicrobial activity of Lactococcus lactis lactic acid bacteria against pathogenic strains, 100 μl of indicator strains cultured for 12 hours per 5 ml of TBS soft agar (0.65% agar) medium were dispensed. The indicator strain is Listeria monocytogenes ( Listeria) monocytogenes ) 2 weeks, Staphylococcus aureus 4 weeks, and other 16 strains of various pathogenic strains (Fig. 3). Figure 3 is a table showing the indicator strain used in the present invention.

 A soft agar medium in which the indicator strain was inoculated on the MRS agar plate was dropped, and 10 μl of the Lactococcus lactis strain was dropped in a spot, followed by incubation for 12 hours to confirm the inhibition zone. At the point where the Lactococcus lactis strain was dropped, it was confirmed that the killing of the indicator strain proceeded.

< Experimental Example  3: pathogenic Fermented Carrot Juice for Strains  Antimicrobial Activity Confirmation>

According to Experimental Example 2, it was confirmed that Lactococcus lactis lactobacillus has antimicrobial activity against the pathogenic strain. In this experimental example, the following experiment was conducted to determine whether the fermented carrot juice of the present invention has antibacterial activity.

First, the lactococcus lactis lactobacillus present in the fermented carrot juice was killed using bacteriophages, so that the antimicrobial activity of the fermented carrot juice was blocked by the lactococcus lactis lactobacillus.

For the experiment, 100 μl of indicator strain (FIG. 3) incubated for 12 hours per 5 ml of TBS soft agar (0.65% agar) medium was dispensed. A soft agar medium inoculated with the indicator strain was inoculated on the MRS agar plate, and 10 µl of the fermented carrot juice of the Example 1 (with Lactococcus lactis lactobacillus killed) was dropped in a spot, followed by culturing for 12 hours. (inhibition zone) was confirmed. It was confirmed that the killing of the indicator strain at the point where the fermented carrot juice (the one in which the Lactococcus lactis lactobacillus was killed) of Example 1 was dropped.

In summary, the present invention is a Lactococcus Lactococcus using the sterilization process and bacteriophage in two stages of sterilization and sterilization lactis ), a two-stage sterilization process that prevents the destruction of useful bioactive substances in carrots and the bacteriophage to remove Lactococcus lactis fermentation strains, resulting in over-fermentation during distribution. It can prevent the product from deterioration. In addition, the fermented carrot juice of the present invention has an antimicrobial activity, high stability against microbial contamination, and also exhibits the effect of increasing the anti-pathology of the human body when ingested.

Claims (5)

Juice of the carrot to obtain carrot juice (a);
Centrifuging the carrot juice obtained above at 6,000 to 10,000 × g for 10 to 20 minutes to obtain a supernatant (b);
(C) autoclaving the supernatant obtained above for 8-12 minutes at 100-120 ° C. temperature;
After the high pressure sterilization, inoculating Lactococcus lactis ( Lactococcus lactis ) and fermentation for 12 to 24 hours (d); And
After the fermentation, the bacteriophage is added to kill Lactococcus lactis (e); to obtain fermented carrot juice, including
The fermented carrot juice production method characterized in that it has an antimicrobial activity.
delete The method of claim 1,
Lactococcus lactis ( Lactococcus lactis ),
Fermented carrot juice production method characterized in that the addition of 0.01 ~ 0.04% (v / v) relative to the volume of carrot juice.
delete delete

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