JPH0751042B2 - Method for producing lactic acid bacteria beverage - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION a. Object of the invention (field of industrial application) The present invention relates to improvement of a method for producing a lactic acid bacterium beverage as a favorite product, and particularly to produce a lactic acid bacterium beverage containing fruit juice such as apple juice. The invention is effective when applied to cases.

(Prior art and this problem) In recent years, various lactic acid bacterium beverages have come to be produced because of diversification of taste and lactic acid bacteria are useful for promoting health.

However, since the conditions for fermenting lactic acid bacteria in beverages are severe, the types of lactic acid bacteria beverages are currently limited, for example, lactic acid bacteria beverages fermented with lactic acid bacteria in fruit juice have not yet been marketed. It is the actual situation.

This is because the PH value of fruit juices such as apples is low (acidic), and the polyphenols contained in the fruit juices inhibit the growth of lactic acid bacteria, so simply adding lactic acid bacteria to the fruit juice kills the lactic acid bacteria. It is believed that.

Also, because the nutritional requirements of lactic acid bacteria are complicated and the growth is slow, even if artificial growth of lactic acid bacteria in fruit juice is possible,
Fermentation time must be lengthened, and the fact that it is not profitable in terms of manufacturing cost is considered to be one reason why it is not commercially available.

As a method for producing a lactic acid bacterium beverage containing natural fruit juice as a main component, a method of separately adding lactic acid bacteria grown in animal milk to the fruit juice, a method of increasing the PH value of the fruit juice, or a polyphenol using a treatment material Although a method for removing lactic acid bacteria is known, it is not possible to obtain a lactic acid bacterium beverage purely composed of fruit juice by using lactic acid bacteria grown in animal milk. In the experiment conducted, it was not possible to produce a lactic acid bacterium beverage containing fruit juice as a main component by any of the above methods.

The method for producing a lactic acid bacterium beverage of the present invention is intended to efficiently produce a lactic acid bacterium beverage containing a juice having a low PH value, so that it is possible to cope with the diversification of tastes in recent years.

b. Structure of the Invention (Means for Solving the Problem) In the method for producing a lactic acid bacteria beverage of the present invention, first, one or two kinds of nutrition extracts such as yeast extract, noodle juice, and grain seed germ extract. The above is added to the fruit juice in an amount that is slightly above the minimum amount required for the growth of lactic acid bacteria and that does not adversely affect the flavor and appearance (color) of the lactic acid bacteria beverage containing fruit juice as a product. When mixed, PH value is 3.0-4.
Make a mixed solution of 5 and use this mixed solution as a stock solution for lactic acid fermentation.

The fermentation stock solution thus prepared is fermented by contact with lactic acid bacteria to give a lactic acid bacterium beverage, but if the fermentation is carried out efficiently, the lactic acid bacteria are included at a rate of 10 ⁸ to 10 ⁹ cells / ml. By continuously contacting the immobilized gel, the lactic acid bacterium is mixed into the mixed solution more quickly than the case where the lactic acid bacterium is fermented in a non-immobilized state, and the lactic acid bacterium is fermented,
A lactic acid bacterium beverage containing fruit juice.

In order to make good contact between the fermentation stock solution and the gel and to accelerate the growth of lactic acid, this fermentation stock solution should be mixed with 10% lactic acid bacteria.
A glass tube with a bottom plate that closes the bottom opening and a lid plate that can be opened and closed at the top opening together with the gel that is comprehensively immobilized at a rate of ⁸ to 10 ⁹ pieces / ml, and this glass tube inside the glass tube. On the inner peripheral edge of the horizontal circular plate provided concentrically with
A straightening plate provided with a cylinder that rises upward from the inner peripheral edge,
A support frame having a large number of small holes fixed by fitting the lower end portion to the cylinder of this straightening plate, and the lower part of the circular plate that constitutes the above straightening plate around the vertical axis by a motor provided above the bottom plate. Put into a culture device consisting of a centrifugal blade that is rotationally driven on the side, rotate the centrifugal blade, and make the stock solution for lactic acid fermentation flow between the gels attached to the outer peripheral surface of the support frame to perform fermentation. To do.

Example Next, an experiment conducted by the present inventor will be described.

I. Preparation of gel beads First, as lactic acid bacteria, Lactobacillus casei IAM 1045 was taken out from the storage medium and subcultured in 10 ml of LPY medium. This LPY medium contains 1% lactose and 0.7% peptone in water,
Yeast extract is added with 0.5%.

Lactic acid bacteria subcultured on such LPY medium were
After culturing for an additional time, inoculate into 250m1 of LPY medium and further at 30 ℃
After culturing for 24 hours, the number of lactic acid bacteria in this medium is (3-10) ×
Proliferated to 10 ⁷ cells / ml.

The above LPY medium in which the number of lactic acid bacteria reached (3 to 10) × 10 ⁷ cells / ml was subjected to centrifugal separation at a speed of 3000 rpm for 5 minutes using a centrifuge to collect lactic acid bacteria.

The collected lactic acid bacteria were further washed with physiological saline that had been sterilized to obtain washed lactic acid bacteria.

The washed lactic acid bacteria were suspended in 20 ml of physiological saline that had been sterilized in advance and suspended, and 30 ml of a 5% sodium alginate solution prepared in advance was added to the suspension to prevent any bacteria. It mixed so that it might not mix.

The mixed solution thus obtained was dropped into a 3% calcium chloride solution and solidified in this solution, and the diameter was 3 to 5 m.
m gel beads were used.

B. In order to grow the lactic acid bacteria contained in the above-mentioned gel beads, the gel beads are put in a fermentation tank as shown in the drawing together with 950 ml of a stock solution for fermentation which has been sterilized in advance, and 48 hours at 30 ° C. Culture work was performed.

The undiluted solution for fermentation here is a mixed solution of PH3.9 apple juice and 0.25% of yeast extract.

In the fermenter used in the experiment, a glass bottom plate 2 having a slightly larger outer diameter than the outer diameter of the glass tube 1 is fixed to the lower end portion of the cylindrical glass tube 1 so that the glass The lower end opening of the cylinder 1 is closed. Lower end portions of a plurality of studs 3 and 3 are respectively screwed and fixed to portions of the bottom plate 2 near the outer periphery of the bottom plate 2 that project outward from the outer peripheral surface of the glass tube 1. The upper ends of the studs 3 and 3 respectively project slightly above the upper edge of the glass tube 1, and the upper ends of the studs 3 and 3 are disc-shaped lids having an outer diameter slightly larger than the outer diameter of the glass tube 1. The plate 4 is inserted into circular holes 5 and 5 formed near the outer periphery of the plate 4, respectively. Insert the circular holes 5 and 5,
Nuts (not shown) can be screwed onto the upper ends of the studs 3 and 3 protruding from the upper surface of the cover plate 4, and the upper end cover plate 4 is openably and closably attached to the upper end opening of the glass tube 1.

Further, inside the glass tube 1, a straightening plate 25 having a cylinder 24 rising upward from the inner peripheral edge of a horizontal circular disk 23 provided substantially concentric with the glass tube is installed. It is fixed by 7, 7.

At the lower end of the straightening plate 25, a lower end of a supporting frame 6 formed by cylindrically forming a metal mesh made by braiding stainless steel filaments is fitted onto the inner side of the glass tube 1. The glass tube 1 is fixed substantially concentrically.

On the other hand, a vertical shaft 8 is planted in the center of the upper surface of the bottom plate 2, and an inverted cylindrical cap 10 fixed to the center of a rotary block 9 is rotatably attached to the vertical shaft 8. There is. An annular magnet 11 is fitted and fixed around the rotary block 9, and the magnet 11 and an output shaft 15 of a motor 14 fixed via a bracket 13 below a base plate 12 to which the bottom plate 2 is fixed. The fixed annular magnet 16 is opposed to the bottom plate 2 via the bottom plate 2.

A plurality of flat blades 17, 17 are provided on the outer periphery of the rotary block 9 rotatably supported by the vertical shaft 8 via the cap 10.
By fixing the radial direction, the sirocco fan-shaped centrifugal blade
It has 18 components.

Further, above the centrifugal blade 18, an air bubble nozzle for blowing a gas containing no oxygen into the liquid stored inside the glass tube 1 is provided inside the cylinder 24 constituting the flow straightening plate 25.
19 are provided. This bubble nozzle 19 has a plurality of small holes 21, 21 at a lower end portion of an air supply pipe 20 fixed in a state of penetrating the lid plate 4 which opens and closes an upper end opening of the glass tube 1.
The nozzle portion 22 having the above is fixed, and the compressed gas fed through the air feeding pipe 20 is configured to be ejected to a position directly above the centrifugal blade 18.

Although illustration and detailed description are omitted, in addition to the air supply pipe 20, a sensor for detecting the state of the culture solution in the glass container 1, a tube for feeding nutrients into the culture solution, etc. Is provided.

When fermenting lactic acid bacteria by the culture device configured as described above to make a lactic acid bacterium beverage mainly composed of fruit juice,
First, a granular gel in which lactic acid bacteria are fixed is put into the inside of the glass tube 1 together with the stock solution for lactic acid fermentation.

Once charging is complete, bubble nozzle 19 through air line 20
Nitrogen (N ₂ ) and carbon dioxide (CO
₂ ) A compressed gas that does not contain oxygen (O ₂ ) that is unsuitable for fermentation of lactic acid bacteria, such as ₂ ), is sent, and the gas is blown into the culture solution stored in the glass tube 1 through the small holes 21, 21 of the nozzle portion 22. The centrifugal blade 18 is rotated by energizing the motor 14.

That is, by energizing the motor 14, the output shaft 15 of the motor 14
When the magnet 16 fixed to the rotor is rotated, the magnet 11 opposed to the magnet 16 via the bottom plate 2 is rotated by the magnet coupling action, and the magnet 11 and the rotation block 9 are rotated together.
Centrifugal blade 18 composed of multiple blades 17 fixed to the
On the lower side of the circular plate 23 that constitutes the flow regulating plate 25, it rotates about the vertical axis 8.

Due to the rotation of the centrifugal blades 18 accompanying the energization of the motor 14, the flow toward the outside from the portion near the center of the glass tube 1 along the lower surface of the circular plate 23 to the bottom of the glass container 1 storing the culture solution. And the cylinder that forms the current plate 25 accordingly.
The pressure inside the support frame 6 communicating inside 24 becomes lower than the pressure outside.

When a pressure difference is generated between the inside and the outside of the support frame 6 as the centrifugal blade 18 rotates in this way, the culture solution existing outside the support frame 6 is caused to flow into the inside of the support frame 6. Is
The culture solution in the glass tube 1 flows down inside the support frame 6 and circulates in the glass tube 1 while rising in a tubular space between the outer peripheral surface of the support frame 6 and the inner peripheral surface of the glass tube 1. The large number of small holes (mesh) formed in the support frame 6 existing in the middle of this flow, the particle size (3 to 5 mm) of the large number of granular gel put into the glass container 1 together with the culture solution. Each of the granular gels, which are smaller than the above, and cannot flow through the small holes, cannot pass through the small holes, remain attached to the outer peripheral surface of the support frame 6, and only the culture liquid flows while passing through the support frame 6. .

As described above, in the culture solution flowing through the support frame 6 having the granular gel adhered to the outer peripheral surface, oxygen is supplied from the bubble nozzle 19 provided inside the cylinder 24 constituting the flow straightening plate immediately above the centrifugal blade 18. Since a gas containing no is sent and the culture solution is always kept in an optimum state for fermentation of lactic acid bacteria which are anaerobic bacteria, the lactic acid bacteria fixed to the granular gel attached to the support frame 6 are: Continuous culture is performed with a large reaction rate, and the number of lactic acid bacteria in the culture solution gradually increases.

Further, by using such a culture device, since the stock solution for fermentation always flows on the surface of the granular gel on which lactic acid bacteria are immobilized, lactic acid, which is a reaction product associated with fermentation, is locally concentrated. This can prevent the decrease in the activity of lactic acid bacteria due to the increase in the concentration of lactic acid.

Further, since the granular gel attached to the outer peripheral surface of the support frame 6 is not hit by the vanes 17 during the operation of the culture device, the granular gel is not destroyed by the operation of the culture device.

The growth of such immobilized cells was carried out at 30 ° C for 48 hours, and the number of lactic acid bacteria contained in the gel beads was 10 ⁸ to 10 ⁹ cells / m 2.
I increased to l.

C. Product removal As described above, reduce the number of lactic acid bacteria contained in gel beads to 10
^{When the} concentration was increased to ⁸ to 10 ⁹ cells / ml, the fermented stock solution was fed into the glass container of the fermenter at a rate of 30 ml / hour, and the same amount of fermented product was taken out from the glass container.

This product was taken out for 30 consecutive days, and the taken out product was tasted by 5 men and women in their 20s, 30s and 40s, for a total of 30 people. Was obtained. Although there was no clear difference in the evaluation with mere apple juice without addition of lactic acid bacteria, there were almost the same number of those who added lactic acid bacteria as those better than those without added lactic acid bacteria, It was found that the lactic acid bacterium drink produced by the method described above can cope with diversified taste.

In addition to apple juice as fruit juice, tangerine juice and grape juice were used, and noodle juice and grain seed germ extract were used in the same experiment as yeast extract in addition to yeast extract, where yeast extract was used. The same result was obtained.

Furthermore, when the same experiment was performed by changing the PH value of the fermentation stock solution up and down, when the PH value was in the range of 3.0 to 4.5, the lactic acid bacterium beverage could be efficiently produced, but PH The value is 3.
Even if it was 0 or less, or conversely, if it was 4.5 or more, the number of lactic acid bacteria did not increase, and it was not possible to efficiently manufacture a lactic acid bacterium beverage.

c. Effects of the invention Since the method for producing a lactic acid bacterium beverage of the present invention is configured and carried out as described above, it is possible to efficiently produce a lactic acid bacterium beverage containing fruit juice, which has been considered difficult in the past. Not only can it respond to changes in consumption, but it can also increase consumption of fruits, whose consumption is sluggish due to changes in taste.

[Brief description of drawings]

The drawing is a longitudinal sectional view showing an example of a fermenter used for carrying out the present invention. 1: Glass tube, 2: Bottom plate, 3: Studs, 4: Lid plate, 5: Circular hole, 6:
Support frame, 7: stay, 8: vertical axis, 9: rotating block, 10: cap, 11: magnet, 12: board, 13: bracket, 14: motor, 15: output shaft, 16: magnet, 17: vane , 18: Centrifugal blade, 19: Bubble nozzle, 20: Air pipe, 21: Small hole, 22: Nozzle part, 23: Ring plate, 24: Cylinder, 25: Rectifier plate.

Claims (2)

[Claims] 1. A PH value is obtained by mixing one or more of yeast extract, noodle juice, rice germ extract, and malt extract in an amount of 0.05 to 1.0%.
A method for producing a lactic acid bacterium beverage, wherein a mixed solution containing fruit juice as a main component, which is 3.0 to 4.5, is used as a stock solution for lactic acid fermentation, and the stock solution for lactic acid fermentation is fermented by contact with lactic acid bacteria. 2. A PH value is obtained by mixing one or more of yeast extract, noodle juice, rice germ extract and malt extract in an amount of 0.05 to 1.0%.
A mixed solution containing 3.0 to 4.5 as the main component of fruit juice is used as a stock solution for lactic acid fermentation, and the stock solution for fermentation is 10 ⁸ to 10 ⁹ cells / ml of lactic acid bacteria.
With the gel fixed comprehensively at the ratio of, the lower end opening was closed by the bottom plate, the upper end opening was provided with a lid plate that could be opened and closed, and the glass tube was provided inside the glass tube and approximately concentrically with this glass tube. A straightening plate having a cylinder that rises upward from the inner circumferential edge of the horizontal circular plate, and a supporting frame having a large number of small holes fixed by fitting the lower end portion to the cylinder of the straightening plate. , Placed on an upper side of the bottom plate by a motor, and put into a culture device consisting of a centrifugal blade that is driven to rotate on the lower side of a circular plate that constitutes the straightening plate around the vertical axis, and the centrifugal blade is rotated. And a method for producing a lactic acid bacterium beverage in which a stock solution for lactic acid fermentation is caused to flow between gels attached to the outer peripheral surface of a support frame.