JPS59125863A - Preparation of seasoning by lactic acid fermentation - Google Patents

Abstract

PURPOSE:To prepare a lactic fermentation seasoning liquid containing lactic acid in high concentration, by culturing a lactobacillus belonging to Lactobacillus genus and resistant to acetic acid. CONSTITUTION:A lactobacillus strain belonging to Lactobacillus genus, resistant to acetic acid having an acidity of >=2%, and capable of carrying out the homo- type lactic acid fermentation, e.g. Lactobacillus acetotolerans MG-1 strain (FERM-P No.6784), is inoculated in a raw material for fermentation which may contain acetic acid having an acidity of >=2%, and cultured at 30-50 deg.C and 3.3-6.6 pH without agitation. After the completion of fermentation, the fermentation, liquid is if necessary subjected to the removal of germs or sterilization to obtain the objective seasoning liquid containing >=3% of lactic acid.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for producing a lactic acid-fermented seasoning solution, and more specifically, a method for producing a lactic acid-fermented seasoning solution containing a high concentration of lactic acid using acetic acid-resistant lactic acid bacteria belonging to the genus Lactobacillus. Relating to a method of manufacturing.

Lactobacillus belonging to the genus Lactobacillus has a higher ability to produce lactic acid than lactic acid cocci belonging to the genus Streptococcus, Leuconostoc, Pediococcus, etc.
To the classification of the lactohashirai (An approach to the c
lassification of the 1acto
bacilli), Journal of Applied Hachte Riology, (Tourna), of
Applied Bacteriology), No. 22
vol. 329-340 (1959), Lactobacillus helveticus (Lacto, bacillus)
When milk is used as a raw material, it has the ability to accumulate 27% of lactic acid, and is said to have the highest lactic acid production ability among known lactic acid bacteria.

Ni IA-1 armor '1 Mukutsu' [Karana 4) Hara ('[wo month 1
If it is ``color'', it is fortune-telling, II) 11.
brueckii) has the ability to store up to 18% of lactic acid.
1-) Miyanami-shi store (1958), No. 2471+
1.

Lactic acid fermentation seasoning liquid containing lactic acid bacteria afi once a month (in the present invention, lactic acid fermentation seasoning liquid refers to the fermented liquid as it is,
When producing I-Y (commonly referred to as 3), a seasoning liquid containing I-Y for sterilization and sterilization, it is natural that it should contain a high concentration of lactic acid. ) or using the conventionally known lactic acid a1 culture (1,
It is said that 27% of lactic acid is accumulated in the above-mentioned amount.
There was t'te. In addition, the 6-lactic acid produced during lactic acid fermentation
If the neutralization operation is delayed, a higher concentration of lactic acid will accumulate.
(It is not possible to obtain the lactic acid seasoning liquid intended by the present invention.

Ma74-1 Quality (1) using previously known lactic acid bacteria
) In order to produce a stable acid-fermented seasoning liquid, it is necessary to take measures against milk intoxication M'i': during fermentation. Of course, the best option is to sterilize the bacteria and perform pure culture using an uninterrupted fermentation facility (Mi'). If possible, it is under fermentation!
It goes without saying that it is very meaningful to try to improve I'iri of equipment and facilities.One example is the use of high-temperature-resistant lactic acid bacteria such as Lactobacillus delprudskii.
0°C e] Near Takafuku, when there is a certain degree of bacterial contamination due to fermentation +J-b) "i] Noyo na natoko, Li, ;j,
It is a well-known fact that when using lactocils as mentioned above, there is no accumulation of lactic acid, and
Regarding the production of lactic acid fermentation seasoning liquid as intended by the present invention d:
Preferably rc.

A selective medium for lactic acid bacteria belonging to the genus Lactobacillus (e.g. "Acetate 1-Agar") has strong antibacterial activity.
a, ga, r”).

Journal of Applied Hatateriorono ~ 1st
9, pp. 95-101 (1956), lactic acid bacteria belonging to the genus Lactobacillus have relatively strong acetic acid tolerance, as is clear from the fact that about 15% acetic acid (including acetate) was not added. Has [2].

■Do/J '-1'j・'1. i, I! +C'4'
How many 1 bacteria/L) 14-. are added to l + °, y r rent 11 U
+l-1-, K) i) j, ・/) (-
be. However, acid resistance concentration of about 15% (vinegar i''・i!
I,,・l k:'+mu）deni 1, many species of Lactobacillus genus 1,・11, zurose acid i'I! Lactic acid fermentation seasoning 771 production ('(Lactic acid ratio of 7.[sugar acid ratio is 1:1, 1 <
, lactic acid may produce other fermentation products and off-flavors).
Pure cultivation of lactic acid bacteria for homozygous lactic acid fermentation II: Production of 3L stable/ζ lactic acid fermentation preparation -
Kazufu 111 Noh-C3j) Tsuda. In particular, lactic acid bacteria that carry out Peter's type lactic acid fermentation often grow as harmful bacteria, causing problems such as a decrease in lactic acid (j), a decrease in sugar acid ratio, and the generation of off-flavors. For (there was no size.

The inventors of the present invention found out through experiments that the known 'i'
'L acid bacteria fl'fi were grown in the presence of acetic acid with an acetic acid acidity of 2% or more, P, and c. Table 1 shows experiments on acetic acid tolerance of Lactohyperus lactic acid bacteria that undergo homofermentation (9 cases are shown in Table 1). , with acetic acid tolerance of 2% or more
, and if a homozygous lactic acid bacterium with strong lactic acid fermentation ability is found, a raw material containing this lactic acid bacterium and acetic acid at 2% or more in terms of acetic acid acidity can be used, thereby eliminating the need for sterilization of the raw material or aseptic fermentation equipment. It is thought that it will be possible to completely prevent bacterial contamination by producing a lactic acid fermentation seasoning liquid by pure culture of lactic acid bacteria.3, na-],,i'i'1. ″″lj acid and 2 or more sizes! 'W'1=can grow in the presence of acid)
)'s microorganisms t1 are strongly aerobic, so lactic acid bacteria Kinen (・
When manufacturing the entire lactic acid fermented seasoning liquid, the surface of the fermented liquid has a golden curve +
To cover with a layer of less or no 1 lJ(',
These microorganisms can be easily controlled3.
However, these microorganisms have a small salt exchange rate 1 (1'), so using frozen soil containing 15% or more salt is still effective in controlling them.

L like this? In this context, the present inventors have discovered that the known lactic acid bacteria are not only more resistant than I but also acid resistant to 14 I, and infants 1,000 to 1,000.

The invention belongs to the genus Lactobacillus,
Lactic acid fermentation is characterized by inoculating lactic acid bacteria that have an acetic acid surface of 12% or more and that performs homobarrier 1-acid fermentation into a fermentation source and culture it. The present invention relates to a method for producing a seasoning liquid.

In carrying out the present invention, by adding glacial acetic acid divine vinegar, etc. to the fermentation raw material so that the acetic acid acidity is 2 or more, the acetic acid-resistant lactic acid bacteria used in the present invention can be selectively grown, and the raw material can be sterilized and Lactic acid fermentation without M contamination becomes possible without using aseptic fermentation equipment.

The acetic acid-resistant lactic acid bacteria used in the present invention have an acetic acid acidity of 2 to 1.
Grows under 0 conditions, performs lactic acid fermentation (・obtains)
Lbacillus aceLoLolerans)
T'j K'i RyoJ is JrJ. The acetotolerance properties of Lactobacillus, a representative strain of lactic acid bacteria, are shown below.

To Lactobacillus acetotolerance 4G-14 Mechanical properties of Vermilion 1 Morphological properties ■ Image shape: Bacillus longacillus □, □: 1 a. \ :tL4 ~ 0
, 5 71m X l , 1 ~ 2
8 μI nol (i1; 1・dou・1'1: otsu・shiku・1) ・l'll+' (two, no i1';
11jF: No 5) I f uniqueness of vesicle 1' (i.1-51, presence or absence of j chromosomes: No 7) Gerano staining = No. 14 (8) Acid-fast f1: negative JT 2Iin "<M acid properties■ Meat juice agar plate culture: No growth (Iri meat i'l Agar plate culture: No growth ([ri Meat juice liquid No. V: No growth (・1) Meat juice agar plate culture: No growth (・1) Puncture culture: no growth, (nocturnal growth (bow)) lidmus milk: no change (growth on J + blinogus agar plate: circular, medium convex shape ~
Convex circular shape, wavy, rough, milky white, opaque, 0.6 to 1.6 m
UI (30'C, I4El culture) (7) Growth on Greenogs agar slant: weak growth, rough.

Milky white, opaque (8) Solinoges agar puncture culture: Growth in two lines (ri) Fininois subiratin puncture culture: Growth in a line.

Liquefied gas [phase] Burinogus River was incubated overnight at -30°C,
It became cloudy in 2 to 4 days and later formed sediment and residue.

3 Physiological properties ■ Reduction of nitrate: negative ■ Denitrification reaction: negative ■ vp test - weakly positive ■ Formation of indole: negative ■ Formation of hydrogen sulfide: negative ■ Hydrolysis of starch'/: negative ■ Citric (use of feces) (Koser's medium and Chr
istensen's medium): Negative ■Pigment production:
shadow. Sex ■Urease: Negative qYuoxitase: Negative @Nonotarase: Negative ■1) Growth PH: PH 3, 3-8.62) Optimal, H: Pll 5, 0-5501) Growth temperature = 2
0-40'C 2) Optimum temperature: 35°C Attitude towards oxygen: Facultative anaerobic (generation of acid and gas from internal carbohydrates = 1) L-arabinose --- 2) D-xylose
--:() l)-glucose
10-4) D-mannose 1-
-5) D-fructose + -6) Each galactose - -~7) Maltose
10-8) Sucrose
-9) Lactose -- H)) l)-Trehalose -11)
l)-Norbit-12) I)-Mannitol-1:
3) ・1)
green rin
-15) Starch - 16) L-rhamnose - 17) I, -norbose - acid gas 18) l) -ribose
± -19) I)-Seven Rochiose -~20) I)-Nori ζ゛ose
---21) Raffinose ---
22) D-meletitose - -23) Aesculin -24) Zarysin
± -25) Amygdalin
--4 Properties of the song ■ Degradation products from glucose: lactic acid and a trace amount of acetic acid, no gas formation ■ Shigure name of lactic acid: 1〕L ■ Decomposition of esculin weakly positive ■ Decomposition of arginine: negative ■ Temperature resistance ■Thorium chloride resistance: No growth at 60°C ■No growth at 12% acetic acid resistance (pH 5,0), No growth at 6th rate (pH 3,5) ■ Auxotrophy 1) Necessary A't・1−/ζ6−1,/1″ To promote growth, it is necessary to promote growth.

liponolahine, carunium pantothenate.

Scolding, uracil, Tween 80 2) Fj m
.

Biotin, dianocobalamin, adenine.

Guanine, Gizuntin, Pebzido 4. citric acid.

Presence or absence of mevalonic acid (...) dichloropropanoic acid (minus fatty acid silk formation)
:I]S+it4+ ill's peptidoglycan type: di1-lysine-D-aspartic acid type full OC2Ji: 34.4% (+2) separation rll;! 'Vinegar moromi J'l, 1- Part aids manual off deterministic bacteriology = (Ber
gey's Manual of Determina
Live Bacter: i,ology) Part 8
Edition (1974,
abora, tory manual) 4th edition (19
77), I-)-Indifference Methods:
・Four microphone Il high A Logisn Bart A (engineering methods demb1.fi, cationmethods fo
r m1crobi, ogists, pa, r
tΔ), Anno J Demik Press In-1-Boreitainod, Niko-York (Academj, c Pre
ss Inc., New York) (] 9
6 G), pp. 65-79, Identification Office, Acid Bacteria (Iden 7j-flca, t), on of the
T, +aC'L;]CAc1. d Bacterj,
a) and the aid of intestinal bacteria, Sobunrei (1:]80) 1
According to Isolation and Identification of Anaerobic Bacteria on pages 03-327, 8J
As a result, this bacterial strain d is a facultative anaerobic Durham-positive non-spore bacillus, negative for catalase, whose main product from glucose is lactic acid, and which contains lactic acid as a bacterial fatty acid. Based on these characteristics, it is thought that it is a bacteria belonging to the genus Lactobacillus.However, I found that the mycological properties of #i ai described in the literature of Juki 4fl match those of this strain.4ε To explain in detail to Katsu/Densa Nno et al., this strain does not grow at 15°C, does not produce scum from glucos, and performs homozygous lactic acid fermentation, so this strain is suitable for the identification method four.・Micronoh Ioronsuno Tsuku-1・A.

Academic Breath Incorporated,
Niko I York (1966) pages 65-79 item) 4 inyo/・issue-7・su・rafcho−nok・annot・bacterium l+, -'(ii)Leamohacterium (1'her+nobac sier) :i,um)
9' (Nihito I1. (-, A1 I)t\kimono de aestu)
Of these, Lactohacilus helvetichus and Lactohazardus neigleti (
Lactobacillus plate punch”).

Lactohadilus acidophilus (Lac; obac
j, 1lus (Namiren Zuizanko).

However, this strain has clearly different properties from the L-] strain shown in Table 2. k (-1-. That is, Lactobacillus suicus, Toro: It has no raw acidity from lactose, D-galactose, does not change lidmus milk, does not require biritoglyl, nicotinic acid and does not require folic acid, 1 〉The temperature is in the appropriate temperature range or lower, and 45°C is l ('1, 7c, -!j':
Suitable 1) H is on the more acidic side and even at pH 3.3/Iiku 1,
It does not grow at pH 7, and its acetic acid tolerance is extremely low.
strong, the size of the bacteria is smaller, and the source of isolation (1.
There are obvious differences, such as the origin of ``411''.

Lactohydralus neigleti is listed in the Burn AIDS Manual, Anatomy, Butaminative Hacteriology, No. 8.
In the edition (1!l7/l), it is considered to be Lactobacillus helhedeix Biowar, which does not have bioacidity from barley sugar, but it is different from Lactobacillus helveticus described in 8J of this strain. In addition, it is clearly different from the fact that it has bioacidity from maltose.
(it's on).

Also, Lactobacillus acidophilus is lactose.

No bioacidity from D-galactose, sucrose, cellohiose, esculin, no change in mass milk, no requirement for nicotinic acid, acetic acid or I71, mevalonic acid, optimum pH is more acidic. Yes, ll
llGrows even at H3.3 and grows at pH 7 [no 7,
They are clearly different in that they are extremely resistant to acetic acid, are smaller than the size of the bacteria, and are isolated from different sources (origins of the bacteria). Based on the above results, this strain 1♀j is considered to belong to a new bacterial species, and is named Lactobacillus a7tolerance I2. It was entrusted to the storage facility (Micho 6) [Kyoyori No. 6784].

Table 2 The raw material E.1 used in the present invention includes the fermentation final liquid (1), or the desired seasoning e.g. by simply sterilizing or sterilizing it. In the context of obtaining all ingredients, it is desirable to use all ingredients, such as those derived from grains, vegetables, fruits, meat, yeast cells, etc. Saccharified liquid, decomposition liquid, extract liquid, juice juice and natural seasonings (specifically HAP, HVI), vegetable extract, seafood extract, yeast extract, flavor liquid, soy sauce, white soy sauce, miso, etc. are preferably used. I know. These natural raw materials d: normally contain sufficient nutrients necessary for the growth of lactic acid bacteria, and from this point of view as well, they are advantageous as the raw materials 1'-1 used in the present invention. In addition,
In order to produce a lactic acid fermented seasoning liquid containing 3 to 10 parts of lactic acid, it is necessary that the raw material contains at least 3 parts or more of fermentable sugars represented by glucose. However, it is expected that these ingredients will be insufficient if only the raw materials listed in -1- are used.In this case, by supplementing all fermentable sugars such as glucose as necessary, the desired lactic acid fermented seasoning solution can be obtained. It is possible to go all the way.

For the purpose of preventing bacterial contamination, raw material 1 containing acetic acid was used in rivers (・
2] In the case of fcl-1, the acetic acid acidity should be 2 or more (2 f1..J), and if the acidity is 1, sufficient effect will not be obtained.

Glacial acetic acid or vinegar are the most commonly used sources of acid resistance.

C; 1. Pll of the month should be within the range where the lactic acid bacteria used for fermentation can grow and lactic acid production can be carried out, but Lacto. I should...l
It is preferable to adjust it to -14.5 to 6.5.

The fermentation temperature may be within a range where the lactic acid bacteria used for fermentation have no growth capacity and can produce lactic acid, but when using Bacillus acetotolerance, l: 30
“C ~ 35°C is tricky.

By performing static fermentation under the above conditions, usually 2 O
Fermentation is completed within Lj, and a fermented liquid containing 3 or more lactic acids is obtained.The fermented liquid obtained here can be used for acid generation or as a seasoning liquid. KotomoichiC
However, it is usually sterilized and sterilized to make a clear liquid, which is then used as a seasoning liquid. The lactic acid fermentation seasoning liquid thus obtained is used as it is or diluted as necessary for seasoning pickles and the like. At this time, it is of course possible to adjust the taste by using salt and other seasonings.

The present invention will be further explained below with reference to Examples.

Example 1 2 parts by weight of glucose and 1 part by weight of yeast extract were added and dissolved in 100 parts by weight of fresh cabbage juice, and the solution was heated to 121°C.
After autoclaving for 0 minutes, inoculate Tobacillus acetotonlans MG-1 strain (FERM P-6784) so that the total number of viable bacteria after inoculation is approximately 107 cells/ml.
Aseptic fermentation was performed at 30°C for 7 days. After the fermentation was completed, bacteria were removed by centrifugation, and the mixture was sterilized by heating at 75°C for 10 minutes to obtain a pale yellow, transparent fermented liquid containing 35 parts of p-lactic acid. The fermented liquid obtained here is
! ．． It could be used as an excellent lactic acid fermentation seasoning liquid.

An example of a seasoning formulation using the lactic acid fermented seasoning liquid produced by A4J is shown below.

Dilute the lactic acid fermentation seasoning liquid 3 times with water, add and dissolve all 7 salts, and then add the same amount of blanched ingredients to the original weight. , By doing so, I don't have any crap! r! A gf product of L was produced. The result of this kimono's self-performance is compared to commercially available sour craft.
``The song received 1 rating.

Example 2 After heating and sterilizing GEO (100 parts) at 120°C for 10 minutes, 01 parts by weight of amylase preparation (trade name: '
: J Crase G2, manufactured by Sankyo ■) was added and heated at 55℃ for 1
After saccharifying for 0 hours, a rice saccharified liquid was obtained by pressing through dj. At the same time, add 100 parts of steamed soybeans to 120'
c. After heat sterilizing for 10 minutes, 01-Φ ridge part [)
``Aase preparation (trade name: ``Amano'' A) manufactured by Ten IJJF Seika Co., Ltd.) was added and decomposed at 2.55''C for 10 hours.

Next, the soybean decomposed liquid is obtained by pressing and passing through the filter. Saccharified rice liquid obtained here 1. 0 0 parts by weight, 40 parts by weight of the decomposed solution and 60 parts by weight of water were mixed together,
] 2 1” After high-pressure sterilization for 10 minutes, rough i.
・-Trus・aceIl・lerans MG-1 strain-t
Contact rl after c7)/l river use? Approximately 107 pieces/m
The seedlings were inoculated so as to be as follows, and fermented aseptically at 30°C for 8 days. After completion of fermentation, bacteria were removed by centrifugation, and the mixture was sterilized by heating at 75°G for 10 minutes to obtain a pale yellow, transparent fermented liquid containing 3.5 parts of lactic acid. The fermented liquid obtained here can be used as an excellent lactic acid fermented seasoning liquid. An example of a seasoning formulation using the obtained lactic acid fermentation seasoning liquid is shown below.

Dilute the lactic acid fermentation seasoning liquid 8 times with water, add and dissolve 6 parts of common salt and 1% gold of sodium glutamate.
By adding Chinese cabbage cut into quarters and preparing it at night, Asamoe produced a green vegetable with a breeze. The results of the sensory evaluation of this pickle were extremely good, with the pickle being characterized by having a mild sour taste and moderate fermented flavor.

Example 3 In the same manner as in Example 2, 110 parts by weight of the saccharified rice, 20 parts by weight of soy sauce, and 70 parts of brewed vinegar with an acetic acid level of 15 parts were mixed, and then 1. Adjusted to Pf I55 using IJ Uno. This was inoculated with lactono-chillless acetotonolance MG-1 strain so that the number of viable bacteria after inoculation was approximately 1 Fl'i/m (F'li), and left standing at 1.30'C for 10 days. The fermented liquid contains 7.2 parts of lactic acid and 4 to 5.2 L of acetic acid, so it can be used as an excellent 17 m acid fermented seasoning liquid even with its age. An example of a seasoning formulation using the lζ lactic acid fermented seasoning liquid is shown in 1. below.

Sprinkle 3% salt on 4 UJJ-ripped Chinese cabbages and marinate overnight/... After that, add lactic acid fermentation seasoning liquid 18, no glutamic acid.
/'3%, 1.7 topiyu 10%, garlic paste 1%, onion powder 0.4 part, Ji'jI fu-1
(Keelin Tangano,) 0.2%, ground chili pepper 0
．． Add a seasoning liquid consisting of 6 parts, 1 part (all brown), 1 part salt, and 3 parts to Chinese cabbage. ?& manufactured things.

The result of the sensory evaluation of this γ11 is d: It is said to have a good Ilf length in that it has a sweet-yet-sour taste and a moderate fermented flavor.

l hi lAf row 4 Rice 11 saccharified liquid obtained in the same manner as in Example 2 5o heavy jI
1.!, 4J decomposition of dried wheat gluten e by the method shown in Example 2 of the specification of Publication No. 55-32344, 80 parts by weight, and 70 parts by weight of brewed vinegar with an acetic acid acidity of 15 parts.
After mixing the heavy phase and 5 parts by weight of glucose, the pH was adjusted to 5.0 using bicarbonate. To this, Lactobacillus acetotolerans MG-] strain (FERM P
-6784), the number of viable bacteria after sampling was approximately 107/me
After inoculation, the liquid surface was covered with a floating lid made of expanded polystyrene, and fermentation was allowed to stand at 30°C for 15 days. After the fermentation was completed, bacteria were removed by centrifugation to obtain a pale yellow, transparent fermentation liquid containing 6.5% lactic acid and 5.5% gold acetate. The fermented liquid obtained here could even be used as an excellent lactic acid fermented seasoning liquid. An example of a seasoning prescription using the obtained lactic acid fermented seasoning liquid is shown below.

By diluting the lactic acid fermentation seasoning liquid 4 times with water, adding 8 parts of common salt and dissolving it in the seasoning liquid, salted cucumbers and eggplants were desalted and pressed under running water and soaked in equal weight overnight. produced living organisms.

The results of the sensory evaluation of this pickle were compared to commercially available fermented Shibasei.
=< 11. There is no harsh fermentation odor, and the sourness is mellow.

Claims (1)

[Scope of Claims] 1. Lactic acid fermentation characterized by inoculating a fermentation raw material with lactic acid bacteria that belong to the genus Lactobacillus, have acetic acid tolerance with an acetic acid level of 2% or more, and perform homozygous lactic acid fermentation, and culture the same. Method for producing seasoning liquid. 2. The production method according to claim 1, wherein the lactic acid bacterium is a new bacterial species Lactobacillus acetotolerans 3. Lactic acid m or Lactobacillus acetotolerans M
The manufacturing method according to claim 1, which is O-1 strain (FERM P-6784,). 4. The fermented raw material is a natural seasoning. Claim 1
Manufacturing method described in section. 5. The production method according to claim 1, wherein the fermentation raw material contains acetic acid with an acetic acidity of 2% or more.