JPH08336355A - Bulgarian yogurt and lactic acid product starters and their production - Google Patents

Abstract

(57) Abstract: A starter for producing other lactic acid products including a Bulgarian yogurt and a product derived from a non-dairy substrate, and a production method thereof. A starter is Lactobacillus
at least one strain and / or St selected from the group delbrueckii subspecies bulgaricus
reptococcus salivarus subspecies t
It consists of at least one strain selected from the group of thermophilus. This starter is produced by culturing in a nutrient medium containing skim milk obtained by hydrolyzing casein and lactose and a biologically active additive.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to the field of manufacturing starters for the production of lactic acid products, including products derived from Bulgarian yogurt and non-dairy substrates.

[0002]

Prior Art and Problems to be Solved by the Invention La
ctobacillus bulgaricus strain N
o. La selected from the group of 5, 26, 37 or 144
C. bacillus bulgaricus strain and Streptococcus thermophi
lus group No. Streptococcus thermophil selected from 12, 14, 17 and 22
A symbiotic starter of Bulgarian yogurt (1) consisting of a 1: 3 to 1:10 ratio of the us strain is known. A particular example is L.L. b. No. 5 + S. t. No. 12; b.
No. 5 + S. t. No. 22; b. No. 37+
S. t. No. 18; b. No. 37 + S. t. N
o. 12; b. No. 144 + S. t. No. 1
2; b. No. 144 + S. t. No. 14;
b. No. 26 + S. t. No. It is a starter consisting of 12.

The starter is produced by a method including Lactobacillus bulgaricus and Streptococcus thermophi containing the above-mentioned strains.
Different combinations of selected strains of lus are inoculated into sterile milk with the combination identities having the usual morphology and normal ratio. It is re-inoculated into sterile sheep milk or a combination of sterile sheep milk and milk, followed by 6 months of re-inoculation,
Then stable morphological properties and lactobaci
A combination with a stable ratio between Ili and Streptococci is used in the manufacture of bulk starters. Inoculation and culturing are carried out at a temperature of 45 ° C. ± 1 ° C., cultivation is continued for 2 hours, and once a week for the next 2 months. The first inoculation is on a 1: 1 ratio of a mixture of sheep and milk.

One or more Lactoba from the National Bank for Industrial Microorganisms and Cell Culture (NBIMCC)
cillus delbrueckii subspecies bulga
rice no. 2183, 2184, 2185, 2
186, 2187, 2188, 2189, 2190, 2
191 and 2192 and strain Streptococ
cus salivarus subspecies thermophi
lus No. 2173, 2174, 2175, 217
6, 2177, 2178, 2179, 2180, 218
A symbiotic starter (2) consisting of 1 and 2182 is known. A nutrient medium consisting of a fermented casein hydrolyzate and a secondary milk product in a 1: 2 ratio obtained by filtering the combination of the above strains according to (2) (the medium was previously neutralized to pH 6.5 to 7.0, Sterilize for 20 minutes at 45 ° C. ± 1 ° C., then periodically neutralize within the limits of pH 5.0 to 6.5, and add 0.05 to 0.2% amount of gelling agent. And freeze-dried to produce the starter as a concentrate. Secondary dairy product refers to a mixture of permeate or whey with degraded lactose and / or sterile skim milk.

[0005]

Starters have been proposed for the production of lactic acid products, including Bulgarian yogurt and non-dairy based products, Lactobacillus delbru.
At least one strain selected from the eckii subspecies bulgaricus group, ━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━ NBIMCC registration Number NBIMCC registration number ────────────────────────────────── LBB B m1 2471 LBB B 31 31 2481 LBB B m2 2472 LBB B 37 286 LBB B m14 2473 LBB B 41 41482 LBB B 5 273 LBB B 120 2483 LBB B 14 B 27 B 28 B B 22 B 4 B 42 B 42 L 4 B 4 B L 2 B 4 B 2 4 B L 2 B 4 B 2 4 L 2 B 4 B 2 L 4 B 2 B 4 L 2 B 4 B 2 L 4 B 2 B 4 B 2 L 4 B 2 B 4 B 2 L 4 B 2 B 4 B 2 B 4 B 2 B 4 B 2 L 4 B 2 B 4 B 2 B 4 B 2 B 4 B 2 B 4 B 2 4 B 4 B 2 4 B 2 B 4 B 4 B 4 B 4 B 2 8 B 4 B 4 B 4 B 2 4 B 4 B 4 B 2 8 B 4 B 4 B 4 B 2 4 B 4 B 4 B 4 9 B 4 B 4 B 4 8 B 4 B 4 9 B 4 B 4 B 4 B 4 B 4 B 4 B 6 B 6 B 6 B 6 B 6 B 6 7 B 6 7 B 6 8 6 6 6 7 6 156 2486 ━━━━━━━━━━━━━━━━━━━━━━━━━ ━━━━━━━━ and / or Streptococcus sali
at least one strain selected from the group varius thermophilus, ━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━ Number NBIMCC registration number ────────────────────────────────── LBB T3 2487 LBB T18 452 LBB T8 2488 LBB T22 453 LBB T10 2489 LBB T28 2492 LBB T12 450 LBB T36 2493 LBB T14 451 LBB T38 2494 LBB T16 2490 LBB Tm5 2474 ━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━ TRB ━━━━━━━━━━━━━━ consists of mixed or co-cultures or monocultures .

The strain is a true culture isolated from the geographical area where Bulgaria yogurt is traditionally produced, according to the classification of Vergie, and according to IDF Standard 1.
46/1991 and IDF Standard 149/1
Classify according to the morphological, physiological and biochemical properties of 991. The preferred variant of the inventive symbiotic starter is NB
Lactobacillus in the table below with IMCC numbers
s delbrueckii subspecies bulgaricus
And Streptococcus salivari
It is a combination of the us subspecies thermophilus. ━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━ Symbiotic starter L.delbrueckii Subspecies S.salivarius Subspecies bulgaricus thermophilus, ─ ───────────────────── NBIMCC registration number NBIMCC registration number ─────────────────────── ─────────── 2484-2491 2484 2491 2482-451 2482 451 2484-453 2484 453 2480-453 2480 453 2478-2489 2478 2489 2479-2488 2479 2488 2479-2490 2479 2490 2485-2490-24 2490 2486-453 2486 453 ━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━

The method for producing the mixed starter is a liquid nutrient medium,
To cultivate a monoculture from the cited strain in an agar nutrient medium. It selects colonies according to their morphological characteristics, selects the technical parameters of the monoculture, and its inoculum has a 5-10% dry matter content, skim milk and meat prehydrolyzed of casein and lactose. Extract (preferably 1-2%), peptone (preferably 0.5)
~ 1%), yeast extract (preferably 0.1-0.5%)
To a liquid nutrient medium (a) consisting of a biologically activated additive consisting of
Incubate to 0.1.

Lactobacillus thus obtained
delbrueckii subspecies bulgaricus and Streptococcus salivariu
Monocultures of the S. subsp.

The method for producing the symbiotic starter is to repeat the re-inoculation for at least 4 months to obtain the strain Lactobac.
illus delbrueckii subspecies bulgar
icus and Streptococcus sali
to maintain the combination of varius subsp. thermophilus. This combination is either skim milk or whole milk, or Lact in the subculture of these strains in the cited nutrient medium (a) in joint cultivation.
obacillus delbrueckii subspecies bu
lgaricus and Streptococcus
It showed a stable correlation within the desired limits between Salivarus subsp. thermophilus.

Methods for producing direct culture (DVS) monoculture concentrates, concentrated mixes or symbiotic starters are: Whey Restorative and CH ₃ COONa (preferably 5-10%),
KH ₂ PO ₄ (preferably 1 to 4%), Na ₂ HPO ₄
(Preferably 1 to 4%), MgSO ₄ (preferably 0.
1-0.5%), MnSO ₄ (preferably 0.01-)
0.06%), yeast extract (preferably 0.1-0.5)
%) Of a nutrient-medium (b) consisting of a buffer-activating mixture, or restored dried milk and CH ₃ COONa.
(Preferably 5 to 10%), KH ₂ PO ₄ (preferably 1 to 4%), Na ₂ HPO ₄ (preferably 1 to 4%),
MgSO ₄ (preferably 0.1 to 0.5%), MnSO
₄ (preferably 0.01 to 0.06%), yeast extract (preferably 0.1 to 0.5%), and lactose and protein concentrate (preferably 0.5) obtained by ultrafiltration.
The nutrient medium (c) consisting of a buffer-activation mixture, which is an aqueous solution of ~ 4%), is inoculated with 3-8% of each culture. The fermentation method of the nutrient media (b) and (c) lasts 4-5 hours. The concentrated culture thus obtained is then mixed with a protective medium and equilibrated at 15-20 ° C. The protective medium is a sterile aqueous solution of sucrose, sodium glutamate and vitamin C or polyethylene oxide (molecular weight 400 or 1000). The concentrated medium thus obtained is frozen or freeze-dried at a temperature of −65 to −80 ° C., or −196 ° C.

The advantage of the strain of the invention is that according to the invention Lac
tobacillus delbrueckii subspecies b
at least one selected from the group ulgaricus and Streptococcus sal according to the invention
It is to provide the opportunity to use mixed starters which represent different combinations of the desired ratios of at least one strain selected from the group Ivarius subsp. thermophilus. This produces essentially the same lactic acid product, but with different amounts depending on different tastes, preferences or specific requirements for production and storage.

Another advantage of the strains according to the invention is that the symbiotic starters obtained thereby are inoculated daily into the nutrient medium for at least 4 months in Lactobacillus delbruecki.
i subspecies bulgaricus and Streptoco
ccus salivarus subspecies thermop
to maintain the desired ratio between ilus. A starter with standardized quality characteristics is obtained.

The method and the nutrient medium created for the purposes of the invention make it possible to obtain monocultures, mixed or symbiotic starters in the form of DVS, which are considerably shortened in comparison with the known methods for obtaining concentrated starters. .

The DVS-cultures (mono or mixed or co-cultivation) obtained according to the invention are used for obtaining bulk starters and for direct application to the production of Bulgarian yogurt both under industrial conditions and at home, and It is suitable for the production of other lactic acid products, including products from non-dairy substrates.

[0015]

【Example】

Example 1 Preparation of mixed starter Lactobacillus delbru incubated in liquid nutrient medium (MRS or M17) for 16 hours at 37 ° C.
eckii subspecies bulgaricus and Strep
tococcus salivarus subspecies ther
A pure culture of M. philus is inoculated into solid nutrient medium (MRS or M17) in the required amount to obtain one colony, and then cultured at 37 ° C. for 48 hours. Colonies with morphological characteristics are selected, inoculated in liquid nutrient medium and then cultured at 37 ° C for 18 hours. Then, using 5 liters of the obtained laboratory culture, 100 liters of prehydrolyzed skim milk of casein and lactose, meat extract 100
g, 50 g of peptone, 10 g of yeast extract and water to a dry matter content of 8% (a), then neutralized to pH 6.5 with ammonia and cultivated at 42 ° C. until pH 4.6. To do. The number of microorganisms obtained is Lactoba
cillus delbrueckii subspecies bulga
In ricus strain 10 ⁷ ~10 ⁸ cfu / ml and S
It is 10 ⁸ to 10 ⁹ for Treptococcus salivarus subsp. thermophilus strain. Reference strain Lactobacillus delbr
ueckii subspecies bulgaricus and Stre
ptococcus salivarus subspecies the
The monoculture thus obtained from rmophilus is 1:
Produce a mixed starter for Bulgarian yogurt using a ratio of 1 to 1:15.

Example 2 100 liters of restored dry whey (6% dry matter) and C
H ₃ COONa (10%), KH ₂ PO ₄ (4%), N
a ₂ HPO ₄ (4%), MgSO ₄ (0.2%), Mn
2.5 liters of buffer-activation mixture, which is an aqueous solution of SO ₄ (0.04%) and yeast extract (0.2%).
Heat treat at 15 ° C. for 10 minutes. Cool the medium to 45 ° C,
Lactobacillus delbrueckii
Subspecies bulgaricus, NBIMCC registration number 24
80 and Streptococcus saliva
rius subspecies thermophilus, NBIMCC
Inoculate 5 liters of fresh laboratory symbiotic starter consisting of registration number 453. Fermenter pH 5.5-6.0, 43
Incubate at 5 ° C for 4 to 5 hours, and automatically adjust pH and temperature. The concentrated starter obtained is cooled to 4 ° C., then 10% sucrose, 2% sodium glutamate and 0.1%.
Mix with 10 liters of protective medium, a sterile aqueous solution of vitamin C, then lyophilize. In another variation, the protective medium uses polyethylene oxide (PEO) in an amount of 1% aqueous solution of PEO-400 or PEO-1000.

Example 3 100 liters of restored skim milk powder (6% dry matter) and 4%
Lactose, 1% concentrated protein, 10% CH ₃ COON
a, 4% KH ₂ PO ₄ , 4% Na ₂ HPO ₄ , 0.2%
2.5 liters of buffer-activation mixture, an aqueous solution of MgSO ₄ , 0.4% MnSO ₄ and 0.2% yeast extract, was heat treated at 115 ° C. for 10 minutes, cooled to 45 ° C., L
actobacillus delbrueckii subspecies bulgaricus, NBIMCC registration number 248
2 and Streptococcus salivar
Ius subsp. thermophilus, 5 liters of laboratory symbiotic starter consisting of NBIMCC accession number 451 or other symbiotic starter according to the invention.
Cultivation and acquisition of the enriched co-cultivation culture continues as described in Example 2.

Example 4 A 100 liter nutrient medium similar to Example 3 consisting of restored dry whey and 2.5% buffer-activating mixture was used in Example 2.
Treat according to the method described. Streptococc
us salivarius subspecies thermophil
Inoculate us with a 5 liter fresh monoculture of NBIMCC accession number 2491. Cultivation is at 42 ° C temperature and pH
5.7-6.0 continues for 6-7 hours, pH value and temperature are automatically adjusted. The concentrated and chilled monoculture is mixed with 10 liters of protective medium as described in Example 2 and freeze dried.

EXAMPLE 5 Lactobac was added to 100 liters of nutrient medium as in Example 2 or 3 treated according to the procedure described in Example 2.
illus delbrueckii subspecies bulgar
icus, inoculated with a 5 liter fresh monoculture of NBIMCC accession number 2479. The method continues in the same manner as described in Example 4.

Example 6 Streptococcus saliva obtained in Example 5
rius subspecies thermophilus, NBIMCC
Registration number 2484 and Streptoco obtained in Example 4
ccus salivarus subspecies thermop
Ilus, NBIMCC Accession No. 2491 lyophilized concentrated culture was mixed in the range of 1: 1 to 1: 5 ratio, and DVS-
Used as a starter. In a similar example to the above example, other strains of both species are used according to the invention to obtain a frozen monoculture or mixed starter in lyophilized form containing a liquid multi-species strain or a symbiotic starter.

The starter thus obtained comprises fresh or reconstituted powdered milk with different fat content from milk such as sheep, cattle, buffalo to the first Bulgarian yogurt and products based on non-dairy substrates such as soybeans and oats. A wide range of lactic acid products can be produced. DVS-starters are used to obtain bulk starters and directly in the production of Bulgarian yogurt or lactic acid products.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification number Internal reference number FI Technical indication C12R 1:46) (72) Inventor Bella Ivanova Andreba Sofia, Bulgaria, App. 33, "Zelkovna" Strasse, No. 75 Buoy (72) Inventor Gepulgii Rieb Georgiev Sofia, Bulgaria, "Opartochenska" Strasebul 37 / Zee (72) Inventor Stefka Zdrabkova Condareba Sofia, Ap 35, Zet. Kei. "Lazadonica", Bull 87,1 (72) Inventor Nevenka Nikolova Loumian Sofia, Bulgaria, App 84, Zet. Kei. "Liulin", Bull 608, 4 (72) Inventor Lumiana Nicolova Blancova Sofia, Bulgaria, App 66, Zet. Kei. "Muldost 2", Bull 204, Bee

Claims (8)

[Claims] 1. Lactobacillus delb
rueckii subspecies bulgaricus group ━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━ NBIMCC registration number NBIMCC registration number ────── ───────────────────────────── LBB B m1 2471 LBB B 31 31 481 LBB B m2 2472 LBB B 37 286 LBB B m 14 2473 LBB B 41 2482 LBB B 5 273 LBB B 120 2483 LBB B 14 2478 LBB B 122 2484 LBB B 24 24 2479 LBB B 129 2485 B 28 B ━ 24 B 242 B 24 B 242 B 24 B 242 B 24 B 242 B 24 B 242 B 24 B 242 B 24 B 242 B 28 B 242 B 24 B 242 B BB BB BB LB 2 ━━━━━━━━━━━━━━━━━━━━━━━━━━━ At least one strain and / or St
reptococcus salivarus subspecies th
ermophilus group ━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━ NBIMCC registration number NBIMCC registration number ────────── ────────────────────────── LBB T3 2487 LBB T18 452 LBB T8 2488 LBB T22 453 LBB T10 2489 LBB T28 2492 LBB 93 B24 T12 450 LBB T14 451 LBB T38 2494 LBB T16 2490 LBB Tm5 2474 LBB T17 2491 LBB TR6 2475 ━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━ Characterized by containing at least one strain selected from the following as a monoculture, mixed culture or co-culture , Bulgaria yogurt and lactic acid products for the starter. 2. The method for producing a monoculture or a mixed starter according to claim 1, wherein the following steps are carried out: a monoculture derived from the strain according to claim 1, which shows a true culture isolated from Bulgarian yogurt. Are cultivated in liquid nutrient medium and then in solid nutrient medium, selected on the basis of morphological properties of the colonies, inoculate sterile skim milk with the same, selected on the basis of technical parameters, and thus selected The resulting monoculture was inoculated into liquid nutrient medium [1] consisting of skim milk hydrolyzed with casein and lactose, meat-peptone broth, water and a biologically active additive selected from lactic acid bacteria species, and then at 42 ° C. pH 4.6 ± 0.1
The method for producing the above-mentioned monoculture or mixed starter, which comprises culturing at 1. 3. A strain Lactobacillus de with normal morphology and a ratio in the range of 1: 1 to 1: 5.
lbrueckii subspecies bulgaricus and S
The method for producing a symbiotic starter according to claim 1, wherein the combination of Treptococcus salivarus subsp. thermophilus is selected by multiple re-inoculation with nutrient medium for at least 4 months. 4. The method comprises restoring dried whey and CH.₃CO
ONa, KH₂PO _Four, Na₂HPO_Four, MgSO_Four,
MnSO_FourAnd yeast extract representing buffer-activated mixture
Of each culture of nutrient medium (b) consisting of
Incubate in a fermenter at pH 5.5-6.0 for 4-5 hours at 45 ° C.
Then cool to 4-6 ° C. and then concentrate the resulting culture.
Loin, sodium glutamate and vitamin C, or
Or PEO-400 or PEO1000 protection
Mix with medium and freeze at -65 to -80 ° C or -196 ° C
The method according to claim 1, which is to bind or freeze-dry.
Direct application (DVS) concentrated mixing or symbiotic starters,
Alternatively, a method for producing a monoculture. 5. The method comprises reconstituted skim milk powder and lactose and protein concentrate obtained by ultrafiltration, CH ₃ COON.
a, KH ₂ PO ₄ , Na ₂ HPO ₄ , MgSO ₄ , Mn
Each culture of nutrient medium (c) consisting of a buffer-activation mixture representing an aqueous solution of SO ₄ and yeast extract was inoculated with 4
Cultivated in a fermenter at pH 5.5-6.0 at 3-45 ° C. for 4-5 hours, cooled to 4-6 ° C., then the concentrated culture obtained was sucrose, sodium glutamate and vitamin C, or PEO. Mixed with a protective medium consisting of -400 or PEO1000, -65 to -80 ° C or -196
2. Freezing at ℃ or freeze-drying.
A process for the production of concentrated mixed or symbiotic starters or monocultures as directly applied as described. 6. Skim milk and meat extract (preferably 1-2) obtained by pre-hydrolyzing casein and lactose for inoculation.
%), Peptone (preferably 0.5-1%) and yeast extract (preferably 0.1-0.5%), a nutrient activating medium comprising 5-10% dry matter. The method for producing a monoculture or a mixed starter according to claim 2, which is carried out in (a). 7. The culture comprises restored dried whey and CH ₃ C.
OONa (preferably 5 to 10%), KH ₂ PO ₄ (preferably 1 to 4%), Na ₂ HPO ₄ (preferably 1 to ₄ )
4%), MgSO ₄ (preferably 0.1-0.5%),
Used for inoculation of a nutrient medium (b) consisting of a buffer-activating mixture representing an aqueous solution of MnSO ₄ (preferably 0.01-0.06%) and yeast extract (preferably 0.1-0.5%). The method for producing a concentrated mixed or symbiotic starter or a monoculture according to claim 4. 8. The culture comprises reconstituted skim milk powder and CH ₃ CO
ONa (preferably 5 to 10%), KH ₂ PO ₄ (preferably 1 to 4%), Na ₂ HPO ₄ (preferably 1 to 4)
%), MgSO ₄ (preferably 0.1-0.5%), M
A buffer-activated mixture representing an aqueous solution of nSO ₄ (preferably 0.01-0.06%) and yeast extract (preferably 0.1-0.5%), and lactose and proteins obtained by ultrafiltration. Concentrate (preferably 0.5-4.
The method for producing a concentrated mixed or symbiotic starter or a monoculture according to claim 5, which is used for inoculation of a nutrient medium (c) consisting of 0%).