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Selection of lactic acid bacteria and yeast for koumiss starter and its impact on quality of koumiss.

Introduction

The purpose of this study was the selection of new strains of lactic acid bacteria and yeast separated from local koumiss for the improvement of koumiss ferment and its impact on quality of koumiss.

The introduction of the leaven koumiss acidophilus was intended to accelerate the formation of the finished product, which meant it had a higher energy of acid than Bulgarian bacteria. In addition, the introduction of Lactobacillus acidophilus in the starter improved nutritional properties of natural koumiss because of it being the only lactic acid bacilli existing in human intestines and because of it having a long positive therapeutic effect on the body (Shigaeva and Ospanova, 1983; Zhilin, 1960; Duysembaev, Seytov, Khasenova et al., 1979)

While studying koumiss prepared using natural starters in different regions of Kazakhstan, acidophilus were selected. These samples of koumiss were recognized as the best due to their taste. Based on production-valuable characteristics of koumiss best strains were selected from the identified cultures of lactic acid bacteria and Lb. acidophilus (74/17, 75/20, 87/75), Lb. bulgaricus (78/30, 79/31), Str. lactis (31/11), Torulopsis sphaerica (105k, 109k, 117k), Torulopsis kefir. var. kumis (114z), Candida mycoderma (125k).

With the purpose of determining the compatibility of testing organisms in joint cultivation, coagulation of milk, acid production, and microscopic examination after 16-18 hour growth were assessed.

A total of 14 combinations of lactic streptococci and bacilli were tested. Strains of Str. lactis monoculture were inferior to acid production strains of Lactobacillus addophilus and Bulgaria, but the joint cultivation with the latter proved the acidity to be much improved. In previous experiments on the combination of acid strains 31/11 (Str'. lactis) 74/17 (Lb. addophilus) and 31/11 (Str. lactis) 78/30 (Lb. bulgaricus) surpassed all other combinations of lactic acid bacteria. It is a well-known fact that yeast plays a key role in preparing koumiss and therefore, the further work concerned the selection of yeast for making sourdough with lactic acid bacteria.

In the laboratory the cultivation of microbial fermentation of koumiss was conducted in an incubator (FHC-12-250) at the Department of Microbiology, Virology, and Immunology (current "Biological Safety") of KazNAU. Five different strains of yeast were tested and they are the following: Torulopsis sphaerica, sht.105k, 109k, 117k Torulopsis kefir. var. kumis, pcs. 114%_ and Candida mycoderma, 125k units isolated from koumiss starters, horse milk in combination with lactic acid bacteria Lb. acidophilus, Lb. bulgaricus, Str. lactis, selected from previous experiments. During cultivation the formation features of starters under consideration were studied. The energy of acid, the growth rate of yeast, and lactic acid bacteria were the points of focus in the experiment.

The master batch koumiss sourdough prepared on sterile milk. Cultures of lactic acid bacteria Lb. acidophilus, Lb. bulgaricus, Str. lactis and yeast Torulopsis sphaerica were grown separately and mixed in 1:1:0.5 ratios just before the fermentation of koumiss on raw horse milk.

The sequence of the experiment is shown in the following combinations, where 20 combinations of koumiss starters were used. Koumiss was prepared with fermentation of the following composition (Table 1):

Natural koumiss produced in the farm "Seysebek" in Talgar district of Almaty region served as the controller. Biochemical parameters were determined after koumiss fermentation. The results of these studies are presented in Table 2.

Results were checked hourly during fermentation based on taste. Koumiss produced with yeast has a sour taste and is free from foreign smell that gives the drink a smooth consistency and a better flavor. Koumiss prepared with these starters organoleptically complies with the standard. Along with this antagonist activity of koumiss with the most optimal combination of pathogens most favorable to make koumiss fermentation starters was also determined. The results of these experiments are shown in Table 3.

The results of the studies in this field show that the finished product obtained through a new method shows high antagonistic activity against the saprophytic, conditionally pathogenic, and pathogenic bacteria (zone of suppression of 12-28 mm).

Thus, among the tested sourdough starters, sourdough No.4 was found to as the best one, which included:

Lb. acidophilus, st.74/17 + Str. lactis, st.31/11 + T. sphaerica, st.117k

The effect of yeast No.4 on the nature and intensity of fermented mare's milk, the quality of koumiss as well as acid and aroma formation in koumiss were studied as well. The results of repeated experiment also confirm the high quality of this product.

Microflora resulting from natural koumiss starter No.4 is shown in Figure 1.

[FIGURE 1 OMITTED]

Having identified the optimal combination of koumiss starter, improving the way of obtaining koumiss, which is to speed up the cooking time of koumiss was the next priority. For this purpose, the experiment, where seven different versions with different mixing rates (in revolutions per minute) under changing temperature conditions on the basis of koumiss was conducted:
1--option   185 rev / min   27-28[degrees]C
2--option   185 rev / min   30-31[degrees]C
3--option   185 rev / min   33-34[degrees]C
4--option   220 rev / min   27-28[degrees]C
5--option   220 rev / min   30-31[degrees]C
6--option   220 rev / min   33-34[degrees]C
7--option   330 rev / min   30-31[degrees]C


In this experiment the best results were obtained while conducting the test in the version No.2. The time necessary for koumiss to be ready decreased from 51 hours to 20 hours or by 2.5 times. The volume of masterbatch starter remained at level of 25-30%, which increased the organoleptic qualities of the product.

DOI: http://dx.doi.org/10.15208/ati.2013.21

References

Duysembaev, K., Seytov, Z., Khasenova, Z., Cherepanov, A., Belokobylenko, V., 1979. Koumiss and Shubat [Koumiss i shubat], in Russian, Alma-Ata

Shigaeva, M., Ospanova, M., 1983. Microflora National Milk Beverages [Mikroflora nacionalnyh kislomolochnyh napitkov], in Russian, Alma-Ata: Nauka

Zhilin, M., 1960 "Antibiotic Properties of Yeast Koumiss," In.: Proceedings of the conference on one horse breeding and dairy koumissodeliyu [Trudy konferencii po molochnomu konevodstvu i koumissodeliyu], in Russian, Moscow, pp.162-74

Zubaira Kozhahmetova, Gulmira Kasenova

Department "Biological safety", Kazakh National Agrarian University, Kazakhstan

e-mail: amirbekkizi_kz@mail.ru

postal address:(KNAU), Abay Ave 8, Almaty 050010, Kazakhstan
TABLE 1. COMBINATIONS OF KOUMISS STARTER

No.           Composition of sourdough

1     Str.lactis, 31/11(10%) + Lb.acidophilus, 74/
      17(10%)+T.sphaerica, 105[kappa] (5%)

2     Str.lactis, 31/11(10%)+ Lb.acidophilus, 74/
      17(10%)+T.sphaerica, 109[kappa] (5%)

3     Str.lactis, 31/11(10%)+ Lb.acidophilus, 74/17(10%)+T.kefir
      var.kumis, 1l43 (5%)

4     Str.lactis, 31/11(10%)+ Lb.acidophilus, 74/
      17(10%)+T.sphaerica, 117[kappa] (5%)

5     Str.lactis, 31/11(10%)+ Lb.acidophilus, 74/
      17(10%)+C.mycoderma, 125[kappa] (5%)

6     Str.lactis, 31/11(10%)+ Lb.acidophilus, 75/
      20(10%)+T.sphaerica, 117[kappa] (5%)

7     Str.lactis, 31/11(10%)+ Lb.acidophilus, 75/
      20(10%)+T.sphaerica, 105[kappa] (5%)

8     Str.lactis, 31/11(10%)+ Lb.acidophilus,.75/
      20(10%)+T.sphaerica, 109[kappa] (5%)

9     Str.lactis, 31/11(10%)+ Lb.acidophilus, 75/20(10%)+T.kefir
      var.kumis, 1143 (5%)

10    Str.lactis, 31/11(10%)+ Lb.acidophilus, 75/
      20(10%)+C.mycoderma, 125[kappa] (5%)

11    Str.lactis, 31/11(10%)+ Lb.acidophilus, 87/
      75(10%)+T.sphaerica, 117[kappa] (5%)

12    Str.lactis, 31/11(10%)+ Lb.acidophilus, 87/
      75(10%)+T.sphaerica, 105[kappa] (5%)

13    Str.lactis, 31/11(10%)+ Lb.acidophilus, 87/
      75(10%)+T.sphaerica, 109[kappa] (5%)

14    Str.lactis, 31/11(10%)+ Lb.acidophilus, 87/75(10%)+T. kefir
      var.kumis, 1143 (5%)

15    Str.lactis, 31/11(10%)+ Lb.acidophilus,.87/
      75(10%)+C.mycoderma, 125[kappa] (5%)

16    Str.lactis, 31/11 (10%)+ Lb.bulgaricus, 78/
      30(10%)+T.sphaerica, 117[kappa] (5%)

17    Str.lactis, 31/11(10%)+ Lb.bulgaricus, 78/
      30(10%)+T.sphaerica, 105[kappa] (5%)

18    Str.lactis, 31/11(10%)+ Lb.bulgaricus, 78/
      30(10%)+T.sphaerica, 109[kappa] (5%)

19    Str.lactis, 31/11(10%)+ Lb.bulqaricus, 78/30(10%)+T. kefir
      var.kumis, 1143 (5%)

20    Str.lactis, 31/11(10%)+ Lb.bulgaricus, 78/
      30(10%)+C.mycoderma, 125[kappa] (5%)

TABLE 2. IMPACT OF DIFFERENT COMBINATIONS OF LACTIC ACID BACTERIA
AND YEASTS ON QUALITY OF KOUMISS

                         Biochemical parameters

Option,   Fermentation   Acidity,     pH    Carbohydrates,   Alcohol,
No           time h      [degrees]T               %             %

1              20           75.0      4.0        4.80          1.20

2              20           90.0      4.0        3.20          2.20

3              20           75.0      4.0        4.31          1.60

4              20          110.0      4.0        3.00          2.60

5              22           60.0      4.5        5.30          0.70

6              22           75.0      4.0        4.25          1.63

7              21           80.0      4.5        4.00          1.90

8              22           85.0      4.0        4.10          1.70

9              20           90.0      4.0        3.20          2.40

10             22           60.0      4.5        5.30          0.80

11             22           60.0      4.5        5.00          1.00

12             22           81.0      4.5        3.50          2.00

13             22           85.0      4.0        3.20          2.40

14             22           67.0      4.5        5.00          1.00

15             22           69.0      4.5        4.40          1.51

16             22           76.0      4.0        4.10          1.70

17             22           62.0      4.5        5.10          0.70

18             22           65.0      4.5        5.00          1.00

19             22           67.0      4.5        5.00          1.00

20             22           60.0      4.6        5.50          0.60

          Biochemical parameters     Organoleptic parameters

Option,   Diacetyl    Acetoin,
No          mg%         mg%

1           0.05        scent      Koumiss non-carbonated, sweet,
                                   fermented, uniform

2           0.12        scent      Koumiss pleasant, fermented,
                                   carbonated, uniform

3            0.1        scent      Koumiss pleasant, sour milk,
                                   fermented, lightly carbonated

4           0.14        0.09       Koumiss pleasant, refreshing,
                                   sparkling, homogeneous,
                                   fermented

5           0.08        scent      Koumiss lightly carbonated,
                                   watery taste

6           0.14        0.08       Koumiss fermented, fermented,
                                   uniform

7           0.08        scent      Koumiss pleasant, cultured
                                   milk, fermented, refreshing

8           0.12        scent      Koumiss pleasant, watery taste

9           0.09        scent      Koumiss pleasant, cultured
                                   milk, fermented, highly
                                   carbonated

10          0.08        scent      Koumiss sour, lightly
                                   carbonated, uniform

11           0.1        scent      Koumiss fermented, lightly
                                   carbonated, sweet, uniform

12          0.05        scent      Koumiss fermented, little
                                   fermented, uniform

13           0.1        scent      Koumiss pleasant, sparkling,
                                   fermented, refreshing, uniform

14          0.08        scent      Koumiss pleasant, lightly
                                   carbonated, fermented

15          0.04        scent      Koumiss fermented, sparkling,
                                   sweet

16          0.09        0.08       Koumiss fermented, sparkling,
                                   sweet

17          0.04        scent      Koumiss fermented, fermented,
                                   lightly carbonated

18          0.09        0.05       Koumiss pleasant, sparkling,
                                   fermented, refreshing, uniform

19          0.05        scent      Koumiss pleasant lightly
                                   carbonated, fermented, uniform

20          0.05        scent      Koumiss fermented, lightly
                                   carbonated, blank taste

TABLE 3. ANTAGONISTIC ACTIVITY OF KOUMISS STARTERS
TO THE SAPROPHYTIC, CONDITIONALLY PATHOGENIC AND
PATHOGENIC BACTERIA

No            Sarcina     Bac.     S. albus,
composition    flava    mycoides    st. 165
sourdough

1               24         12         23
2               24         12         23
3               20         10         20
4               24         14         24
5               16         10         10
6               24         12         23
7               16         12         14
8               14         12         16
9               16         10         20
10              16         12         14
11              17         10         18
12              16         10         20
13              16         10         10
14              13         10         14
15              16         10         10
16              24         12         23
17              16         10         10
18              20         10         20
19              16         10         20
20              14         12         16

No            E. coli,   E.coli,     D.
composition   st. O.5     st. f    septicus
sourdough

1                22        22         22
2                22        22         22
3                20        20         23
4                28        26         24
5                20        20         22
6                22        22         22
7                17        15         17
8                17        15         20
9                20        20         22
10               17        15         17
11               16        16         16
12               20        20         22
13               20        20         22
14               17        15         15
15               20        20         22
16               22        22         22
17               20        20         22
18               20        20         23
19               20        20         22
20               17        15         20

No            Proteus    S.choleraesus,    S.abortus
composition   vulgaris       st.177       equi, st 841
sourdough                                     841

1                12            18              18
2                12            18              18
3                10            18              20
4                12            22              22
5                8             18              18
6                12            18              18
7                10            10              10
8                10            10              10
9                10            16              16
10               10            10              10
11               10            10              10
12               10            16              16
13               8             18              18
14               8             14              14
15               8             18              18
16               12            18              18
17               8             18              18
18               10            18              20
19               10            16              16
20               10            10              10

No            S. abortus       S.          S.
composition      ovis      typhimurium   dublin
sourdough

1                 16           12          12
2                 16           12          12
3                 10           12          10
4                 22           18          12
5                 12           10          11
6                 16           12          12
7                 11           10          10
8                 11           10          10
9                 16           10          10
10                11           10          10
11                14           10          10
12                16           10          10
13                12           10          11
14                15           10          10
15                12           10          11
16                16           12          12
17                12           10          11
18                10           12          10
19                16           10          10
20                11           10          10

No               S.           P.            L.
composition   gallinarum   multocida   monocytogenes
sourdough

1                 14          25            15
2                 14          25            15
3                 14          19            12
4                 20          23            18
5                 10          20            13
6                 14          25            15
7                 10          15            16
8                 10          15            13
9                 10          20            13
10                10          15            16
11                10          17            12
12                10          20            13
13                10          20            13
14                10          12            13
15                10          20            13
16                14          25            15
17                10          20            13
18                14          19            12
19                10          20            13
20                10          15            13

No                 E.          C.       Bac.
composition   rhusipathiae   faetus   Anthracis
sourdough

1                  15          16        12
2                  15          16        12
3                  12          12        10
4                  18          16        16
5                  13          13        10
6                  15          16        12
7                  16          12        12
8                  13          12        12
9                  13          12        10
10                 16          12        12
11                 12          13        10
12                 13          12        10
13                 13          13        10
14                 13          12        10
15                 13          13        10
16                 15          16        12
17                 13          13        10
18                 12          12        10
19                 13          12        10
20                 13          12        12
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Author:Kozhahmetova, Zubaira; Kasenova, Gulmira
Publication:Applied Technologies and Innovations
Date:Oct 1, 2013
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