Research of the adaptation process of LBM. аcidophilus К-3 vegetable juices

All kinds of vegetable juice along with a low calorie contain many vitamins, minerals, pectin and other biologically active substances. They can be attributed to the functional products due to wide range of therapeutic and prophylactic properties. It was shown that the fermentation process of vegetable juice using lactic acid bacteria enhances their antioxidant properties and also reduces the nitrate content in the fermented juice compared with natural [1].

Food industries in developed countries produce juices lactic fermentation of cabbage, carrots, beets, celery, tomatoes, red pepper [2-4].

The fermented carrot juice contains a large quantity of mineral substances, particularly potassium and iron, as well as trace elements and vitamins, and can therefore be used to compensate for deficiency of these substances in food. It is recommended to use the juice for kidney, liver, cardiovascular system.

All vegetable juices can be used for lactic fermentation. Fermented juices are effective therapeutic and prophylactic agents for goiter, as microbiocenosis normalize bowel regulate its motor function. These drinks are recommended to patients with impaired capillary permeability by reducing the venous congestion. The soluble pectin contained in juices and beverages entering the digestive tract swell and promote intestinal absorption of toxins and heavy metal salts.

For producing juices with stable quality and shortening the fermentation process is carried out using pure cultures of lactic acid bacteria. Pure штаммы, cultures are added to fermentable product before or after heat treatment intended to inactivate the enzymes and microflora suppress unwanted materials. In juices introduced various strains of lactic acid bacteria B.bifidumsp, L. plantarum; L. plantarum and Str. faecium; Leuc. mesen-teroides, L. plantarum and L. brevis; L. xylosus; L. casei; L. acidophilus; L. brevis; L. bifudus; Str. faecium and Str. lactis; L. farciminis. The choice of starter cultures of lactic acid is important for a successful course of fermentation [5].

In Poland method of manufacturing fermented vegetable juice with pulp was used. According to method the washed vegetables (carrots, spinach), whole or sliced pour boiling water (1: 1.5), blanch a few minutes up to 95 ° C to remove tissue, partial sterilization and mitigation. Water was added and cooled. For fermenting mix probiotic bacteria (L.plantarum, L.brevis, Leuc. mesenteroides) are used to pH 3,4-3,7. Fermentation process was interrupted by heating the suspension to 90 ° C. then sugar and salt were added, heated to 70-80 ° C for removing air, bottling and pasteurizing at 85° C during 25 minutes.

In Germany for fermented vegetable juice raw juice is heated to 32-36 C and mixed with dosed amounts of sorbitol and lactic acid bacteria and fermented by L. Casei 18 h under anaerobic conditions. Then fermented juice heated to 100 ° C was bottled. Use of soft mode pasteurization selected lactic ferments and enrichment juice sorbitol allow to cook perfect drink for diabetics [6].

Noteworthy is a method of producing an improved vegetable juice flavor in Japan. It eliminates unpleasant "heat" flavor in tomato and other vegetable juices, which they acquire during heat treatment. This is achieved by adding to the juice of lactic acid bacteria and fermentation L.brevis it at 15-25 ° C for 2-3 days, followed by cold sterilization. The finished product has a pleasant taste.

Unfortunately, in Kazakhstan fermented with lactic acid bacteria vegetable juice industry is almost not made. Therefore, improvement of the technology of vegetable juices lactic fermentation with the possible actions canneries is one of the priority areas of research.

Analysis of global trends in food shows that promising are these types of beverages that not only quench your thirst, but also contain a complex of biologically active substances. It is proved that a valuable raw material for the production of such products is whey, which has a unique chemical composition, medical and dietary properties. Bioavailability whey contains the protein nitrogenous compounds, carbohydrates, lipids, mineral salts, vitamins, organic acids, enzymes, immune bodies, trace elements. The functional activity of s whey and the products based on it are effective in the treatment and prevention of diseases such as dysentery, colitis, gastritis with low acidity, varicose veins, headaches, bronchitis, etc .

The influence of the individual components of whey on consumer properties and nutritional value of drinks are researched in many papers of our and foreign researchers. A major component of the whey solids is lactose. Lactose slowly absorbed in the body and reaching of the colon stimulates the vital functions of bacteria, lactic acid [7]. Important in the composition of milk - whey protein have a nitrogen compound content reaches 1%. Whey proteins (albumin and globulins) contain an optimal of sulfur and other essential amino acids. In terms of nutritional physiology it closes to the amino acid scale "ideal" protein that determines their use by the human body for the structural metabolism, mainly proteins of liver regeneration, the formation of hemoglobin and blood plasma [8].

Table 1 - Contents of essential amino acids and whey proteins in the "ideal" protein g per 100 g protein

Amino acid	Whey proteins	"Ideal" protein
Isoleucine	6,2	4
Leucine	12,3	7
Lysine	9,1	5,5
Methionine	2,3 3,4 к	3,5
Cystine	4,4 3,8	6
Phenylalanine	5,2 J          1	4
Tyrosine	2,2	1
Threonine	5,7	5

In addition to the rich chemical composition of whey-based drinks have the following advantages: cheap, a readily available at any time of the year as raw materials; simple processes and their instrumentation solution; reduces the load on the wastewater treatment system of dairy industry enterprises, improving the environment of milk production. Mass producing whey season (summer) coincides with the maximum consumption of beverages, which creates favorable conditions for their marketing [9]. In Germany the popular drinks are beverages containing 80-90% whey and 1020% strawberry and peach juice or grapefruit 7-20%. In the beverage «Frusighurt» in addition to whey contains 10% apple juice; a beverage enriched with vitamins

«Multivitamin» except whey added fruit juices ten items; beverage «Sanoghurt» is a mixture of whey and the addition of fruit juice andwhey proteins [10]. At the same time, the specific flavor and aroma whey acceptable only for a limited range of consumers, this hinders its widespread use. This can be fragrances, herbal supplements and other substances that have a strong pleasant aroma, masking tone of whey or flavor blends well with them and enables consumer products must be quality.

Currently, in our country and abroad developed considerable variety of beverage, depending on the kind which use a natural whey (fresh acidified fermented), as well as products thereof (clarified whey, protein concentrate in liquid or dry form). One of the versions of the classification of beverages are used of the main raw material - whey, is shown in the figure 1.

Beverages based on whey

1--------------------	1  —~	1      ~	1  —~	-----I-	1
Drinks from	Drinks from a	Drinks from	Drinks made	Drinks	Concentrated
whey	natural whey	the	of natural	from the	whey
drinks from the	using	natural	whey	clarified	beverages
natural	biotechnology	whey	removing one	whey	with added
whey without removing components (clarification)		adding taste and flavors	part of whey	using biotechno logy	flavoring and aromatic additives

I .. I ..     —Г

Concentrated whey concentrate with the release of proteins and spices and flavors and additives

Concentrated whey concentrate with the release of proteins and biotechnological processing

Concentrated whey concentrate with the release of proteins and biotechnologic al processing

Figure 1- Classification of beverages based on whey.

Concentrated	Dry whey
whey	concentrate
concentrate	with special
(base and other	treatment and
beverages.	the addition
Products	of other
including alcohol)	juices

Dry whey concentrates tablet. enriched whey protein

In Switzerland produce beverage "Fresh" containing 50% whey, sugar and natural flavors of citrus. Sterilized at 110 ° C and aseptically packaged beverage can be stored for 6 months without refrigeration [8].

The development of new types of whey drinks combining beneficial properties with a pleasant taste, flavor and attractive appearance, containing the minimum amount of additives and adapted to the existing conditions of production is an important scientific and technical problem. In addition, the technology of drinks from whole whey is quite simple. To improve the taste and improve their nutritional and biological value, biological treatment can be by contributing fruit and vegetable.

Objects and Methods of Research

25 strains of lactic bacteria were isolated and identified from different kinds of national sour-milk products made by crude and industrial methods in the South Kazakhstan region such as koumiss, katyk, kaimac, ayran, kurt, sozbe, irimshik, kymyran. Among them Lactobacillus acidophilus K-3 was used for research of the adaptation process to vegetable juices. Bifidobacteriumbifidum sp.strains belongs to the preparation "dry Bifidumbakterin ".

Object of study used plant material grown and widespread in our country - Carrot. We used the carrot kind of Karotel, Nantes.

Analysis of raw material components juice was conducted in accordance with the following conventional methods and their modifications: used for the extraction of juice extractor type laboratory juicer with a hole diameter of 0.5 mm sieve. Determination of dry matter content by drying to constant weight; active acidity measured electrometrically for pH pH meter 121 for measuring range from 4 to 9 units. pH measurement with an accuracy of 0.05 units, pH; determination of titratable acidity potentiometrically; determination of reducing substances iodometric method; determination of nitrite by the Griess method; determining the amino acid composition by chromatography on a column; determining the amount of organic acids by ionexchange chromatography [11, 12].

Results and Discussion

For adapt to the carrot strains Lactobacillus acidophilus K-3 was used. The characteristics of strains comply with the requirements of the starter culture except for the ability to ferment carrot juice.

Rules for the production of fermented vegetable juices installed for pH and acidity that should be followed in this technology: the duration of the fermentation of 10-20 hours, acidity in terms of malic acid should be at least 0.5%, pH less than 4.4.

To determine the ability of Lactobacillus acidophilus K-3 to ferment carrot juice regulated values of Lactobacillus acidophilus K-3 ferments inoculated daily at 5, 10, 15%. Fermentation was carried out at 37 0C.

The experiments showed that none of the proposed doses leaven Lactobacillus acidophilus K-3 can be used for the fermentation of carrot juice. Regulated pH and acidity is not reached after 50 hours of fermentation even at a dose of 15% of inoculum.

Regardless of inoculum juice acquired by the 20th hour of fermentation sour vegetable smell and taste, and to the 50th hour became unpleasantly sour and even in some cases with a slightly bitter taste.

For research the influence of leaven Lactobacillus acidophilus K-3 on the main characteristics of the fermented juice one-day and two-day leaven are used in the amount of 10% (Table 2). In juices with one-day leaven Lactobacillus acidophilus K-3 rapidly accumulating acid, faster and more completely consumed by reducing agents, so further research was selected one-day starter.

For study the effect of solids concentration on the growth of Lactobacillus acidophilus K-3 carrot juice used with a dry substance of 6% and 8%.

During the fermentation the significant differences in pH and acidity in juices with different solids was not observed, but the highest level of acidity reaches in juice having a dry substance content of 8%.

Consumption of reducing agents per 50 hours of fermentation carrot juice goes to dry substance of 8% in about 2 times more efficient in comparison with other juices used in the experiment, however further fermentation of vegetable juice used carrot juice with a dry substance of 8%.

Table 2 - The dependence of the main indicators of fermented carrot juice by culture Lactobacillus acidophilus K-3

Time. h	Age ferment	pH of the juice		Acidity% juicewith		Reducing agent%
		6% DS	8% DS	6% DS	8% DS	6% DS	8% DS
Feedjuice		6,10	6,02	0,05	0,06	2,37	2,95
0	1-Day	5,86	5,81	0,09	0,09	2,37	2,95
	2-Day	5,54	5,55	0,09	0,08	2,37	2,95
5	1-Day	5,31	5,25	0,10	0,11	2,14	2,35
	2-Day	5,48	5,50	0,09	0,09	2,27	2,55
20	1-Day	4,62	4,58	0,20	0,21	1,93	1,95
	2-Day	5,20	5,19	0,12	0,13	2,15	2,19
25	1-Day	4,60	4,49	0,23	0,25	1,81	1,90
	2-Day	4,53	4,55	0,15	0,19	2,05	2,08
40	1-Day	4,43	4,41	0,28	0,29	1,73	1,87
	2-Day	4,50	4,48	0,19	0,21	2,00	2,03
50	1-Day	4,40	4,39	0,31	0,31	1,68	1,80
	2-Day	4,41	4,38	0,25	0,27	1,98	2,00

We carry out all experiences 3 - 5 th repeating for reception reliable result. The Difference in experience forms + 0,01.

To increase the efficiency of fermentation in juices made by various growth factors in the different amounts of Lactobacillus acidophilus K-3 with inoculum dose 5% of Lactobacillus acidophilus K-3. Selection of the optimal growth factor analysis conducted on the basis of pH and acidity in fermented juice.

In 20 hours of fermentation carrot juice acidity were highest value achieved by adding to the juice: yeast autolysate, whey and riboflavin. However, with the addition of yeast autolysate juice acquired an unpleasant taste and smell of yeast and unattractive color. Therefore, the optimum additive in carrot juice was selected whey in an amount of 20%. Dependence on growth factors Lactobacillus acidophilus K-3 acid-forming ability during fermentation of carrot juice is shown in Table 3.

The main characteristic of juice fermented with the addition may be regulated. In carrot juice with added whey (20%) after 15 hours of fermentation, the acidity reached in terms of 0.51% malic acid and pH 3.85. Juice has light orange colour, pleasant sour taste and light sour vegetable smell.

Table 3 - Effect of growth factors on the acid-forming ability of bifidobacteria in the fermentation of carrot juice

Growth factors	The Addition amount,%	The duration of fermentation, h
		0		5		20
		pH	acidity, %	pH	acidity, %	pH	acidity, %
1. Yeast	10,0	4,9 + 0,1	0,17	3,50 + 0,1	0,48	3,20 + 0,1	0,58
autolysate	20,0	4,6 + 0,1	0,17	3,70 + 0,1	0,53	3,70 + 0,1	0,66
2. Whey	10,5	5,8 + 0,1	0,17	4,50 + 0,1	0,29	4,40 + 0,1	0,33
	20,0	4,9 + 0,1	0,20	3,85 + 0,1	0,51	3,80 + 0,1	0,56
3. Riboflavin	10,5	5,2 + 0,1	0,10	4,40 + 0,1	0,31	3,90 + 0,1	0,40
	20,0	5,3 + 0,1	0,11	4,10 + 0,1	0,33	3,50 + 0,1	0,41

Carrot juices was inoculated with starter cultures composed of B. bifidum sp. and Lbm. аcidophilusК-3 in the ratio 3: 1, 1: 1 and 1: 3. Juices fermented for 24 hours with daily ferments in an amount of 5%. It was shown that the strains in a ratio of 1: 1 the residual content of reducing substances is smallest, and the pH and acidity reaches values in carrot juice to 4.45 and 0.38% by weight of malic acid (Table 4).

At a ratio of 1: 3 juice has a lower pH and higher acidity but microscopic studies have shown that in the case of B. bifidum sp. poorly developed. At a ratio of 3: 1 juice is fermented very slowly and the pH level after 24 hours is low.

Table 4 - Influence of the correlation of the cultures in leaven on features fermented carrot juice

The ratio of cultures	рН	Acidity,%	Reducing agent,%
Juice	6,10 + 0,01	0,05 + 0,01	2,86 + 0,01
3:1	4,55 + 0,01	0,23 + 0,01	2,12 + 0,01
1:1	4,45 + 0,01	0,38 + 0,01	1,97 + 0,01
1:3	4,20 + 0,01	0,52 + 0,01	2,02 + 0,01

Levels were determined in carrot juice fermented with whey additives. The results are shown in Table 5.

In carrot juice fermented with added whey content vitamin C increased in 4 times the carotene content is increased 1.2 times.

During the fermentation the total acidity in carrot juice increased in 9 times, the total organic acid content - 49.7%.

During the fermentation by the association of B. bifidum sp. and Lbm. Аcidophilus К-3 the level of lactic acid is increased in carrot juice to 36.7%, acetic acid is increased 74.4%. The level of citric acid is increased in carrot juice to 17.2%.

Table 5- Comparative characteristics of the vitamin content in carrot juice

Carrot juice	Vitamin C content, mg / 100 ml	β-carotene content in mg / 100ml	The vitamin B1, mg%	The vitamin B 2 mg%
Before fermentation	0,45 + 0,01	8,60 + 0,01	0,050 + 0,005	0,070 + 0,005
Fermented without additives	0,59 + 0,01	9,10 + 0,01	0,060 + 0,005	0,073 + 0,005
supplemented with the fermented milk whey	1,80 + 0,01	10,30 + 0,01	0,065 + 0,005	0,075 + 0,005

All experiences are carried out 3 - 5 th repeating for reception reliable result. Qualitative analysis of the amino acid composition of fermented carrot juice showed that the samples contained seven essential amino acids. It is proved that during the culturing of bacteria B. bifidum sp. and Lbm.

acidophilus К-3 in the carrot juice total sugar level is reduced from 35.0% to 24.7%.

B. bifidum sp. and Lbm. acidophilus К-3 are known as microorganisms with high antagonistic ability to many pathogenic microorganism. Therefore B. bifidum sp. and Lbm. acidophilus К-3 and their metabolic products in vegetable juices can inhibit the existence of pathogenic microflora.

Conclusion

Vegetable juice is good subject for lactic fermentation, comprising organic acids (malic, citric, tartaric, lactic, etc.). Carrot contains sufficient volume of natural juice. Carrot juice is rich in vitamins C, B1, B2, PP and carotene, potassium, phosphorus, iron and other minerals. Therefore development of technology fermented vegetable juices is one of the priority areas of research.

The highest level of acidity was reached in carrot juice with contents 8% of dry substance. Comparative characteristics of juice supplemented with milk whey and fermented juice without additives showed increasing in three tame Vitamin C. The optimum additive in carrot juice was chosen whey in amount 20%.

In our research the strains Lbm. аcidophilus К-3 and Bifidobacterium bifidum sp in a ratio of 1: 1 were chosen. B. bifidum sp. and Lbm. аcidophilus К-3 and their metabolic products in vegetable juices can inhibit the existence of pathogenic microflora. The residual content of reducing substances are smallest, pH in fermented carrot juice reached to 4.45 and acidity reached to 0.38% by count into malic acid.