CN103131654B - Application of Lactobacillus casei KL1 for producing bile salt hydrolase in functional sour milk - Google Patents
Application of Lactobacillus casei KL1 for producing bile salt hydrolase in functional sour milk Download PDFInfo
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Abstract
The invention relates to an application of Lactobacillus casei KL1 for producing bile salt hydrolase in functional sour milk, and is suitable for assimilate cholesterol functional sour milk and microecology preparation production in fermented dairy product. The application provided by the invention employs a straight casting mode to add Lactobacillus casei KL1 (CGMCC No.1809) dry powder leavening agent used for effectively reducing cholesterol and a common sour milk dry powder leavening agent to prepare the functional sour milk with excellent quality. On a base that a common sour milk fermentation technology is not changed, the characteristic of KL1 bacterial strain for producing bile salt hydrolase can be employed to prepare the sour milk for reducing serum cholesterol, the sour milk whey precipitation problem can be solved, the post-acidifying degree of the sour milk can be reduced, the sour milk viscosity is enhanced, the product is stored for 21 days at the temperature of 4 DEG C, the live bacteria quantity of Lactobacillus casei KL1 can reach more than 106CFU/g, and the Lactobacillus casei KL1 has health care function. The prepared functional sour milk has simple fermentation technology, short fermentation period and low cost, and is adapted to large scale industrial production.
Description
Technical field
The present invention relates to a kind of product bile salt hydrolase (Bile Salt Hydrolase, BSH) lactobacterium casei (Lactobacillus casei) KL1 dry powder leaven and common sour milk dry powder leaven mixed fermentation functional yoghourt production technology, be applicable to the functional yoghourt of decreasing cholesterol and the production of probiotics in cultured milk prod.
Background technology
Probiotic bacterium (Probiotics) refers to that a class is improved host's privileged site microecological balance by enteron aisle colonized action and has the microorganism of other beneficial physiological functions some concurrently.They are by improving the enteric microorganism eubiosis, control intestinal tract infections, reduce serum cholesterol level, improve lactose intolerance people to the utilization of lactose, and can stimulate specificity or nonspecific immune reaction, have the effect of immunostimulant.The viable bacteria of probiotic composition should metabolic stability, and activity is comparatively strong, by large number of viable after digestive tube, plays probiotic action after entering enteron aisle.The current excellent species for the production of probiotic agent has the Lactococcus lactis subsp.lactis etc. in genus bifidobacterium, lactobacillus, lactococcus.
The present invention's lactobacterium casei used KL1 bacterial strain is separation and purification a kind of probiotic lactobacillus out from Kefir grain, the bile salt hydrolase of a large amount of high vigor can be produced in growth metabolism process, prove there is the effect reducing body serum cholesterol content through external Oxford cup test and experimentation on animals.It reduces cholesterol mechanism is bile salt hydrolase (the Bile Salt Hydrolase produced because of the metabolism of lactobacterium casei KL1 bacterial strain, BSH), BSH be genus bifidobacterium in enteron aisle, the meta-bolites that produces in process of growth of multiple lactic-acid-bacterium in lactobacillus, lactococcus.Experimental results demonstrate, bile salt hydrolase will can decompose in conjunction with cholate in hepato-enteric circulation, produces amino acid and the lower free cholate of solubleness.The latter can be combined with cholesterol and forms sediment composite and excrete, thus reduces the content of serum total cholesterol.
Just confirm that the Yoghourt of picked-up containing certain Bacterium lacticum or bifidobacterium fermentation has the effect reducing serum cholesterol content as far back as people in 1963.Up to now, the research of Chinese scholars to the milk-acid bacteria decreasing cholesterol mechanism of action remains in different viewpoints, mainly concentrates on following 3 points: the direct assimilation cholesterol of (1) milk-acid bacteria somatic cells; (2) the bile salt hydrolase activity of milk-acid bacteria makes combined cholate be degraded to free state cholate, and the latter's solubleness declines and cholesterol generation coprecipitation effect; (3) other are theoretical, as assimilation and co-precipitation combined action.
The domestic research about lactobacterium casei is mainly on the health-care effecies such as reducing blood-fat, hypertension, treatment anaphylactic disease, as " lactobacterium casei Bd-II bacterial strain and the application at reduction lipid aspects thereof " of Shanghai Bright Dairy & Food Co., Ltd's application, number of patent application is: CN03128995.9; " lactobacterium casei LC2W bacterial strain and the application in hypertension thereof ", number of patent application is: CN03129450.2; " purposes of new microbial strain lactobacillus paracasei GM-080 and treatment allergy-related disorder thereof " of Jingyue Biological Science and Technology Co., Ltd.'s application, number of patent application is: CN200410038566.X.
Although the research abroad about lactobacterium casei reduction serum cholesterol effect also has a patent report: Lactobacillus casei bd-II stain and used to reduce blood cholesterol (20060127380), but the content that external relevant lactobacterium casei produces bile salt hydrolase at present there is no patent report.Although have the patent report of " functional yoghourt and preparation method thereof (200310112715.8) containing lactobacillus reuteri " domestic, the domestic production about utilizing the lactobacterium casei of decreasing cholesterol and high yield bile salt hydrolase to be applied to functional yoghourt there is not yet pertinent literature report and patent report.
Summary of the invention
The object of this invention is to provide a kind of product bile salt hydrolase lactobacterium casei KL1 dry powder leaven and common sour milk dry powder leaven mixed fermentation functional yoghourt production technology.
It is obtained by lactobacterium casei (Lactobacillus casei) the KL1 dry powder leaven and common sour milk dry powder leaven with efficient reduction cholesterol and high yield bile salt hydrolase that the present invention prepares functional yoghourt.
A kind of method utilizing the lactobacterium casei KL1 producing bile salt hydrolase to apply in functional yoghourt, the technique adopting method of delivering directly to add Yoghourt dry powder leaven is: lactobacterium casei KL1 bacterial strain dry powder leaven inoculum size is 0.5 ‰ (W/V, mass volume ratio concentration), the inoculative proportion of lactobacterium casei KL1 bacterial strain dry powder leaven and common sour milk dry powder leaven is 1: 2 (W/W, massfraction).
In described method, lactobacterium casei (Lactobacillus casei) KL1 separation screening from Jilin, the Kefir grain (also known as Kefir granule) of Huhehaote City's average family, and being preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on September 11st, 2006, its preserving number is: CGMCC No.1809 (bacterial strain KL1).On the MRS selective medium containing calcium carbonate and tennecetin, the bacterium colony size 2 ~ 3mm of lactobacterium casei (Lactobacillus casei) KL1 bacterial strain, surface irregularity is as flakes, edge is irregular, matt, flat, canescence, translucent, bacterium colony has comparatively high viscosity; Individual morphology is shaft-like, and varying length, short catenation, thalline is spherical in shape sometimes, G
+sporeless bacterium.
In described method, common sour milk dry powder leaven comes from the Jiajiale direct putting type ferment agent for sour milk produced by Harbin Meihua Biologic Technology Co., Ltd..
In described method, functional yoghourt prepared by the dry powder leaven of adding lactobacterium casei KL1 in common sour milk is produced, has good reduction serum cholesterol effect.Do not changing on common sour milk zymotechnique basis, both KL1 bacterial strain can have been utilized to produce the Yoghourt of bile salt hydrolase characteristic preparation reduction serum cholesterol, solve again soured milk whey and separate out problem, also reduce post-acidification of yoghurt degree, improve Yoghourt viscosity, the number of viable that product stores lactobacterium casei KL1 after 21 days under 4 DEG C of conditions can reach 10
6more than CFU/g, still has nourishing function.
A kind of method utilizing product bile salt hydrolase lactobacterium casei KL1 to apply in functional yoghourt obtained above and products thereof belongs to scope with special preparing strain.
Accompanying drawing explanation
Application lactobacterium casei (Lactobacillus casei) KL1 and common sour milk dry powder leaven are produced functional yoghourt process flow sheet and are described as follows:
(1) raw dairy: the raw dairy making Yoghourt requires that bacterium number is generally lower than 10
4individual/mL, not containing microbiotic and Herb of Common violet, should not select trouble Bovine Mastitis Milk.(2) purify: with the white corpuscle in whizzer removing cow's milk and other macroscopic impurity.(3) stdn: the main component index of raw dairy should meet National standards of food sanitation GB5408-85.Its total dry matter should be not less than 11.5%, lipid content is adjusted to 4 kinds substantially according to product difference: 3.2%, 2.5%, 1.0% and < 0.1%, by remove cream or add 1% ~ 3% skim-milk or rare cream to adjust total dry matter or lipid content.(4) preheating, batching and filtration: raw dairy is heated to about 60 DEG C, add the sucrose of 6% (W/V, mass volume ratio concentration), filtering and impurity removing after dissolving.(5) homogeneous: under 8 ~ 10MPa pressure, homogenization treatment is carried out to the raw dairy of preheating in clarifixator.Object makes curdling admittedly even, and quality is finer and smoother, level and smooth, and Oil globule also can be made to diminish prevent fat floating.(6) sterilization: raw dairy is heated to 90 DEG C, insulation 5 ~ 10min.(7) cool: the raw dairy after sterilization is cooled to rapidly 43 DEG C, to be seeded.(8) inoculate: method interpolation lactobacterium casei KL1 bacterial strain dry powder leaven is delivered directly in employing and common sour milk dry powder leaven technique is: lactobacterium casei KL1 bacterial strain dry powder leaven inoculum size is 0.5 ‰ (W/V, mass volume ratio concentration), the inoculative proportion of lactobacterium casei KL1 bacterial strain dry powder leaven and common sour milk dry powder leaven is 1: 2 (W/W, massfraction).(9) packing: for avoiding miscellaneous bacteria to invade, the operation being sub-packed in plastic small container should be carried out fast in sterilisable chamber.(10) heat-preservation fermentation: plastic small container is placed in proving room and keeps leavening temperature 43 DEG C.When the acidity of fermented-milk reaches 38.75 ° of T, when curdling solidity shape is good, i.e. fermenting-ripening.Fermentation time is 3h.(11) cool: the container containing Yoghourt is taken out from proving room, is cooled to rapidly less than 10 DEG C with cold wind.(12) refrigeration and after-ripening: through the Yoghourt of cooling process, be stored in 0 ~ 5 DEG C of refrigerating chamber and preserve, until drink.Object prevents from Yoghourt from continuing fermentation and acid cause pH too low and affect mouthfeel, and prevent living contaminants from breeding.After-ripening under refrigerated condition is conducive to the formation of yoghourt-flavored material, the functional finished product Yoghourt that final acquisition local flavor is soft, strong.
The raw material sources that the present invention makes functional yoghourt are convenient, and zymotechnique is simple, and fermentation period is short, simple to operate, low for equipment requirements, and cost is lower, is suitable for suitability for industrialized production.
Embodiment
The invention will be further described and do not limit the scope of the invention for following embodiment.
Experimental technique in following embodiment, if no special instructions, is ordinary method.
Percentage composition in following embodiment, if no special instructions, is mass volume ratio concentration (W/V).
The lactobacterium casei KL1 that embodiment 1, utilization produce bile salt hydrolase applies in common sour milk
1, lactobacterium casei KL1 dry powder leaven addition is on the impact of fermented yogurt acidity
Under common sour milk dry powder leaven inoculum size is the prerequisite of 1.0 ‰, the acidity that lactobacterium casei KL1 dry powder leaven addition is 0.5 ‰, 1.0 ‰, 1.5 ‰ fermented yogurts is measured with 0.1mol/L NaOH solution, and with the common sour milk not adding lactobacterium casei KL1 dry powder leaven for control group, unit represents with gill Nie Erdu (° T).Get 10mL sample, use 20mL distilled water diluting, add the phenolphthalein indicator 3 of 0.5%, be titrated to blush with the NaOH solution of demarcating, 30s does not fade for terminal, the results are shown in Table 1.
Acidity calculation formula: acidity (° T)=c × V × 10/0.1
C in formula---NaOH concentration (mol/L) after demarcating
V---consume and demarcate NaOH volume number
Table 1 different dry Lactobacillus paracasei KL1 dry powder leaven addition is on the impact of fermented yogurt acidity
Note:
*represent and compare P < 0.05 with control group
As shown in Table 1, compared with control group, minimum on the impact of Yoghourt acidity in fermenting process when lactobacterium casei KL1 dry powder leaven addition is 0.5 ‰, reduce the Yoghourt acidity in fermenting process; When addition is 1.0 ‰, add the Yoghourt acidity in fermenting process; When addition is 1.5 ‰, the Yoghourt acidity in fermenting process is significantly higher than control group (P < 0.05).Therefore illustrate in fermented yogurt acidity index, the addition of lactobacterium casei KL1 dry powder leaven 0.5 ‰ is better than the addition of 1.0 ‰ and 1.5 ‰.
2, lactobacterium casei KL1 dry powder leaven addition is on the impact of post-acidification of yoghurt
Under common sour milk dry powder leaven inoculum size is the prerequisite of 1.0 ‰, be that 0.5 ‰, 1.0 ‰, 1.5 ‰ storages are to the Yoghourt acidity of 1d, 7d, 14d, 21d with 0.1mol/LNaOH measured in solution lactobacterium casei KL1 dry powder leaven addition, and with the common sour milk not adding lactobacterium casei KL1 dry powder leaven for control group, measuring method is the same, the results are shown in Table 2.
Table 2 different dry Lactobacillus paracasei KL1 dry powder leaven addition is on the impact of post-acidification of yoghurt
Note:
*represent and compare P < 0.05 with control group
As shown in Table 2, compared with control group, when lactobacterium casei KL1 dry powder leaven addition is 1.0 ‰, significantly reduces the impact (P < 0.05) on post-acidification of yoghurt, and decline at most; Addition takes second place when being 0.5 ‰, when addition is 1.5 ‰, adds post-acidification of yoghurt degree.Therefore illustrate in post-acidification of yoghurt index, the addition of lactobacterium casei KL1 dry powder leaven 1.0 ‰ is better than the addition of 0.5 ‰ and 1.5 ‰.
3, lactobacterium casei KL1 dry powder leaven addition is on the impact of Yoghourt viscosity
Under common sour milk dry powder leaven inoculum size is the prerequisite of 1.0 ‰, measuring lactobacterium casei KL1 dry powder leaven addition respectively with viscometer is the Yoghourt viscositys of 0.5 ‰, 1.0 ‰, 1.5 ‰ storages to 1d, 7d, 14d, 21d, not add the common sour milk of lactobacterium casei KL1 dry powder leaven for control group.Get 50mL sample during mensuration, detect parameters: sp3 rotor, rotating speed 30r/min, the results are shown in Table 3.
Table 3 different dry Lactobacillus paracasei KL1 dry powder leaven addition is on the impact of Yoghourt viscosity
Note:
*represent and compare P < 0.05 with control group
As shown in Table 3, lactobacterium casei KL1 dry powder leaven addition significantly improves Yoghourt viscosity (P < 0.05) when being 0.5 ‰, and significantly reduces Yoghourt viscosity (P < 0.05) when addition is 1.0 ‰ and 1.5 ‰.Therefore illustrate in Yoghourt viscosity index, the addition of lactobacterium casei KL1 dry powder leaven 0.5 ‰ is better than the addition of 1.0 ‰ and 1.5 ‰.
4, lactobacterium casei KL1 dry powder leaven addition is on the impact of KL1 bacterial strain viable count in Yoghourt
Under common sour milk dry powder leaven inoculum size is the prerequisite of 1.0 ‰, be that 0.5 ‰, 1.0 ‰, 1.5 ‰ storage to the Yoghourts of 1d, 7d, 14d, 21d carry out plate count by lactobacterium casei KL1 dry powder leaven addition.Get 5g Yoghourt respectively with 45mL stroke-physiological saline solution dilution 10
-1, then get 1mL Yoghourt diluent, with stroke-physiological saline solution dilution 10
-2to 10
-6, get 10 simultaneously
-4to 10
-6dilution bacterium liquid 1mL, adopt modified MRS selective medium (in substratum, to add the bovine bile of 0.2% ~ 0.3%; KL1 bacterial strain has the characteristic of tolerance cholate, and its growth is not suppressed by cholate; And the growth of common sour milk bacterial classification can be suppressed by cholate) with the number of viable of KL1 bacterial strain in pour plate culture method detection Yoghourt, each extent of dilution establishes 3 repetitions, and count after 37 DEG C of cultivation 48h, the results are shown in Table 4.
Table 4 lactobacterium casei KL1 dry powder leaven addition is on the impact of KL1 bacterial strain viable count in Yoghourt
As shown in Table 4, add the Yoghourt of lactobacterium casei KL1 dry powder leaven fermentation, in storage after 21 days, the number of viable of KL1 bacterial strain can reach 10
6more than CFU/g, still has nourishing function.Therefore illustrate in Yoghourt viable count index, when lactobacterium casei KL1 dry powder leaven addition is 0.5 ‰, 1.0 ‰, 1.5 ‰, all can make functional yoghourt.But consider production cost problem, determine that lactobacterium casei KL1 dry powder leaven addition is 0.5 ‰.
5, lactobacterium casei KL1 dry powder leaven addition is on the impact of soured milk whey amount of precipitation
Under common sour milk dry powder leaven inoculum size is the prerequisite of 1.0 ‰, measuring lactobacterium casei KL1 dry powder leaven addition respectively with graduated cylinder is the soured milk whey amount of precipitations of 0.5 ‰, 1.0 ‰, 1.5 ‰ storages to 1d, 7d, 14d, 21d, not add the common sour milk of lactobacterium casei KL1 dry powder leaven for control group, the results are shown in Table 5.
Table 5 different dry Lactobacillus paracasei KL1 addition is on the impact of soured milk whey amount of precipitation
Note:
*represent and compare P < 0.05 with control group
As shown in Table 5, significantly reduce soured milk whey amount of precipitation (P < 0.05) when lactobacterium casei KL1 dry powder leaven addition is 0.5 ‰, when addition is 1.0 ‰, make soured milk whey amount of precipitation decrease; When lactobacterium casei KL1 dry powder leaven addition is 1.5 ‰, significantly increase soured milk whey amount of precipitation (P < 0.05).Therefore illustrate in soured milk whey amount of precipitation index, the addition of lactobacterium casei KL1 dry powder leaven 0.5 ‰ is better than the addition of 1.0 ‰, 1.5 ‰.
In sum, in order to obtain colory functional yoghourt, for lactobacterium casei KL1 dry powder leaven Different adding amount on the impact of Quality of Yoghourt, determine its optimum addition.By analyzing the acidity measuring fermented yogurt, and viscosity, KL1 bacterial strain number of viable, the whey amount of precipitation of storage Yoghourt, determine that functional yoghourt lactobacterium casei KL1 dry powder leaven optimum addition is 0.5 ‰, the inoculative proportion of lactobacterium casei KL1 bacterial strain dry powder leaven and common sour milk dry powder leaven is 1: 2.Finished product functional yoghourt viscosity is higher, and the curdled milk time is shorter, and curdled milk is solid, and minute quantity whey is separated out, and mouthfeel is sticky, fine and smooth and children is sliding, and sweet mild acidity, has strong ester Flavor, and the KL1 bacterial strain viable count refrigerated after 21 days in Yoghourt can reach 10
6more than CFU/g, still with health role.
The experimentation on animals of embodiment 2, functional yoghourt serum cholesterol-lowering
The present invention's production functional yoghourt starter used is lactobacterium casei (Lactobacilluscasei) KL1 dry powder leaven and common sour milk dry powder leaven.Wherein lactobacterium casei KL1 is the functional strain of high yield bile salt hydrolase, and bacterial strain stomach juice-resistant, resistance to biliary salts ability are stronger, there is GIT Conditions Tolerance Ability, intestinal secretion bile salt hydrolase can be arrived smoothly, the degraded of combined cholate is produced amino acid and the lower free state cholate of solubleness.The latter can be combined with cholesterol and forms sediment composite and excrete, thus reduces the content of serum total cholesterol.Utilize big white mouse do functional yoghourt per os gavage test, its method and result as follows:
1, animal grouping and process
40 Wistar rats (tieing up laboratory animal technology company of tonneau China by Beijing to provide) are divided into 4 groups at random, are hyperlipidemia model group, low dose group, middle dosage group, high dose group respectively.All groups of high lipid foods of all feeding (basal feed 78.8%, cholesterol 1%, yolk powder 10%, lard 10%, cholate 0.2%, laboratory animal technology company of dimension tonneau China provides by Beijing), adaptability is raised 1 week.After 1 week, all groups are continued high lipid food of feeding, and by low dose group, middle dosage group, high dose group, by 0.5g/kgd, 1.5g/kgd and 2.5g/kgd dosage gavage functional yoghourt, (KL1 bacterial strain viable count reaches 10 respectively simultaneously
7more than CFU/g), hyperlipidemia model group gavage, with dosage distilled water, is fed 7 weeks so continuously.
2, serum preparation and detection
Test the 1st, 3,5,7 weekends, respectively to the blood sampling of rat orbital venous plexus, after the centrifuge tube room temperature filling whole blood is left standstill 10min, the centrifugal 10min of 3000r/min, Aspirate supernatant is serum sample.Adopt rCO-peroxidase-4-AA and phenol method to measure the content of serum total cholesterol (TC), the results are shown in Table 6; Adopt GPO-peroxidase-4-AA and phenol method to measure the content of serum levels of triglyceride (TG), the results are shown in Table 7.
Table 6 functional yoghourt reduces the experimental result of Serum TC content
Note:
*represent and compare P < 0.01 with control group
From table 6, after experimentation on animals proceeds to the 7th week, serum TC content and the hyperlipidemia model group of high, medium and low dosage group rat contrast, all there is pole significant difference (P < 0.01), and middle dosage group effect is the most obvious, show that functional yoghourt has the effect extremely significantly reducing hypercholesterolemicrats rats serum total cholesterol content.
Table 7 functional yoghourt reduces the experimental result of rat blood serum TG content
Note:
*represent and compare P < 0.01 with control group
From table 7, after experimentation on animals proceeds to the 7th week, serum TG content and the hyperlipidemia model group of high, medium and low dosage group rat contrast, all extremely significantly reduce (P < 0.01), and low dose group effect is the most obvious, show that functional yoghourt has the effect extremely significantly reducing hypercholesterolemicrats rats serum levels of triglyceride content.
In sum, functional yoghourt prepared by the dry powder leaven of adding lactobacterium casei KL1 in common sour milk is produced, there is good reduction serum cholesterol effect, develop serum cholesterol-lowering functional yoghourt with it and probiotics has wide market outlook.
Project 1 belonging to this patent: Beijing's Natural Science Fund In The Light " Kefir grain probiotic bacterium produces the research of active substances and physiological function "
Item number: 5062006
The project beginning and ending time: 2006.01-2008.12
Project leader: Liu Hui
Claims (1)
1. utilize the method for producing bile salt hydrolase lactobacterium casei KL1 bacterial strain and applying in functional yoghourt, it is characterized in that: the preserving number of lactobacterium casei (Lactobacillus casei) KL1 is CGMCC No.1809; The technique adopting method of delivering directly to add Yoghourt dry powder leaven is: lactobacterium casei KL1 bacterial strain dry powder leaven inoculum size is 0.5 ‰, and the inoculative proportion of lactobacterium casei KL1 bacterial strain dry powder leaven and common sour milk dry powder leaven is 1: 2.
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| CN104357359A (en) * | 2014-11-14 | 2015-02-18 | 北京北农汇邦生物科技有限公司 | Production technology of bile salt hydrolase-yielding lactobacillus paracasei dry powder living bacterium preparation |
| CN104774783A (en) * | 2015-02-28 | 2015-07-15 | 北京和美科盛生物技术有限公司 | Lactobacillus casei capable of reducing cholesterol and method for producing probiotic yogurt from same |
| CN104726380B (en) * | 2015-03-30 | 2017-11-07 | 成都市食品药品检验研究院 | One plant of Lactobacillus casei and its application |
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| RU2290435C1 (en) * | 2005-08-05 | 2006-12-27 | Борис Александрович Татаров | Enzyme for production of fermented milk product |
| CN101333505A (en) * | 2007-06-26 | 2008-12-31 | 内蒙古农业大学 | Applications of lactobacillus casei for antioxidation |
| CN101331900A (en) * | 2007-06-28 | 2008-12-31 | 北京农学院 | Four kinds of lactobacillus for generating bilesalt hydrolase and extracellular polysaccharide and functionality yoghourt production technique thereof |
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| RU2290435C1 (en) * | 2005-08-05 | 2006-12-27 | Борис Александрович Татаров | Enzyme for production of fermented milk product |
| CN101333505A (en) * | 2007-06-26 | 2008-12-31 | 内蒙古农业大学 | Applications of lactobacillus casei for antioxidation |
| CN101331900A (en) * | 2007-06-28 | 2008-12-31 | 北京农学院 | Four kinds of lactobacillus for generating bilesalt hydrolase and extracellular polysaccharide and functionality yoghourt production technique thereof |
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