CN104774783A - Lactobacillus casei capable of reducing cholesterol and method for producing probiotic yogurt from same - Google Patents
Lactobacillus casei capable of reducing cholesterol and method for producing probiotic yogurt from same Download PDFInfo
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- CN104774783A CN104774783A CN201510090444.3A CN201510090444A CN104774783A CN 104774783 A CN104774783 A CN 104774783A CN 201510090444 A CN201510090444 A CN 201510090444A CN 104774783 A CN104774783 A CN 104774783A
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Abstract
The invention provides Lactobacillus casei J1 capable of reducing cholesterol. According to a technical scheme in the invention, the Lactobacillus casei J1 is separated and screened from Tibetan kefir (also called as kefir grain) cultured in a common household and is preserved in China General Microbiological Culture Collection Center on September 11, 2006, with an accession number of CGMCC No. 1808. The invention also discloses a method for producing probiotic yogurt by using the Lactobacillus casei J1. On the basis of no change of common yoghourt fermentation process, the yoghourt capable of reducing serum cholesterol can be prepared by utilizing the characteristic that the J1 strain produces bile salt hydrolase, and the problem of precipitation of whey of the yoghourt is overcome. The method for producing the probiotic yogurt has the advantages of simple process, a short fermentation period, low cost and applicability to large-scale industrial production.
Description
Technical field
The present invention relates to a kind of method reducing cholesterol lactobacterium casei (Lactobacillus casei) J1 and production probiotics yogurt thereof, dry powder leaven and the mixed fermentation of common sour milk dry powder leaven generate probiotics yogurt, are applicable to the production of the probiotics yogurt of decreasing cholesterol in cultured milk prod, probiotics tablets, probiotics.
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, it is by improving the enteric microorganism eubiosis, control intestinal tract infections, reduce serum cholesterol level, improve lactose intolerance people to the utilization of lactose, and specificity or nonspecific immune reaction can be stimulated, there is 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 J1 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 J1 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 probiotics yogurt 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 method reducing cholesterol lactobacterium casei J1 and produce probiotics yogurt.
Technical scheme of the present invention is: a kind of lactobacterium casei (Lactobacillus casei) J1 reducing cholesterol, be separated in the Kefir grain (also known as Kefir granule) of average family, there is excellent product bile salt hydrolase and tolerate gi tract against environmental characteristics, can the balance of regulating intestinal canal flora, and have prebiotic health-care effecies such as reducing serum cholesterol, this bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on September 11st, 2006, preservation address is: Datun Road, Chaoyang District, Beijing City, preserving number: CGMCC No.1808.
It is obtained by lactobacterium casei (Lactobacillus casei) the J1 dry powder leaven and common sour milk dry powder leaven with efficient reduction cholesterol and product bile salt hydrolase that the present invention prepares probiotics yogurt.
A kind of method utilizing the lactobacterium casei J1 of decreasing cholesterol to produce probiotics yogurt, adopt method of delivering directly to add dry powder leaven to produce the technique of probiotics yogurt and be: be 37 ~ 43 DEG C at leavening temperature, fermentation time is 6 ~ 3h, sucrose addition is 5% ~ 8% (W/V, mass volume ratio concentration) under prerequisite, lactobacterium casei J1 bacterial strain dry powder leaven inoculum size is 1 ‰ (W/V, mass volume ratio concentration), the inoculum size of common sour milk dry powder leaven is 1 ‰ (W/V, mass volume ratio concentration), the inoculative proportion of lactobacterium casei J1 bacterial strain dry powder leaven and common sour milk dry powder leaven is 1:1 (W/W, massfraction).
In described method, common sour milk dry powder leaven is the mixed bacterium powder of commercially available lactobacillus bulgaricus and thermophilus streptococcus.
Effect of the present invention is: probiotics yogurt prepared by the dry powder leaven of adding lactobacterium casei J1 has good reduction serum cholesterol effect.Do not changing on common sour milk zymotechnique basis, both J1 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 J1 after 21 days under 4 DEG C of conditions can reach 10
6more than CFU/g, still has nourishing function.
The raw material sources that the present invention makes probiotics yogurt 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.
Below in conjunction with drawings and Examples, the present invention is described further.
Accompanying drawing explanation
Accompanying drawing is that application lactobacterium casei J1 (Lactobacillus casei) J1 and common sour milk dry powder leaven produce probiotics yogurt process flow sheet.
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).
A kind of lactobacterium casei (Lactobacillus casei) J1 reducing cholesterol, be separated in the Kefir grain (also known as Kefir granule) of average family, there is excellent product bile salt hydrolase and tolerate gi tract against environmental characteristics, can the balance of regulating intestinal canal flora, and have prebiotic health-care effecies such as reducing serum cholesterol, this bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preserving number on September 11st, 2006: CGMCC No.1808.
It is obtained by lactobacterium casei (Lactobacillus casei) the J1 dry powder leaven and common sour milk dry powder leaven with efficient reduction cholesterol and product bile salt hydrolase that the present invention prepares probiotics yogurt.
A kind of method utilizing the lactobacterium casei J1 of decreasing cholesterol to produce probiotics yogurt, adopt method of delivering directly to add dry powder leaven to produce the technique of probiotics yogurt and be: be 37 ~ 43 DEG C at leavening temperature, fermentation time is 6 ~ 3h, sucrose addition is 5% ~ 8% (W/V, mass volume ratio concentration) under prerequisite, lactobacterium casei J1 bacterial strain dry powder leaven inoculum size is 1 ‰ (W/V, mass volume ratio concentration), the inoculum size of common sour milk dry powder leaven is 1 ‰ (W/V, mass volume ratio concentration), the inoculative proportion of lactobacterium casei J1 bacterial strain dry powder leaven and common sour milk dry powder leaven is 1:1 (W/W, massfraction).
In described method, common sour milk dry powder leaven is the mixed bacterium powder of commercially available lactobacillus bulgaricus and thermophilus streptococcus.
In accompanying drawing, application lactobacterium casei J1 (Lactobacillus casei) J1 and common sour milk dry powder leaven produce probiotics yogurt technical process, comprise the following steps:
(1) raw dairy: the raw dairy making Yoghourt requires that bacterium number is lower than 10 not containing microbiotic and Herb of Common violet
4individual/mL.
(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 GB 5408-85.Its total dry matter should be not less than 11.5%, lipid content 1.0% ~ 3.2%.
(4) preheating, batching and filtration: raw dairy is heated to about 60 DEG C, add the sucrose of 5% ~ 8% (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.
(6) sterilization: raw dairy is heated to 90 ~ 95 DEG C, insulation 5 ~ 10min.
(7) cool: the raw dairy after sterilization is cooled to rapidly 37 ~ 43 DEG C, to be seeded.
(8) inoculate: lactobacterium casei J1 bacterial strain dry powder leaven inoculum size is 1.0 ‰ (W/V, mass volume ratio concentration), the inoculum size of common sour milk dry powder leaven is 1.0 ‰ (W/V, mass volume ratio concentration), the inoculative proportion of lactobacterium casei J1 bacterial strain dry powder leaven and common sour milk dry powder leaven is 1:1 (W/W, massfraction).
(9) packing: 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 37 ~ 43 DEG C, fermentation time is 6 ~ 3h.
(11) cool: take out from proving room, be cooled to less than 10 DEG C.
(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.
1, lactobacterium casei J1 application experiment in probiotics yogurt of decreasing cholesterol is utilized
(1), lactobacterium casei J1 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 ‰, lactobacterium casei J1 dry powder leaven addition is respectively 1.0 ‰, 1.5 ‰, 2.0 ‰, after 37 ~ 43 DEG C of fermentation 6 ~ 3h, be that 1.0 ‰, 1.5 ‰, 2.0 ‰ storages are to the Yoghourt acidity of 1d, 7d, 14d, 21d with 0.1mol/LNaOH measured in solution lactobacterium casei J1 dry powder leaven addition, and with the common sour milk not adding lactobacterium casei J1 dry powder leaven for control group, unit represents with gill Nie Erdu (° T), the results are shown in Table 1.
Table 1 different dry Lactobacillus paracasei J1 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 1, compared with control group, when lactobacterium casei J1 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 1.5 ‰, when addition is 2.0 ‰, adds post-acidification of yoghurt degree.Therefore illustrate in post-acidification of yoghurt index, the addition of lactobacterium casei J1 dry powder leaven 1.0 ‰ is better than the addition of 1.5 ‰ and 2.0 ‰.
(2), lactobacterium casei J1 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 ‰, lactobacterium casei J1 dry powder leaven addition is respectively 1.0 ‰, 1.5 ‰, 2.0 ‰, after 37 ~ 43 DEG C of fermentation 6 ~ 3h, measuring lactobacterium casei J1 dry powder leaven addition respectively with viscosmeter is the Yoghourt viscositys of 1.0 ‰, 1.5 ‰, 2.0 ‰ storages to 1d, 7d, 14d, 21d, not add the common sour milk of lactobacterium casei J1 dry powder leaven for control group, the results are shown in Table 2.
Table 2 different dry Lactobacillus paracasei J1 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 2, lactobacterium casei J1 dry powder leaven addition significantly improves Yoghourt viscosity (P<0.05) when being 1.0 ‰, and when addition is 1.5 ‰ and 2.0 ‰, Yoghourt viscosity does not significantly improve (P<0.05).Therefore illustrate in Yoghourt viscosity index, the addition of lactobacterium casei J1 dry powder leaven 1.0 ‰ is better than the addition of 1.5 ‰ and 2.0 ‰.
(3), lactobacterium casei J1 dry powder leaven addition is on the impact of J1 bacterial strain number of viable in Yoghourt
Under common sour milk dry powder leaven inoculum size is the prerequisite of 1.0 ‰, lactobacterium casei J1 dry powder leaven addition is respectively 1.0 ‰, 1.5 ‰, 2.0 ‰, after 37 ~ 43 DEG C of fermentation 6 ~ 3h, be that 1.0 ‰, 1.5 ‰, 2.0 ‰ storage to the Yoghourts of 1d, 7d, 14d, 21d carry out plate count by lactobacterium casei J1 dry powder leaven addition.(J1 bacterial strain has the characteristic of tolerance cholate to adopt the modified MRS selective medium adding cholate, its growth is not suppressed by cholate, and the growth of common sour milk bacterial classification can be suppressed by cholate, therefore bacterium colony can not be grown on modified MRS selective medium flat board), the number of viable of J1 bacterial strain in Yoghourt is detected with pour plate culture method, count after 37 DEG C of cultivation 48h, the results are shown in Table 3.
Table 3 lactobacterium casei J1 dry powder leaven addition is on the impact of J1 bacterial strain viable count in Yoghourt
From table 3, add the Yoghourt of lactobacterium casei J1 dry powder leaven fermentation, in storage after 21 days, the number of viable of J1 bacterial strain can reach 10
6more than CFU/g, still has nourishing function.Therefore illustrate in Yoghourt viable count index, when lactobacterium casei J1 dry powder leaven addition is 1.0 ‰, 1.5 ‰, 2.0 ‰, all can make probiotics yogurt.But consider production cost problem, determine that lactobacterium casei J1 dry powder leaven addition is 1.0 ‰.
In sum, in order to obtain colory probiotics yogurt, for lactobacterium casei J1 dry powder leaven Different adding amount on the impact of Quality of Yoghourt, determine its optimum addition.It is 37 ~ 43 DEG C at leavening temperature, fermentation time is 6 ~ 3h, sucrose addition is 5% ~ 8% (W/V, mass volume ratio concentration) under prerequisite, by the acidity of acidifying after analysis mensuration fermented yogurt, and the viscosity of storage Yoghourt, J1 bacterial strain number of viable, determine that probiotics yogurt lactobacterium casei J1 dry powder leaven optimum addition is 1.0 ‰ (W/V, mass volume ratio concentration), common sour milk dry powder leaven inoculum size is 1.0 ‰ (W/V, mass volume ratio concentration), the inoculative proportion of lactobacterium casei J1 bacterial strain dry powder leaven and common sour milk dry powder leaven is 1:1 (W/W, massfraction).Finished product probiotics yogurt 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 J1 bacterial strain viable count refrigerated after 21 days in Yoghourt can reach 10
6more than CFU/g, still with health role.
2, the experimentation on animals of probiotics yogurt serum cholesterol-lowering
The present invention's production probiotics yogurt starter used is lactobacterium casei J1 (Lactobacillus caseiJ1) dry powder leaven and common sour milk dry powder leaven.Wherein lactobacterium casei J1 is the prebiotic bacterial classification 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 small white mouse do probiotics yogurt per os gavage test, its method and result as follows:
(1), animal grouping and process
By the healthy adult male ICR mouse of 60 SPF levels, body weight 20 ± 2g (Beijing Vital River Experimental Animals Technology Co., Ltd. provides), being divided into 5 groups at random, is low dose group, middle dosage group, high dose group, blank group, hyperlipidemia model group respectively.Probiotic acid Feeding volume 300mL/d60kg body weight (Body weight is recommended according to human body, bw) calculate, be equivalent to 5mL/d/kgbw every day, press 1 times of human body recommended amounts, 5 times, 10 times basic, normal, high three the dosage groups of design, namely probiotics yogurt awards examination agent amount is 5mL/kgbw, 25mL/kgbw, 50mL/kgbw, and blank group and hyperlipidemia model group are filled with and compared with distilled water.Each gavage capacity is 0.2mL/10gbw, feeds so continuously 8 weeks.
(2), serum preparation and detection
At test the 4th, 8 weekends, after fasting (can't help water) 12h, orbital venous plexus blood sampling respectively, 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 4; Adopt GPO-peroxidase-4-AA and phenol method to measure the content of serum levels of triglyceride (TG), the results are shown in Table 5.
Table 4 probiotics yogurt is to the test-results of mice serum TC
Note: * *: compare P<0.01 with blank group; Δ: compare P<0.05 with model group
From table 4, after 4 weeks, hyperlipidemia model group and blank group mice serum TC content have pole significant difference (p<0.01), show that model is successfully established.4th week and 8 weekends, the serum TC content of low dose group mouse all significantly (p<0.05) lower than hyperlipidemia model group, and the mice serum TC content of middle and high dosage group with compared with model group, substantially unchanged; Show that the probiotics yogurt of low dosage has the effect significantly reducing hypercholesterolemia mice serum TC content.
Table 5 probiotics yogurt is to the test-results of mice serum TG
Note: *: compare P<0.1 with model group
From table 5, in whole process of the test, hyperlipidemia model group mice serum TG content, compared with blank group, slightly raises, but without significant difference.4th week and 8 weekends, the serum TG content of low dose group mouse, compared with model group, has obvious reduction (p<0.1), and reduction amplitude is 17.9% and 18.4% respectively, all lower than blank group of basis level; The serum TG content of middle dosage group mouse, compared with hyperlipidemia model group, slightly reduces, and reduction amplitude is 2.0% and 8.9% respectively; The mice serum TG content of high dose group is without reduction trend; Indicate probiotics yogurt and have the effect reducing hypercholesterolemia mice serum TG content, the effect of low dose group is better than middle and high dosage group.
In sum, probiotics yogurt prepared by the dry powder leaven of adding lactobacterium casei J1 in common sour milk is produced, there is good reduction serum cholesterol effect, develop serum cholesterol-lowering probiotics yogurt, probiotics tablets and probiotics with it and there are wide market outlook.
Claims (3)
1. one kind is reduced lactobacterium casei (Lactobacillus casei) J1 of cholesterol, be separated in the Kefir grain of average family, there is excellent product bile salt hydrolase and tolerate gi tract against environmental characteristics, can the balance of regulating intestinal canal flora, and have prebiotic health-care effecies such as reducing serum cholesterol, this bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preserving number on September 11st, 2006: CGMCC No.1808.
2. one kind utilizes lactobacterium casei (Lactobacillus casei) J1 reducing cholesterol as claimed in claim 1 to produce the method for probiotics yogurt, it is characterized in that: leavening temperature is 37 ~ 43 DEG C, fermentation time is 6 ~ 3h, sucrose addition is 5% ~ 8%, prebiotic lactobacterium casei J1 bacterial strain dry powder leaven inoculum size is 1 ‰, the inoculum size of common sour milk dry powder leaven is 1 ‰, and the inoculative proportion of lactobacterium casei J1 bacterial strain dry powder leaven and common sour milk dry powder leaven is 1:1.
3. the method utilizing lactobacterium casei (Lactobacillus casei) J1 reducing cholesterol to produce probiotics yogurt as claimed in claim 2, it is characterized in that: lactobacterium casei J1 probiotics yogurt, confirm that there is good reduction serum cholesterol effect through experimentation on animals, can be used as development of functional food application.
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Cited By (4)
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| CN105062929A (en) * | 2015-09-06 | 2015-11-18 | 天津科技大学 | Water kefir composite flora and manufacturing method of fermented beverage by using same |
| CN110384129A (en) * | 2019-08-16 | 2019-10-29 | 大连工业大学 | A kind of low cholesterol acidified milk and preparation method thereof |
| CN110973247A (en) * | 2019-12-12 | 2020-04-10 | 新派(上海)餐饮管理有限公司 | Lactobacillus casei Haidilao-1-containing lactobacillus beverage and preparation method thereof |
| CN111771975A (en) * | 2020-07-28 | 2020-10-16 | 浙江大学 | Preparation method of low-cholesterol egg yoghourt |
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| CN103131654A (en) * | 2013-03-05 | 2013-06-05 | 北京农学院 | Application of Lactobacillus casei KL1 for producing bile salt hydrolase in functional sour milk |
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| CN103131654A (en) * | 2013-03-05 | 2013-06-05 | 北京农学院 | Application of Lactobacillus casei KL1 for producing bile salt hydrolase in functional sour milk |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105062929A (en) * | 2015-09-06 | 2015-11-18 | 天津科技大学 | Water kefir composite flora and manufacturing method of fermented beverage by using same |
| CN110384129A (en) * | 2019-08-16 | 2019-10-29 | 大连工业大学 | A kind of low cholesterol acidified milk and preparation method thereof |
| CN110973247A (en) * | 2019-12-12 | 2020-04-10 | 新派(上海)餐饮管理有限公司 | Lactobacillus casei Haidilao-1-containing lactobacillus beverage and preparation method thereof |
| CN111771975A (en) * | 2020-07-28 | 2020-10-16 | 浙江大学 | Preparation method of low-cholesterol egg yoghourt |
| CN111771975B (en) * | 2020-07-28 | 2023-03-21 | 浙江大学 | Preparation method of low-cholesterol egg yoghourt |
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