CN106676026A - Lactobacillus bulgaricus strain with quorum sensing and application of lactobacillus bulgaricus strain - Google Patents
Lactobacillus bulgaricus strain with quorum sensing and application of lactobacillus bulgaricus strain Download PDFInfo
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- CN106676026A CN106676026A CN201510756046.0A CN201510756046A CN106676026A CN 106676026 A CN106676026 A CN 106676026A CN 201510756046 A CN201510756046 A CN 201510756046A CN 106676026 A CN106676026 A CN 106676026A
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- bulgaricus
- lactobacillus bulgaricus
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Abstract
The invention discloses a lactobacillus bulgaricus strain with quorum sensing and application of the lactobacillus bulgaricus strain. The lactobacillus delbrueckii subsp. bulgaricus ljj-6 strain provided by the invention has a collection registration number of CGMCC NO. 11346, and is called ljj-6 for short. The invention further discloses a method for preparing a yogurt product. The method comprises the following steps: inoculating lactobacillus ljj-6 to liquid milk, and fermenting, so as to obtain the yogurt product. According to the lactobacillus ljj-6 provided by the invention, the flora density can be regulated in an autolysis manner in the later fermentation stage, so that the postacidification phenomenon of the yogurt is reduced, and the lactobacillus ljj-6 can serve as a high-quality yogurt fermentation agent and has major application and popularization values.
Description
Technical field
The invention belongs to biological technical field, and in particular to one plant has the Lactobacillus bulgaricus of quorum sensing and its answers
With.
Background technology
Yoghourt is the milk product containing viable bacteria, after normal fermentation terminates, product storage, transport, sale, it is edible before this
During one, thalline is still to growth and breeding, the pH of acidification phenomenon (postacidification), i.e. Yoghourt after generation
Value continues to decline, so that occur that consumer is unacceptable to be crossed tart flavour and aesthetic quality and decline.Post-acidification of yoghurt phenomenon is led
The result of cause is that Yoghourt cannot be stored for a long time, and Yoghourt storage period short defect annoyings the consumption of Yoghourt.
Quorum sensing (Quorum Sensing, QS) refers to that the expression of its gene of certain micro-organisms colony is subject to close with colony
The phenomenon of the related signaling molecule regulation and control of degree.QS systems participate in a series of biological functions of regulation and control, such as：Thalline is luminous,
The biosynthesiss of antibiotic, the generation of virulence factor, the synthesis of extracellular polysaccharide, antibacterial are gathered together, biofilm formation, matter
Grain engagement transfer etc..
QS systems are considered as participating in the multinomial physiological function of regulation and control antibacterial, and the system includes self-induction peptide
(autoinducingpeptide, AIP), Histidine kinase and sensing regulatory protein, are also called three-component system.
AIP for indicator cellses density, when AIP reaches a certain critical concentration threshold, just can be swashed as signaling molecule
A series of expression of specific genes of work.Observe the presence of quorum sensing phenomenon in various bacteria at present, than
Such as staphylococcus aureuses, Streptococcus mutans.
Some antibacterials of nature have intervention school-based, can be by colony induction signaling point in monitoring surrounding
Sub- concentration to perceive itself and surrounding in other floras number change, when signaling molecule concentration reaches certain threshold
Just start thalline related gene expression, regulation and control bacteria concentration and related biological function during value.
The content of the invention
It is an object of the invention to provide one plant of Lactobacillus bulgaricus with quorum sensing and its application.
The lactobacillus delbruockii subspecies bulgaricus that the present invention is provided
(Lactobacillusdelbrueckiisubsp.bulgaricus) ljj-6, protected on 09 09th, 2015
(abbreviation CGMCC, address is to be hidden in China Committee for Culture Collection of Microorganisms's common micro-organisms center：Beijing
The institute 3 of Chaoyang District North Star West Road 1, Institute of Microorganism, Academia Sinica), preservation registration number is CGMCC
NO.11346.Lactobacillus delbruockii subspecies bulgaricus (Lactobacillus delbrueckiisubsp.bulgaricus)
Ljj-6, abbreviation lactobacilluss ljj-6.
On the one hand, the culture of same time is carried out in MRS fluid mediums, when 12 hours, lactobacilluss ljj-6's
Strain density is similar with existing Lactobacillus bulgaricus, and the strain density of lactobacilluss ljj-6 is substantially less than existing after 24 hours
Lactobacillus bulgaricus.On the other hand, the culture of same time is carried out, the Yoghourt for adding lactobacilluss ljj-6 to obtain
Acidity value be not significantly different from the acidity value of the Yoghourt for adding existing Lactobacillus bulgaricus to obtain, but Yoghourt is entered
After row shelf life is preserved, the acidity value of the Yoghourt that addition lactobacilluss ljj-6 is obtained is substantially less than and adds existing Bulgaria
The acidity value of the Yoghourt that lactobacilluss are obtained.As a result show, compared with existing Lactobacillus bulgaricus, the present invention is provided
Lactobacilluss ljj-6 can solve post-acidification of yoghurt phenomenon.As a result show, the lactobacilluss ljj-6 that the present invention is provided is
Lactobacillus bulgaricus bacterial strain with colony's perceptual phenomenon.
The present invention also protects applications of the lactobacilluss ljj-6 in food is prepared.The food concretely Yoghourt.
The present invention also protects a kind of preparation method of Yoghourt product, comprises the steps：Lactobacilluss ljj-6 is seeded to
Liquid milk simultaneously ferments, and obtains Yoghourt product.In methods described, lactobacilluss ljj-6 is initial close in the liquid milk
Degree concretely 104cfu/ml.The condition of the fermentation is concretely：37 DEG C of quiescent cultures 12 hours.The liquid
Milk concretely milk and/or sheep milk.
The Yoghourt product that any of the above methods described is prepared falls within protection scope of the present invention.
After fermentation the phase can regulate and control population density to the lactobacilluss ljj-6 that the present invention is provided by way of self-dissolving, so as to
The rear acidification phenomenon of Yoghourt is reduced, can be as good yoghurt leaven, with great application and promotional value.
Description of the drawings
Fig. 1 is the acidity value result in embodiment 3.
Specific embodiment
Below example facilitates a better understanding of the present invention, but does not limit the present invention.Experiment in following embodiments
Method, if no special instructions, is conventional method.Test material used in following embodiments, if no special instructions,
It is what is be commercially available from routine biochemistry reagent shop.Quantitative test in following examples, is respectively provided with three repetitions real
Test, results averaged.
The preparation method of MRS fluid mediums is as follows：Take 10.0 grams of casein peptone, beef 10.0 grams of thing of leaching, ferment
5.0 grams of female extracting solution, 5.0 grams of glucose, 5.0 grams of sodium acetate, 2.0 grams of Fructus Citri Limoniae acid diamine, Tween 80 1.0
Gram, 2.0 grams of dipotassium hydrogen phosphate, 0.2 gram of Magnesium sulfate heptahydrate, 0.05 gram of seven water manganese sulfate and 20.0 grams of Calcium Carbonate,
With distillation water dissolution and it is settled to 1.0 liters, adjusts pH to 6.8.
Lactobacillus bulgaricus LJJ：Cui Wenming etc., " analysis of Influential Factors of Lactobacillus bulgaricus LJJ self-dissolvings ",
Food and fermentation industries, the 7th phase (total 307th phase) of volume 39 in 2013,6-12.
Embodiment 1, the identification of lactobacilluss ljj-6 and preservation
08 month 2012, isolated one plant of bacterium, was named as ljj-6 from the traditional zymotic dairy product of Inner Mongol pastoral area.
The morphological characteristic of ljj-6：Gram-positive bacillus, thalline is thick and grow, it is 2 μm～9 μm of thalline length, wide by 0.15
μm～0.18 μm, single body is slightly justified in elongated rod shape or chaining, thalline two ends, generally in single, parallel or short chain arrangement.
The physiological and biochemical property of ljj-6：Facultative anaerobic bacteria, most suitable cultivation temperature is 3 DEG C, and the pH of optimum growth is
6.5-7.0, optimum culture medium is MRS culture medium；Lactose that can be in fermented milk generates lactic acid, while producing
Special fragrance.
The coded sequence (see the sequence 1 of sequence table) of the 16sRNA of ljj-6 adds with the Deshi Lactobacilluss guarantor in GenBank
The coding sequence homology of the 16sRNA of Leah subspecies ATCC11842 is 98%.
Lactobacillus delbruockii subspecies bulgaricus (the Lactobacillusdelbrueckii that the present invention is provided
Subsp.bulgaricus) ljj-6, was preserved in Chinese microorganism strain preservation management on 09 09th, 2015
(abbreviation CGMCC, address is committee's common micro-organisms center：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3,
Institute of Microorganism, Academia Sinica), preservation registration number is CGMCC NO.11346.Lactobacillus delbrueckii is sub-
Plant (Lactobacillus delbrueckiisubsp.bulgaricus) ljj-6, abbreviation lactobacilluss ljj-6.
The culture (MRS culture medium) of embodiment 2, lactobacilluss ljj-6
1st, packet transaction following (carrying out three repetitions to test, per group arranges 5 repetition process during each repetition is tested)：
First group：Lactobacilluss ljj-6 is seeded to into MRS fluid mediums and makes its initial concentration be 104Cfu/ml, 37 DEG C
Quiescent culture 20 hours；
Second group：Lactobacillus bulgaricus LJJ is seeded to into MRS fluid mediums and makes its initial concentration be
104Cfu/ml, 37 DEG C of quiescent cultures 20 hours.
2nd, in step 1, during quiescent culture 12 hours, the OD of cultivating system is detected600nmValue.First group of OD600nmValue
For 2.4 ± 0.1, second group of OD600nmIt is worth for 2.4 ± 0.15.
3rd, complete after step 1, detect the OD of cultivating system600nmValue.
First group of OD600nmIt is worth for 2.8 ± 0.1, second group of OD600nmIt is worth for 3.45 ± 0.15.
4th, complete after step 1, detect the pH value of cultivating system.
First group of pH value 4.4 ± 0.2, second group of pH value 4.3 ± 0.2, there was no significant difference between two groups.
As a result show：In incubation, the initial growth speed of lactobacilluss ljj-6 and Lactobacillus bulgaricus LJJ
It is not significantly different from；With the increase of incubation time, the bacteria concentration more and more higher in cultivating system, lactobacilluss ljj-6
Due to possessing quorum sensing phenomenon so as to the speed of growth is slack-off, and Lactobacillus bulgaricus LJJ is not due to possessing colony's sense
Property phenomenon is not slack-off so as to the speed of growth.
The culture (milk) of embodiment 3, lactobacilluss ljj-6
1st, packet transaction following (carrying out three repetitions to test, per group arranges 5 repetition process during each repetition is tested)：
First group：Lactobacilluss ljj-6 is seeded to into ternary plain chocolate and makes its initial concentration be 104Cfu/ml, 37 DEG C quiet
Put culture 12 hours；
Second group：Lactobacillus bulgaricus LJJ is seeded to into ternary plain chocolate and makes its initial concentration be 104Cfu/ml,
37 DEG C of quiescent cultures 12 hours.
It is vaccinated with after the plain chocolate quiescent culture of lactobacilluss and is referred to as Yoghourt.
2nd, complete after step 1, detect the acidity value of Yoghourt.
The acidity of first group, second group Yoghourt is 60 ° of T-80 ° of T.
3rd, complete after step 1, Yoghourt is carried out into shelf life preservation (shelf life preserves i.e. 4-6 DEG C preservation), take daily
Sample simultaneously detects acidity.
As a result see that Fig. 1 (preserves from shelf life and starts timing, be 1 day per 24 hours, abscissa is preservation natural law；It is vertical
Coordinate is acidity value, and unit is ° T).With the prolongation of holding time, the difference between first group and second group starts
Manifest, the acidity value of the Yoghourt of lactobacilluss ljj-6 fermentations rises relatively slowly, when preservation to the 10th day, lactobacilluss
Yoghourt of the acidity value of the Yoghourt of ljj-6 fermentations significantly lower than Lactobacillus bulgaricus LJJ fermentations.
Claims (8)
1. lactobacillus delbruockii subspecies bulgaricus (Lactobacillus delbrueckiisubsp.bulgaricus)
Ljj-6, its deposit number is CGMCC NO.11346.
2. application of the lactobacillus delbruockii subspecies bulgaricus described in claim 1 in food is prepared.
3. food as claimed in claim 2, it is characterised in that：The food is Yoghourt.
4. a kind of preparation method of Yoghourt product, comprises the steps：Deshi Lactobacilluss described in claim 1 are protected and is added
Leah subspecies are seeded to liquid milk and ferment, and obtain Yoghourt product.
5. method as claimed in claim 4, it is characterised in that：In methods described, De Shi described in claim 1 is newborn
Initial density of the bacillus subspecies bulgaricus in the liquid milk is 104cfu/ml。
6. the method as described in claim 4 or 5, it is characterised in that：The condition of the fermentation is：37 DEG C stand training
Support 12 hours.
7. the method as described in arbitrary in claim 4 to 6, it is characterised in that：The liquid milk be milk and/or
Sheep milk.
8. the Yoghourt product that arbitrary methods described is prepared in claim 4 to 7.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101472481A (en) * | 2006-06-23 | 2009-07-01 | 科.汉森有限公司 | Low post-acidifying lactic acid bacteria |
| CN103215199A (en) * | 2013-03-13 | 2013-07-24 | 内蒙古农业大学 | Lactobacillus bulgaricus bacterial strain with post-acidification delaying effect and application thereof |
| CN104694418A (en) * | 2015-01-28 | 2015-06-10 | 东北农业大学 | Lactobacillus delbrueckii subsp. bulgaricus strain with low beta-galactase activity and application thereof |
-
2015
- 2015-11-09 CN CN201510756046.0A patent/CN106676026A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101472481A (en) * | 2006-06-23 | 2009-07-01 | 科.汉森有限公司 | Low post-acidifying lactic acid bacteria |
| CN103215199A (en) * | 2013-03-13 | 2013-07-24 | 内蒙古农业大学 | Lactobacillus bulgaricus bacterial strain with post-acidification delaying effect and application thereof |
| CN104694418A (en) * | 2015-01-28 | 2015-06-10 | 东北农业大学 | Lactobacillus delbrueckii subsp. bulgaricus strain with low beta-galactase activity and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| XIAO-YANG PANG等: "Gene Knockout and Overexpression Analysis Revealed the Role of N-Acetylmuramidase in Autolysis of Lactobacillus delbrueckii subsp. bulgaricus Ljj-6", 《PLOS ONE》 * |
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