CN105820967B - One plant of Lactobacillus brevis and its application - Google Patents
One plant of Lactobacillus brevis and its application Download PDFInfo
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- CN105820967B CN105820967B CN201510005242.4A CN201510005242A CN105820967B CN 105820967 B CN105820967 B CN 105820967B CN 201510005242 A CN201510005242 A CN 201510005242A CN 105820967 B CN105820967 B CN 105820967B
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/90—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in food processing or handling, e.g. food conservation
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Abstract
One plant of Lactobacillus brevis, is named as Lactobacillus brevis AR-123, and deposit number is CGMCC 9761.The Lactobacillus brevis AR-123 can effectively degrading nitrite can be very good to keep the original flavor of vegetables and nutrition, content of nitrite is only 1.6mg/L in fermented vegetable after fermentation as fermented vegetable agent application.
Description
Technical field
The present invention relates to microorganisms, in particular to one plant of Lactobacillus brevis and its application.
Technical background
Fermented vegetable refers to that through everfermentation can be the Typical Representative of China's traditional zymotic food with the vegetables of long-term storage
One of, there are unique flavor characteristics.Fermented vegetable mainly by the vegetables such as Chinese cabbage, radish, cucumber certain density salt,
Under the action of water and condiment, using leavenings such as the microorganisms or artificial infection lactic acid bacteria for being naturally attached to vegetable surface, detest
The nutriments such as the fermentable saccharide in vegetables are decomposed under the conditions of oxygen and generate the process of a large amount of lactic acid.During the fermentation, exist
Under a large amount of lactic acid and condition of salt stress, some putrefactive microorganisms and the growth of other anaerobes in vegetables will receive strong
Inhibit, while the flavor substances such as ethyl alcohol, acetaldehyde, mannitol can be generated, to form the fermented food of peculiar taste.Fermented vegetable
Primary raw material be various fresh vegetables full of nutrition, rich in nutrients and the lactic acid such as vitamin, calcium, iron, phosphorus.It is delicious tender
It is crisp, appetite can not only be promoted, and there is certain health-care efficacy, for example preventing hypertension, adjust cholesterol level and slow
Slow aging.Some researches show that fermented vegetable also has certain prophylactic activity to cancer.
Fermented vegetable is that China eats time-honored lactic fermentation vegetable product, has good flavor and health care function
Can, but content of nitrite excessive problem limits the fast development of such cure foods always.In recent years, due to nitrogenous fertilizer
It is a large amount of to use, cause Nitrate in Vegetable to increase significantly, and vegetables are a kind of plant foods for being easy to be enriched with nitrate,
In fermented vegetable curing process, the nitrate in vegetables can be by the part bacterial reduction in Pickle at nitrite.
Studies have shown that the content of nitrite in Pickle is higher by several times even as many as tens times than fresh vegetables of the same race, moreover, by
In Pickle in storage, that there may be preservation conditions in transportational process is improper, cause the pollution microbes in product to increase so that
Content of nitrite therein further increases.Therefore, how strict control pickle pickles in nitrite content, it is ensured that
The safety in utilization of this traditional food of pickles, making it, preferably circulation is urgent problem on the market.
Summary of the invention
The purpose of the present invention, aiming at the lactic acid bacteria for lacking safe and efficient degrading nitrite in the prior art
Deficiency, the Lactobacillus brevis and its application, bacterial strain of the invention for providing one plant of energy effectively degrading nitrite send out under optimum conditions
Fermented pickled vegetable can be very good to keep the original flavor of vegetables and nutrition, and Nitrite in Pickles content is only after fermentation
1.6mg/L。
In order to achieve the object of the present invention, the invention adopts the following technical scheme: one plant of Lactobacillus brevis
(Lactobacillus brevis), is named as Lactobacillus brevis AR-123, and the bacterial strain is micro- in China on October 13rd, 2014
Biological inoculum preservation administration committee common micro-organisms center (address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3) preservation,
Its deposit number is CGMCC 9761, and gene order is as shown in SEQ ID NO.1.
Lactobacillus brevis AR-123 bacterial strain screening of the invention is from conventional Kimchi.Separation screening is carried out to pickles sample, is utilized
Sample liquid after dilution is coated on MRS plate by gradient dilution method, and 37 DEG C of constant temperature incubations obtain separation after scribing line separation
The lactic acid bacteria obtained carries out morphologic observation, is determined as effectively degrading nitrite bacterial strain in conjunction with degrading nitrite fermenting experiment,
Degradation efficiency is up to 99.5%.
Application of the above-mentioned Lactobacillus brevis AR-123 as fermented vegetable agent.
Above-mentioned Lactobacillus brevis AR-123 is as follows as the application method of fermented vegetable agent:
(1) Lactobacillus brevis AR-123 is activated as inoculation liquid；
(2) 300mL salt water, condiment and cleaned, stripping and slicing vegetables are added into sterilized container；
(3) inoculation liquid is inoculated in the above-mentioned container for being placed with vegetables by the inoculum concentration of 5-15% (v/v), 25 DEG C of standing hairs
Ferment 7 days, obtain the fermented vegetable of low nitrite content.
Activation described in step (1) refers to: Lactobacillus brevis AR-123 being inoculated in MRS culture medium, 37 DEG C of standing trainings
It supports and carries out secondary expansions culture for 24 hours, under similarity condition, 4000rpm is centrifuged 8min collection thallus, then with physiological saline suspended bacteria
Body, being modulated into Lactobacillus brevis AR-123 concentration is 108The inoculation liquid of cfu/mL.
The weight percent concentration of salt is 3-10% in salt water described in step (2), and the amount that vegetables are added is 75-
200g。
The Lactobacillus brevis AR-123 that the present invention filters out can effectively degrading nitrite, under optimum conditions fermentation bubble
Dish can be very good to keep the original flavor of vegetables and nutrition, and Nitrite in Pickles content is only 1.6mg/ after fermentation
L.Using the strain fermentation vegetables, there is unique flavor, fresh perfume is tender crisp, and it is in good taste, and content of nitrite is extremely low.The present invention
Lactobacillus brevis AR-123 to be used to prepare fermented vegetable process simple, heavy industrialization application may be implemented.
Detailed description of the invention
Fig. 1 is the growth curve of Lactobacillus brevis AR-123.
Specific implementation method
The present invention is further explained combined with specific embodiments below.For specific method or material used in embodiment
Material, those skilled in the art can carry out conventional replacement according to existing technology and select on the basis of the technology of the present invention thinking
It selects, is not limited solely to the specific record of the embodiment of the present invention.
Experimental method used in following embodiments is conventional method unless otherwise specified；Used material,
Reagent etc., is commercially available unless otherwise specified.
MRS solid medium: peptone 10g/L, beef extract 10g/L, yeast extract 5g/L, K2HPO42g/L, citric acid
Diammonium 2g/L, sodium acetate 5g/L, glucose 20g/L, Tween 80 1mL, MgSO4·7H2O0.58g/L, MnSO4·4H2O
0.25g/L, agar 20g/L adjust pH to 6.4.
The screening and identification of 1 Lactobacillus brevis AR-123 of embodiment
The preparation of 1.1 sample diluting liquids and strain isolation
Acquisition comes from the homemade fermented vegetable sample of Sichuan farmers', shreds, and 15mL sterile water is added, acutely shakes 20min.
Using concentration gradient dilution method (10-1-10-4), take dilution 100uL to be coated on MRS agar medium plate, 37 DEG C of anaerobism
48h is cultivated, the bacterial strain that choosing colony is big, growth is fast, further scribing line isolates and purifies.
1.2 bacterial strain screening
By the strain inoculated in step 1.1, into MRS fluid nutrient medium, 37 DEG C of stationary cultures for 24 hours, are prepared into seed liquor, press
According in 5% MRS fluid nutrient medium of the inoculum concentration access containing 0.2mg/mL nitrite, 37 DEG C of stationary culture 48h, culture is tied
Shu Hou, 4000rpm, 10min centrifugation medium, take supernatant.Using hydrochloric acid how ethylenediamine method measurement culture solution in it is remaining
NaNO2Content.Experimental result is as shown in table 1.
The screening of 1 primary dcreening operation strains for degrading nitrite of table
The identification of 1.3 bacterial strains
One plant of isolated bacterium is named as AR-123.Bacterium colony ellipse or similar round, neat in edge, bacterium colony size one
As be 2~3mm, milky.By 16SrDNA sequence comparative analysis, short newborn bar in the sequence and ncbi database as the result is shown
The homology of bacterium can identify that the bacterium is Lactobacillus brevis (Lactobacillusbrevis), be named as Lactobacillus brevis up to 99%
AR-123。
1.4 strain growth characteristics
By activated Lactobacillus brevis AR-123 by 1% (v/v) inoculum concentration access MRS fluid nutrient medium, 37 DEG C are stood
Culture is for 24 hours, dense in the bacterium of 600nm measurement culture solution every 2h.It is plotted against time to obtain strains A R-123 in MRS so that bacterium is dense
Growth curve, result are as shown in Figure 1.The experimental results showed that Lactobacillus brevis AR-123 grown in MRS culture medium rapidly,
6h or so enters logarithmic phase, and 12h or so enters stationary phase.
2 Lactobacillus brevis AR-123 of embodiment prepares low nitrite fermented vegetable
(1) Sterile Saline and condiment that 300mL concentration is 3% are added into sterilized container, takes fresh Chinese cabbage, clearly
It washes, drains excessive moisture, the fritter that length and width is 5*3cm is cut under aseptic condition and is fitted into container, vegetables weight is put into
Amount is 75g；
(2) the activated Lactobacillus brevis AR-123 of 5% (v/v) is accessed into container, water-stop is added to be placed at 25 DEG C, is sent out
Ferment 7d is to get fermented vegetable.Through detecting, the content of nitrite of the present embodiment products obtained therefrom is 8.4mg/L.The result shows that institute
The content of nitrite for obtaining fermented vegetable is lower, and product has peculiar flavour, and the fresh perfume of mouthfeel is tender crisp.
3 Lactobacillus brevis AR-123 of embodiment prepares low nitrite fermented vegetable
(1) Sterile Saline and condiment that 300mL concentration is 7% are added into sterilized container, takes fresh Chinese cabbage, clearly
It washes, drains excessive moisture, the fritter that length and width is 5*3cm is cut under aseptic condition and is fitted into container, vegetables weight is put into
Amount is 150g；
(2) the activated Lactobacillus brevis AR-123 of 10% (v/v) is accessed into container, and water-stop is added to be placed at 25 DEG C,
7d ferment to get fermented vegetable.Through detecting, the content of nitrite of the present embodiment products obtained therefrom is 1.6mg/L.The result shows that
The content of nitrite of gained fermented vegetable is lower, and product has peculiar flavour, and the fresh perfume of mouthfeel is tender crisp.
4 Lactobacillus brevis AR-123 of embodiment prepares low nitrite fermented vegetable
(1) Sterile Saline and condiment that 300mL concentration is 7% are added into sterilized container, takes fresh Chinese cabbage, clearly
It washes, drains excessive moisture, the fritter that length and width is 5*3cm is cut under aseptic condition and is fitted into container, vegetables weight is put into
Amount is 200g；
(2) the activated Lactobacillus brevis AR-123 of 15% (v/v) is accessed into container, and water-stop is added to be placed at 25 DEG C,
7d ferment to get fermented vegetable.Through detecting, the content of nitrite of the present embodiment products obtained therefrom is 1.8mg/L.The result shows that
The content of nitrite of gained fermented vegetable is lower, and product has peculiar flavour, and the fresh perfume of mouthfeel is tender crisp.
5 Lactobacillus brevis AR-123 of embodiment prepares low nitrite fermented vegetable
(1) Sterile Saline and condiment that 300mL concentration is 10% are added into sterilized container, takes fresh Chinese cabbage, clearly
It washes, drains excessive moisture, the fritter that length and width is 5*3cm is cut under aseptic condition and is fitted into container, vegetables weight is put into
Amount is 200g；
(2) the activated Lactobacillus brevis AR-123 of 15% (v/v) is accessed into container, and water-stop is added to be placed at 25 DEG C,
7d ferment to get fermented vegetable.Through detecting, the content of nitrite of the present embodiment products obtained therefrom is 2.6mg/L.The result shows that
The content of nitrite of gained fermented vegetable is lower, and product has peculiar flavour, and the fresh perfume of mouthfeel is tender crisp.
Claims (5)
1. one plant of Lactobacillus brevis (Lactobacillus brevis), is named as Lactobacillus brevis AR-123, the bacterial strain is in 2014
On October 23, in is in China Committee for Culture Collection of Microorganisms's common micro-organisms center's preservation, address: Chaoyang District, Beijing City
The institute 3 of North Star West Road 1, deposit number are CGMCC NO.9761,16s rna gene sequence such as SEQ ID NO.1 institute
Show.
2. application of the Lactobacillus brevis AR-123 described in claim 1 as fermented vegetable agent.
3. application of the Lactobacillus brevis AR-123 according to claim 2 as fermented vegetable agent, it is characterised in that:
Application method is as follows:
(1) Lactobacillus brevis AR-123 is activated as inoculation liquid；
(2) 300mL salt water, condiment and cleaned, stripping and slicing vegetables are added into sterilized container；
(3) inoculation liquid is inoculated in the above-mentioned container for being placed with vegetables by the inoculum concentration of 5-15% (v/v), 25 DEG C of standing for fermentation 7
It, obtains the fermented vegetable of low nitrite content.
4. application of the Lactobacillus brevis AR-123 according to claim 3 as fermented vegetable agent, it is characterised in that: step
(1) activation described in refers to: Lactobacillus brevis AR-123 is inoculated in MRS culture medium, 37 DEG C of stationary cultures for 24 hours, same to batten
Carry out secondary expansion culture under part, 4000rpm is centrifuged 8min and collects thallus, then with physiological saline suspension thalline, is modulated into short cream
Bacillus AR-123 concentration is 108The inoculation liquid of cfu/mL.
5. application of the Lactobacillus brevis AR-123 according to claim 3 as fermented vegetable agent, it is characterised in that: step
(2) weight percent concentration of salt is 3-10% in the salt water described in, and the amount that vegetables are added is 75-200g.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106962741A (en) * | 2017-03-16 | 2017-07-21 | 江南大学 | The method of microbial compound preparation efficient degradation salted fish nitrite |
| CN109645403B (en) * | 2018-11-30 | 2022-05-31 | 四川东坡中国泡菜产业技术研究院 | Low-temperature-resistant and bacteriophage-resistant lactobacillus brevis and application thereof in pickle |
| CN113416668B (en) * | 2021-06-08 | 2023-06-06 | 大连工业大学 | Lactobacillus brevis for reducing nitrite content, and method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101326972A (en) * | 2008-07-28 | 2008-12-24 | 东北农业大学 | Directly thrown leaven for effectively degrading nitrite in pickled vegetables and preparation thereof |
| CN102018154A (en) * | 2010-10-26 | 2011-04-20 | 浙江大学 | Method for inoculating lactobacillus to rapidly reduce nitrite content in pickles |
| CN104187519A (en) * | 2014-09-05 | 2014-12-10 | 浙江正味食品有限公司 | Method for effectively inhibiting browning |
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2015
- 2015-01-04 CN CN201510005242.4A patent/CN105820967B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101326972A (en) * | 2008-07-28 | 2008-12-24 | 东北农业大学 | Directly thrown leaven for effectively degrading nitrite in pickled vegetables and preparation thereof |
| CN102018154A (en) * | 2010-10-26 | 2011-04-20 | 浙江大学 | Method for inoculating lactobacillus to rapidly reduce nitrite content in pickles |
| CN104187519A (en) * | 2014-09-05 | 2014-12-10 | 浙江正味食品有限公司 | Method for effectively inhibiting browning |
Non-Patent Citations (3)
| Title |
|---|
| Lactobacillus brevis strain KLDS1.0727 16S ribosomal RNA gene, partial sequence;Zhang Y et al;《GenBank：EU626012.1》;20080506;origin * |
| 乳酸菌对泡菜发酵过程中亚硝酸盐含量变化及泡菜品质的影响研究;周光燕;《西南农业学报》;20091231;第19卷(第2期);摘要，第2.1节，图1 * |
| 几种发酵蔬菜中乳酸菌的分离与筛选;商军 等;《食品科学》;20071231;第28卷(第4期);摘要 * |
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