CN104531587B - One lactobacillus plantarum and its application in active fermentation breast - Google Patents
One lactobacillus plantarum and its application in active fermentation breast Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING OR TREATMENT THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
- A23C9/1234—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt characterised by using a Lactobacillus sp. other than Lactobacillus Bulgaricus, including Bificlobacterium sp.
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
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- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
- C12R2001/25—Lactobacillus plantarum
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Abstract
The invention discloses a lactobacillus plantarum and its application in active fermentation breast, for Lactobacillus plantarum LP ONLLY on December 6th, 2004 in China Committee for Culture Collection of Microorganisms's common micro-organisms center's preservation, preservation registration number is CGMCC NO.1258.The Lactobacillus plantarum has high aminopeptidase activity, high beta galactosidase activity and high glucose glycosides enzymatic activity, and it is freeze-dried the preparation that viable bacteria powder can be applied to active fermentation breast as independent leavening.The active fermentation breast not only contains high probiotics viable bacteria, and viable bacteria therein has high stability energy, can probiotic supplemented, play Lactobacillus plantarum LP ONLLY healthy functions.
Description
Invention field
The present invention relates to one plant of lactobacillus new strains and its application in active fermentation breast, more precisely, this hair
A bright new strains LP-ONLLY, CGMCC for describing Lactobacillus plantarum (Lactobacillus plantarum)
No.1258。
Background technology
Lactobacillus plantarum (Lactobacillus plantarum) is distributed in a variety of natural environments, is human body ancestral home bacterium.
2005, using the neonate of 66 births 1 to 7 day in China as research object, its enteric bacteria DNA PCR amplification clonal diversities
Analysis shows, Lactobacillus plantarum be Chinese neonates enteron aisle in advantage lactobacillus, account for it is all detection lactobacillus 74% (and
The baby intestinals bacterium such as spring, Yang Guo, Wang Jingsong, Hang Xiaomin and lactobacillus population Study on Diversity China microecology magazine,
2005,l7(4):276~277).Over the past thousands of years, Lactobacillus plantarum is widely used in fermented dairy product, meat products, plant production
In product and bakery product.Its main function is that lactic acid can be produced, reduces pH, prevents spoilage organisms from breeding, improves product special flavour
And quality, extend shelf life of products.In recent years, research is found, Lactobacillus plantarum has good probiotics function, can suppress to wrap
Include the gastrointestinal tract pathogens such as helicobacter pylori, improvement gastrointestinal function and metabolism, enhancing gut barrier function, reduce infection risk
Deng (ZL 2004100933121.8, a kind of composition containing Lactobacillus plantarum；
ZL200410093213.X, a kind of Lactobacillus plantarum).
Fermented dairy product is healthy food widely loved by consumers, the fermented dairy product containing profitable probliotics, particularly energy
The product of probiotics function is enough played, i.e., the active fermentation breast with high viable bacteria and high stability energy, turns into market development focus.
But existing research shows that Lactobacillus plantarum is applied to dairy products as independent leavening, grown in cow's milk extremely slowly, production
Sour ability is weak, and its acidified milk all makes it difficult for consumer to receive from texture and mouthfeel.Therefore, generally, plant breast
Bacillus and streptococcus thermophilus are united and applied in acidified milk system with ferment agent for sour milk (streptococcus thermophilus and lactobacillus bulgaricus)
Product (Yu Zhihui, Zhang Xue, Li Shengyu etc., application of the Lactobacillus plantarum in Yoghourt, China Dairy Industry, 2013,41 (7):27-
30), but because streptococcus thermophilus or ferment agent for sour milk grow comparatively fast in cow's milk, it is suppressed that the growth of Lactobacillus plantarum, fermentation
Lactobacillus plantarum viable count is low in dairy products, can not play the healthy functions of probiotics.
The content of the invention
In view of the foregoing, it is an object to a lactobacillus plantarum is provided and its applied in active fermentation breast.
Realize that the solution of foregoing invention purpose is as follows：
One lactobacillus plantarum (Lactobacillus plantarum) LP-ONLLY, the bacterial strain were isolated from 1989
Healthy babies, opportunistic pathogen strain code name are 8901, and its 16S rRNA Gene Partials sequence is AY590777 in GeneBank accession number.
Respectively on December 6th, 2004 and on March 28th, 2011 in China Committee for Culture Collection of Microorganisms's common micro-organisms
The heart (China General Microbiological Culture Collection Center, abbreviation CGMCC, address：North
One No. 13 Institute of Microorganism, Academia Sinica in Jing Shi Haidian District Zhong Guan-cun north, postcode：And French microorganism fungus kind 100080)
Collection (Collection Nationale De Cultures De Microorganisms, CNCM) preservation, preservation are stepped on
Mark is respectively CGMCC NO.1258 and CNCM I-4459.
Concrete technical scheme is as follows：
One lactobacillus plantarum, the Lactobacillus plantarum is in China Committee for Culture Collection of Microorganisms's common micro-organisms
The deposit number at center is CGMCC NO.1258.
The Lactobacillus plantarum includes aminopeptidase, beta galactosidase, glucuroide, and the aminopeptidase includes leucine
Amidase, Valinamide enzyme and cystine amidase,
The leucine amide enzyme, Valinamide enzyme, cystine amidase, beta galactosidase, glucuroide
Activity reaches more than 40nmols.
The curdled milk ability of the Lactobacillus plantarum is：In cow's milk, curdled milk in 16 hours.
Application of the Lactobacillus plantarum in active fermentation breast.
The Lactobacillus plantarum uses separately as leavening, and form is freeze-drying viable bacteria powder, and its viable count is more than 1.0
×1011CFU/g。
The preparation method of the active fermentation breast is as follows：Dispensing, homogeneous sterilizing, product fermentation, cooling and refrigeration；Wherein,
For temperature control at 37~42 DEG C, it is 70 ° of T~90 ° T that fermentation termination, which controls titratable acidity, when product ferments；
Raw material weight percentage in the dispensing, which forms, is：Cow's milk 84%~94%, nitrogen source 0.25%~1.0%, carbon
Source 5.75%~15%；The addition of the Lactobacillus plantarum is 1.0 × 106CFU/ml~1.0 × 107CFU/ml。
The nitrogen source is the one or more in casein hydrolysate, yeast extract, peptone, and the carbon source is white sand
One or more in sugar, glucose, fructose syrup.
The active fermentation breast is under cold chain preservation condition, and viable bacteria can stablize in 1,000,000,000 CFU/g in 1 month.
The Lactobacillus plantarum of the present invention and its active fermentation breast tool of preparation have the advantage that：With high aminopeptidase activity,
High betagalactosidase activity and high glucose glycosides enzymatic activity, it is freeze-dried viable bacteria powder, can be applied to as independent leavening
The preparation of active fermentation breast.The active fermentation breast not only contains high probiotics viable bacteria, and viable bacteria therein has high stability
Can, can probiotic supplemented, play Lactobacillus plantarum LP-ONLLY healthy functions.
Embodiment
The Lactobacillus plantarum LP-ONLLY of the present invention was isolated from healthy babies in 1989, and opportunistic pathogen strain code name is 8901, its
16S rRNA Gene Partials sequences are AY590777 in GeneBank accession number.Respectively on December 6th, 2004 and 2011 3
The moon 28 is in China Committee for Culture Collection of Microorganisms's common micro-organisms center (China General
Microbiological Culture Collection Center, CGMCC) and French Culture Collection
(Collection Nationale De Cultures De Microorganisms, CNCM) preservation, preservation registration number difference
For CGMCC NO.1258 and CNCM I-4459.
The Lactobacillus plantarum LP-ONLLY of embodiment 1 enzyme activity evaluation
Enzyme activity is the important indicator of leavening, using API ZYM kits (French Mei Liai companies) to Lactobacillus plantarum
LP-ONLLY and Laboratories Accession bacterial strain --- Lactobacillus plantarum LP6009 enzyme activity carry out semi-quantitative analysis.Grasped according to kit
Explain, collect the somatic cells grown in MRS nutrient solutions, prepare bacteria suspension with sterilized water analyzes for the enzyme activity of next step.
After adding thalline and reagent, the depth of color of reagent strip is divided into 0~5 totally 6 grades from shallow to deep, and 0 equivalent to negative reaction, and 5
For most strong reaction.Approximate thalline enzyme activity is understood by depth of color, color and luster 1~5 is respectively equivalent to the enzyme activity concentration of release
5nmoles, 10nmols, 20nmols, 30nmols and 40nmols.
As shown in Table 1, Lactobacillus plantarum LP-ONLLY has higher aminopeptidase and grape compared with Lactobacillus plantarum LP6009
Glycosidase activity, and both of which has higher betagalactosidase activity.
Aminopeptidase activity is in close relations to the maturation and flavor effect of dairy products；Beta galactosidase is mainly produced using lactose
Lactogenesis acid, reduces lactose content, promotes curdled milk；And glucuroide is then using glucose or Portugal's glucose class oligosaccharide etc., separately
Outside, also studies have found that, glucuroide is advantageous to lactobacillus synthesis exocellular polysaccharide, and makes acidified milk quality moist.
Conclusion：Lactobacillus plantarum LP-ONLLY high aminopeptidase is living, high beta galactosidase is living and high glucose glycosides enzyme activity,
The preparation of active fermentation dairy products can be applied to, so as to bring flavor and all good probiotic active of quality to consumer
Acidified milk.
The API ZYM kits enzyme activity of table 1 analyzes (representing enzyme activity with color and luster 0~5)
The Lactobacillus plantarum LP-ONLLY curdled milk abilities of embodiment 2 are evaluated
Curdled milk ability is another important performance of Dairy fermentation agent.Lactobacillus plantarum LP-ONLLY and LP6009 are pressed
107CFU/ml make an addition to respectively 10% skimmed milk power, the yeast extract of 10% skimmed milk power+0.25%, 10% skimmed milk power+
In 0.25% casein hydrolysate, the peptone of 10% skimmed milk power+0.25% and the glucose of 10% skimmed milk power+1%, in
37 DEG C of cultures, assess the curdled milk time.
As shown in Table 2, Lactobacillus plantarum LP6009 is no matter in any nutrient solution, can not in 16 hours curdled milk,
And Lactobacillus plantarum LP-ONLLY can be added with nitrogen source (such as addition yeast extract, casein hydrolysate and peptone)
Curdled milk in 10% skimmed milk power, there is quick curdled milk ability.
Conclusion：The more common bacterial strains of Lactobacillus plantarum LP-ONLLY, there is quick curdled milk ability, leavening can be further used as
Applied in active fermentation breast.And common bacterial strain Lactobacillus plantarum LP6009 can not be applied individually to any in active fermentation breast.
The curdled milk ability of table 2 is evaluated
(16 hours interior energy curdled milks, show quick curdled milk, with+represent；Otherwise it is curdled milk at a slow speed, with-represent)
The Lactobacillus plantarum LP-ONLLY of embodiment 3 is freeze-dried the preparation of viable bacteria powder
Lactobacillus plantarum LP-ONLLY produces bacterial strain, after primary seed solution and secondary seed solution culture, makes an addition to fermentation
Cultivation and fermentation in tank, Lactobacillus plantarum LP-ONLLY bacterium powders are made after centrifuging, being freeze-dried.Prepared suitable for active fermentation breast
Lactobacillus plantarum LP-ONLLY bacterium powders, viable count be more than 1.0 × 1011CFU/g。
The preparation (one) of the Lactobacillus plantarum LP-ONLLY active fermentations of embodiment 4 breast
Raw material forms：Cow's milk 84%, casein hydrolysate 1.0%, white granulated sugar 8%, glucose 7%, Lactobacillus plantarum
LP-ONLLY addition is 1.0 × 106CFU/ml。
After adding casein hydrolysate, white granulated sugar and glucose etc. in cow's milk, stir, homogeneous sterilizing is cooled to 37
After DEG C, by 1.0 × 106CFU/ml inoculum concentration adds Lactobacillus plantarum LP-ONLLY, after 37 DEG C of fermentations to 80 ° of T of titratable acidity,
It is cooled to 20 DEG C.Active fermentation breast after cooling is filling to packing container, and freezer preservation is transferred to rapidly.
Active fermentation breast, product are in the milky of uniformity, and structural state is uniform, and delicate mouthfeel is smooth, plant breast
Bacillus viable bacteria content 1.6 × 109CFU/g.And after being preserved 1 month under the conditions of 4 DEG C, viable bacteria content is 1.1 × 109CFU/g。
The preparation (two) of the Lactobacillus plantarum LP-ONLLY active fermentations of embodiment 5 breast
Raw material forms：Cow's milk 94%, peptone 0.25%, white granulated sugar 5.75%, Lactobacillus plantarum LP-ONLLY's adds
Dosage is 1.0 × 107CFU/ml。
After adding white granulated sugar in cow's milk, stir, after homogeneous sterilizing is cooled to 37 DEG C, by 1.0 × 107CFU/ml's
Inoculum concentration adds Lactobacillus plantarum LP-ONLLY, after 37 DEG C of fermentations to 90 ° of T of titratable acidity, is cooled to 20 DEG C.By the work after cooling
Property acidified milk it is filling to packing container, and be transferred to rapidly freezer preservation.
Active fermentation breast, product are in the milky of uniformity, and structural state is uniform, and delicate mouthfeel is smooth, plant breast
Bacillus viable bacteria content 2.1 × 109CFU/g.And after being preserved 1 month under the conditions of 4 DEG C, viable bacteria content is 1.3 × 109CFU/g。
The preparation (three) of the Lactobacillus plantarum LP-ONLLY active fermentations of embodiment 6 breast
Raw material forms：Cow's milk 93.25%, yeast extract 0.25%, peptone 0.5%, white granulated sugar 4%, high fructose corn
Slurry 2%, Lactobacillus plantarum LP-ONLLY addition is 5.0 × 106CFU/ml。
After adding yeast extract, peptone, white granulated sugar and fructose syrup in cow's milk, stir, homogeneous sterilizing is cold
But to after 37 DEG C, by 5.0 × 106CFU/ml inoculum concentration adds Lactobacillus plantarum LP-ONLLY, 37 DEG C of fermentations to titratable acidity
After 70 ° of T, 20 DEG C are cooled to.Active fermentation breast after cooling is filling to packing container, and freezer preservation is transferred to rapidly.
Active fermentation breast, product are in the milky of uniformity, and structural state is uniform, and delicate mouthfeel is smooth, plant breast
Bacillus viable bacteria content 1.9 × 109CFU/g.And after being preserved 1 month under the conditions of 4 DEG C, viable bacteria content is 1.2 × 109CFU/g。
Conclusion：Embodiment 4~6 shows that Lactobacillus plantarum LP-ONLLY can be used as independent leavening, applied to active fermentation
The preparation of breast, the active fermentation breast number of live bacteria of probiotics is more than 1,000,000,000 CFU/g, and is preserved one month under the conditions of cold chain, viable bacteria
Number can also be stablized in 1,000,000,000 CFU/g.The active fermentation breast can probiotic supplemented, play Lactobacillus plantarum LP-ONLLY healthy work(
Energy.
Claims (4)
1. a kind of applications of Lactobacillus plantarum (Lactobacillus plantarum) LP-ONLLY in active fermentation breast, its
It is characterised by, Lactobacillus plantarum (Lactobacillus plantarum) LP-ONLLY uses separately as leavening, shape
State is freeze-drying viable bacteria powder, and its viable count is more than 1.0 × 1011CFU/g；Lactobacillus plantarum (the Lactobacillus
Plantarum) LP-ONLLY is in the deposit number of China Committee for Culture Collection of Microorganisms's common micro-organisms center
CGMCC NO.1258。
2. application according to claim 1, it is characterised in that the preparation method of the active fermentation breast includes following step
Suddenly：Dispensing, homogeneous sterilizing, product fermentation, cooling and refrigeration；Wherein, temperature control is at 37~42 DEG C when product ferments, and fermentation is eventually
It is 70~90 ° of T that point, which controls titratable acidity,；
Raw material weight percentage in the dispensing, which forms, is：Cow's milk 84~94%, nitrogen source 0.25~1.0%, carbon source 5.75~
15%；The addition of Lactobacillus plantarum (Lactobacillus plantarum) LP-ONLLY is 1.0 × 106CFU/ml
~1.0 × 107CFU/ml。
3. application according to claim 2, it is characterised in that the nitrogen source is casein hydrolysate, yeast extract, egg
One or more in white peptone, the carbon source is the one or more in white granulated sugar, glucose, fructose syrup.
4. application according to claim 1 or 2, it is characterised in that the active fermentation breast is under cold chain preservation condition, and 1
Viable bacteria can stablize in 1,000,000,000 CFU/g in individual month.
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| CN104789511B (en) * | 2015-05-08 | 2016-02-03 | 内蒙古农业大学 | One strain has plant lactobacillus AB-2 and the application thereof of broad-spectrum antibacterial characteristic |
| CN104830731B (en) * | 2015-05-08 | 2016-02-03 | 内蒙古农业大学 | One strain has plant lactobacillus AB-1 and the application thereof of broad-spectrum antibacterial characteristic |
| CN105733996B (en) * | 2016-04-27 | 2019-05-17 | 中南民族大学 | A kind of extracellular polysaccharide and the lactobacillus plantarum CZ401 bacterial strain of beta galactosidase and application thereof |
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| CN108041385A (en) * | 2017-12-28 | 2018-05-18 | 上海交大昂立股份有限公司 | A kind of compound probiotic medicinal and edible plant fermented beverage and preparation method thereof |
| CN111035661A (en) * | 2018-10-15 | 2020-04-21 | 深圳市华大农业应用研究院 | Application of lactobacillus plantarum |
| CN109402000A (en) * | 2018-10-31 | 2019-03-01 | 南京师范大学 | One plant of production beta-glucosidase lactic acid bacteria and its screening technique and the preparation method rich in active flavone aglycone Yoghourt |
| CN110250270B (en) * | 2019-07-08 | 2022-03-04 | 北京工商大学 | A kind of method that utilizes Lactobacillus plantarum to improve fermented milk folic acid content |
| CN110760464B (en) * | 2019-11-15 | 2022-05-10 | 湖南农业大学 | Lactobacillus plantarum and application thereof |
| AU2023324639A1 (en) * | 2023-06-08 | 2025-01-02 | Bright Dairy & Food Co., Ltd | A Lactiplantibacillus plantarum strain, uses and dairy products thereof |
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