CN106399205A - Lactobacillus plantarum capable of realizing high-yield acetaldehyde and application thereof - Google Patents
Lactobacillus plantarum capable of realizing high-yield acetaldehyde and application thereof Download PDFInfo
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- IKHGUXGNUITLKF-UHFFFAOYSA-N Acetaldehyde Chemical compound CC=O IKHGUXGNUITLKF-UHFFFAOYSA-N 0.000 title claims abstract description 66
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- 238000000855 fermentation Methods 0.000 claims description 62
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- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 42
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- A23C9/00—Milk preparations; Milk powder or milk powder preparations
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- 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
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Abstract
The invention discloses lactobacillus plantarum capable of realizing high-yield acetaldehyde, with the preservation number of CCTCC M2016514. The invention also provides a working leavening agent containing the above lactobacillus plantarum. The invention also provides use of the lactobacillus plantarum capable of realizing the high-yield acetaldehyde in fermented food. Through detection and sensory evaluation on the acetaldehyde content in fermented milk, a result indicates that the lactobacillus plantarum disclosed by the invention has high yield of the acetaldehyde, with a higher score of sensory evaluation; flavor quality of the fermented milk is improved; and quality of the fermented milk is improved.
Description
Technical field
The invention belongs to microbiological art, it is related to a kind of Lactobacillus plantarum, specifically a kind of plant of high yield acetaldehyde
Thing lactobacilluss (Lactobacillus plantarum) and its application.
Background technology
Lactic acid bacteria (Lactic acid bacteria, LAB) is that a class can produce a large amount of lactic acid using fermentable sugar
Antibacterial general name, belongs to at least 23 on taxonomy on this kind of antibacterial that nature has been found that at present.In food, medicine and other fields
More lactic acid bacteria is applied mainly to have Lactobacillus, Streptococcus, Enterococcus, Lactococcus, Pediococcuses and leukonid
Belong to etc..Lactic acid bacteria is the topmost source of probiotic bacteria, and many lactic acid bacterias are the intrinsic probiotic bacterias of human body intestinal canal, have and improve people
Body intestinal microbial population, adjusts immunity of organisms, inhibiting cancer, reduce serum cholesterol, adjusts the important physiologically actives such as blood pressure.
Local flavor is one of key factor of fermentation milk quality responses, is also the key to fermentation milk degree of recognition for the impact consumer
Index.In fermentation milk, most representational flavor substance is acetaldehyde, but the utilization rate of acetaldehyde is relatively low, and its content is in 5-
In the range of 21ppm, the higher local flavor to fermentation milk of content is more favourable.Some researchs just think sub-standard weak fragrance fermentation milk
It is because wherein second ferment content is less than 4.0ppm.This content is also considered as up to standard critical of acetaldehyde in fermentation milk
Content, and the acetic acid content with the fermentation milk of excellent flavor can reach 8.0ppm (list of references Sandine W E, Daly
C,Elliker P R,et al.Causes and control of culture-related flavor defects in
cultured dairy products[J].Journal of Dairy Science,1972,55(7):1031-1039.).Cause
This improves the ability that lactic acid bacteria produces acetaldehyde, can improve the local flavor of yogurt from source, improves fermentation milk flavouring essence quality, is milk work
The production of industry provides certain directive significance.
Content of the invention
For above-mentioned technical problem of the prior art, the invention provides a kind of Lactobacillus plantarum of high yield acetaldehyde and its
Application, the described Lactobacillus plantarum of this high yield acetaldehyde and its application will solve the energy that lactic acid bacteria of the prior art produces acetaldehyde
Hypodynamic technical problem.
The invention provides a kind of Lactobacillus plantarum of high yield acetaldehyde, its deposit number is CCTCC M2016514.
Present invention also offers a kind of working stock culture, above-mentioned Lactobacillus plantarum strain is inoculated in culture in sterilized milk
To curdled milk, in continuous culture two generations of activation, are as mother culture；Mother culture is inoculated in interpolation milk surum by percent by volume 3-5%
Cultivate to curdled milk in the sterilized milk of albumen, obtain final product working stock culture.
Further, during culture is to curdled milk, cultivate 10-12h under the conditions of 35-37 DEG C.
Present invention also offers a kind of working stock culture, above-mentioned Lactobacillus plantarum strain is inoculated in fluid medium
In two generations of continuous activation, then activation culture thing is inoculated in culture, solid-liquid separation in fluid medium by percent by volume 3-5%
Obtain cell precipitation, precipitation is suspended with aseptic breast, obtains final product working stock culture, wherein, viable count is 109More than cfu/mL.
Further, during activating in liquid medium within, cultivate 10-12h under the conditions of 35-37 DEG C.
Present invention also offers a kind of purposes in fermented food for the Lactobacillus plantarum of above-mentioned high yield acetaldehyde.
Further, described fermented food is lactic acid bacteria milk beverage or fermentation milk.
Present invention also offers a kind of preparation method of above-mentioned lactobacillus beverage, cool down after raw milk is sterilized, then
Add above-mentioned working stock culture mix homogeneously, make above-mentioned Lactobacillus plantarum concentration reach 106More than cfu/mL, that is, contained
The lactic acid bacteria milk beverage of the Lactobacillus plantarum having the right described in requirement 1.
Present invention also offers a kind of preparation method of above-mentioned fermentation milk, cool down after raw milk is sterilized, add
The working stock culture stated and customary commercial leaven, the volume that described working stock culture accounts for fermentation milk is 3-5%, and described is normal
The volume that rule commodity leaven accounts for fermentation milk is 3-5%, ferments, to titratable acidity 70- after mixing under conditions of 35-37 DEG C
80 ° of T, that is, obtain the fermentation milk of above-mentioned Lactobacillus plantarum.
Further, described customary commercial leaven is Lactobacillus bulgaricus, bacillus bifiduss, streptococcus thermophiluss or thermophilic
Any one in Lactobacillus lactiss or two or more combinations.
Wherein, described sterilizing methods are conventional raw milk sterilizing methods, preferably as instantaneous ultrahigh-temperature sterilization, more preferably
As at 95 DEG C heating sterilization 20min.Described Lactobacillus plantarum working stock culture, cooling is added after breast cooling subject to sterilization
Conventional Temperature, is preferably cooled to 37-40 DEG C.Fermentation temperature is conventional, preferably 37 DEG C.
The assay method of acetaldehyde is that conventional headspace solid-phase microextraction combines gas chromatography combined with mass spectrometry (HS-SPME-GC-
MS method) is detected, preferably includes for the fermentation milk of Lactobacillus plantarum CCTCC M2016514 being placed in head space solid phase micro-
To be measured in extraction bottle.
Wherein, the condition determination of headspace solid-phase microextraction is：The above-mentioned fermentation milk of 5g-10g is placed in 20ml ml headspace bottle, and 55
30-40min is extracted, extracting head is placed in parsing 5min on GC-MS under DEG C -60 DEG C of waters bath with thermostatic control.
Wherein, GC-MS condition determination is：60m × 0.25mm × 0.25 μm HP-INNOWX capillary chromatograph (U.S. J＆W
Company)；50/30 μm of DVB/CAR/PDMS solid phase micro-extracting head (Supelco company of the U.S.)；60 DEG C of initial temperature, holding 3min, 5
DEG C/min is warming up to 230 DEG C, keeps 10min.
Lactobacillus plantarum (Lactobacillus plantarum) the bacterial strain CCTCC M2016514 of the present invention, it is biological
Scientific name is referred to as：Lactobacillus plantarum Z02 (Lactobacillus plantarum Z02), it belongs to Lactobacillus Lactobacillus plantarum
Kind.It is preserved in China typical culture collection center, preservation address in September in 2016 within 22nd：Wuhan City, Hubei Province Wuchang Luo Ka
Mountain Wuhan University collection.Postcode：430072, its deposit number is CCTCC M2016514.
The present invention compares with prior art, and its technological progress is significant.The Lactobacillus plantarum of the high yield acetaldehyde of the present invention
CCTCC M2016514, by being detected and subjective appreciation to the acetaldehyde in its fermentation milk, result shows the present invention's
Lactobacillus plantarum acetaldehyde yield is higher, and sensory evaluation scores are higher, improves the flavouring essence quality of fermentation milk, promotes carrying of fermentation milk quality
Height, has broad prospects in fermentation milk field.
Brief description
The feature of the explanation present invention and beneficial effect in conjunction with the following drawings.
Fig. 1 shows the colonial morphology of Lactobacillus plantarum CCTCC M2016514 of the present invention.
Fig. 2 shows the cellular morphology (× 1000) of Lactobacillus plantarum CCTCC M2016514 of the present invention.
Fig. 3 shows the growth curve of Lactobacillus plantarum CCTCC M2016514 of the present invention.
Fig. 4 shows the optimum growth temperature of Lactobacillus plantarum CCTCC M2016514 of the present invention.
Fig. 5 shows the fermentation milk GC-MS total ion current figure of Lactobacillus plantarum CCTCC M2016514 of the present invention.
Fig. 6 shows the fermentation yogurt of Lactobacillus plantarum CCTCC M2016514 of the present invention and the sense organ of blank yogurt
Score radar map.
Specific embodiment
The present invention gathers sample, the lactobacilli strain of screening high yield acetaldehyde from lactic acid bacteria habitat, studies it and produces acetaldehyde
Ability, develops new probiotic bacteria.Raw material used by the present invention or reagent are in addition to special instruction, all commercially available.
The present invention filters out a strains of lactic acid bacteria 1-34 from tibet koumiss, using the life of morphological characteristic, cultural colony and physiology
Change the Microbiological Characteristics such as feature and its inherited character 16s rDNA is accredited as Lactobacillus plantarum to this lactic acid bacteria 1-34
(Lactobacillus plantarum), this bacterial strain is preserved in China typical culture collection center on 22nd in September in 2016
(abbreviation CCTCC), its deposit number is CCTCC M2016514.
The morphological feature of Lactobacillus plantarum CCTCC M2016514 of the present invention：
Colony characteristicses：Bacterial strain separates in the flat lining out of MRS, 37 DEG C of culture 48h, and strain growth is good, its colonial morphology
As shown in Figure 1.Bacterium colony size 0.2-2mm, bacterium colony is circular, neat in edge, front microprotrusion, and the milky white middle band point of color is slightly yellow,
Opaque, surface wettability is smooth.
Thalline feature：Thalline is in shaft-like (Fig. 2), and thalline size is generally 1.5 μm of 0.6 μ m, does not produce spore, and gram contaminates
Color is positive.
The cultural characteristic of Lactobacillus plantarum CCTCC M2016514 of the present invention：Lactobacillus plantarum CCTCC M2016514 is
Low growth temperature is 15 DEG C, and maximum growth temperature is 45 DEG C, optimal in 30-40 DEG C of growth temperature；Bacterial strain CCTCC M2016514
Period of delay relatively short, 2h enters exponential phase, and 10h reaches stable phase.
The Lactobacillus plantarum CCTCC M2016514 of the present invention derives from traditional fermented food, belongs to generally recognized as safe
(Generally Recognized As Safe, GRAS) strain, can be used in lactobacillus food.
Therefore, the invention still further relates to purposes in fermented food for the described Lactobacillus plantarum CCTCC M2016514.Institute
State the fermented food containing Lactobacillus plantarum CCTCC M2016514 and include lactic acid bacteria milk beverage and fermentation milk.
The present invention also provides described Lactobacillus plantarum CCTCC M2016514 working stock culture.
Working stock culture of the present invention preferably adopts following preparation method preparations：By Lactobacillus plantarum CCTCC
M2016514 strain is inoculated in the skimmed milk of 12% (w/v), and under the conditions of 37 DEG C, culture 10-12h to curdled milk, live by continuous culture
Changed for two generations, use as mother culture；Mother culture is inoculated in above-mentioned sterilized milk by 3-5% (v/v), cultivates 10-12h
To curdled milk, now in curdled milk viable count about 109Cfu/mL, obtains described working stock culture；Or by Lactobacillus plantarum
CCTCC M2016514 strain is inoculated in MRS fluid medium, and under the conditions of 37 DEG C, culture 10-12h is activated, and continuously lives
Changed for two generations, then activation culture thing is inoculated in MRS culture medium by 3-5% (v/v), cultivate 10-12h, under the conditions of 4 DEG C
6000r/min is centrifuged 10min, removes supernatant, obtains cell precipitation, precipitation is suspended with a certain amount of sterile absorbent breast, obtains
To described working stock culture.
Preferably described lactic acid bacteria milk beverage in the present invention prepares as steps described below：Raw milk is at 95 DEG C
Lower heating sterilization 20min, is then cooled to 40 DEG C, adds described Lactobacillus plantarum CCTCC M2016514 work fermentation
Agent is so as to concentration reaches 106More than cfu/mL, obtains containing Lactobacillus plantarum CCTCC M2016514 in 4 DEG C of stored refrigerated
Lactic acid bacteria milk beverage.
In the present invention, preferably described fermentation milk prepares as steps described below：Raw milk heats at 95 DEG C
Sterilization 20min or at 140 DEG C high temperature thermal sterilization 2s, be then cooled to 37 DEG C, add described plant according still further to 3-5% (V/V)
Lactobacilluss CCTCC M2016514, adds 3-5% (V/V) fermentation milk commodity leaven, after mixing at 37 DEG C mixed fungus fermentation
To 70-80 ° of T of titratable acidity, it is subsequently cooled to 40 DEG C, then carries out stored refrigerated obtain containing Lactobacillus plantarum CCTCC
The fermentation milk of M2016514.
Further illustrate the present invention with embodiment below, but the present invention is not intended to be limited thereto.Do not note in the following example
The experimental technique of bright actual conditions, generally according to normal condition, or according to the condition proposed by manufacturer.Described in embodiment
" room temperature " refer to the temperature of operation room tested, generally 25 DEG C.
The collection of embodiment 1 Lactobacillus plantarum CCTCC M2016514, separation
(1), sample pretreatment
Take 3g tibet koumiss several pieces respectively, be respectively put into equipped with the 50ml conical flask of 15 sterilized water, after concussion, stand
20min, standby.
(2), lactobacilli strain separates
Using sterilized water by volume, according to 1：10 pairs of above-mentioned samples carry out serial dilution, take 0.1mL in each dilution factor
Dilute sample, is respectively coated MRS agar plate and M17 agar plate, constant temperature culture 24-48h under 37 DEG C of anaerobic conditions, with no
Bacterium toothpick picking is of different sizes, raised, micro white, moistens, neat in edge, and the bacterium colony back side is the single bacterium colony of yellow.Then in phase
On the agar plate answered, line point is pure, obtains pure single bacterium colony, carries out Gram’s staining, and catalase is tested.Purification bacterial strain preservation
In corresponding isolation medium, as protective agent, -20 DEG C frozen for the glycerol of interpolation 30%.
Used in it, culture medium prescription is as follows：
MRS culture medium (Bifidobacterium selective culture medium, Beijing overpass technical concern company limited buys).
M17 culture medium (Lactococcus selective medium, Beijing overpass technical concern company limited buys).
Different batches sample isolates 100 plants of bacterium on MRS agar culture medium and M17 agar culture medium.These bacterial strains
Show projection, micro white on separating flat board, moisten, neat in edge, the bacterium colony back side is yellow.
(3) different strains produce acetaldehyde ability and measure
The separation strains obtaining on flat board are inoculated into culture 12h in MRS fluid medium, then the inoculation by 5% (V/V)
Amount is inoculated into re-activation in fluid medium containing MRS, in 37 DEG C of 10h that ferment.By under fermentation liquid 6000rpm/min when 4 DEG C
Centrifugation 10min, takes thalline.With aseptic water washing 3 times, sterilized water is added to mix, standby.
Raw milk heat at 95 DEG C sterilization 20min or at 140 DEG C high temperature thermal sterilization 2s, be then cooled to 40 DEG C, then
Add bacteria suspension, add 5% (V/V) can symbiosis prepare fermentation milk commodity leaven, after mixing at 37 DEG C mixed fungus fermentation
To 70-80 ° of T of titratable acidity, it is subsequently cooled to room temperature, then carry out 4 DEG C of stored refrigerated.
The fermentation milk of above-mentioned cold preservation is weighed 5g, is placed in headspace solid-phase microextraction bottle (20ml).Ml headspace bottle is placed in
In 55 DEG C of thermostat water bath, insert aging in advance good extracting head (50/30 μm of DVB/CAR/PDMS), headspace absorption
40min, extraction terminates, and extracting head is transferred at GC-MS injection port, parses 5min, completes sample introduction.
Wherein, the fermentation milk of described Lactobacillus plantarum is conventional fermentation milk, preferably 37 DEG C condition bottom fermentation 10h and
Obtain fermentation milk.
Wherein, the condition determination of headspace solid-phase microextraction is：The above-mentioned fermentation milk of 5g is placed in 20ml ml headspace bottle, 55 DEG C of constant temperature
40min is extracted, extracting head is placed in parsing 5min on GC-MS under water-bath.
GC-MS, with 60 DEG C of initial temperature, keeps 3min, 5 DEG C/min to be warming up to 230 DEG C, keeps the ramp of 10min, complete
The detection of fermentation milk volatile compound, thus measure the content drawing acetaldehyde in fermentation milk.Experimental result is listed in Table 1 below.From table
In as can be seen that bacterial strain 1-34 acetaldehyde yield of a relatively high, choose this plant of bacterium, be named as CCTCC M2016514.
Table 1. produces the initial gross separation of acetaldehyde lactic acid bacteria
The identification of embodiment 2 Lactobacillus plantarum CCTCC M2016514
(1) physiological and biochemical test
Bacterial strain CCTCC M2016514 is Gram-positive, negative, the no motion of bacillus of peroxidase, at 15 DEG C and 45 DEG C
Can grow.
(2) the 16s rDNA sequence analysis of bacterial strain CCTCC M2016514
Bacterial strain CCTCC M2016514 genome DNA extracting method：The CCTCC M2016514 single bacterium colony of picking purification connects
Plant in 10mL MRS fluid medium, after 37 DEG C of culture 8h, bacterium solution is centrifuged (5000g, 10min) collects thalline.Using gene
Group DNA extraction agent box (Sangon Biotech (Shanghai) Co., Ltd.) is extracted.
PCR amplification is using the universal primer of two kinds of synthesis：
16s 27F:GAGAGTTTGATCCTGGCTCAG；
16s 1492R：CGGCTACCTTGTTACGACTT；
PCR primer adopts pillar PCR primer purification kit (Sangon Biotech (Shanghai) Co., Ltd.) to reclaim,
Sangon Biotech (Shanghai) Co., Ltd. is sent to be sequenced after purification.The 16s DNA core of obtained strains CCTCC M2016514
Nucleotide sequence is 1669bp (the SEQ ID NO in sequence table：1), send GenBank (GenBank accetion number：
CP002222) do Blast analysis.Bacterial strain CCTCC M2016514 homology highest bacterial strain is L.plantarum ST- III
(Sequence ID：CP002222.1), homology is 99%.
The G+C (mol%)≤10%～12% of the DNA according to described in Goodfellow and O ' Donnell and 16S rRNA
The kind of sequence homology >=95% can be classified as a genus, and Embley and Stackebrangdt think when 16S rRNA's
A kind is may be considered during sequence homology >=97%.Thus may infer that：Bacterial strain CCTCC M2016514 with
L.plantarum ST- III belongs to same kind.Bacterial strain CCTCC M2016514 is accredited as Lactobacillus plantarum.
According to the Microbiological Characteristics such as morphological characteristic, physiological and biochemical property and its inherited character 16s rDNA to lactic acid bacteria
CCTCC M2016514 is accredited as Lactobacillus plantarum (Lactobacillus plantarum), and this bacterial strain is in September 22 in 2016
Day is preserved in China typical culture collection center (abbreviation CCTCC), and its deposit number is CCTCC M2016514.
The growth characteristics of embodiment 3 CCTCC M2016514 bacterial strain
(1) drafting of CCTCC M2016514 strain growth curve
The Lactobacillus plantarum having activated CCTCC M2016514 is accessed MRS fluid medium by 5% (V/V) inoculum concentration
In, 37 DEG C of constant temperature culture 16h, measure the OD value of culture fluid every 1-2h in 600nm, be plotted against time with OD value and obtain bacterial strain
Growth curve in MRS for the CCTCC M2016514, its result (Fig. 3) shows：Lactobacillus plantarum CCTCC M2016514 is in MRS
Grow rapid in culture medium, in 2h about enter logarithmic (log) phase, 10h about enter stable phase.
(2) CCTCC M2016514 bacterial strain optimum growth temperature measures
The Lactobacillus plantarum having activated CCTCC M2016514 is connected to 10mL MRS liquid respectively by 5% (V/V) inoculum concentration
In body culture medium, it is respectively placed in constant temperature culture 8h under the conditions of 15 DEG C, 25 DEG C, 32 DEG C, 37 DEG C, 40 DEG C and 45 DEG C, with nonvaccinated
MRS fluid medium compares, and measures the OD value of the culture fluid of culture under different temperatures in 600nm, and the size according to OD value is true
Determine optimum growth temperature.Result shows：(Fig. 4) growth temperature range of Lactobacillus plantarum CCTCC M2016514 is wider, from 15
DEG C all grow to 45 DEG C, in 30 DEG C of -40 DEG C of well-growns, optimum growth temperature is 37 DEG C.
Embodiment 4 Lactobacillus plantarum CCTCC M2016514 produces acetaldehyde ability and measures
(1) actication of culture：Lactobacillus plantarum CCTCC M2016514 strain is inoculated in MRS fluid medium, 37
Under the conditions of DEG C, culture 10h is activated, two generations of continuous activation.
(2) preparation of working stock culture：Activation culture thing is inoculated in MRS culture medium by 5% (v/v), cultivates 10h,
Under the conditions of 4 DEG C 6000r/min centrifugation 10min, remove supernatant, obtain cell precipitation, by precipitation with a certain amount of aseptic take off
Fat breast suspends, and obtains described working stock culture.
(3) fermentation milk preparation：Raw milk heat at 95 DEG C sterilization 20min or at 140 DEG C high temperature thermal sterilization 2s, then
It is cooled to 40 DEG C, be subsequently adding working stock culture and the volume hundred of percent by volume 5% Lactobacillus plantarum CCTCC M2016514
Divide ratio 5% commodity leaven, mix after fermentation to 70-80 ° of T of titratable acidity, that is, obtain the fermentation containing described Lactobacillus plantarum
The fermentation milk of newborn CCTCC M2016514.
(4) acetaldehyde measures：The fermentation milk of above-mentioned cold preservation is weighed 5g, is placed in headspace solid-phase microextraction bottle (20ml)
In.Ml headspace bottle is placed in 55 DEG C of thermostat water bath, inserts aging in advance good extracting head (50/30 μm of DVB/CAR/
PDMS), headspace absorption 40min, extraction terminates, and extracting head is transferred at GC-MS injection port, parses 5min, completes sample introduction.
GC-MS, with 60 DEG C of initial temperature, keeps 3min, 5 DEG C/min to be warming up to 230 DEG C, keeps the ramp of 10min, complete the Olibanum that ferments
The detection of gasification compound, thus measuring the content drawing acetaldehyde in fermentation milk, total ion current figure is shown in Fig. 5.
Application Example 1 Lactobacillus plantarum CCTCC M2016514 working stock culture
12% (w/v) that Lactobacillus plantarum CCTCC M2016514 strain is inoculated in interpolation 1% lactalbumin (WPC) exists
In the skimmed milk of 115 DEG C of sterilizing 15min, under the conditions of 37 DEG C, to curdled milk, in continuous culture two generations of activation, as mother for culture 10-12h
Leaven uses；Mother culture is inoculated in above-mentioned sterilized milk by 5% (v/v), 10h is to curdled milk, now in curdled milk for culture
Viable count is about 109Cfu/mL, obtains the working stock culture (1) of the present invention.
Lactobacillus plantarum CCTCC M2016514 strain is inoculated in MRS fluid medium, cultivates under the conditions of 37 DEG C
10h is activated, and in two generations of continuous activation, is then inoculated in MRS fluid medium activation culture thing by 5% (v/v), culture
10h, under the conditions of 4 DEG C, 6000r/min centrifugation 10min, removes supernatant, obtains cell precipitation, and precipitation is used a certain amount of nothing
Bacterium skimmed milk suspends, and obtains the working stock culture (2) of the present invention.
The lactic acid bacteria milk beverage containing Lactobacillus plantarum CCTCC M2016514 for the Application Example 2
Raw milk heat at 95 DEG C sterilization 20min or at 140 DEG C high temperature thermal sterilization 2s, be then cooled to 40 DEG C, with
The amount of 5% (v/v) adds the Lactobacillus plantarum CCTCC M2016514 working stock culture of Application Example 1 gained【Work fermentation
Agent (1) or (2)】So as to concentration reaches 106More than cfu/m, obtains containing Lactobacillus plantarum CCTCC in 4 DEG C of stored refrigerated
The lactic acid bacteria milk beverage of M2016514.
The fermentation milk containing Lactobacillus plantarum CCTCC M2016514 for the Application Example 3
By raw milk fresh milk 95 DEG C of heat sterilization 20min or at 140 DEG C high temperature thermal sterilization 2s, be cooled to 40 DEG C,
Add the Lactobacillus plantarum CCTCC M2016514 working stock culture of Application Example 1 gained with the amount of 5% (v/v)【Work is sent out
Ferment agent (1) or (2)】, and commercial fermentation agent Lactobacillus bulgaricus, this mixed bacterium is fermented to titratable acidity at 37 DEG C and is 70 °
T, to 4 DEG C and stored refrigerated obtains the fermentation milk containing Lactobacillus plantarum CCTCC M2016514 for cold preservation.
The preparation of comparative example 1 blank run breast
By raw milk fresh milk after 95 DEG C of heat sterilization 20min, it is cooled to 37 DEG C, business is added with the amount of 5% (v/v)
Leaven Lactobacillus bulgaricus, fermenting to titratable acidity at 37 DEG C is 70 ° of T, and to 4 DEG C and stored refrigerated obtains blank for cold preservation
Fermentation milk.
The fermentation milk containing Lactobacillus plantarum CCTCC M2016514 for the effect example 1 and blank run breast acetaldehyde
Contrast
Fermentation milk in above-mentioned Application Example 3 and comparative example 1 is carried out second according to the method in embodiment 4 respectively
Aldehyde measures, and result shows that the acetaldehyde of the fermentation milk containing Lactobacillus plantarum CCTCC M2016514 is 8.75mg/L,
And the acetaldehyde in the blank run breast in comparative example 1 is 3.44mg/L, acetaldehyde improves about twice it was demonstrated that planting
Thing lactobacilluss CCTCC M2016514 has the ability of higher product acetaldehyde.
The fermentation milk containing Lactobacillus plantarum CCTCC M2016514 for the effect example 2 and blank run breast sensory evaluation
Contrast.
Carry out sensory evaluation by 8 valuation officers (3 men, 5 female), need before evaluation, to their professional trainings, to make them complete
Understand organoleptic attribute, know the evaluation criterion of fermentation milk.According to [list of references (Chen Jie. mesophilic digestion yogurt product Quality Research
[D]. Southern Yangtze University, 2008.)/(Kang Huan. the pre-test [D] of flavor substance and enzyme activity and organoleptic attribute relation in Yoghourt. northeast agriculture
Sparetime university is learned, and 2013.)], 9 organoleptic properties that may have significant contribution to the acceptability of product of initial option.Sensory evaluation
The corresponding definition of term is shown in Table 2.
Table 2 sensory evaluation term and standards of grading
First, take out the sample (sample in Application Example 2 and comparative example 1) of mark random code from refrigerator,
It is presented to valuation officer at random.After they open lid, be gently agitated for yogurt with spoon first, by smelling determine odor types and
Other properties are evaluated by intensity successively.Using 9 points of scaling laws.All of evaluation procedure is all having white light and controllable environment
Sensory evaluation laboratory in carry out.In evaluation procedure, prepare one glass of pure water and gargle.Each sample is in triplicate.Result is shown in
Fig. 6.
We can see that the fermentation milk of Lactobacillus plantarum CCTCC M2016514 in typical Yoghourt taste, creamy taste from Fig. 6
Higher than blank run breast, this is had relatively with Lactobacillus plantarum CCTCC M2016514 in effect example 1 with overall acceptance
The ability of high product acetaldehyde is consistent, and acetaldehyde has the fragrance of butter, and its presence can significantly improve the overall wind of fermentation milk
Taste feature, brings the sensory experience of more pleasure to consumer, can also draw this conclusion from sensory evaluation figure.
It should be understood that after the above having read the present invention, those skilled in the art can make various to the present invention
The present invention is made various changes or modifications, these equivalent form of values equally fall within the model that the application appended claims are limited
Enclose.
Sequence table
<110>Shanghai application technology university
<120>A kind of Lactobacillus plantarum of high yield acetaldehyde and its application
<160> 3
<170> PatentIn version 3.5
<210> 1
<211> 1669
<212> DNA
<213> lactobacillus plantarum
<400> 1
aaactggtcg taaatacgac tcactatagg gcgacatatg atcgatgata tcccatgggc 60
ggccgcctgc agaccaggtc tggttacctt gttacgactt caccctaatc atctgtccca 120
ccttaggcgg ctggttccta aaaggttacc ccaccgactt tgggtgttac aaactctcat 180
ggtgtgacgg gcggtgtgta caaggcccgg gaacgtattc accgcggcat gctgatccgc 240
gattactagc gattccgact tcatgtaggc gagttgcagc ctacaatccg aactgagaat 300
ggctttaaga gattagctta ctctcgcgag ttcgcaactc gttgtaccat ccattgtagc 360
acgtgtgtag cccaggtcat aaggggcatg atgatttgac gtcatcccca ccttcctccg 420
gtttgtcacc ggcagtctca ccagagtgcc caacttaatg ctggcaactg ataataaggg 480
ttgcgctcgt tgcgggactt aacccaacat ctcacgacac gagctgacga caaccatgca 540
ccacctgtat ccatgtcccc gaagggaacg tctaatctct tagatttgca tagtatgtca 600
agacctggta aggttcttcg cgtagcttcg aattaaacca catgctccac cgcttgtgcg 660
ggcccccgtc aattcctttg agtttcagcc ttgcggccgt actccccagg cggaatgctt 720
aatgcgttag ctgcagcact gaagggcgga aaccctccaa cacttagcat tcatcgttta 780
cggtatggac taccagggta tctaatcctg tttgctaccc atactttcga gcctcagcgt 840
cagttacaga ccagacagcc gccttcgcca ctggtgttct tccatatatc tacgcatttc 900
accgctacac atggagttcc actgtcctct tctgcactca agtttcccag tttccgatgc 960
acttcttcgg ttgagccgaa ggctttcaca tcagacttaa aaaaccgcct gcgctcgctt 1020
tacgcccaat aaatccggac aacgcttgcc acctacgtat taccgcggct gctggcacgt 1080
agttagccgt ggctttctgg ttaaataccg tcaatacctg aacagttact ctcagatatg 1140
ttcttcttta acaacagagt tttacgagcc gaaacccttc ttcactcacg cggcgttgct 1200
ccatcagact ttcgtccatt gtggaagatt ccctactgct gcctcccgta ggagtttggg 1260
ccgtgtctca gtcccaatgt ggccgattac cctctcaggt cggctacgta tcattgccat 1320
ggtgagccgt taccccacca tctagctaat acgccgcggg accatccaaa agtgatagcc 1380
gaagccatct ttcaaacttg gaccatgcgg tccaagttgt tatgcggtat tagcatctgt 1440
ttccaggtgt tatcccccgc ttctgggcag gtttcccacg tgttactcac cagttcgcca 1500
cttactcaaa tgtaaatcat gatgcaagca ccaatcaata ccagagttcg ttcgacttgc 1560
atgtattagg cacgccgcca gcgttcgtcc tgagccagga tcaaactcta agactggaga 1620
tctggatccc tcgagtctag agtcgacctg caggcatgca aaggtttgg 1669
<210> 2
<211> 21
<212> DNA
<213>Primer 16s 27f
<400> 2
gagagtttga tcctggctca g 21
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<212> DNA
<213>Primer 16s 1492r
<400> 3
cggctacctt gttacgactt 20
Claims (10)
1. a kind of Lactobacillus plantarum of high yield acetaldehyde it is characterised in that：Its deposit number is CCTCC M2016514.
2. a kind of working stock culture it is characterised in that：Lactobacillus plantarum strain described in claim 1 is inoculated in sterilized milk
In cultivate to curdled milk, in continuous culture two generations of activation, are as mother culture；Mother culture is inoculated in by percent by volume 3-5% and adds
Plus cultivate to curdled milk in the sterilized milk of lactalbumin, obtain final product working stock culture.
3. a kind of working stock culture according to claim 2 it is characterised in that：During culture is to curdled milk, in 35-
10-12h is cultivated under the conditions of 37 DEG C.
4. a kind of working stock culture it is characterised in that：Lactobacillus plantarum strain described in claim 1 is inoculated in liquid culture
In two generations of continuous activation in base, then activation culture thing is inoculated in culture, solid-liquid in fluid medium by percent by volume 3-5%
Separate and obtain cell precipitation, precipitation is suspended with aseptic breast, obtains final product working stock culture, wherein, viable count is 109Cfu/mL with
On.
5. a kind of working stock culture according to claim 4 it is characterised in that：The process of activation in liquid medium within
In, cultivate 10-12h under the conditions of 35-37 DEG C.
6. purposes in fermented food for the Lactobacillus plantarum of a kind of high yield acetaldehyde described in claim 1.
7. the purposes according to claim 4 it is characterised in that：Described fermented food is lactic acid bacteria milk beverage or sends out
Kefir milk.
8. a kind of preparation method of lactobacillus beverage it is characterised in that：Cool down after raw milk is sterilized, be subsequently adding claim
Working stock culture mix homogeneously described in 2 or 4, makes the Lactobacillus plantarum concentration described in claim 1 reach 106cfu/mL
More than, that is, obtain the lactic acid bacteria milk beverage containing the Lactobacillus plantarum described in claim 1.
9. a kind of preparation method of fermentation milk it is characterised in that：After raw milk is sterilized cool down, add claim 2 or
Working stock culture described in person 4 and customary commercial leaven, the volume that described working stock culture accounts for fermentation milk is 3-5%, described
Customary commercial leaven account for fermentation milk volume be 3-5%, after mixing under conditions of 35-37 DEG C ferment, to titratable acidity
70-80 T, that is, obtain the fermentation milk containing Lactobacillus plantarum described in claim 1.
10. a kind of fermentation milk according to claim 9 preparation method it is characterised in that：Described commodity leaven is
In conventional Lactobacillus bulgaricus, bacillus bifiduss, streptococcus thermophiluss or bacillus acidophilus any one or two or more
Combination.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102453689A (en) * | 2011-12-06 | 2012-05-16 | 光明乳业股份有限公司 | Lactobacillus plantarum strain producing extracellular polysaccharide, and application thereof |
| CN106085901A (en) * | 2016-06-08 | 2016-11-09 | 江南大学 | A kind of Lactobacillus plantarum with biacetyl synthesis capability and application thereof |
-
2016
- 2016-11-28 CN CN201611063111.2A patent/CN106399205A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102453689A (en) * | 2011-12-06 | 2012-05-16 | 光明乳业股份有限公司 | Lactobacillus plantarum strain producing extracellular polysaccharide, and application thereof |
| CN106085901A (en) * | 2016-06-08 | 2016-11-09 | 江南大学 | A kind of Lactobacillus plantarum with biacetyl synthesis capability and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| MARYAM GHOBADI DANA1: "High-level acetaldehyde production by an indigenous Lactobacillus strain obtained from traditional dairy products of Iran", 《AFRICAN JOURNAL OF MICROBIOLOGY RESEARCH》 * |
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