CN108949645A - Lactobacillus plantarum CQ02-108 and its application in ferment sausage preparation - Google Patents
Lactobacillus plantarum CQ02-108 and its application in ferment sausage preparation Download PDFInfo
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- CN108949645A CN108949645A CN201811111254.5A CN201811111254A CN108949645A CN 108949645 A CN108949645 A CN 108949645A CN 201811111254 A CN201811111254 A CN 201811111254A CN 108949645 A CN108949645 A CN 108949645A
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- 238000002360 preparation method Methods 0.000 title description 9
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- 238000001035 drying Methods 0.000 claims description 7
- 230000005070 ripening Effects 0.000 claims description 7
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- 239000003795 chemical substances by application Substances 0.000 claims description 5
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- 244000203593 Piper nigrum Species 0.000 claims description 4
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- 229930006000 Sucrose Natural products 0.000 claims description 4
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- 241000295644 Staphylococcaceae Species 0.000 description 1
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Classifications
-
- 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
-
- A—HUMAN NECESSITIES
- A22—BUTCHERING; MEAT TREATMENT; PROCESSING POULTRY OR FISH
- A22C—PROCESSING MEAT, POULTRY, OR FISH
- A22C11/00—Sausage making ; Apparatus for handling or conveying sausage products during manufacture
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B4/00—General methods for preserving meat, sausages, fish or fish products
- A23B4/14—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12
- A23B4/18—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12 in the form of liquids or solids
- A23B4/20—Organic compounds; Microorganisms; Enzymes
- A23B4/22—Microorganisms; Enzymes; Antibiotics
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
- A23L13/70—Tenderised or flavoured meat pieces; Macerating or marinating solutions specially adapted therefor
- A23L13/72—Tenderised or flavoured meat pieces; Macerating or marinating solutions specially adapted therefor using additives, e.g. by injection of solutions
- A23L13/74—Tenderised or flavoured meat pieces; Macerating or marinating solutions specially adapted therefor using additives, e.g. by injection of solutions using microorganisms or enzymes
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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
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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
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- 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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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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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- Polymers & Plastics (AREA)
- Nutrition Science (AREA)
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Abstract
The invention discloses a lactobacillus plantarum CQ02-108 and its preparing the application in ferment sausage.Lactobacillus plantarum (Lactobacillus plantarum) CQ02-108 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on July 17th, 2018, and deposit number is CGMCC No.16121.The lactobacillus plantarum CQ02-108 can produce protease, quickly produce acid, do not produce it is viscous, do not produce gas, non-generation amine, do not produce H2O2, do not produce H2S, not chromogenic element, tolerance high salt concentration, tolerance highly acidity, growth, the bile tolerance for inhibiting Escherichia coli and staphylococcus aureus, are resistant to gastric juice.When applied to ferment sausage, the quantity of viable lactic acid bacteria in ferment sausage can be significantly improved, reduces the pH value of sausage, inhibits the growth of harmful bacteria, the quality of ferment sausage can be obviously improved.
Description
Technical field
The invention belongs to microorganisms technical fields, and in particular to lactobacillus plantarum and its answering in ferment sausage preparation
With.
Background technique
Ferment sausage, which refers to, to be filled after the mixing such as minced steak and animal tallow, salt, sugar, leavening and spice into intestines
Clothing undergoes microbial fermentation and manufactured fermentation meat product under the conditions of artificial or natural contral.The traditional Fermented Sausages in China are not
The step of carrying out artificial addition leavening, completes to be fermented into a natural environment mainly by the microbial flora of raw meat itself
It is ripe, there is unique flavor.Therefore fermentation period is long, and is easy to be limited by weather, and the safety and quality of product are difficult to
It is protected.
The microbial flora of traditional zymotic sausage is mainly by lactic acid bacteria, coagulase-negative staphylococci, yeast and mould group
At wherein lactic acid bacteria plays an important role in meat fermentation and preservation, is the major microorganisms for participating in fermentation.Pathogenic bacteria are to influence
The one of the major reasons of Safety of Fermented Sausage, and lactic acid bacteria can generate the pH value that lactic acid reduces rapidly product, and part cream
Sour bacterium can generate bacteriocin, this plays inhibiting effect to the growth of pathogenic bacteria and spoilage organisms.Use the lactic acid bacteria of bacteriocinogeny
Extend the shelf-life of ferment sausage as leavening and guarantees that its safety has become a kind of trend.The generation of lactic acid can also
Accelerate conversion of the nitrite to nitric oxide myoglobins, reduce the residual quantity of nitrite in product, while promoting good
The formation in lustful pool.On the other hand, lactic acid bacteria can assign the distinctive flavor of product and solid quality.Therefore, lactic acid bacteria
Screening study is of great significance to the exploitation of fermentation meat product.
In addition, usually salt and nitrite are added in fermentation meat product to guarantee matter when making ferment sausage
Amount improves flavor etc..And during the fermenting-ripening of sausage, with the continuous reduction of pH, moisture content, Aw also constantly drop
Low, salinity then constantly increases.Therefore, screen one plant and be resistant to high salt and highly acidity, can bacteriocinogeny to inhibiting to cause
The lactic acid bacteria of pathogen growth, and be applied in ferment sausage, have great importance to the processing of sausage.
Summary of the invention
The present invention is in view of the above problems, provide a lactobacillus plantarum and its application in ferment sausage.It will be from biography
What is screened in system spontaneous fermentation sausage is resistant to high salt concentration, highly acidity, inhibits pathogenic bacteria growing plants lactobacillus
It is applied in ferment sausage, improves the color of sausage, improve the quality of sausage.
The purpose of the present invention is be achieved through the following technical solutions:
The first purpose of the invention is to provide a lactobacillus plantarum CQ02-108, classification naming is lactobacillus plantarum
It is general to be preserved in China Committee for Culture Collection of Microorganisms on July 17th, 2018 by Lactobacillus plantarum
Logical microorganism center, deposit number are CGMCC No.16121.
In one embodiment of the invention, the lactobacillus plantarum CQ02-108 produces protease, quickly produces acid, do not produce
It glues, does not produce gas, non-generation amine, do not produce H2O2, do not produce H2S, not chromogenic element.
In one embodiment of the invention, the lactobacillus plantarum CQ02-108 is resistant to high salt concentration, peracid
Degree.
In one embodiment of the invention, the lactobacillus plantarum CQ02-108 is able to suppress Escherichia coli, golden yellow
The staphylococcic growth of color.
In one embodiment of the invention, the lactobacillus plantarum CQ02-108 is resistant to cholate and gastric juice.
A second object of the present invention is to provide a kind of microbial bacterial agents comprising afore-mentioned plants lactobacillus CQ02-108.
Further, the microbial bacterial agent is solid-state microbial inoculum or liquid microbial inoculum.
Third object of the present invention is to provide a kind of function leavenings comprising afore-mentioned plants lactobacillus CQ02-108.
Further, the function leavening is 37 DEG C of trainings in MRS meat soup by lactobacillus plantarum CQ02-108 above-mentioned
It supports for 24 hours, 6000g is centrifuged 10min at 4 DEG C, collects precipitating sterile water washing 3 times and obtained bacteria suspension is resuspended.
Fourth object of the present invention is to provide afore-mentioned plants lactobacillus CQ02-108 or function leavening above-mentioned is being made
Make the application in fermentation meat product.
Further, the fermentation meat product is ferment sausage.
Further, it is described application the following steps are included:
S1: being added seasoning in raw meat and auxiliary material is placed in 4 DEG C and pickles；
S2: function leavening above-mentioned being inoculated into raw meat and is mixed, and records sausage, and the inoculum concentration is 105-
108Cfu/g, preferably 107cfu/g；
S3: it fermentation: 25 DEG C~35 DEG C, ferments 1 day under the conditions of RH 80%；
S4: it drying and ripening: 15 DEG C, places 4 days under the conditions of RH 75%；16 days are dried under the conditions of 12 DEG C, RH 72% extremely
Sausage fermentation is mature；
Further, above-mentioned steps further include
S5: sausage is trimmed, and finished product is obtained after vacuum packaging.
Further, raw meat described in S1 is fresh pig back leg lean meat and pig back fat, by the fascia on its surface and big
The positions such as tendon are rinsed after rejecting, and mix for 8:2 according to the ratio of pig back leg lean meat and back fat after meat is blended
It arrives；
The seasoning and auxiliary material are added according to the following mass percent of raw meat: salt 2%, sucrose 1%, grape
Sugared 1%, sodium nitrite 0.015%, sodium isoascorbate 0.05%, ginger powder 0.1%, white pepper powder 0.1%, five-spice powder 0.1%.
The proportion of the seasoning and auxiliary material combines the use of lactobacillus plantarum CQ02-108 leavening of the present invention, ferment sausage can be made to have
Excellent organoleptic attribute.
Further, fermentation temperature described in S3 is 30 DEG C.
Further, the function leavening is able to suppress Escherichia coli, staphylococcus aureus breeds in food, and
It can be bred in cholate, simulate the gastric juice.
Further, afore-mentioned plants lactobacillus CQ02-108 or function leavening above-mentioned are in production fermentation meat product
Using being to improve product quality using the characteristics such as lactobacillus plantarum CQ02-108 salt tolerant, acidproof, antibacterial, quick production acid.
Beneficial effects of the present invention:
(1) lactobacillus plantarum of the invention not only meets except the basic standard of starter of meat products, moreover it is possible to be resistant to highly concentrated
Salt, tolerance highly acidity, bacteriocinogeny effectively inhibit the growth of pathogenic bacteria, while also with the feature of bile tolerance resistant to gastric juice, to function
The exploitation of energy property starter of meat products provides certain theoretical foundation.
(2) present invention is by addition lactobacillus plantarum, and lactic acid bacteria number is still up to 10 at the end of sausage maturation7, significantly mention
The quantity of lactic acid bacteria in high ferment sausage inhibits the growth of Escherichia coli and staphylococcus aureus.
(3) lactobacillus plantarum of the invention makes ferment sausage as leavening, effectively improves the wind of ferment sausage
Taste and quality.
(4) it with traditional zymotic sausage ratio, is inoculated with lactobacillus plantarum CQ02-108 of the invention sausage is effectively reduced and producing
PH in the process reduces the risk of sausage corruption and mildew, improves the safety of product.
Detailed description of the invention
Fig. 1 is the growth curve of lactobacillus plantarum CQ02-108
Fig. 2 is lactobacillus plantarum CQ02-108 growing state under different salinity
Fig. 3 is lactobacillus plantarum CQ02-108 growing state under different pH degree
Fig. 4 is ferment sausage pH situation of change
Fig. 5 is lactic acid bacteria number situation of change in ferment sausage
Fig. 6 is total plate count situation of change in ferment sausage
Biological deposits information explanation
One lactobacillus plantarum (Lactobacillus plantarum) CQ02-108, in preservation on July 17 in 2018
In China Committee for Culture Collection of Microorganisms's common micro-organisms center, classification naming is lactobacillus plantarum
Lactobacillus plantarum, deposit number are CGMCC No.16121.Depositary institution address: Chaoyang District, Beijing City north
The institute 3 of occasion West Road 1, research institute of the Chinese Academy of Sciences；Postcode: 100101.
Specific embodiment
The present invention will be further described combined with specific embodiments below:
Embodiment 1: the screening of bacterial strain
The specific implementation step of screening technique is as follows:
1, sample acquires
Primary sample picks up from the traditional natural ferment sausage of Chongqing City food market purchase.
2, the separating lactic acid bacterium from sample
25g sausage sample is aseptically shredded, the mixing of 225mL sterile saline is dissolved in, stands several minutes, it is broken
After meat precipitates completely, takes supernatant to be gradually incremented by dilution by 1:10, the bacterium solution 100ul of suitable dilution is selected to be spread evenly across
It is added to the MRS solid medium of calcium carbonate, cultivates 48h in 37 DEG C of anaerobic culture boxes.On picking plate different shape, have
Molten calcium circle and the independent single bacterium colony for meeting lactic acid bacteria colony morphology characteristic, cross on MRS culture medium, until obtaining pure
Until culture.Picking colonies typical carries out Gram's staining and catalase experiment, obtains 20 plants of Gram-positives and connects
The bacterial strain of catalase feminine gender.
3, the primary dcreening operation of bacterial strain
20 plants of bacterium of above-mentioned separation are carried out to produce acid respectively, protease, resistance to nitrite, produce hydrogen sulfide, generation amine, nitre
Sour reductase, produce gas, produce mucus, it is chromogenic element test, finally obtain can quickly produce acid, produce protease, resistance to nitrite and
It produces hydrogen sulfide, biogenic amine, nitrate reductase, produce totally 3 plants of bacterial strain that gas, mucus, pigment are negative, number is CQ02- respectively
108、CQ02-117、CQ02-120。
4, the secondary screening of bacterial strain
3 plants of bacterium of above-mentioned gained are subjected to bacteriostatic test, are measured using bacteriostasis of the Odontothrips loti to bacterial strain.Instruction
Bacterium is Escherichia coli and staphylococcus aureus.The supernatant bacterium solution of lactic acid bacteria stationary phase is collected, supernatant reuses 0.22 μm
Filtering with microporous membrane degerming.By the indicator bacteria (10 after 3 generations of activation6Cfu/mL) it is spread evenly across nutrient agar table
Face places 4 sterile Oxford cups in culture dish moderate distance, 200 μ L test strain supernatants, blank is added in each Oxford cup
Control is fluid nutrient medium, after 4 DEG C of diffusion 6h around 37 DEG C of culture 48h, observation Oxford cup inhibition zone situation and antibacterial to its
Loop diameter, which measures, calculates its average value.Final result is as shown in table 1, and CQ02-108 is relative to CQ02-117 and CQ02-120
Two plants of bacterium fungistatic effects are best, therefore finally obtain CQ02-108.
The antibacterial result of 1 different strains of table
Note: result is expressed as average ± standard deviation, n=4；It is significant difference (p < that same column, which marks different lowercases,
0.05)。
5, the identification of strain
By the CQ02-108 filtered out in the flat lining out of MRS, picking single colonie utilizes universal amplification 16SrRNA gene,
Use the universal primer of amplification bacteria 16 S rRNA genes: upstream primer 27F, downstream primer 1492R.PCR condition are as follows: 94
DEG C initial denaturation 5min；94 DEG C of denaturation 1min, 55 DEG C of annealing 1min, 72 DEG C of extension 90s, 30 recycle；72 DEG C of final extensions
10min。
PCR product send Sangon Biotech (Shanghai) Co., Ltd. to be sequenced, sequence after the detection of 1% agarose gel electrophoresis
Column acquired results are compared with known array in ncbi database.Determine that the bacterial strain is lactobacillus plantarum (Lactobacillus
Plantarum), similarity 100%.The bacterium has been preserved in Chinese microorganism strain preservation management committee on July 17th, 2018
Member's meeting common micro-organisms center, deposit number are CGMCC No.16121.Depositary institution address: BeiChen West Road, Chaoyang District, BeiJing City
No. 1 institute 3, research institute of the Chinese Academy of Sciences.
Embodiment 2: lactobacillus plantarum CQ02-108 bile tolerance test
The bacterial suspension of strain to be tested is adjusted to 109CFU/mL.It is inoculated into and contains respectively according to the ratio of 1% (V/V)
In the MRS meat soup for having 0.1%, 0.3% cholate, control group is made of the MRS meat soup without cholate.The Anaerobic culturel 24 at 37 DEG C
Hour, by spectrophotometer with 0.5h interval measurement OD620.The Bile salt resistance of each bacterial strain is based on OD620 value and increases by 0.3
Time needed for a unit.
The results are shown in Table 2, and CQ02-108 OD620 in 0.1% cholate and 0.3% cholate increases by 0.3 unit
The required time is substantially less than remaining two plants of bacterium, shows that the tolerance cholate ability of CQ02-108 is most strong.
The cholate resistance results of 2 different strains of table
Note: same column marks different lowercase letter indication differences significantly (p < 0.05), n=3.
Embodiment 3: lactobacillus plantarum CQ02-108 resistant to gastric juice test
The preparation of simulate the gastric juice: 0.35g pepsin is added in 0.2% Sterile Saline of 100mL, with dense HCl by pH
It is spare to be adjusted to 3.0 filtration sterilizations.
By strain to be tested in 37 DEG C of overnight incubations in MRS meat soup, then it is inoculated into 5mL gastric juice simultaneously according to 1% amount
It is incubated for 3 hours at 37 DEG C.After digesting in the gastric juice of simulation, the viable count (log of every kind of bacterial strain is measured by colony counting method
10cfu/mL), and survival rate, survival rate (%)=3h viable count/0h viable count × 100% are calculated.
The results are shown in Table 3, and three plants of bacterium all have good gastric fluid-resistant, by 3h simulation digest, viable count according to
10 can so be reached7Cfu/ml, wherein the survival rate of CQ02-108 is significantly higher than remaining two plants of bacterium, shows the tolerance stomach of CQ02-108
The ability of liquid is most strong.
The gastric fluid-resistant situation of 3 different strains of table
Note: result is expressed as average ± standard deviation, n=3；Same column marks significant (the p < of different lowercase letter indication differences
0.05)。
Embodiment 4: the Salt tolerance of lactobacillus plantarum CQ02-108
1, the measurement of lactobacillus plantarum CQ02-108 growth curve
Lactobacillus plantarum CQ02-108 is inoculated in MRS fluid nutrient medium, 37 DEG C of culture 48h, every 2h light splitting light
Degree meter measures its light absorption value at wavelength 600nm, while being compared with blank MRS fluid nutrient medium.As a result as shown in Figure 1.
2, Salt tolerance:
Lactobacillus plantarum is inoculated into the MRS liquid that NaCl concentration is 3%, 6%, 9%, 12%, 15% by 2% respectively to train
It supports in base, is cultivated at 37 DEG C for 24 hours, repeat 6 using blank cultures as control with light absorption value at spectrophotometric determination 600nm
It is secondary.
As a result as shown in Fig. 2, salt additive amount is generally 2%~3% in ferment sausage, with the reduction of moisture content,
The concentration of salt can be increased constantly, will receive inhibition to the growth of the lactic acid bacteria of salt density value.Lactobacillus plantarum CQ02- in this experiment
108 when NaCl concentration is 3%, 6%, 9% well-grown, and can also be given birth in 12%, 15% MRS fluid nutrient medium
Long, lactobacillus plantarum CQ02-108 of the invention has very strong salt-resistant type, it is made to become dominant bacteria during the fermentation.
Embodiment 5: the acid resistance test of lactobacillus plantarum CQ02-108
Lactobacillus plantarum is inoculated into the MRS fluid nutrient medium that pH is 2.5,3.5,4.5,5.5,6.5 by 2%, 37 DEG C
For 24 hours, with light absorption value at spectrophotometric determination 600nm, the blank cultures to correspond to pH are repeated 6 times as control for lower culture.
As a result as shown in figure 3, lactobacillus plantarum CQ02-108 can be grown at different pH, and in pH4.5,5.5,6.5
Lower well-grown.At fermentation initial stage, lactic acid bacteria can produce rapidly acid, so that the pH of product declines to a great extent, therefore lactobacillus plantarum energy
Enough tolerance highly acidity make it become dominant bacteria during the fermentation.
Embodiment 6: the preparation (test group) of ferment sausage
The preparation of 1 leavening: for 24 hours, 6000g is centrifuged lactobacillus plantarum CQ02-108 at 4 DEG C for 37 DEG C of cultures in MRS meat soup
10min collects precipitating sterile water washing 3 times and is resuspended that obtain bacteria suspension stand-by.The lactobacillus plantarum CQ02-108
The deposit number of (Lactobacillus plantarum CQ02-108) is CGMCC No.16121.
The preparation of 2 ferment sausages
The fresh pig back leg lean meat of 2.1 selections and pig back fat, the positions such as the fascia on its surface and big tendon are rejected laggard
Row rinsing mixes after blending meat according to the ratio of pig back leg lean meat and back fat for 8:2, and seasoning is added and auxiliary material is placed in
4 DEG C marinated.The seasoning and auxiliary material are re-added by meat: salt 2%, sucrose 1%, glucose 1%, sodium nitrite 0.015%,
Sodium isoascorbate 0.05%, ginger powder 0.1%, white pepper powder 0.1%, five-spice powder 0.1%.
2.2 mix bacterial suspension inoculation described in 2.1 into meat, record sausage, and the inoculum concentration is 107cfu/g。
2.3 fermentations: it 30 DEG C, ferments 1 day under the conditions of RH 80%.
2.4 drying and ripenings: it 15 DEG C, places 4 days under the conditions of RH 75%；16 days are dried under the conditions of 12 DEG C, RH 72% extremely
Sausage fermentation is mature.
2.5 trim sausage, and finished product is obtained after vacuum packaging.
Embodiment 7: the preparation (control group) of ferment sausage
(1) fresh pig back leg lean meat and pig back fat are selected, the positions such as the fascia on its surface and big tendon are rejected laggard
Row rinsing mixes after blending meat according to the ratio of pig back leg lean meat and back fat for 8:2, and seasoning is added and auxiliary material is placed in
4 DEG C marinated.The seasoning and auxiliary material are re-added by meat: salt 2%, sucrose 1%, glucose 1%, sodium nitrite 0.015%,
Sodium isoascorbate 0.05%, ginger powder 0.1%, white pepper powder 0.1%, five-spice powder 0.1%.
(2) it ferments: 30 DEG C, fermenting 1 day under the conditions of RH 80%.
(3) it drying and ripening: 15 DEG C, places 4 days under the conditions of RH 75%；16 days are dried under the conditions of 12 DEG C, RH 72% extremely
Sausage fermentation is mature.
(4) sausage is trimmed, finished product is obtained after vacuum packaging.
Embodiment 8: the measurement of ferment sausage pH value
Ferment sausage prepared by embodiment 6 and embodiment 7 respectively fermentation, drying and ripening during the 1st day, the 7th
It, the 14th day, the 21st day measurement pH value: after test group and control group ferment sausage are rubbed respectively, accurately weigh 10.00g sample
Product are added 90mL distilled water, filter after homogeneous, supernatant is taken to be measured with pH meter.
As a result as shown in figure 4, in entire fermentation period, compared with control group sausage, inoculated plant lactobacillus CQ02-
108 (Lactobacillus plantarum CQ02-108) significantly reduce the pH value of sausage, can incite somebody to action at fermentation first day
PH is down to 4.62, i.e., lactobacillus plantarum CQ02-108 of the invention has very strong characteristic of lactic acid production, and the ferment sausage of preparation can
The growth for inhibiting harmful bacteria, extends the shelf-life of product, improves the safety of product.
Embodiment 9: the measurement of ferment sausage lactic acid bacteria number
Ferment sausage prepared by embodiment 6 and embodiment 7 respectively fermentation, drying and ripening during the 1st day, the 7th
It, the 14th day, the 21st day measurement lactic acid bacteria number: test group and control group ferment sausage are aseptically removed into sausage respectively
Casing takes 25g sample to shred, and is added in 225mL sterile saline, and gradient dilution after homogeneous mixes selects appropriate dilution
Bacterium solution be coated on MRS solid medium, carry out plate count after 37 DEG C of culture 48h.
As a result as shown in figure 5, fermenting the 0th~1 day, the viable count of lactobacillus in test group and control group ferment sausage is aobvious
It writes and rises, test group reaches 8.67 (the log cfu/g) that 9.46 (log cfu/g) are significantly higher than control group.Then gradually under
Drop, when same measured number of days, the lactic acid bacteria number of test group ferment sausage is all remarkably higher than control group, until at the 21st day, test group
The lactic acid bacteria number of sausage is 6.67 (the log cfu/g) that 7.66 (log cfu/g) are significantly higher than control group, and test group is fermented at this time
Lactobacillus plantarum CQ02-108 has become absolute predominance flora in sausage.Illustrate lactobacillus plantarum CQ02-108 energy of the invention
The highly acidity being enough resistant in fermentation process.
Embodiment 10: the measurement of ferment sausage total plate count
Ferment sausage prepared by embodiment 6 and embodiment 7 respectively fermentation, drying and ripening during the 1st day, the 7th
It, the 14th day, the 21st day measurement total plate count: test group and control group ferment sausage are aseptically removed into sausage respectively
Casing takes 25g sample to shred, and is added in 225mL sterile saline, and gradient dilution after homogeneous mixes selects appropriate dilution
Bacterium solution be coated on PCA solid medium, carry out plate count after 37 DEG C of culture 48h.
As a result as shown in fig. 6, fermenting the 0th~1 day, the total plate count in two groups of sausages is significantly risen, and reaches 9 (log
Cfu/g it more than), is then gradually reduced, tended to be steady since 7 days to 21 days.At 21 days, the total plate count in experimental group reaches
8.09 (log cfu/g), lower than the total plate count 8.43 (log cfu/g) of control group, this may be in control group sausage in addition to
Except the lactic acid bacteria of itself, there are also other miscellaneous bacterias.This also illustrates inoculated plant lactobacillus CQ02-108 ferment sausages can be more
The growth for inhibiting remaining miscellaneous bacteria well improves the safety of product.
Embodiment 11: the measurement of ferment sausage coloration
Ferment sausage prepared by embodiment 6 and embodiment 7 is measured into coloration respectively: by test group and control group ferment sausage
Peel off sausage sample casing respectively, meat rubbed and is mixed, is pressed into diameter 2cm, the thin slice of thickness about 1cm, choose 6 it is different
Position uses brightness value (L*), red scale value (a*), the yellow value degree (b*) of color difference meter measurement sample.The results are shown in Table 4, and right
It is compared according to a group sausage, addition lactobacillus plantarum CQ02-108 can significantly improve the brightness and redness of ferment sausage, significantly reduce
The yellow value degree of sausage effectively improves the color of sausage.
4 ferment sausage color difference analysis of table
Note: result is expressed as average ± standard deviation, n=6；Same column marks significant (the p < of different lowercase letter indication differences
0.05)。
Embodiment 12: the measurement of ferment sausage texture
Ferment sausage prepared by embodiment 6 and embodiment 7 is measured into texture respectively: by test group and control group ferment sausage
Sausage casing foil is peelled off respectively, and sausage is cut into high 10mm diameter about 20mm size with two-edged knife, uses T-type metal pressure head, machine
Parameter setting are as follows: induction source 1000N, starting force 0.3N, pressure head diameter 41mm, 50% compression ratio, finding speed 50mm/min two
Secondary circulation.Measurement result takes hardness, sticks 4 poly- property, elasticity, chewiness indexs.
The results are shown in Table 5, and compared with blank control group sausage, addition lactobacillus plantarum CQ02-108 can be significantly improved
Hardness, chewiness and the elasticity of ferment sausage assign sausage preferably quality, improve the quality of sausage.
The analysis of 5 ferment sausage texture of table
Note: result is expressed as average ± standard deviation, n=6；Same column marks significant (the p < of different lowercase letter indication differences
0.05)
Embodiment 13: the subjective appreciation of sausage
Ferment sausage prepared by embodiment 6 and embodiment 7 is measured sense organ respectively: subjective appreciation is by 12 Majors of Food people
The subjective appreciation group of member's composition completes, and group member gives training according to GB/T 22210-2008, standards of grading such as 6 institute of table
Show, four color of sausage, smell, structural state and flavour aspects are evaluated respectively.The results are shown in Table 7, can by table
Know, at four color of sausage, smell, structural state and flavour aspects, the ferment sausage of inoculated plant lactobacillus group is significant
Higher than the sausage of blank control group, this shows that lactobacillus plantarum CQ02-108 can be effectively improved the flavor and product of ferment sausage
Matter.
6 ferment sausage sensory evaluation criteria of table
The subjective appreciation of 7 ferment sausage of table
Note: result is expressed as average ± standard deviation；Same column marks significant (the p < of different lowercase letter indication differences
0.05)。
Claims (10)
1. a lactobacillus plantarum CQ02-108, classification naming is lactobacillus plantarum (Lactobacillus plantarum),
It is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on July 17th, 2018, deposit number is
CGMCC No.16121。
2. a kind of microbial bacterial agent comprising lactobacillus plantarum CQ02-108 described in claim 1.
3. microbial bacterial agent described in claim 2, which is characterized in that the microbial bacterial agent is solid-state microbial inoculum or liquid microbial inoculum.
4. a kind of function leavening comprising lactobacillus plantarum CQ02-108 described in claim 1.
5. function leavening as claimed in claim 4, which is characterized in that the function leavening is will be described in claim 1
For 24 hours, 6000g is centrifuged 10min to lactobacillus plantarum CQ02-108 at 4 DEG C for 37 DEG C of cultures in MRS meat soup, collects precipitating sterile water
It washs 3 times and obtained bacteria suspension is resuspended.
6. function leavening described in lactobacillus plantarum CQ02-108 or claim 4 or claim 5 described in claim 1 exists
Make the application in fermentation meat product.
7. application according to claim 6, which is characterized in that the fermentation meat product is ferment sausage.
8. application according to claim 7, which is characterized in that it is described application the following steps are included:
S1: being added seasoning in raw meat and auxiliary material is placed in 4 DEG C and pickles；
S2: function leavening described in claim 4 or claim 5 being inoculated into raw meat and is mixed, sausage is recorded, described
Inoculum concentration is 105-108Cfu/g, preferably 107cfu/g；
S3: it fermentation: 25 DEG C~35 DEG C, ferments 1 day under the conditions of RH80%；
S4: it drying and ripening: 15 DEG C, is placed 4 days under the conditions of RH75%；16 days are dried under the conditions of 12 DEG C, RH 72% to sausage hair
Ferment is mature.
9. application according to claim 8, which is characterized in that
Raw meat described in S1 is fresh pig back leg lean meat and pig back fat, and the positions such as the fascia on its surface and big tendon are rejected
After rinsed, after meat is blended according to the ratio of pig back leg lean meat and back fat be 8:2 be mixed to get；
The seasoning and auxiliary material are added according to the following mass percent of raw meat: salt 2%, sucrose 1%, glucose
1%, sodium nitrite 0.015%, sodium isoascorbate 0.05%, ginger powder 0.1%, white pepper powder 0.1%, five-spice powder 0.1%.
10. application according to claim 8, which is characterized in that fermentation temperature described in S3 is 30 DEG C；The application is also wrapped
It includes S5: sausage is trimmed, finished product is obtained after vacuum packaging.
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| WO2023159960A1 (en) * | 2022-02-24 | 2023-08-31 | 中国肉类食品综合研究中心 | Probiotic lactobacillus plantarum and application thereof in preparation of low-salt fermented meat food product |
| CN114591872B (en) * | 2022-04-12 | 2023-04-25 | 东北农业大学 | Air-dried sausage starter, application thereof and preparation method of air-dried sausage |
| CN114591872A (en) * | 2022-04-12 | 2022-06-07 | 东北农业大学 | Air-dried sausage leavening agent, application thereof and preparation method of air-dried sausage |
| CN114836347A (en) * | 2022-04-27 | 2022-08-02 | 东北农业大学 | Lactobacillus plantarum SL1 and fermentation method thereof |
| NL2033420B1 (en) * | 2022-04-27 | 2023-06-09 | Univ Northeast Agricultural | Lactobacillus plantarum sl1 and fermentation method thereof |
| CN115369055B (en) * | 2022-06-20 | 2023-06-23 | 河北经贸大学 | Antioxidant lactobacillus plantarum and application thereof in low-salt fermented sausage |
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