CN109097312A - Lactobacillus pentosus HN127 and its application in ferment sausage preparation - Google Patents
Lactobacillus pentosus HN127 and its application in ferment sausage preparation 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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- 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
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- 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
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- 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
- A23L13/40—Meat products; Meat meal; Preparation or treatment thereof containing additives
- A23L13/45—Addition of, or treatment with, microorganisms
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- 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
- A23L13/60—Comminuted or emulsified meat products, e.g. sausages; Reformed meat from comminuted meat product
- A23L13/65—Sausages
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- 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
Abstract
The invention discloses one plant of Lactobacillus pentosus HN127 and its preparing the application in ferment sausage.Lactobacillus pentosus (Lactobacillus pentosus) HN127 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.16122.The Lactobacillus pentosus HN127 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 pentosus and its answering in ferment sausage preparation
With.
Background technique
Traditional Chinese ferment sausage is well-known because of its unique flavor, fragrance, quality and taste.China's traditional zymotic intestines
In winter through spontaneous fermentation in a few days, then mature several weeks at room temperature make it have distinctive strong flavor and solid
Quality.However, traditional zymotic be mainly lean on the microbial flora of raw meat itself to complete fermentation and mature, therefore process time compared with
It grows and is easy to be limited by weather, the safety and quality of product are difficult to be protected.
The microbial flora of traditional zymotic sausage is mainly by lactic acid bacteria, coagulase-negative staphylococci, yeast and mould group
At.The microbial flora constitutes participation acidification, denitrification, the most active endogenous microbes of steatolysis and proteolysis process.
Wherein lactic acid bacteria plays an important role in meat fermentation and preservation, therefore is considered as the major microorganisms for participating in fermentation.One side
Face, lactic acid bacteria can generate the pH value that lactic acid reduces rapidly product, and part lactic acid bacteria energy using the carbohydrate in raw material
Bacteriocin is generated, this plays inhibiting effect to the growth of pathogenic bacteria and spoilage organisms.On the other hand, lactic acid bacteria can assign product
Distinctive flavor and solid quality.In addition, the generation of lactic acid can also accelerate nitrite to turn to nitric oxide myoglobins
Change, reduce the residual quantity of nitrite in product, while promoting the formation of good colour.Therefore, the screening study of lactic acid bacteria
The exploitation of fermentation meat product is of great significance to
When making ferment sausage, usually salt and nitrite are added in fermentation meat product to guarantee quality, changed
Kind flavor etc..And during the fermenting-ripening of sausage, with the continuous reduction of pH, moisture content is also constantly reduced, salinity
Then constantly increase.Therefore, it screens one plant and is resistant to high salt and highly acidity, be able to suppress the lactic acid bacteria of pathogenic bacteria growth, and
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 one plant of Lactobacillus pentosus 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, the Lactobacillus pentosus for inhibiting pathogenic bacteria growth
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 one plant of Lactobacillus pentosus HN127, classification naming is Lactobacillus pentosus
It is common to be preserved in China Committee for Culture Collection of Microorganisms on July 17th, 2018 by Lactobacillus pentosus
Microorganism center, deposit number are CGMCC No.16122.
In one embodiment of the invention, the Lactobacillus pentosus HN127 produce protease, quickly produce acid, do not produce it is viscous,
Gas is not produced, non-generation amine, does not produce H2O2, do not produce H2S, not chromogenic element.
In one embodiment of the invention, the Lactobacillus pentosus HN127 is resistant to high salt concentration, highly acidity.
In one embodiment of the invention, the Lactobacillus pentosus HN127 is able to suppress Escherichia coli, golden yellow Portugal
The growth of grape coccus.
In one embodiment of the invention, the Lactobacillus pentosus HN127 is resistant to cholate and gastric juice.
A second object of the present invention is to provide a kind of microbial bacterial agents comprising aforementioned Lactobacillus pentosus HN127.
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 aforementioned Lactobacillus pentosus HN127.
Further, the function leavening is 37 DEG C of cultures in MRS meat soup by Lactobacillus pentosus HN127 above-mentioned
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 aforementioned Lactobacillus pentosus HN127 or function leavening above-mentioned and sends out in production
Application in ferment meat products.
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 described in claim 4 or claim 5 being inoculated into raw meat and is mixed, sausage is recorded,
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.
Raw meat described in further S1 is fresh pig back leg lean meat and pig back fat, by the fascia on its surface and big muscle
The positions such as tendon are rinsed after rejecting, and are mixed to get after meat is blended according to the ratio of pig back leg lean meat and back fat for 8:2；
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 pentosus HN127 leavening of the present invention, ferment sausage can be made to have excellent
Different 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, aforementioned Lactobacillus pentosus HN127 or function leavening above-mentioned answering in production fermentation meat product
With being to improve product quality using the characteristics such as Lactobacillus pentosus HN127 salt tolerant, acidproof, antibacterial, quick production acid.
Beneficial effects of the present invention:
(1) Lactobacillus pentosus of the invention not only meets the basic standard of starter of meat products, moreover it is possible to be resistant to highly concentrated salt, resistance to
By highly acidity, the growth of effectively inhibition pathogenic bacteria, while also with the feature of bile tolerance resistant to gastric juice, to functional starter of meat products
Exploitation certain theoretical foundation is provided.
(2) present invention is by addition Lactobacillus pentosus, 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 pentosus of the invention makes ferment sausage as leavening, effectively improves the wind of ferment sausage
Taste and quality.
(4) fabrication cycle of traditional zymotic sausage is longer, is inoculated with Lactobacillus pentosus of the invention as leavening, effectively
Fermentation period is shortened, the fermentation period of product is 21 days.
(5) it with traditional zymotic sausage ratio, is inoculated with Lactobacillus pentosus HN127 of the invention and sausage is effectively reduced in production process
In pH, reduce the risk of sausage corruption and mildew, improve the safety of product.
Detailed description of the invention
Fig. 1 is the growth curve of Lactobacillus pentosus HN127
Fig. 2 is Lactobacillus pentosus growing state under different salinity
Fig. 3 is Lactobacillus pentosus 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
It is micro- to be preserved in China on July 17th, 2018 by Lactobacillus pentosus (Lactobacillus pentosus) HN127
Biological inoculum preservation administration committee common micro-organisms center, classification naming are Lactobacillus pentosus Lactobacillus
Pentosus, deposit number are CGMCC No.16122.Depositary institution address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3,
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 Hunan Province Yiyang 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 27 plants of Gram-positives and connects
The bacterial strain of catalase feminine gender.
3, the primary dcreening operation of bacterial strain
27 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
Produce hydrogen sulfide, biogenic amine, nitrate reductase, produce gas, mucus, pigment are negative totally 3 plants of bacterial strain, respectively number be HN106,
HN118、HN127。
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 HN127 is antibacterial relative to two plants of bacterium of HN106, HN118
Effect is best, therefore finally obtains HN127.
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 HN127 filtered out in the flat lining out of MRS, picking single colonie universal amplification 16SrRNA gene, using amplification
The universal primer of bacteria 16 S rRNA genes: upstream primer 27F, downstream primer 1492R.PCR condition are as follows: 94 DEG C of initial denaturations
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 extension 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 pentosus (Lactobacillus
Pentosus), similarity 100%.The bacterium is preserved in Chinese microorganism strain preservation conservator on July 17th, 2018
Meeting common micro-organisms center, deposit number are CGMCC No.16122.Depositary institution address: BeiChen West Road, Chaoyang District, BeiJing City 1
Number institute 3, research institute of the Chinese Academy of Sciences.
Embodiment 2: Lactobacillus pentosus HN127 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 HN127 OD620 in 0.1% cholate and 0.3% cholate increases needed for 0.3 unit
Time be substantially less than remaining two plants of bacterium, show that the tolerance cholate ability of HN127 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 pentosus HN127 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 3 plants of bacterium all have good gastric fluid-resistant, digests by the simulation of 3h, viable count is still
10 can be reached7Cfu/ml, wherein the survival rate of HN127 is significantly higher than remaining two plants of bacterium, shows the ability of the tolerance gastric juice of HN127
It 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 pentosus HN127
1, the measurement of Lactobacillus pentosus HN127 growth curve
Lactobacillus pentosus HN127 is inoculated in MRS fluid nutrient medium, 37 DEG C of culture 48h, every 2h spectrophotometer
Its light absorption value is measured at wavelength 600nm, while being compared with blank MRS fluid nutrient medium.
As a result as shown in Figure 1.
2, Salt tolerance:
Lactobacillus pentosus 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, and will receive inhibition to the growth of the lactic acid bacteria of salt density value, and HN127 is dense in NaCl in this experiment
Degree makes it become advantage during the fermentation to grow in 3%, 6%, 9%, 12%, 15% MRS fluid nutrient medium
Bacterium.
Embodiment 5: the acid resistance test of Lactobacillus pentosus HN127
Lactobacillus pentosus 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 it as shown in figure 3, Lactobacillus pentosus HN127 can be grown at different pH, and is given birth under pH4.5,5.5,6.5
It is long good.At fermentation initial stage, lactic acid bacteria can produce rapidly acid, so that the pH of product declines to a great extent, therefore Lactobacillus pentosus can be resistant to
It is set to become dominant bacteria during the fermentation by highly acidity.
Embodiment 6: the preparation (test group) of ferment sausage
The preparation of 1 leavening: for 24 hours, 6000g is centrifuged Lactobacillus pentosus HN127 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 pentosus HN127
The deposit number of (Lactobacillus pentosus HN127) is CGMCC No.16122.
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 separately added into 90mL distilled water, filter after homogeneous, supernatant is taken to be measured with pH meter.
As a result as shown in figure 4, compared with the control group sausage of non-inoculating strain, it is inoculated with Lactobacillus pentosus HN127
(Lactobacillus pentosus HN127) significantly reduces the pH value of sausage, can be down to pH within first day in fermentation
4.72, and the pH of controllable sausage is lower than 5.2 in entire fermentation period, i.e., Lactobacillus pentosus HN127 of the invention has
The ferment sausage of very strong characteristic of lactic acid production, preparation is able to suppress the growth of harmful bacteria, extends the shelf-life of product, improves product
Safety.
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 separately added into 225mL sterile saline, and gradient dilution after homogeneous mixes selects appropriate dilutions
The bacterium solution of degree is coated on MRS solid medium, carries 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, be then gradually reduced, when same measured number of days, the lactic acid bacteria number of test group ferment sausage is all remarkably higher than control
Group, until the lactic acid bacteria number of test group sausage is the 6.67 (log that 7.73 (log cfu/g) are significantly higher than control group at the 21st day
Cfu/g), Lactobacillus pentosus HN127 has become absolute predominance flora in test group ferment sausage at this time.Illustrate the present invention simultaneously
Lactobacillus pentosus HN127 be resistant to the highly acidity during sausage fermentation.
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, the total plate count after fermentation 1 day in two groups of sausages significantly rises, reach 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 7.9
(log cfu/g), lower than the total plate count 8.4 (log cfu/g) of control group, this may be in control group sausage in addition to itself
Except lactic acid bacteria, there are also other miscellaneous bacterias.This also illustrates inoculation Lactobacillus pentosus HN127 ferment sausages can preferably inhibit it
The growth of remaining miscellaneous bacteria 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 compared with control group sausage, it is fragrant that addition Lactobacillus pentosus HN127 can significantly improve fermentation
The brightness and redness of intestines, significantly reduce the yellow value degree of sausage, effectively improve 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 25mm diameter about 20mm size with two-edged knife, is popped one's head in using P/50A, machine ginseng
Number setting are as follows: 50kg load cell, the compression of speed 2.0mm/s, test speed 5mm/s, speed 5mm/s after survey, 50% before surveying
Rate.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 pentosus HN127 can significantly improve hair
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 measurement of sausage sense organ
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, respectively to the face of sausage
Four color, smell, structural state and flavour aspects are evaluated.
The results are shown in Table 7, as seen from table, in four color of sausage, smell, structural state and flavour aspects, inoculation
The ferment sausage of Lactobacillus pentosus group is all remarkably higher than the sausage of blank control group, this shows that Lactobacillus pentosus HN127 can have
Effect improves the flavor and quality of ferment sausage.
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. one plant of Lactobacillus pentosus HN127, classification naming is Lactobacillus pentosus (Lactobacillus pentosus), in 2018
On July 17, in is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number CGMCC
No.16122。
2. a kind of microbial bacterial agent comprising Lactobacillus pentosus HN127 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 pentosus HN127 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 pentosus HN127 at 4 DEG C for 37 DEG C of cultures in MRS meat soup, collects and precipitates sterile washing
It washs 3 times and obtained bacteria suspension is resuspended.
6. function leavening described in Lactobacillus pentosus HN127 or claim 4 or claim 5 described in claim 1 is being made
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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