CN108949645B - Lactobacillus plantarum CQ02-108 and application thereof in preparation of fermented sausages - Google Patents
Lactobacillus plantarum CQ02-108 and application thereof in preparation of fermented sausages Download PDFInfo
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- 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
- 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
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- 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
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- 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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Abstract
The invention discloses lactobacillus plantarum CQ02-108 and application thereof in preparing fermented sausages. The Lactobacillus plantarum CQ02-108 is preserved in the general microorganism center of China general microbiological culture Collection center in 2018, 7 and 17 months, and the preservation number is CGMCC No. 16121. The Lactobacillus plantarum CQ02-108 can produce protease, rapidly produce acid, no viscosity, no gas, no biogenic amine and no H2O2Failure to produce H2S, no pigment production, high salt concentration tolerance, high acidity tolerance, growth inhibition of escherichia coli and staphylococcus aureus, bile salt tolerance and gastric juice tolerance. When the fermented sausage is applied, the number of live lactobacillus in the fermented sausage can be obviously increased, the pH value of the sausage is reduced, the growth of harmful bacteria is inhibited, and the quality of the fermented sausage can be obviously improved.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to lactobacillus plantarum and application thereof in preparation of fermented sausages.
Background
The fermented sausage is a fermented meat product prepared by mixing minced meat, animal fat, salt, sugar, leavening agent, spices and the like, filling the mixture into a casing and performing microbial fermentation under artificial or natural control conditions. The traditional fermented sausage in China does not need the step of artificially adding a leavening agent, is mainly fermented and matured under the natural environment by the microbial flora of raw meat, and has unique flavor. Therefore, the fermentation period is long, and the method is easily limited by the climate, and the safety and the quality of the product are difficult to guarantee.
The microbial flora of the traditional fermented sausage mainly comprises lactic acid bacteria, coagulase negative staphylococcus, yeast and mould, wherein the lactic acid bacteria play an important role in meat fermentation and preservation and are main microorganisms participating in fermentation. Pathogenic bacteria are one of the important reasons influencing the safety of the fermented sausage, and the lactic acid bacteria can generate lactic acid to rapidly reduce the pH value of the product, and part of the lactic acid bacteria can generate bacteriocin, thereby inhibiting the growth of the pathogenic bacteria and spoilage bacteria. It has become a trend to use bacteriocin-producing lactic acid bacteria as leavening agents to extend the shelf life and ensure the safety of fermented sausages. The generation of lactic acid can also accelerate the conversion of nitrite to nitric oxide myoglobin, reduce the residual amount of nitrite in the product and promote the formation of good color. On the other hand, lactic acid bacteria can impart a characteristic flavor and firm texture to the product. Therefore, the screening research of the lactic acid bacteria has very important significance for the development of the fermented meat products.
In addition, in the manufacture of fermented sausages, salt and nitrite are often added to fermented meat products to ensure quality, improve flavour etc. In the process of fermenting and maturing the sausage, the water content and the Aw are continuously reduced along with the continuous reduction of the pH value, and the salt concentration is continuously increased. Therefore, the lactobacillus capable of tolerating high salt and high acidity and producing bacteriocin to inhibit the growth of pathogenic bacteria is screened, and the lactobacillus is applied to fermented sausages and has important significance for processing the sausages.
Disclosure of Invention
Aiming at the problems, the invention provides a lactobacillus plantarum and application thereof in fermented sausages. The lactobacillus plantarum which is screened from the traditional naturally fermented sausage and can tolerate high salt concentration and high acidity and inhibit the growth of pathogenic bacteria is applied to the fermented sausage, so that the color and luster of the sausage are improved, and the quality of the sausage is improved.
The purpose of the invention is realized by the following technical scheme:
the invention aims to provide a Lactobacillus plantarum CQ02-108 which is classified and named as Lactobacillus plantarum and already preserved in China general microbiological culture Collection center on 7-17 th month in 2018 with the preservation number of CGMCC No. 16121.
In one embodiment of the invention, the Lactobacillus plantarum CQ02-108 produces protease, produces acid rapidly, does not produce viscosity, does not produce gas, does not produce biogenic amine, does not produce H2O2Failure to produce H2S, no pigment is produced.
In one embodiment of the present invention, the lactobacillus plantarum CQ02-108 is capable of tolerating high concentrations of salt, high acidity.
In one embodiment of the invention, the lactobacillus plantarum CQ02-108 is capable of inhibiting the growth of escherichia coli, staphylococcus aureus.
In one embodiment of the present invention, the lactobacillus plantarum CQ02-108 is resistant to bile salts and gastric juices.
The second purpose of the invention is to provide a microbial agent containing the lactobacillus plantarum CQ 02-108.
Further, the microbial agent is a solid microbial agent or a liquid microbial agent.
The third purpose of the invention is to provide a functional leaven containing the lactobacillus plantarum CQ 02-108.
Further, the functional starter is a bacterial suspension obtained by culturing the lactobacillus plantarum CQ02-108 in MRS broth at 37 ℃ for 24h, centrifuging at 4 ℃ for 10min at 6000g, collecting the precipitate, washing with sterile water for 3 times, and resuspending.
The fourth purpose of the invention is to provide the application of the lactobacillus plantarum CQ02-108 or the functional leavening agent in the preparation of fermented meat products.
Further, the fermented meat product is fermented sausage.
Further, the application comprises the following steps:
s1: adding flavoring and adjuvants into raw meat, and pickling at 4 deg.C;
s2: inoculating the functional leavening agent into raw meat, mixing uniformly, and filling sausage, wherein the inoculation amount is 105-108cfu/g, preferably 107cfu/g；
S3: fermentation: fermenting for 1 day at 25-35 ℃ and RH 80%;
s4: drying and maturing: standing at 15 deg.C and RH 75% for 4 days; drying at 12 deg.C and RH 72% for 16 days until the sausage is fermented;
further, the above steps further comprise
S5: and (4) trimming the sausage, and carrying out vacuum packaging to obtain a finished product.
Further, the raw material meat in the S1 is fresh pig hind leg lean meat and pig backfat, the fascia, large tendons and other parts on the surface of the raw material meat are removed and rinsed, and the meat is stirred and mixed according to the ratio of the pig hind leg lean meat to the backfat of 8:2 to obtain the mixture;
the seasoning and the auxiliary materials are added according to the following mass percentage with the raw meat: 2% of salt, 1% of cane sugar, 1% of glucose, 0.015% of sodium nitrite, 0.05% of sodium erythorbate, 0.1% of ginger powder, 0.1% of white pepper powder and 0.1% of five spice powder. The mixture ratio of the seasoning and the auxiliary materials is combined with the use of the lactobacillus plantarum CQ02-108 leavening agent, so that the fermented sausage has excellent sensory characteristics.
Further, the fermentation temperature in S3 was 30 ℃.
Furthermore, the functional leavening agent can inhibit propagation of escherichia coli and staphylococcus aureus in food, and can propagate in bile salt and simulated gastric juice.
Furthermore, the lactobacillus plantarum CQ02-108 or the functional leavening agent is applied to the preparation of fermented meat products by utilizing the characteristics of the lactobacillus plantarum CQ02-108 such as salt resistance, acid resistance, bacteriostasis, rapid acid production and the like, so that the product quality is improved.
The invention has the beneficial effects that:
(1) the lactobacillus plantarum not only meets the basic standard of the meat starter, but also can tolerate high-concentration salt, tolerate high acidity and produce bacteriocin to effectively inhibit the growth of pathogenic bacteria, and simultaneously has the characteristics of cholate resistance and gastric juice resistance, thereby providing a certain theoretical basis for the development of functional meat starter.
(2) According to the invention, lactobacillus plantarum is added, so that the number of lactic acid bacteria is still as high as 10 at the end of sausage maturation7The method can obviously improve the quantity of lactic acid bacteria in the fermented sausage and inhibit the growth of escherichia coli and staphylococcus aureus.
(3) The lactobacillus plantarum provided by the invention is used as a leavening agent to prepare the fermented sausage, so that the flavor and quality of the fermented sausage are effectively improved.
(4) Compared with the traditional fermented sausage, the lactobacillus plantarum CQ02-108 inoculated in the invention effectively reduces the pH value of the sausage in the production process, reduces the risks of sausage putrefaction and mildew, and improves the safety of the product.
Drawings
FIG. 1 is a growth curve of Lactobacillus plantarum CQ02-108
FIG. 2 shows the growth of Lactobacillus plantarum CQ02-108 at various salt concentrations
FIG. 3 shows the growth of Lactobacillus plantarum CQ02-108 at various pH levels
FIG. 4 shows the pH change of fermented sausage
FIG. 5 shows the change of the number of lactic acid bacteria in the fermented sausage
FIG. 6 shows the change of the total number of colonies in the fermented sausage
Biological preservation information description
A strain of Lactobacillus plantarum CQ02-108, which has been deposited in the China general microbiological culture Collection center on 7 th and 17 th months in 2018, is classified and named as Lactobacillus plantarum, and the deposition number is CGMCC No. 16121. The address of the depository: xilu No.1 Hospital No. 3, national academy of sciences, North Kyoho, Beijing, Chaoyang; and E, postcode: 100101.
Detailed Description
The invention is further illustrated by the following specific examples:
example 1: screening of strains
The screening method comprises the following specific implementation steps:
1. sample collection
The original sample is taken from the traditional naturally fermented sausage purchased in the dish market of Chongqing.
2. Isolation of lactic acid bacteria from samples
Shearing 25g of sausage sample under aseptic condition, dissolving in 225mL of sterile normal saline, mixing uniformly, standing for several minutes, after the ground meat is completely precipitated, taking supernatant, gradually diluting in an incremental manner according to a ratio of 1:10, selecting 100ul of bacterial liquid with proper dilution degree, uniformly coating the bacterial liquid on MRS solid culture medium added with calcium carbonate, and culturing for 48 hours in an anaerobic incubator at 37 ℃. Selecting independent single colonies which have different forms, calcium-dissolving rings and meet the morphological characteristics of the lactobacillus colonies on the flat plate, and streaking on an MRS culture medium until a pure culture is obtained. Typical colonies were picked for gram stain and catalase experiments to give 20 gram stain positive and catalase negative strains.
3. Preliminary screening of bacterial strains
And (3) respectively carrying out acid production, protease, nitrite resistance, hydrogen sulfide production, amine product production, nitrate reductase production, gas production, mucus production and pigment production tests on the 20 separated strains to finally obtain 3 strains which are negative in acid production, protease production, nitrite resistance, hydrogen sulfide production, biogenic amine, nitrate reductase production, gas production, mucus production and pigment production, and are respectively numbered CQ02-108, CQ02-117 and CQ 02-120.
4. Rescreening of bacterial strains
And (3) carrying out a bacteriostasis test on the obtained 3 strains, and determining the bacteriostasis capacity of the strains by adopting an Oxford cup method. The indicator bacteria are Escherichia coli and Staphylococcus aureus. Collecting the supernatant bacterial liquid in the lactic acid bacteria stationary phase, and filtering and sterilizing the supernatant by using a 0.22 mu m microporous filter membrane. The activated indicator bacterium (10) after 3 generations6cfu/mL) is evenly coated on the surface of a nutrient agar culture medium, 4 aseptic oxford cups are placed in a culture dish at equal distance, 200 mu L of test strain supernatant is added into each oxford cup, a blank control is a liquid culture medium, the blank control is cultured for 48h at 37 ℃ after diffusion for 6h at 4 ℃, the situation of a bacteriostatic circle around the oxford cup is observed, and the diameter of the bacteriostatic circle is measured to calculate the average value. The final results are shown in Table 1, CQ02-108 has the best bacteriostatic effect compared with two strains CQ02-117 and CQ02-120, so CQ02-108 is finally obtained.
TABLE 1 results of bacteriostasis of different strains
Note: results are expressed as mean ± standard deviation, n is 4; the same column notes different lower case letters as significantly different (p < 0.05).
5. Identification of strains
Streaking the selected CQ02-108 on MRS plate, picking single colony to amplify 16SrRNA gene by using general primer for amplifying 16SrRNA gene: the upstream primer was 27F and the downstream primer was 1492R. The PCR conditions were: pre-denaturation at 94 ℃ for 5 min; denaturation at 94 ℃ for 1min, annealing at 55 ℃ for 1min, extension at 72 ℃ for 90s, 30 cycles; final extension at 72 ℃ for 10 min.
The PCR product is detected by 1% agarose gel electrophoresis, and then sent to the biological engineering (Shanghai) company Limited for sequencing, and the result obtained by the sequence is compared with the known sequence in the NCBI database. The strain is determined to be Lactobacillus plantarum (Lactobacillus plantarum), and the similarity is 100%. The strain is preserved in China general microbiological culture Collection center (CGMCC) in 2018, 7 and 17 months, and the preservation number is CGMCC No. 16121. The address of the depository: xilu No.1 Hospital No. 3, Beijing, Chaoyang, the institute of Chinese academy of sciences.
Example 2: lactobacillus plantarum CQ02-108 bile salt resistance test
The bacterial suspension of the test strain is adjusted to 109CFU/mL. The control group consisted of MRS broth without bile salts, inoculated at a ratio of 1% (V/V) into MRS broth containing 0.1%, 0.3% bile salts, respectively. The cells were incubated anaerobically at 37 ℃ for 24 hours, and OD620 was measured by spectrophotometer at 0.5h intervals. The bile salt tolerance of each strain was based on the time required for an increase of 0.3 units in the OD620 value.
Results as shown in table 2, the time required for OD620 to increase 0.3 units in both 0.1% and 0.3% bile salts of CQ02-108 was significantly less than the remaining two strains, indicating that CQ02-108 was most resistant to bile salts.
TABLE 2 results of bile salt tolerance of different strains
Note: the same column is labeled with different lower case letters indicating significant difference (p < 0.05) and n is 3.
Example 3: lactobacillus plantarum CQ02-108 gastric juice resistance test
Preparation of simulated gastric fluid: 0.35g pepsin was added to 100mL of 0.2% sterile saline, the pH was adjusted to 3.0 with concentrated HCl and filter sterilized for use.
The test strains were cultured overnight in MRS broth at 37 ℃ and then inoculated in 5mL gastric juice at 1% and incubated for 3 hours at 37 ℃. After digestion in simulated gastric fluid, the viable cell count (log 10cfu/mL) of each strain was determined by plate counting method, and the survival rate (%) -3 h viable cell count/0 h viable cell count × 100% was calculated.
The results are shown in Table 3, the three strains have good gastric juice tolerance, and the viable count can still reach 10 after 3h simulated digestion7cfu/ml, in which the survival rate of CQ02-108 was significantly higher than the remaining two strains, indicating that CQ02-108 was most resistant to gastric juice.
TABLE 3 gastric juice tolerance of different strains
Note: results are expressed as mean ± standard deviation, n is 3; the same column is labeled with different lower case letters indicating significant difference (p < 0.05).
Example 4: salt tolerance test of lactobacillus plantarum CQ02-108
1. Determination of growth curves of Lactobacillus plantarum CQ02-108
Lactobacillus plantarum CQ02-108 was inoculated into MRS liquid medium, cultured at 37 ℃ for 48 hours, and absorbance was measured every 2 hours at a wavelength of 600nm using a spectrophotometer, while blank MRS liquid medium was used as a control. The results are shown in FIG. 1.
2. Salt tolerance test:
inoculating Lactobacillus plantarum 2% into MRS liquid culture medium with NaCl concentration of 3%, 6%, 9%, 12%, 15%, respectively, culturing at 37 deg.C for 24h, measuring absorbance at 600nm with spectrophotometer, and repeating for 6 times with blank culture medium as control.
As shown in FIG. 2, the amount of salt added to the fermented sausage was generally 2% to 3%, and as the water content decreased, the salt concentration increased and the growth of the salt-sensitive lactic acid bacteria was inhibited. In the experiment, the lactobacillus plantarum CQ02-108 grows well when the NaCl concentration is 3%, 6% and 9%, and can also grow in MRS liquid culture media of 12% and 15%, and the lactobacillus plantarum CQ02-108 has strong salt tolerance, so that the lactobacillus plantarum CQ02-108 becomes a dominant bacterium in the fermentation process.
Example 5: acid resistance test of Lactobacillus plantarum CQ02-108
Inoculating Lactobacillus plantarum 2% into MRS liquid culture medium with pH of 2.5, 3.5, 4.5, 5.5, and 6.5, culturing at 37 deg.C for 24h, measuring absorbance at 600nm with spectrophotometer, and repeating for 6 times with blank culture medium with corresponding pH as control.
As shown in FIG. 3, the Lactobacillus plantarum CQ02-108 grew at various pH values, and grew well at pH values of 4.5, 5.5, and 6.5. In the initial stage of fermentation, lactic acid bacteria can rapidly produce acid, so that the pH value of the product is greatly reduced, and therefore, the lactobacillus plantarum can tolerate high acidity to become dominant bacteria in the fermentation process.
Example 6: preparation of fermented sausages (test group)
1, preparation of a leavening agent: lactobacillus plantarum CQ02-108 was cultured in MRS broth at 37 ℃ for 24h, centrifuged at 4 ℃ at 6000g for 10min, the pellet was collected, washed 3 times with sterile water and resuspended to obtain a bacterial suspension for use. The preservation number of the Lactobacillus plantarum CQ02-108(Lactobacillus plantarum CQ02-108) is CGMCC No. 16121.
2 preparation of fermented sausage
2.1 selecting fresh pork back leg lean meat and pork back fat, removing fascia, large tendons and other parts on the surface of the pork back leg lean meat and the pork back fat, rinsing, stirring the meat, mixing according to the ratio of the pork back leg lean meat to the pork back fat of 8:2, adding seasonings and auxiliary materials, and pickling at 4 ℃. The seasoning and the auxiliary materials are added according to the weight of meat: 2% of salt, 1% of cane sugar, 1% of glucose, 0.015% of sodium nitrite, 0.05% of sodium erythorbate, 0.1% of ginger powder, 0.1% of white pepper powder and 0.1% of five spice powder.
2.2 inoculating the bacterial suspension of 2.1 into meat, mixing uniformly, and filling the sausage, wherein the inoculation amount is 107cfu/g。
2.3, fermentation: fermenting at 30 deg.C and RH 80% for 1 day.
2.4 drying and ripening: standing at 15 deg.C and RH 75% for 4 days; drying at 12 deg.C and RH 72% for 16 days until the sausage is fermented and matured.
2.5 pruning the sausage, and carrying out vacuum packaging to obtain a finished product.
Example 7: preparation of fermented sausage (control group)
(1) Selecting fresh pig hind leg lean meat and pig backfat, removing fascia, large tendon and other parts on the surface of the pig hind leg lean meat and the pig backfat, rinsing, stirring the meat, mixing according to the ratio of the pig hind leg lean meat to the pig backfat of 8:2, adding seasonings and auxiliary materials, and pickling at 4 ℃. The seasoning and the auxiliary materials are added according to the weight of meat: 2% of salt, 1% of cane sugar, 1% of glucose, 0.015% of sodium nitrite, 0.05% of sodium erythorbate, 0.1% of ginger powder, 0.1% of white pepper powder and 0.1% of five spice powder.
(2) Fermentation: fermenting at 30 deg.C and RH 80% for 1 day.
(3) Drying and maturing: standing at 15 deg.C and RH 75% for 4 days; drying at 12 deg.C and RH 72% for 16 days until the sausage is fermented and matured.
(4) And (4) trimming the sausage, and carrying out vacuum packaging to obtain a finished product.
Example 8: determination of pH value of fermented sausage
The pH of the fermented sausages prepared in example 6 and example 7 were measured at day 1, day 7, day 14 and day 21 during fermentation, drying and maturation, respectively: respectively mincing the fermented sausages of the test group and the control group, accurately weighing 10.00g of sample, adding 90mL of distilled water, homogenizing, filtering, and measuring the supernatant by using a pH meter.
As shown in the figure 4, in the whole fermentation period, compared with the sausage of the control group, the pH value of the sausage is obviously reduced by inoculating the Lactobacillus plantarum CQ02-108(Lactobacillus plantarum CQ02-108), and the pH value can be reduced to 4.62 in the first day of fermentation, namely the Lactobacillus plantarum CQ02-108 has strong acid-producing characteristics, and the prepared fermented sausage can inhibit the growth of harmful bacteria, prolong the shelf life of the product and improve the safety of the product.
Example 9: determination of lactic acid bacteria number of fermented sausage
The fermented sausages prepared in example 6 and example 7 were subjected to fermentation, drying and maturation for the following days 1, 7, 14 and 21: removing sausage casings of the fermented sausages of the test group and the control group respectively under the aseptic condition, taking 25g of samples, cutting into pieces, adding the samples into 225mL of aseptic normal saline, homogenizing and uniformly mixing, then diluting in a gradient manner, selecting bacterial liquid with proper dilution degree to coat on an MRS solid culture medium, culturing for 48h at 37 ℃, and then counting plates.
The results are shown in fig. 5, the viable count of the lactic acid bacteria in the fermented sausages of the test group and the control group is obviously increased after 0-1 day of fermentation, and the viable count of the lactic acid bacteria in the fermented sausages of the test group and the control group is 9.46(log cfu/g) which is obviously higher than 8.67(log cfu/g) of the control group. Then the number of the lactobacillus in the fermented sausage in the test group is obviously higher than that in the control group on the same measuring days, and the number of the lactobacillus in the sausage in the test group is 7.66(log cfu/g) obviously higher than 6.67(log cfu/g) in the control group by the 21 st day, at the moment, the lactobacillus plantarum CQ02-108 in the fermented sausage in the test group becomes an absolute dominant flora. It is demonstrated that the Lactobacillus plantarum CQ02-108 of the invention is able to tolerate high acidity during fermentation.
Example 10: determination of the Total bacterial count of fermented sausages
The total number of colonies was determined on days 1, 7, 14 and 21 during fermentation, drying and maturation of the fermented sausages prepared in example 6 and example 7, respectively: removing sausage casings of the fermented sausages of the test group and the control group respectively under the aseptic condition, taking 25g of samples, cutting into pieces, adding the samples into 225mL of aseptic normal saline, homogenizing and uniformly mixing, then diluting in a gradient manner, selecting bacterial liquid with proper dilution degree to coat on a PCA solid culture medium, culturing for 48h at 37 ℃, and then counting plates.
The results are shown in fig. 6, the total number of colonies in the two groups of sausages is remarkably increased to more than 9(log cfu/g) at 0-1 day of fermentation, and then gradually decreased, and the number of colonies is stable from 7 days to 21 days. At 21 days, the total number of colonies in the experimental group reaches 8.09(log cfu/g) and is lower than the total number of colonies in the control group by 8.43(log cfu/g), which is probably that the sausages in the control group contain other mixed bacteria besides the lactobacillus of the control group. The fact that the fermented sausage inoculated with the lactobacillus plantarum CQ02-108 can better inhibit the growth of other mixed bacteria and improve the safety of products is also shown.
Example 11: measurement of color of fermented sausage
The fermented sausages prepared in example 6 and example 7 were each tested for color: peeling sausage sample casings of the fermented sausages of the test group and the control group respectively, mincing and uniformly mixing meat, pressing into slices with the diameter of 2cm and the thickness of about 1cm, selecting 6 different positions, and measuring the brightness value (L), the red value (a) and the yellow value (b) of the samples by using a color difference meter. As shown in Table 4, compared with the sausage of the control group, the added Lactobacillus plantarum CQ02-108 can obviously improve the brightness and the redness of the fermented sausage, obviously reduce the yellowness value of the sausage and effectively improve the color of the sausage.
TABLE 4 color difference analysis of fermented sausages
Note: results are expressed as mean ± standard deviation, n is 6; the same column is labeled with different lower case letters indicating significant difference (p < 0.05).
Example 12: determination of the texture of fermented sausages
The fermented sausages prepared in example 6 and example 7 were each subjected to texture determination: the sausage casings of the fermented sausages of the test group and the control group are respectively stripped, the sausages are cut into the size of 10mm in height and 20mm in diameter by a double-sided knife, a T-shaped metal pressure head is used, and the machine parameters are set as follows: the induction source is 1000N, the initial force is 0.3N, the diameter of the pressure head is 41mm, the compression ratio is 50 percent, and the measuring speed is 50mm/min for two cycles. The measurement results are 4 indexes of hardness, cohesiveness, elasticity and chewiness.
The results are shown in table 5, and compared with the sausage of the blank control group, the addition of lactobacillus plantarum CQ02-108 can significantly improve the hardness, chewiness and elasticity of the fermented sausage, impart better texture to the sausage, and improve the quality of the sausage.
TABLE 5 texture analysis of fermented sausages
Note: results are expressed as mean ± standard deviation, n is 6; the same column marks different lower case letters to indicate significant difference (p < 0.05)
Example 13: sensory evaluation of sausages
The fermented sausages prepared in example 6 and example 7 were separately tested for sensory: sensory evaluation was performed by a sensory evaluation panel consisting of 12 food professionals, trained in accordance with GB/T22210-. The results are shown in table 7, and it can be seen from the table that the fermented sausages inoculated with lactobacillus plantarum group were significantly higher than the sausages of the blank control group in all four aspects of color, smell, tissue state and taste of the sausages, which indicates that lactobacillus plantarum CQ02-108 can effectively improve the flavor and quality of the fermented sausages.
TABLE 6 sensory evaluation criteria for fermented sausages
TABLE 7 sensory evaluation of fermented sausages
Note: results are expressed as mean ± standard deviation; the same column is labeled with different lower case letters indicating significant difference (p < 0.05).
Claims (11)
1. A Lactobacillus plantarum CQ02-108, classified and named as Lactobacillus plantarum (Lactobacillus plantarum), has been deposited in China general microbiological culture Collection center (CGMCC) in 2018, 7 and 17, with the deposit number of CGMCC No. 16121.
2. A microbial inoculant comprising lactobacillus plantarum CQ02-108 of claim 1.
3. The microbial agent according to claim 2, wherein the microbial agent is a solid microbial agent or a liquid microbial agent.
4. A functional starter culture comprising lactobacillus plantarum CQ02-108 of claim 1.
5. The functional starter of claim 4, wherein the functional starter is a bacterial suspension obtained by culturing lactobacillus plantarum CQ02-108 of claim 1 in MRS broth at 37 ℃ for 24h, centrifuging at 4 ℃ at 6000g for 10min, collecting the precipitate, washing with sterile water 3 times, and resuspending.
6. Use of lactobacillus plantarum CQ02-108 according to claim 1 or the functional starter according to claim 4 or claim 5 for the manufacture of a fermented meat product.
7. Use according to claim 6, wherein the fermented meat product is a fermented sausage.
8. The application according to claim 7, characterized in that it comprises the following steps:
s1: adding flavoring and adjuvants into raw meat, and pickling at 4 deg.C;
s2: inoculating the functional starter of claim 4 or claim 5 into raw meat, mixing, and making sausage, wherein the inoculation amount is 105-108cfu/g；
S3: fermentation: fermenting for 1 day at 25-35 ℃ and RH 80%;
s4: drying and maturing: standing at 15 deg.C and RH 75% for 4 days; drying at 12 deg.C and RH 72% for 16 days until the sausage is fermented and matured.
9. The use according to claim 8,
the raw material meat in the S1 is fresh pig hind leg lean meat and pig backfat, fascia, big tendons and other parts on the surface of the raw material meat are removed and rinsed, and the meat is stirred and mixed according to the ratio of the pig hind leg lean meat to the backfat of 8:2 to obtain the mixture;
the seasoning and the auxiliary materials are added according to the following mass percentage with the raw meat: 2% of salt, 1% of cane sugar, 1% of glucose, 0.015% of sodium nitrite, 0.05% of sodium erythorbate, 0.1% of ginger powder, 0.1% of white pepper powder and 0.1% of five spice powder.
10. The use of claim 8, wherein the amount of inoculation in S2 is 107cfu/g。
11. The use according to claim 8, wherein the fermentation temperature in S3 is 30 ℃; the application further comprises S5: and (4) trimming the sausage, and carrying out vacuum packaging to obtain a finished product.
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