Method for reducing biogenic amine in air-dried goose product through combination of pulsed electric field and inoculation fermentation

Abstract

The invention discloses a method for reducing biogenic amine in an air-dried goose product by combining a pulsed electric field with inoculation fermentation, which is characterized by comprising the following steps: treating fresh goose meat at 0.5-1kv / cm for 50-180s, pickling the goose meat in a 8wt% sodium chloride solution for 24h, hanging the pickled goose meat in an air dryer for 1h, draining off water, and drying the goose meat in the air dryer to obtain dried goose meat; the method comprises the following steps: uniformly spraying a compound bacterial solution consisting of lactobacillus plantarum pdd-1 and lactococcus lactis according to the inoculation amount of 1 * 10 < 5 >-1 * 10 < 7 > CFU / mL, and air-drying for 3-5 days at 10-20 DEG C to obtain the air-dried goose product with low biogenic amine, which has the advantages of obviously reducing the content and variety of biogenic amine, inhibiting the degradation of protein in the air-dried goose and obviously increasing flavor substances.

Description

Technical Field

[0001] The invention relates to a method for reducing biogenic amines in air-dried poultry products, in particular to a method for reducing biogenic amines in air-dried goose products by combining pulse electric field with inoculated fermentation. Background Art

[0002] Goose meat is an ideal high-protein, low-fat, low-cholesterol nutritious and healthy food. It is not only tender, delicious and juicy, but also rich in a variety of essential amino acids for the human body, and has a high content of unsaturated fatty acids, which is very beneficial to human health. In addition, goose meat also has the function of medicinal diet therapy. As a unique traditional Chinese meat product, Feng goose is favored by consumers because of its fresh taste, unique flavor and easy preservation. However, due to the high content of protein and amino acids in goose meat, it may cause microbial contamination in a certain suitable environment, or different types and quantities of biogenic amines may be produced under the condition of strain fermentation. The production of biogenic amines in the production process of Feng goose is extremely complex. Different microorganisms may cause the accumulation of different biogenic amines. At the same time, it may also be related to the residual decarboxylase activity produced by microorganisms in the raw materials. Therefore, according to the production process of Feng goose, strains with the function of degrading biogenic amines are screened to provide effective measures for the degradation of biogenic amines in meat products.

[0003] Biogenic amines are low molecular weight organic, alkaline, nitrogen-containing compounds that exist in plants, microorganisms and animal cells. A certain amount of monoamine compounds has a significant effect on the relaxation and contraction of blood vessels and muscles; it can also regulate the cerebral cortex and nervous system of organisms; polyamines can promote the synthesis of proteins, nucleotides and hormones in the body, and play an important role in the growth process of the body. Although biogenic amines have multiple physiological functions, consuming foods containing a large amount of such compounds may have toxicological effects and may cause adverse reactions to special consumers. Among them, histamine is the most toxic biogenic amine. Excessive content can cause symptoms such as headache, dizziness, nausea, palpitations or difficulty breathing. With the development of technology, packaging forms, irradiation, plant extract blocking and other methods of controlling biogenic amines have emerged one after another. These technologies are indeed effective in the theoretical research process, but a single technology cannot effectively control biogenic amines. For example, irradiation and microbial degradation technology have production cost problems for meat product companies in actual production, making it difficult to apply them to actual production.

[0004] As a non-thermal food processing technology, pulsed electric field technology uses short electric pulses under high voltage to keep the thermal effect at a low level, so that the food material does not undergo large temperature changes during the processing process, which is beneficial to maintaining the color, aroma, taste and nutritional components of the food. The electroporation effect produced by pulsed electric field technology can improve the quality of materials and the transport of energy, and will also affect the quality characteristics of food materials. In addition to assisting pickling, pulsed electric field-assisted processing technology is currently mainly used in the field of meat processing to improve tenderness, sterilize, assist freezing and thawing, extract substances, etc. There are currently no reports on its assisted degradation of biogenic amines in meat products. Summary of the invention

[0005] The technical problem to be solved by the present invention is to provide a method for reducing biogenic amines in air-dried goose meat products by combining pulsed electric field with inoculation and fermentation, which can significantly reduce the content and types of biogenic amines, inhibit protein degradation in air-dried goose, and significantly increase flavor substances.

[0006] The technical solution adopted by the present invention to solve the above technical problems is: a method for reducing biogenic amines in air-dried goose meat products by pulse electric field combined with inoculated fermentation, characterized by comprising the following steps: treating fresh goose meat with 0.5-1 kV / cm for 50-180 seconds, placing it in 8wt% sodium chloride solution for 24 hours, hanging the marinated goose meat in an air dryer for 1 hour, draining the water, and pressing 1×10 5 ~1×10 7 The composite bacterial liquid composed of Lactobacillus plantarum PDD-1 and Lactococcus lactis is evenly sprayed at an inoculum amount of CFU / mL, and air-dried at 10-20°C for 3-5 days to obtain an air-dried goose meat product with low biogenic amines.

[0007] Further, the method comprises the following steps: treating fresh goose meat in a pulse electric field treatment chamber at 0.7 kV / cm for 150 seconds, then marinating the goose meat in an 8 wt % sodium chloride solution at 4°C for 24 hours, hanging the marinated goose meat in an air dryer for 1 hour, draining the water, and drying the goose meat at 1×10 6 The composite bacterial liquid consisting of Lactobacillus plantarum PPD-1 and Lactococcus lactis in a volume ratio of 1:1 was evenly sprayed at a CFU / mL inoculum amount, and after air-drying at 16°C for 4 days, an air-dried goose meat product with low biogenic amines was obtained.

[0008] Furthermore, the OD values ​​of the Lactobacillus plantarum PDD-1 bacterial liquid and the Lactococcus lactis bacterial liquid are both 1.

[0009] Further, the plant lactobacillus is classified and named as plant lactobacillus (Lactobacillus plantarum subsp. plantarum) PDD-1 strain, which was deposited in the General Microbiology Center of China National Microbiological Culture Collection Committee on June 19, 2018, with a deposit number of CGMCC No. 15953; the Lactococcus lactis subsp. lactis strain was purchased from China National Microbiological Culture Collection Center (CGMCC) and is numbered JCM5805.

[0010] Compared with the prior art, the advantages of the present invention are: a method for reducing biogenic amines in air-dried goose meat products by combining pulse electric field with inoculation and fermentation, compared with pulse electric field alone and inoculation and fermentation alone, this method can be used to significantly reduce the content and types of biogenic amines in goose meat, and can significantly inhibit protein degradation in air-dried goose, and its flavor substances are also significantly increased. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Figure 1 is the standard curve of different biogenic amines;

[0012] Figure 2 The degradation rates of total biogenic amines in MRS medium by 15 strains;

[0013] Figure 3 The single factor influences the content of biogenic amines in wind geese; A is the influence of electric field intensity on the content of total biogenic amines in wind geese; B is the influence of electric field treatment time on the content of total biogenic amines in wind geese; C is the influence of air-drying time on the content of total biogenic amines in wind geese; D is the influence of different volume ratios of composite strains on the content of total biogenic amines in wind geese;

[0014] Figure 4 The effects of different treatment methods on the quality of low-biogenic amine process wind goose; A is various biogenic amines of wind goose; B is the total biogenic amines of wind goose;

[0015] Figure 5 is the TVB-N value of wind goose with different treatment methods;

[0016] Figure 6 The electronic noses of wind geese were subjected to different treatments; A is the radar chart of the odor difference analysis of wind geese; B is the principal component analysis (PCA) score chart of the odor difference analysis of wind geese. DETAILED DESCRIPTION

[0017] The present invention is further described in detail below with reference to the accompanying drawings. Specific embodiment 1

[0019] 1. Method for determination of biogenic amines in wind goose by high performance liquid chromatography (HPLC)

[0020] (1) Preparation of standard solution and pre-column derivatization

[0021] Weigh 10 mg each of tryptamine, phenylethylamine, putrescine, cadaverine, histamine, tyramine, spermidine and spermine, dilute to 10 mL with 0.1 mol / L hydrochloric acid, and dilute to prepare mixed standard solutions with final concentrations of 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, and 50.0 mg / L, respectively. Take 1 mL of the mixed solution of the standard, add 100 μL of 2 mol / L NaOH to make it alkaline, then add 300 μL of saturated NaHCO3 solution for buffering, then add 2 mL of 10 g / L dansyl chloride acetone solution, react the dansyl chloride solution at 40 ° C in the dark for 45 min, then add 100 μL of ammonia water to stop the reaction, react in the dark at room temperature for 30 min, remove the residual dansyl chloride solution, and add acetonitrile to make the final volume 5 mL. After derivatization, filter with a 0.22 μm filter membrane for analysis and detection.

[0022] (2) Extraction of biogenic amines

[0023] Take 5g of goose meat product and add 20mL 0.1mol / L HCL, homogenize thoroughly on a homogenizer, centrifuge in a refrigerated centrifuge (4℃ 4000rpm / min) for 10min, and re-extract the precipitate as before. Take the supernatant from the second extraction and make it up to 50mL with 0.1mol / L HCL, and take 1mL of the sample solution for pre-column derivatization as the standard solution.

[0024] (3) Chromatographic conditions

[0025] The chromatographic column is Waters C 18 (4.6×250mm, 5μm), flow rate of 1mL / min, UV detection wavelength of 254nm, injection volume of 10μL, column temperature of 30℃, mobile phase A is water, mobile phase B is acetonitrile, gradient elution program is used, the elution program is:

[0026] Table 1 Elution program

[0027]

[0028] The biogenic amine standard solutions with concentrations of 1.0 mg / L, 2.5 mg / L, 5.0 mg / L, 10.0 mg / L, 15.0 mg / L, 25.0 mg / L, and 50.0 mg / L were subjected to HPLC detection, and the obtained peak area was used as the ordinate and the concentration of the standard solution was used as the abscissa to draw the biogenic amine standard curve. Figure 1 It can be seen that there is a good linear relationship between the peak areas of the eight biogenic amines and their corresponding concentrations.

[0029] 2. Degradation ability of strains on biogenic amines

[0030] Fifteen test strains were taken from the strain bank of the laboratory of the Animal Processing Team of the School of Food and Pharmaceutical Sciences of Ningbo University. They were inoculated into MRS liquid culture medium at a volume ratio of 4% and activated for 3 generations. The medium was centrifuged at 8000rpm / min for 10 minutes, washed twice with phosphate buffer, and the bacteria were collected and inoculated with the bacterial suspension at a volume ratio of 2% in an inoculum ratio into MRS culture medium containing 8 biogenic amines (tryptamine, phenylethylamine, putrescine, cadaverine, histamine, tyramine, spermidine, and spermine, with a concentration of 100mg / L for each biogenic amine). After culturing at 37°C for 48h, 1mL of the sample was taken, 1mL of 0.1M hydrochloric acid was added and mixed, and 1mL of the mixed solution was taken for pre-column derivatization. The MRS culture medium containing biogenic amines without any bacteria was stored under the same conditions for 48h as a blank control. The samples were measured in parallel three times and the average value was taken. L1 is Levilactobacillus brevis X10; L2 is Levilactobacillus brevis X11; L3 is Levilactobacillus brevis 54; L4 is Lactobacillus fermentum 55; L5 is Lactobacillus acidophilus 6074; L6 is Lactobacillus fermentum JF6; L7 is Lactobacillus fermentum JF9; L8 is Lactobacillus fermentum JF10; L9 is Lactiplantibacillus plantarum c8; L10 is Lactiplantibacillus plantarum PDD-1; L11 is Lactobacillus reuteri reuteri)max; L12 is Lactobacillus reuteri 3k; L13 is Lactobacillus reuteri 4k; L14 is Lactococcus lactis subsp. lactis; L15 is Lactobacillus casei. Results are shown in Figure 2, a total of 5 strains had a total biogenic amine degradation rate of more than 26%. The total biogenic amine degradation rates of Lactobacillus acidophilus 6074, Lactobacillus plantarum PDD-1, Lactobacillus plantarum C8, Lactococcus lactis and Lactobacillus casei were 28.68%, 30.69%, 30.42%, 31.93% and 30.32% respectively, which were significantly higher than the remaining 10 strains (p < 0.05).

[0031] 3. Effects of environmental factors on strain growth

[0032] The five strains screened were renumbered, where L1 was Lactobacillus acidophilus 6074, L2 was Lactobacillus plantarum PDD-1, L3 was Lactobacillus plantarum C8, L4 was Lactococcus lactis subsp. lactis; and L5 was Lactobacillus casei.

[0033] (1) Effect of temperature on the degradation of four biogenic amines by five strains

[0034] After the test strain was activated for 3 generations, it was transferred to MRS liquid culture medium containing 100 mg / L of each of the four biogenic amines (putrescine, cadaverine, histamine, and tyramine) at a 4% inoculation rate. Since the temperature of the air-dried goose was 16°C, after culturing at 16°C for 24 hours, it was centrifuged at 4000 rpm / min for 10 minutes, and 1 mL of the supernatant was taken for derivatization and testing. The effect of temperature on the growth of the five biogenic amine-degrading bacteria is shown in Table 2.

[0035] Table 2 Effect of temperature on the degradation of biogenic amines by strains

[0036]

[0037] Note: Different lowercase letters in the same column indicate significant differences (p<0.05), and different uppercase letters in the same row indicate significant differences (p<0.05);

[0038] The results showed that the ability of Lactobacillus plantarum PDD-1 and Lactococcus lactis to degrade tyramine, putrescine and cadaverine was significantly stronger than that of the other three strains, among which Lactococcus lactis had a degradation capacity of 24.62% for tyramine and Lactobacillus plantarum PDD-1 had a degradation capacity of 23.81%. Overall, Lactobacillus plantarum PPD-1 and Lactococcus lactis had a strong ability to degrade the four biogenic amines at 16°C.

[0039] (2) Effect of initial pH on the degradation of four biogenic amines by five strains

[0040] After the test strain was activated for 3 generations, it was transferred to MRS liquid culture medium with pH of 5.4, 5.7, and 6.0 containing 100 mg / L of 4 biogenic amines (putrescine, cadaverine, histamine, and tyramine) at a 4% inoculation amount. After culturing at 16°C for 24 hours, it was centrifuged at 4000 rpm / min for 10 minutes, and 1 mL of supernatant was taken for derivatization and testing. The pH of wind goose is generally above 5.6, so strains with effective biogenic amine degradation ability under different pH conditions were selected. The effects of different initial pH on the growth of 5 biogenic amine degrading bacteria are shown in Table 3.

[0041] Table 3 Effect of initial pH on biodegradation of biogenic amines by strains

[0042]

[0043] Note: Different lowercase letters in the same column indicate significant differences (p<0.05), and different uppercase letters in the same row indicate significant differences (p<0.05);

[0044] As shown in Table 3, under different initial pH conditions, the five strains have different abilities to degrade various biogenic amines. When the pH is low, the strains have weaker abilities to degrade the four biogenic amines. As the pH increases, the strains' abilities to degrade various biogenic amines increase, especially the degradation of tyramine. Among them, the degradation rates of tyramine by Lactobacillus plantarum and Lactococcus lactis in MRS medium with an initial pH of 6.0 reached more than 60%. Overall, the strains have the strongest ability to degrade tyramine, followed by putrescine, cadaverine and histamine. Among them, Lactobacillus plantarum PDD-1 and Lactococcus lactis have the best effects in degrading the four biogenic amines at different pH.

[0045] In summary, Lactobacillus plantarum PDD-1 and Lactococcus lactis

[0046] (Lactococcus lactis subsp. lactis) was significantly better than the other three strains in its ability to degrade four biogenic amines under different environmental factors, so these two strains were selected for subsequent experiments.

[0047] (3) Effect of initial pH on the degradation of four biogenic amines by two bacterial strains

[0048] In order to explore whether there is antagonism between the two strains, the degradation ability of the two strains at different initial pH to four biogenic amines was determined. After the test strains were activated for 3 generations, Lactobacillus plantarum PDD-1 with an OD value of 1 and Lactococcus lactis subsp. lactis with an OD value of 1 were mixed in a volume ratio of 1:1, and then inoculated in an inoculum volume ratio of 4% to MRS liquid culture medium with pH values ​​of 5.4, 5.7, and 6.0, respectively, containing 100 mg / L of four biogenic amines (putrescine, cadaverine, histamine, and tyramine). After culturing at 16°C for 24 hours, the mixture was centrifuged at 4000 rpm / min for 10 minutes, and 1 mL of the supernatant was taken for derivatization and testing. The results are shown in Table 4 below.

[0049] Table 4 Effect of initial pH on degradation of biogenic amines by composite bacteria

[0050]

[0051] Note: Different capital letters in the same row indicate significant differences (p<0.05);

[0052] The results showed that the ability of the composite strain to degrade putrescine, cadaverine, histamine and tyramine at different pH values ​​was stronger than that of the single strain, indicating that the composite strain was better than the single strain in degrading the four biogenic amines and had a synergistic effect.

[0053] In summary, Lactobacillus plantarum PDD-1 and Lactococcus lactis subsp. lactis JCM 5805 have good ability to degrade putrescine, cadaverine, histamine and tyramine under different environmental factors, and the interaction between the two is more effective than that of a single strain. Specific embodiment 2

[0055] 1. Effect of electric field intensity on the content of biogenic amines in wind geese

[0056] The goose meat was placed in the treatment chamber of the pulse electric field extraction equipment, and was subjected to pulse electric field treatment for 60 seconds under the conditions of pulse field strength of 0.5 kV / cm, 1 kV / cm, 1.5 kV / cm, and 2 kV / cm, pulse width of 2 ms, and pulse frequency of 100 Hz. The goose meat was then soaked in 8 wt % sodium chloride solution and marinated at 4 ° C for 24 hours. After being air-dried in an air dryer for 3 days, the goose breast meat was taken out, the fat and fascia were removed, and the meat was minced and packaged. 5 g of goose meat was taken from each group and the content of biogenic amines was determined by HPLC after pre-treatment. The results are shown in Table 5.

[0057] Table 5 Biogenic amine content in wind goose under different pulse electric field intensities (mg / 100g)

[0058]

[0059] Note: Different lowercase letters in the same column indicate significant differences (p<0.05), and different uppercase letters in the same row indicate significant differences (p<0.05).

[0060] As shown in Table 5, under different electric field intensities, a total of 4 biogenic amines were detected, among which the contents were spermine, spermidine, phenylethylamine and cadaverine from high to low. The contents of cadaverine, spermidine and spermine in the wind goose treated with an electric field intensity of 0.5 kv / cm were significantly lower than those in other treatment groups.

[0061] 2. Effect of electric field treatment time on the content of biogenic amines in wind geese

[0062] The goose meat was placed in the treatment chamber of the pulse electric field extraction equipment, and the pulse electric field treatment was carried out under the conditions of pulse field strength of 1kv / cm, pulse width of 2ms, and pulse frequency of 100HZ. After treatment for 30s, 60s, 90s, 120s, 150s, and 180s, the goose meat was soaked in 8wt% sodium chloride solution at 4°C for 24h, and air-dried in an air dryer for 3 days. The goose breast meat was taken, the fat and fascia were removed, minced and packaged, and 5g of goose meat was taken from each group for pre-treatment and the content of biogenic amines was determined by HPLC. With the increase of treatment time, the total biogenic amine content in the air-dried goose showed dynamic changes. A total of 5 biogenic amines were detected under the treatment of 60s, among which the content of spermine was the highest, and the content of phenylethylamine and cadaverine was lower. A total of 4 biogenic amines were detected under the remaining treatment time, and the content of spermine was also the highest. When the treatment was 120s, the content of cadaverine was second only to spermine, reaching 3.01mg / 100g. Overall, the content of biogenic amines in wind geese was the lowest when the electric field intensity was treated for 180s.

[0063] Table 6 Biogenic amine content in wind geese under different pulse electric field treatment time (mg / 100g)

[0064]

[0065] Note: Different lowercase letters in the same column indicate significant differences (p < 0.05), and different uppercase letters in the same row indicate significant differences (p < 0.05)

[0066] 3. Effect of air-drying time on the content of biogenic amines in wind goose

[0067] The goose meat was soaked in 8wt% sodium chloride solution at 4℃ for 24h, and then put into the air dryer for air drying. The goose breast meat was taken at 0d, 1d, 2d, 3d, 4d, 5d, 6d, 7d, and 8d of air drying, and the fat and fascia were removed. The goose breast meat was minced and packaged. 5g of goose meat was taken from each group and the content of biogenic amines was determined by HPLC after pretreatment. A total of 6 biogenic amines were detected during the air drying process of the air-dried goose, among which the spermine content was the highest at 0 to 6d of air drying, the tyramine content was the highest at 7d of air drying, and the cadaverine content was the highest at 8d of air drying. Overall, except for 0d, the content of biogenic amines in the air-dried goose for 3d was the lowest.

[0068] Table 7 Biogenic amine content in air-dried geese at different air-drying times (mg / 100g)

[0069]

[0070] Note: Different lowercase letters in the same column indicate significant differences (p < 0.05), and different uppercase letters in the same row indicate significant differences (p < 0.05)

[0071] 4. Effect of inoculation amount on the content of biogenic amines in Feng Geese

[0072] The goose meat was soaked in 8 wt % sodium chloride solution at 4 °C for 24 h, air-dried in an air dryer for 1 h to drain the water, and then sprayed with a composite bacterial solution of Lactobacillus plantarum PDD-1 and Lactococcus lactis in a volume ratio of 1:1, 1:2, 1:3, 2:1, and 3:1, respectively. The inoculation amount was 1×10 6 CFU / g, goose breast meat was taken on the 3rd day of air drying, fat and fascia were removed, minced and packaged, and 5g of goose meat was taken from each group for pre-treatment and the content of biogenic amines was determined by HPLC. Four biogenic amines were detected in the air-dried goose at different volume ratios, and tyramine was also detected when the volume ratio was 2:1. Among them, the content of biogenic amines from high to low were spermine, spermidine, phenylethylamine and cadaverine. The content of spermine and spermidine in the wind goose treated with a volume ratio of 1:1 of bacterial solution (OD value = 1) was the lowest. The cadaverine, spermidine and spermine in the wind goose treated with a volume ratio of 3:1 of bacterial solution were significantly higher than those in the wind goose treated with bacterial solution of other volume ratios. Overall, the content of biogenic amines in the wind goose treated with a volume ratio of 1:1 of bacterial solution was the lowest.

[0073] Table 8 Biogenic amine content in wind goose under different composite inoculation amounts (mg / 100g)

[0074]

[0075] Note: Different lowercase letters in the same column indicate significant differences (p<0.05), and different uppercase letters in the same row indicate significant differences (p<0.05); the above ratios are all Lactobacillus plantarum: Lactococcus lactis.

[0076] 4. Response surface experimental design

[0077] like Figure 3 A. The total biogenic amine content in the wind goose showed a trend of first increasing and then decreasing with the increase of electric field intensity, but overall, the total biogenic amine content in the wind goose treated with an electric field intensity of 0.5 kv / cm was the lowest. Figure 3 B. The total biogenic amine content in the wind goose changes dynamically with the electric field treatment time, reaching the highest value at 120s. The total amine content at 150s is lower than that at 30s, and there is a downward trend. Figure 3 The content of biogenic amines in C. goose increased slowly during the air-drying process, but decreased slightly on the third day. Figure 3 D. The content of biogenic amines in Feng Geese was different under the treatment of composite strains with different volume ratios. Generally speaking, the content of biogenic amines in Feng Geese with a volume ratio of 1:1 was the lowest. Based on the above single-factor experimental results, three indicators were selected: A: electric field intensity (kv / cm), B: electric field time (s), and C: air-drying time (d). A three-factor three-level Box-Behnken test was designed, and the total biogenic amine content (mg / 100g) was used as the response value to determine the optimal process of pulsed electric field combined with inoculation and fermentation to control biogenic amines in the processing of Feng Geese.

[0078] Table 9 Response surface experimental design scheme and results

[0079]

[0080] Table 10 Analysis of variance of regression model

[0081]

[0082]

[0083] Note: **p<0.01; *p<0.05

[0084] As can be seen from Table 10, the F value of each factor is ranked as B>C>A, that is, electric field time>air drying time>electric field strength. The F value of the model's lack of fit is 1.79, and the p value is 0.2883, which is greater than 0.05. The lack of fit is not significant, indicating that the model can be used to predict the optimal low-biogenic amine process of wind goose. The experimental data were fitted and analyzed using Design-Expert 13 software, and the quadratic polynomial regression equation was obtained: Y=81.68-118.37A+0.50B-47.00C+0.21AB+6.45AC+0.18BC+44.59A2-0.003B2+1.81C2.

[0085] Considering the interaction between electric field intensity, electric field time and air-drying time on total biogenic amines, taking total biogenic amine content as the optimization target, according to the results of Design-Expert 13 software operation, the optimal low biogenic amine process conditions for air-dried goose are electric field intensity of 0.674kv / cm, electric field time of 150.000s, and air-drying time of 4.193d. Under these conditions, the model predicted the biogenic amine content of 7.954mg / 100g. According to the prediction results of response surface software and the feasibility of actual process conditions, the optimal low biogenic amine process conditions for air-dried goose are electric field intensity of 0.7kv / cm, electric field time of 150s, and air-drying time of 4d. Under these conditions, three repeated experiments were carried out, and the average total biogenic amine was 7.71mg / 100g, which is close to the model prediction results, indicating that the method of optimizing the low biogenic amine air-dried goose meat process based on the response surface model is feasible.

[0086] 3. Application Examples

[0087] Effect of the low biogenic amine process obtained in the above specific example 2 on the quality of air-dried goose meat

[0088] CK-control group: fresh goose meat was marinated in 8wt% sodium chloride solution for 24h. The marinated goose meat was hung in an air dryer at 4℃ for 1h. After draining the water, the goose breast meat was air-dried at 16℃ for 4 days, and then minced and packaged, vacuumed, and stored at -80℃ for testing.

[0089] F-pulsed electric field group: fresh goose meat was treated at 0.7 kv / cm for 150 s, then marinated in 8 wt% sodium chloride solution at 4 °C for 24 h. The marinated goose meat was hung in an air dryer for 1 h, and after draining the water, it was air-dried at 16 °C for 4 days. The goose breast meat was minced, packaged, and vacuumed, and stored at -80 °C for testing.

[0090] PL-inoculation and fermentation group: Fresh goose meat was marinated in 8wt% sodium chloride solution for 24h. The marinated goose meat was hung in an air dryer for 1h at 4℃. After draining the water, 1×10 6 The CFU / mL was a composite bacterial solution composed of Lactobacillus plantarum PDD-1 and Lactococcus lactis in a volume ratio of 1:1. After air-drying at 16°C for 4 days, the goose breast meat was minced, packaged, vacuumed, and stored at -80°C for testing.

[0091] FPL-pulsed electric field combined with inoculation and fermentation group: fresh goose meat was treated with 0.7 kv / cm for 150 s, then marinated in 8 wt% sodium chloride solution at 4 °C for 24 h, and the marinated goose meat was hung in an air dryer for 1 h. After draining the water, 1×10 6The CFU / mL was a composite bacterial solution composed of Lactobacillus plantarum PDD-1 and Lactococcus lactis in a volume ratio of 1:1. After air-drying at 16°C for 4 days, the goose breast meat was minced, packaged, vacuumed, and stored at -80°C for testing.

[0092] 1. Effect of low biogenic amine process on biogenic amines in air-dried goose meat

[0093] like Figure 4 A, Five biogenic amines, including phenylethylamine, putrescine, histamine, tyramine and spermidine, were detected in air-dried goose meat, of which only three biogenic amines, including phenylethylamine, putrescine and tyramine, were detected in the FPL group. Compared with the control group, the content of putrescine, histamine, tyramine and spermidine in the group treated with pulsed electric field alone was significantly reduced, and the content of phenylethylamine increased but the difference was not significant. Compared with the control group, the content of putrescine, tyramine and spermidine in the group treated with inoculation and fermentation alone was significantly reduced, the content of histamine increased significantly, and the content of phenylethylamine decreased but the difference was not significant. In addition, after the pulsed electric field combined with inoculation and fermentation treatment of air-dried goose, the types of biogenic amines in goose meat decreased, the content of putrescine and tyramine decreased significantly, and the content of phenylethylamine was significantly different from that of the control group. Figure 4 B is the total biogenic amine content in air-dried goose under four different treatments. The results show that the biogenic amine content in the pulsed electric field group and the inoculated fermentation group is significantly lower than that in the control group, and under the synergistic effect of the two, the total biogenic amine content is significantly reduced to 7.54 mg / 100g. This shows that the pulsed electric field and inoculated fermentation can effectively inhibit the formation of biogenic amines in air-dried goose meat, and the pulsed electric field combined with inoculated fermentation has a more significant effect.

[0094] 2. Effect of low biogenic amine process on the texture of air-dried goose meat

[0095] Cut the air-dried goose breast into 1cm×1cm×1cm cubes, and use TA-XT2i texture analyzer with P50 probe to measure the texture of the sample. The TPA parameters are: pre-test speed 2mm / s, test speed 1mm / s, post-test speed 2mm / s, compression ratio 30%. Test items: hardness; elasticity; cohesion; viscosity; chewiness; resilience.

[0096] Table 11 Texture of low biogenic amine wind goose

[0097]

[0098]

[0099] Note: CK-control group, F-pulsed electric field group, PL-inoculation and fermentation group, FPL-pulsed electric field combined with inoculation and fermentation group.

[0100] The results are shown in Table 11. The pulsed electric field improves the tenderness of goose meat. Therefore, the hardness, stickiness and chewiness of goose meat decreased significantly after pulsed electric field treatment, while the hardness, stickiness and chewiness of goose meat increased significantly after inoculation and fermentation. This is because the growth and reproduction of the fermentation strain and its metabolic activity reduce the moisture content in the wind goose, which increases the hardness of the sample. The study shows that pulsed electric field and inoculation and fermentation have little effect on the elasticity, cohesion and resilience of meat products.

[0101] 3. Effect of low biogenic amine process on total volatile basic nitrogen (TVB-N) in air-dried goose meat

[0102] TVB-N is an important indicator for measuring the freshness of meat. The nitrogen-containing compounds in goose meat are degraded by microorganisms and enzymes, which often leads to an increase in TVB-N content. Take 6g of air-dried goose meat, add 30mL of sterile water, homogenize, let stand for 30 minutes, filter with filter paper, take 10mL of liquid, add 5mL of magnesium oxide suspension (10g / L), put it in the digestive tube, and distill it through a Kjeldahl nitrogen analyzer for 5 minutes. Add 2-3 drops of mixed indicator to the distillate, and titrate with 0.01M hydrochloric acid. The titration is terminated when the color changes from light yellow-green to light pink. Mixed indicator: 2 parts of methyl red-ethanol solution (1g / L) + 1 part of methylene blue-ethanol solution (1g / L), mix well, and use immediately after preparation. If Figure 5 As shown, the TVB-N of the CK group was significantly higher than that of the F, PL and FPL groups, among which the TVB-N value of the pulsed electric field combined with inoculation and fermentation group was the lowest, at 15.75 mg / 100 g. The results showed that the pulsed electric field and inoculation and fermentation could effectively inhibit the increase of TVB-N in Feng Geese, and the combination of the two had a better effect, that is, the pulsed electric field combined with inoculation and fermentation could effectively inhibit the degradation of Feng Geese protein.

[0103] 4. Effect of low biogenic amine process on flavor substances in air-dried goose meat

[0104] Weigh 5g of minced meat sample into a beaker, seal it with a sealing film, place it in a 50℃ water bath for 30min, and put it on the machine. Electronic nose instrument parameters: cleaning time 70s, data detection time 150s. Volatile components will generate signals when passing through 10 highly sensitive metal oxide detectors. The electronic nose sensors and their performance are shown in Table 12.

[0105] Table 12 Sensors and performance of PEN3 electronic nose

[0106]

[0107]

[0108] The electronic nose tends to be stable after 90 seconds from the beginning of detection. To ensure the accuracy and stability of the data, the data from 143 to 145 seconds were selected as the feature value extraction time point and PCA analysis was performed. The electronic nose radar chart and PCA analysis results of the air-fried goose breast samples processed by different processes are shown in Figure 6 A and Figure 6 B. The four wind goose samples produced strong responses on the W1S (methyl compounds) and W1W (organic sulfur compounds) sensors, indicating that the presence of these compounds may have a greater impact on the volatile compounds in the wind goose samples. The results show that pulsed electric field and inoculation fermentation can significantly increase the content of aromatic components, organic sulfides and nitrogen oxides in wind goose and improve the flavor. In addition, the W1S, W5S, W2S, W2W and W1W response values ​​of the FPL samples were the highest, indicating that the FPL samples may contain more nitrogen oxides, hydrides, methyls, short-chain alkane aromatics, alcohols, aldehydes and ketones, sulfides and pyrazines than other samples. At the same time, the results showed that the aroma of the FPL samples was very different from the other samples. The principal component analysis can reflect the characteristics of the volatile odors of wind geese treated differently. In the figure, PC1 and PC2 are 94.5% and 0.312%, respectively, and the cumulative is 94.812%. The results show that the odors of samples in different groups are significantly different.

[0109] The above description is not intended to limit the present invention, and the present invention is not limited to the above examples. Any changes, modifications, additions or substitutions made by a person skilled in the art within the spirit and scope of the present invention shall also fall within the protection scope of the present invention.

Claims

1. A method for reducing biogenic amines in air-dried goose meat products by combining pulsed electric field with inoculated fermentation, characterized in that The following steps are involved: Fresh goose meat was treated with 0.5-1 kv / cm for 50-180 s, then marinated in 8 wt% sodium chloride solution for 24 h. The marinated goose meat was hung in an air dryer for 1 h. After draining the water, the goose meat was dried at 1×10 5 ~1×10 7 The composite bacterial liquid composed of Lactobacillus plantarum PDD-1 and Lactococcus lactis is evenly sprayed at an inoculum amount of CFU / mL, and air-dried at 10-20°C for 3-5 days to obtain an air-dried goose meat product with low biogenic amines.

2. The method for reducing biogenic amines in air-dried goose meat products by combining pulsed electric field with inoculated fermentation according to claim 1, characterized in that The following steps are involved: Fresh goose meat was treated in a pulse electric field treatment chamber at 0.7 kV / cm for 150 s, and then marinated in 8 wt% sodium chloride solution at 4 °C for 24 h. The marinated goose meat was hung in an air dryer for 1 h, and after draining the water, the goose meat was dried at 1 × 10 6 The composite bacterial liquid consisting of Lactobacillus plantarum PPD-1 and Lactococcus lactis in a volume ratio of 1:1 was evenly sprayed at a CFU / mL inoculum amount, and after air-drying at 16°C for 4 days, an air-dried goose meat product with low biogenic amines was obtained.

3. The method for reducing biogenic amines in air-dried goose meat products by combining pulsed electric field with inoculated fermentation according to claim 2, characterized in that: The OD values ​​of the Lactobacillus plantarum PPD-1 bacterial liquid and the Lactococcus lactis bacterial liquid are both 1.

4. The method for reducing biogenic amines in air-dried goose meat products by combining pulsed electric field with inoculated fermentation according to claim 3, characterized in that: The plant lactobacillus is classified and named as plant lactobacillus (Lactiplantibacillus plantarum) PPD-1 strain, which is preserved in the General Microbiological Center of China Microorganism Culture Collection Administration Committee and has a preservation number of CGMCC No.15953; the lactococcus lactis (Lactococcus lactis subsp. lactis) strain is purchased from China Microorganism Culture Collection Center and has a number of JCM5805.

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