CN107217019B - Lactobacillus plantarum and application thereof in preparation of Chinese pork fermented sausage - Google Patents
Lactobacillus plantarum and application thereof in preparation of Chinese pork fermented sausage Download PDFInfo
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- 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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- 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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- 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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- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
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- C12R2001/00—Microorganisms ; Processes using microorganisms
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Abstract
Lactobacillus plantarum and application thereof in preparation of Chinese pork fermented sausage. The lactobacillus plantarum is a new strain selected from food and is applied to the production of the sausage, the fermentation period of the sausage is shortened, and the oxidation degree of fat and protein is reduced. Lactobacillus plantarum NJAU-01(Lactobacillus plantarum NJAU-01) with the preservation number: CGMCC No. 14194. The fermentation strain has strong antioxidant activity, the quality and the antioxidant property of the produced pork fermented sausage are superior to those of the traditional fermented sausage, the fermentation period is short, the fat oxidation degree is low, and the protein oxidation degree is obviously reduced. In addition, the pH value of the fermented sausage added with the leavening agent is lower than that of the traditional fermented sausage, so that the growth and propagation of harmful bacteria are effectively inhibited, the sausage putrefaction and mildew can be inhibited to a certain extent, the quality guarantee period of the Chinese-style fermented sausage is prolonged, and the economic loss caused by rapid product deterioration is reduced for enterprises.
Description
Technical Field
The invention relates to the technical field of food processing, in particular to lactobacillus plantarum and application thereof in preparation of Chinese pork fermented sausages.
Background
The meat product processing industry in China is in the transition period from traditional to finish processing. The traditional fermented sausage depends on natural fermentation of microorganisms in the environment, has long fermentation time and single product, has low oxidation resistance, particularly generates a plurality of substances harmful to human bodies due to oxidation of fat, and is not beneficial to the health after being eaten. During the oxidative metabolism of life activities, various free radicals are continuously generated, and the free radicals can attack polyunsaturated fatty acids to trigger lipid peroxidation, so that the body is damaged. At present, in order to reduce the oxidation degree, a chemically synthesized antioxidant is generally added in the preparation process, but the safety problem exists, so that the search for a natural antioxidant is one of effective ways for solving the problem. Lactic acid bacteria reduce the production of ROS and scavenge free radicals in vivo, and studies have shown that lactic acid bacteria cells and cell-free extracts have similar antioxidant activity in vitro. The strain with excellent oxidation resistance is screened from meat products and applied to the fermented sausage, and a foundation is laid for the development of the functional fermented sausage.
Disclosure of Invention
Aiming at the problems, the invention provides the lactobacillus plantarum and the application thereof in the preparation of the Chinese pork fermented sausage, and the lactobacillus plantarum of a new strain screened from food is applied to the production of the sausage, so that the fermentation period of the sausage is shortened, and the oxidation degree of fat and protein is reduced.
The technical scheme of the invention is as follows: lactobacillus plantarum NJAU-01(Lactobacillus plantarum NJAU-01) with the preservation number: CGMCC No. 14194.
The application of Lactobacillus plantarum NJAU-01(Lactobacillus plantarum NJAU-01) in preparation of Chinese sausage is characterized in that the Lactobacillus plantarum NJAU-01(Lactobacillus plantarum NJAU-01) is deposited with the following accession number: CGMCC No. 14194.
A preparation method of a Chinese pork fermented sausage comprises the following steps:
1) and preparing a Lactobacillus plantarum NJAU-01(Lactobacillus plantarum NJAU-01) seed solution: culturing Lactobacillus plantarum strain with MRS broth at 37 deg.C for 18h, centrifuging at 4 deg.C in 6000g centrifuge for 10min, washing with deionized water twice, and resuspending in deionized water to obtain bacterial suspension, adjusting bacterial count of bacterial suspension cell precipitate to 105-109cfu/ml, wherein the Lactobacillus plantarum NJAU-01(Lactobacillus plantarum NJAU-01) is deposited with a deposit number of: CGMCC No. 14194;
2) and preparing the sausage:
2.1) separating fat from lean of fresh pork, cutting into blocks, washing in water at 3-10 ℃, removing blood clots and pigskin, and mincing for later use, wherein the weight of fat meat accounts for 30% of the total weight of the pork;
2.2) weighing the following ingredients according to 100 parts of the total weight of the meat: 6-7 parts of white sugar, 3-4 parts of salt, 0.1-0.2 part of monosodium glutamate, 0.1-0.2 part of five spice powder, 0.1-0.2 part of ginger and 10-20ml/kg of Daqu, and then adding water accounting for 9-12% of the total weight of the meat and the ingredients;
2.3) inoculating the bacterial suspension prepared in the step 1) into the pickled meat paste and mixing uniformly, wherein the inoculation amount is 109cfu/g, and then filling the sausage;
2.4), 30 +/-0.5 ℃ and RH 90-95 percent for 1 day;
2.5), 16 +/-0.5 ℃, RH 85% -90% for 6 days, and then 12 +/-0.5 ℃, RH 75% -80% for 14 days to ferment and mature the sausage;
2.6) trimming the fermented sausage, and vacuum packaging to obtain the finished product of the fermented sausage.
And 2.2) mixing the pork and the ingredients in the step 2.2), and pickling for 1-2h at normal temperature.
And (3) removing the mould in the fermentation period in the step 2.4).
The invention has the beneficial effects that: the fermentation strain has strong antioxidant activity, the quality and the antioxidant property of the produced pork fermented sausage are superior to those of the traditional fermented sausage, the fermentation period is short, the fat oxidation degree is low, and the protein oxidation degree is obviously reduced. In addition, the pH value of the fermented sausage added with the leavening agent is lower than that of the traditional fermented sausage, so that the growth and propagation of harmful bacteria are effectively inhibited, the sausage putrefaction and mildew can be inhibited to a certain extent, the quality guarantee period of the Chinese-style fermented sausage is prolonged, the economic loss caused by rapid product deterioration is reduced for enterprises, and the fermented sausage is specifically embodied in the following aspects:
(1) the fermentation period of the product is 21 days, the fermentation period of the traditional fermented sausage is 28-45 days, and the fermentation period is shortened.
(2) The product has the advantages of rapid color development, compact texture, smooth and fine cut surface, jujube red color, white fat, luster, moderate saltiness, and obviously enhanced hardness, chewiness and elasticity.
(3) Degree of fat oxidation, represented by TBA value, inoculated with bacteriaThe oxidation degree of the fat is low, and the inoculation amount is 109The malondialdehyde content of cfu/g group is significantly lower than that of the control group 1.46 × 10-5mg MDA/kg; effectively inhibit lipid oxidation, and improve taste and nutritional safety of the product.
(4) The protein oxidation resistance of the product is represented by carbonyl number, and the inoculation amount is 109The carbonyl content of cfu/g group is lower than that of the control group by 0.12nmol carbonyl/mg protein.
(5) The oxidation resistance of the protein of the product is represented by mercapto value, and the inoculation amount is 109The content of sulfydryl in cfu/g group is higher than that in the control group by 2.57nmol sulfydryl/mg protein.
(6) Compared with the traditional fermented sausage, the product has lower pH value, slows down the putrefaction and the mildew of the sausage caused by the action of harmful microorganisms such as staphylococcus aureus, salmonella and the like, prolongs the shelf life of the product and ensures the safety of the product.
The Lactobacillus plantarum NJAU-01(Lactobacillus plantarum NJAU-01) screened by the method is preserved in China general microbiological culture Collection center (CGM C) in 2017, 5 months and 26 days, and is classified and named as follows: lactobacillus plantarum (Lactobacillus plantarum), accession number: CGMCC No. 14194. The address of the depository: xilu No.1 Hospital No. 3, national academy of sciences, North Kyoho, Beijing, Chaoyang; and E, postcode: 100101.
drawings
FIG. 1 is a graph comparing DPPH clearance;
FIG. 2 is a graph comparing carbonyl values;
FIG. 3 is a comparison of total thiol groups.
Detailed Description
The present invention will be described in detail with reference to examples.
First, screening of strain and research of antioxidant activity
EXAMPLE 1 screening of bacterial species
(1) Separation and purification: randomly selecting Jinhua ham, randomly determining 4 points on the point-signed muscle and biceps femoris, cutting off a part with the thickness of about 0.5cm and the side length of about 4cm on the surface of the ham by using a sterile scissors and a scalpel, and discarding the ham; cutting into meat blocks with thickness of about 2cm and similar size; the method comprises the steps of firstly carrying out surface sterilization on meat blocks on an alcohol lamp, then placing the meat blocks into a sterilized plastic bottle, carrying out aseptic operation, shearing about 2.0g of deep muscle from a meat sample by using an aseptic scissors, placing the meat sample into a sterilized mortar for crushing, then adding sterilized sea sand for grinding, adding sterilized water after grinding, uniformly mixing, diluting into 1:10 diluent, and then gradually diluting by 10 times. Spreading on MRS culture medium, culturing in biochemical incubator at 37 deg.C for 18-24 hr, and repeatedly streaking to separate and purify single colony until single strain is obtained.
(2) Screening: gram-positive, non-mucus-producing, NaCl (8%) resistant, non-gas-producing, non-H-producing2S, non-productive H2O2Without amino acid decarboxylase as the primary screening index; and (3) screening out bacterial strains suitable for being inoculated in the fermented sausage by taking the acid production speed, the acid production capacity and the aroma production as the secondary screening indexes of the lactic acid bacteria.
(3) And (3) identification: according to the phenotypic characteristics and the physiological and biochemical characteristics of colonies, the strain is determined to be Lactobacillus plantarum (Lactobacillus plantarum) by combining 16S sequencing.
(4) And (4) preservation: mixing bacterial liquid and glycerol according to the proportion of 1:1 and preserving at-70 ℃.
The strain is preserved in China general microbiological culture Collection center (CGMCC) and Lactobacillus plantarum NJAU-01(Lactobacillus plantarum NJAU-01) in 2017, 5 and 26 months (the preservation number is CGMCC No. 14194).
Example 2 study of antioxidant Activity of Lactobacillus plantarum NJAU-01(Lactobacillus plantarum NJAU-01)
The cells of the strain obtained in example 1 were centrifuged (6000g, 10min, 4 ℃), washed twice with deionized water and resuspended in deionized water. Bacterial count of the cell pellet was adjusted to 107、108、109cfu/ml to obtain a sample solution.
(1) DPPH clearance assay: 2mL of sample solution and 1mL of 0.2mmol/L DPPH-absolute ethanol solution, mixing and performing dark treatment for 30min, centrifuging at 6000 rpm for 10min, taking supernatant (adjusted to zero by equal volume of mixed solution of distilled water and absolute ethanol and positioned at 517 nm), and measuring absorbance A1The absorbance A was measured in the same manner by using an equal volume of absolute ethanol instead of DPPH0Steaming with equal volumeMeasuring absorbance A by the same method using distilled water instead of sample solution2. DPPH clearance ═ 1- (A)1-A0)/A2]× 100% Vc as control, results are shown in figure 1, the highest DPPH free radical clearance rate of Vc in control group reaches 57.36%, and then the concentration of Lactobacillus plantarum is 109The DPPH free radical clearance rate of the intracellular cell-free extract of cfu/ml is higher, the clearance rate reaches 42.62 percent and is obviously higher than the strain concentration of 108cfu/ml and 107DPPH radical scavenging Rate (p) of cfu/ml<0.05), the lactobacillus plantarum of the invention has good ability to scavenge DPPH radicals.
(2) Hydrogen peroxide tolerance test: an overnight culture of Lactobacillus plantarum Strain was inoculated with 1% (V/V) to MRS broth containing 0.4, 0.7, 1.0mM H2O2(30%) Vc was used as a control and incubated at 37 ℃ for 8 h. OD was measured with a spectrophotometer at a wavelength of 600 nm. The results are shown in table 1 and show that: the Lactobacillus plantarum NJAU-01(Lactobacillus plantarum NJAU-01) strain can tolerate a certain concentration of hydrogen peroxide (H)2O2) The tolerance of the strain to hydrogen peroxide is in a concentration-dependent relationship.
TABLE 1 tolerance of Lactobacillus plantarum to hydrogen peroxide
Preparation of sausage
Example 3
1) And preparing a Lactobacillus plantarum NJAU-01(Lactobacillus plantarum NJAU-01) seed solution: culturing Lactobacillus plantarum strain with MRS broth at 37 deg.C for 18h, centrifuging at 4 deg.C in 6000g centrifuge for 10min, washing with deionized water twice, and resuspending in deionized water to obtain bacterial suspension, adjusting bacterial count of bacterial suspension cell precipitate to 105cfu/ml, wherein the Lactobacillus plantarum NJAU-01(Lactobacillus plantarum NJAU-01) is deposited with a deposit number of: CGMCCNo.14194;
2) and preparing the sausage:
2.1) separating fresh pork and fat and lean meat, cutting into blocks, cleaning in water at 3 ℃, removing blood clots and pigskin, and mincing for later use, wherein the weight of fat meat accounts for 30% of the total weight of meat;
2.2) weighing the following ingredients according to 100 parts of the total weight of the meat: 6 parts of white sugar, 3 parts of salt, 0.1 part of monosodium glutamate, 0.1 part of five spice powder, 0.1 part of ginger and 10ml/kg of Daqu, adding water accounting for 9 percent of the total weight of meat and the ingredients, mixing pork and the ingredients, and pickling for 1 hour at normal temperature;
2.3) inoculating the bacterial suspension prepared in the step 1) into the pickled meat paste and mixing uniformly, wherein the inoculation amount is 105cfu/g, and then filling the sausage;
2.4), 29.5 ℃, RH 90 percent for 1 day, and removing mold in the fermentation period;
2.5), 15.5 ℃, RH 85% for 6 days, and then 11.5 ℃ and RH 75% for 14 days to ferment and mature the sausage;
2.6) trimming the fermented sausage, and vacuum packaging to obtain the finished product of the fermented sausage.
Example 4
1) And preparing a Lactobacillus plantarum NJAU-01(Lactobacillus plantarum NJAU-01) seed solution: culturing Lactobacillus plantarum strain with MRS broth at 37 deg.C for 18h, centrifuging at 4 deg.C in 6000g centrifuge for 10min, washing with deionized water twice, and resuspending in deionized water to obtain bacterial suspension, adjusting bacterial count of bacterial suspension cell precipitate to 107cfu/ml, wherein the Lactobacillus plantarum NJAU-01(Lactobacillus plantarum NJAU-01) is deposited with a deposit number of: CGMCCNo.14194;
2) and preparing the sausage:
2.1) separating fresh pork and fat and lean meat, cutting into blocks, washing in water at 10 ℃, removing blood clots and pigskin, and mincing for later use, wherein the weight of fat meat accounts for 30% of the total weight of the pork;
2.2) weighing the following ingredients according to 100 parts of the total weight of the meat: 7 parts of white sugar, 4 parts of salt, 0.2 part of monosodium glutamate, 0.2 part of five spice powder, 0.2 part of ginger and 20ml/kg of Daqu, adding water accounting for 12 percent of the total weight of meat and the ingredients, mixing pork and the ingredients, and pickling for 2 hours at normal temperature;
2.3) inoculating the bacterial suspension prepared in the step 1) into the pickled meat paste and mixing uniformly, wherein the inoculation amount is 107cfu/g, then filling with incenseA bowel;
2.4), 30.5 ℃, RH 95 percent for 1 day, and removing mould in the fermentation period;
2.5), 16.5 ℃, RH 90 percent for 6 days, and then 12.5 ℃ and RH 80 percent for 14 days to ferment and mature the sausage;
2.6) trimming the fermented sausage, and vacuum packaging to obtain the finished product of the fermented sausage.
Example 5
1) And preparing a Lactobacillus plantarum NJAU-01(Lactobacillus plantarum NJAU-01) seed solution: culturing Lactobacillus plantarum strain with MRS broth at 37 deg.C for 18h, centrifuging at 4 deg.C in 6000g centrifuge for 10min, washing with deionized water twice, and resuspending in deionized water to obtain bacterial suspension, adjusting bacterial count of bacterial suspension cell precipitate to 109cfu/ml, wherein the Lactobacillus plantarum NJAU-01(Lactobacillus plantarum NJAU-01) is deposited with a deposit number of: CGMCCNo.14194;
2) and preparing the sausage:
2.1) separating fresh pork and lean meat, cutting into blocks, washing in water at 6 ℃, removing blood clots and pigskin, and mincing for later use, wherein the weight of fat meat accounts for 30% of the total weight of meat;
2.2) weighing the following ingredients according to 100 parts of the total weight of the meat: 6.5 parts of white sugar, 3.5 parts of salt, 0.15 part of monosodium glutamate, 0.15 part of five spice powder, 0.15 part of ginger and 15ml/kg of Daqu, adding water accounting for 10 percent of the total weight of meat and the ingredients, mixing pork and the ingredients, and pickling for 1.5 hours at normal temperature;
2.3) inoculating the bacterial suspension prepared in the step 1) into the pickled meat paste and mixing uniformly, wherein the inoculation amount is 109cfu/g, and then filling the sausage;
2.4), 30 ℃, RH 92 percent for 1 day, and removing mould in the fermentation period;
2.5), 16 ℃, RH 88 percent for 6 days, and then 12 ℃ and RH 78 percent for 14 days to ferment and mature the sausage;
2.6) trimming the fermented sausage, and vacuum packaging to obtain the finished product of the fermented sausage.
EXAMPLE 6 control group (CK)
2.1) separating fresh pork and lean meat, cutting into blocks, washing in water at 6 ℃, removing blood clots and pigskin, and mincing for later use, wherein the weight of fat meat accounts for 30% of the total weight of meat;
2.2) weighing the following ingredients according to 100 parts of the total weight of the meat: 6.5 parts of white sugar, 3.5 parts of salt, 0.15 part of monosodium glutamate, 0.15 part of five spice powder, 0.15 part of ginger and 15ml/kg of Daqu, adding water accounting for 10 percent of the total weight of meat and the ingredients, mixing pork and the ingredients, and pickling for 1.5 hours at normal temperature;
2.3), 30 ℃, RH 92 percent for 1 day, and removing mould in the fermentation period;
2.4), 16 ℃, RH 88 percent for 6 days, and then 12 ℃ and RH 78 percent for 14 days to ferment and mature the sausage;
2.5) trimming the fermented sausage, and vacuum packaging to obtain the finished product of the fermented sausage.
Third, performance detection
Example 7pH value
10g of the sample was weighed, 90mL of distilled water was added, and the mixture was homogenized and the pH was measured with an acidimeter. The results are shown in table 2, and it can be seen from table 2 that the addition of Lactobacillus plantarum (Lactobacillus plantarum NJAU-01) significantly lowers the pH of the fermented sausage, the low pH value can effectively inhibit the growth and propagation of harmful bacteria, the sausage spoilage and mildew can be inhibited to a certain extent, and the shelf life of the Chinese style fermented sausage is prolonged.
TABLE 2 pH values of the examples
| Example 3 | Example 4 | Example 5 | Example 6(CK) | |
| pH value | 5.07±0.068 | 4.99±0.064 | 4.53±0.015 | 5.27±0.027 |
Example 8TBA value
1g of meat was thoroughly minced (5 s twice at 5000 rpm), 10ml of distilled water was added and homogenized 2 times at 12000rpm for 30s each time. 0.2ml of homogenate was added to 0.2ml of 8.1% SDS solution, 1.5ml of 20% acetic acid buffer solution of pH 3.5, 1.5ml of 0.8% TBA solution, and 0.6ml of distilled water, and the mixture was shaken and mixed, subjected to water bath at 95 ℃ for 60min, and cooled with flowing water for 10 min. Then adding 1ml of distilled water and 4ml of a mixture of pyridine and n-butanol (1: 15) into the test tube, uniformly mixing, vortexing, centrifuging at 4000rpm for 10min, and taking the supernatant and measuring the absorbance at 532 nm.
Taking 1 mu g/ml TEP standard solution 0, 0.1, 0.2, 0.3, 0.4, 0.5 and 0.6ml, adding the standard solution into a test tube, adding 0.2ml of 8.1 percent SDS solution, 1.5ml of 20 percent acetic acid buffer solution (pH 3.5) and 1.5ml of 0.8 percent TBA solution, fixing the volume to 4ml to ensure that the TEP amount in the system is respectively 0, 0.1, 0.2, 0.3, 0.4, 0.5 and 0.6mg, carrying out water bath at 95 ℃ for 60min, cooling with flowing water for 10min, adding 1ml of distilled water and a mixture of 4ml pyridine and n-butanol (1: 15), centrifuging at 4000rpm for 10min after uniformly mixing and vortexing, and measuring the absorbance at 532 nm.
The results are shown in table 3, the content of Malondialdehyde (MDA) in the inoculation group is significantly lower than that in the blank group, the lipid oxidation of the product is effectively inhibited, the content of MDA increases with the fermentation time, and the increasing trend of MDA is reduced in the later period of fermentation.
TABLE 3 TBA values in the examples
Example 9 carbonyl number
Weighing 1.5g of meat sample in a 50mL centrifuge tube (only taking lean meat and meat to stir), adding 15mL of pyrophosphate buffer solution, taking homogenate liquid in the centrifuge tube (each tube is 3mL), dividing into a control group and an experimental group, adding 3mL of 20% TCA, shaking and mixing uniformly, centrifuging 12000g for 5min, taking precipitate, adding 3mL of 10% TCA, shaking and mixing uniformly, centrifuging (12000g for 5min), and taking precipitate. Adding 3mL of 2M HCl into a control group, adding 3mL of DNPH into an experimental group, standing in the dark for 30min, shaking once every 10min for 10s, adding 20% TCA, shaking, mixing uniformly, centrifuging (12000g, 5min), and taking a precipitate. Adding 7.5mL ethanol/ethyl acetate mixture, mixing, centrifuging (12000g, 5min), collecting precipitate, and repeating for 3 times. 3mL of guanidine hydrochloride was added, the mixture was stored at 4 ℃ overnight and centrifuged (12000g, 5min), and the supernatant was collected to measure the absorbance (370 nm).
As shown in FIG. 2, it can be seen from FIG. 2 that the degree of protein oxidation (represented by carbonyl value) of the product prepared by the present invention is significantly lower than that of the control group, which indicates that Lactobacillus plantarum NJAU-01(Lactobacillus plantarum NJAU-01) effectively reduces the generation of carbonyl groups and inhibits the oxidation of proteins.
Example 10 mercapto number
Taking 0.2ml of 2mg/ml protein solution, adding 1.8ml of urea buffer solution and 80 mu L of DTNB, carrying out water bath reaction at 40 ℃ for 25min, taking out the reaction solution, cooling, and measuring the absorbance value of the reaction solution at 412nm, wherein the molar absorption coefficient is 13,600M-1/cm-1. Total thiol units are expressed in nanomoles of thiol per milligram of protein.
As shown in FIG. 3, it can be seen from FIG. 3 that the degree of protein oxidation (represented by a thiol value) of the product prepared by the present invention is significantly higher than that of the control group, which indicates that Lactobacillus plantarum NJAU-01(Lactobacillus plantarum NJAU-01) effectively inhibits the oxidation of thiol, i.e., inhibits the oxidation of protein.
Example 11 sensory quality
Cutting the sample into a cylinder with the thickness of 1cm by using a double-sided knife, connecting a T-shaped metal pressure head on a material instrument, sensing a source 1000N, an initial force 0.3N, the diameter of the pressure head 4l mm, a compression ratio 50%, and measuring the speed: 50mm/min, two cycles, measurement items: hardness (N); elasticity (mm); chewiness (mJ); cohesiveness.
The texture analysis is shown in Table 4, the color difference analysis is shown in Table 5,
TABLE 4 texture analysis in the examples
TABLE 5 analysis of color difference in examples
| Example 3 | Example 4 | Example 5 | Example 6(CK) | |
| L* | 46.73±1.11 | 49.49±0.45 | 49.51±0.83 | 42.98±2.66 |
| a* | 13.57±0.94 | 11.47±0.33 | 9.85±0.27 | 12.36±0.04 |
| b* | 10.43±0.77 | 8.95±0.36 | 10.26±0.18 | 9.01±1.19 |
L: represents the shade (black and white), a: representing red and green, b: represents a yellow-blue color
As can be seen from tables 4 and 5, the pork fermented sausage added with Lactobacillus plantarum NJAU-01(Lactobacillus plantarum NJAU-01) has the advantages of compact muscle texture, smooth and fine cut surface, red color, moderate salinity and strong fermentation flavor. The strains used by the invention are all separated from Jinhua dry-cured ham and are subjected to safety analysis, so the use is safe and reliable.
Claims (5)
1. Lactobacillus plantarum NJAU-01(Lactobacillus plantarum NJAU-01), which is characterized by being deposited with a accession number of: CGMCC No. 14194.
2. The application of Lactobacillus plantarum NJAU-01(Lactobacillus plantarum NJAU-01) according to claim 1 to preparation of Chinese pork fermented sausage, wherein the Lactobacillus plantarum NJAU-01(Lactobacillus plantarum NJAU-01) is deposited with the following accession number: CGMCC No. 14194.
3. A preparation method of a Chinese pork fermented sausage is characterized by comprising the following steps:
1) and preparing a Lactobacillus plantarum NJAU-01(Lactobacillus plantarum NJAU-01) seed solution: culturing Lactobacillus plantarum strain with MRS broth at 37 deg.C for 18h, centrifuging at 4 deg.C in 6000g centrifuge for 10min, washing with deionized water twice, and resuspending in deionized water to obtain bacterial suspension, adjusting bacterial count of bacterial suspension cell precipitate to 105-109cfu/ml, wherein the Lactobacillus plantarum NJAU-01(Lactobacillus plantarum NJAU-01) is deposited with a deposit number of: CGMCCNo.14194;
2) and preparing the sausage:
2.1) separating fat from lean of fresh pork, cutting into blocks, washing in water at 3-10 ℃, removing blood clots and pigskin, and mincing for later use, wherein the weight of fat meat accounts for 30% of the total weight of the pork;
2.2) weighing the following ingredients according to 100 parts of the total weight of the meat: 6-7 parts of white sugar, 3-4 parts of salt, 0.1-0.2 part of monosodium glutamate, 0.1-0.2 part of five spice powder, 0.1-0.2 part of ginger and 10-20ml/kg of Daqu, and then adding water accounting for 9-12% of the total weight of the meat and the ingredients;
2.3) inoculating the bacterial suspension prepared in the step 1) into the pickled meat paste and mixing uniformly, wherein the inoculation amount is 109cfu/g, and then filling the sausage;
2.4), 30 +/-0.5 ℃ and RH 90-95 percent for 1 day;
2.5), 16 +/-0.5 ℃, RH 85% -90% for 6 days, and then 12 +/-0.5 ℃, RH 75% -80% for 14 days to ferment and mature the sausage;
2.6) trimming the fermented sausage, and vacuum packaging to obtain the finished product of the fermented sausage.
4. The preparation method of the Chinese pork fermented sausage of claim 3, wherein the pork in the step 2.2) is mixed with the ingredients and then is cured at normal temperature for 1-2 h.
5. The preparation method of the Chinese pork fermented sausage of claim 3, wherein the mould is removed in the fermentation period in the step 2.4).
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