CN113416676A - Lactobacillus plantarum DC2 and application of lactobacillus plantarum DC2 in fermented sour pork - Google Patents
Lactobacillus plantarum DC2 and application of lactobacillus plantarum DC2 in fermented sour pork Download PDFInfo
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- A—HUMAN NECESSITIES
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- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
- A23L13/70—Tenderised or flavoured meat pieces; Macerating or marinating solutions specially adapted therefor
- A23L13/72—Tenderised or flavoured meat pieces; Macerating or marinating solutions specially adapted therefor using additives, e.g. by injection of solutions
- A23L13/74—Tenderised or flavoured meat pieces; Macerating or marinating solutions specially adapted therefor using additives, e.g. by injection of solutions using microorganisms or enzymes
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- 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
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/90—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in food processing or handling, e.g. food conservation
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Abstract
The invention belongs to the technical field of strain screening and application, and provides lactobacillus plantarum DC2 and application of lactobacillus plantarum DC2 in fermented sour pork. The lactobacillus plantarum DC2 provided by the invention, named Lactobacillus plantarum DC2 by Latin, is preserved in China center for type culture Collection with the preservation address: china, Wuhan university, the preservation number: CCTCC NO: m2021427, date of deposit: 22/4/2021. The lactobacillus plantarum DC2 provided by the invention is used for fermenting sour pork, and the sour pork produced by the lactobacillus plantarum DC2 through a low-temperature fermentation technology has good appearance and color, so that the aim of reducing the cholesterol content in the pork is fulfilled on the premise of not influencing the pork sense.
Description
Technical Field
The invention belongs to the technical field of strain screening and application, and particularly relates to lactobacillus plantarum DC2 and application of lactobacillus plantarum DC2 in fermented sour pork.
Background
For a long time, pork is a basic and important meat food for meat production and consumption in China, accounts for 60 percent of the total meat amount, and accounts for over 5000 ten thousand tons each year. With the change of life style, the annual consumption of pork in average population shows a trend of steadily increasing, from 12.5 kg in 1995 to 21.0 kg in recent years. Pork is the main food source of animal protein (21.4g/100g pork) and lipid (3.5g/100g pork), and the quality of the pork is closely related to the health of residents. According to the dietary guidelines, healthy intake of lipids is recommended to be 20.0% to 30.0% of total energy. However, lipid intake of over 67.9% of residents is too high, over 30%.
Cholesterol is a sterol substance widely present in various tissues and cells of the human body, plays an important physiological role in the body, and is an essential nutrient component of the human body, but accumulation of excessive cholesterol in the body is accompanied by various health problems. The daily intake of cholesterol of residents in China sharply rises from 165.8 mg/day thirty years ago to 300 mg/day, and pork lipid is one of the main sources of cholesterol. High cholesterol diets are risk factors for developing chronic diseases such as metabolic and cardiovascular diseases, in part by activating inflammation and relying on regulation by low density lipoproteins. Therefore, the deep processing method for reducing the cholesterol content in pork is one of important ways for improving the availability of nutrient substances in pork and making the pork a healthy diet for human beings.
Pork processed products are various in types, including fermented sour meat, dried meat, preserved meat, roasted meat, pickled meat and the like, and have a long history in China. At present, the sour pork is mainly prepared by traditional natural fermentation. However, it is still difficult for natural sour pork to meet quality standards due to potential food safety problems caused by various microorganisms and unstable fermentation processes. In pork processing, high temperature technology is still the most important processing mode at present, but has disadvantages compared with low temperature processing of pork. The low-temperature fermentation of pork by using probiotics is a promising method, and sour pork with unique flavor, texture, color and luster and low cholesterol content can be commercialized to balance dietary nutrition and facilitate digestion and absorption. However, the pork fermentation industry has many difficulties such as lack of advantageous probiotic resources and corresponding mature fermentation technology.
Disclosure of Invention
The invention aims to reduce the cholesterol content in pork on the premise of not influencing the pork sense.
In order to solve the technical problems, the invention provides the following technical scheme:
the invention provides Lactobacillus plantarum DC2 with the Latin name of Lactobacillus plantarum DC2, which is preserved in China center for type culture Collection with the preservation address: china, Wuhan university, the preservation number: CCTCC NO: m2021427, date of deposit: 22/4/2021.
The invention also provides application of the lactobacillus plantarum DC2 in fermenting sour pork.
Preferably, the lactobacillus plantarum DC2 is used for fermenting low-cholesterol sour pork.
The invention also provides a method for fermenting sour pork by using the lactobacillus plantarum DC2, wherein the lactobacillus plantarum DC2 is inoculated into the pork, and the salt, the corn flour and the pepper are added in an amount which is 8-12 wt% of the pork, 10-12 wt% of the pork and 3-5 wt% of the pork, and the mixture is sealed and fermented.
Preferably, the inoculation amount of the lactobacillus plantarum DC2 is (1-32). times.106cfu/g。
Preferably, the fermentation temperature is 10-45 ℃.
Preferably, the pH value of the fermentation is 3.6-9.6.
Preferably, the fermentation time is 18-25 days.
The invention also provides the sour pork fermented according to the method.
The invention utilizes probiotics to ferment the pork, can improve the flavor of the pork, reduce the cholesterol content in the pork and improve the bioavailability of nutrient substances. The dietary sour pork produced by the lactobacillus plantarum DC2 through the low-temperature fermentation technology has good appearance and color, and the purpose of reducing the cholesterol content in the pork on the premise of not influencing the pork sense is achieved. The prepared dietary sour pork is beneficial to reducing blood fat, and has important significance for people with metabolism impairment or high cholesterol intake syndrome.
Drawings
FIG. 1 is a colony characteristic of DC 2;
FIG. 2 is the gram stain result of the DC2 strain;
FIG. 3 is a PCR amplification band of 16S rRNA of DC2 strain;
FIG. 4 is a molecular phylogenetic diagram of the strain DC 2;
FIG. 5 shows sour pork prepared in example 2 of the present invention;
FIG. 6 is a standard curve of cholesterol plotted in example 2 of the present invention;
FIG. 7 shows the cholesterol content of pre-fermented pork and post-fermented sour pork;
FIG. 8 shows the body weight changes of mice in the pre-fermented pork group and the fermented sour pork group;
FIG. 9 shows serum Lipopolysaccharide Binding Protein (LBP) content of mice in pre-fermented pork groups and fermented sour pork groups;
FIG. 10 shows Total Cholesterol (TC) levels in pre-fermented pork and fermented sour pork mice;
FIG. 11 shows the Low Density Lipoprotein (LDL) content of mice in the pre-fermented pork group and the fermented sour pork group;
FIG. 12 is Triglyceride (TG) content of mice in pre-fermented pork groups and fermented sour pork groups;
fig. 13 is the High Density Lipoprotein (HDL) content of mice in the pre-fermented pork group and the fermented sour pork group.
Deposit description
Lactobacillus plantarum DC2, Latin with the name Lactobacillus plantarum DC2, deposited in the China center for type culture Collection with the deposition address: china, Wuhan university, the preservation number: CCTCC NO: m2021427, date of deposit: 22/4/2021.
Detailed Description
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
Example 1
The DC2 strain provided by the invention is subjected to morphological observation, physiological and biochemical identification and 16S rRNA sequence analysis.
1. Morphological observation
The colony characteristics of DC2 are shown in FIG. 1, and the colony diameter is 1.5 mm. + -. 1mm, and appears circular, swollen, irregular edge, and turns from off-white to light pink in color. Gram staining results (see FIG. 2) were gram positive, without Bacillus.
2. Physiological and biochemical identification
The physiological and biochemical identification adopts a catalytic enzyme test, a gelatin liquefaction test, a nitrate reduction test and H2S production test, gluconic acid and gas production test, exercise test, litmus milk test and sugar fermentation test.
TABLE 1 physiological and biochemical Properties of DC2 Strain
As can be seen from Table 1, the strain DC2 showed positive results in the gelatin liquefaction test, the gluconic acid and gas production test, the exercise test, the litmus milk test and the sugar fermentation test, and in the catalytic enzyme test, the nitrate reduction test and the H2The gas production test shows negative.
3. 16S rRNA sequence analysis
A sequence of 1509bp was amplified from 16S rRNA of DC2 strain (PCR amplified band is shown in FIG. 3), and its nucleotide sequence is shown in SEQ ID NO: 1 is shown. Molecular phylogenetic analysis was performed using the maximum likelihood method, and the tree (-9870.6765) with the highest log-likelihood involved 101 nucleotide sequences (see fig. 4), which showed close homology (99.93%) between the DC2 strain and the lactobacillus plantarum strain (NR 115605.1). Therefore, the DC2 strain was identified as lactobacillus plantarum.
Example 2
Taking 1000g of pork loin square meat, scalding pork into feather, scraping and washing, passing through boiling water, draining, slicing, and cutting according to the proportion of 107cfu/g of pork is inoculated with lactobacillus plantarum DC2, then 100g of salt, 120g of corn flour and 40g of pepper are added and fully mixed, the pH value is measured to be neutral, the mixture is placed into a sealed jar and fermented for 20 days at 37 ℃, and the sour pork is prepared. The fermented sour pork showed good appearance and color (see fig. 5).
And measuring the cholesterol content of the unfermented pork and the fermented sour pork by taking the unfermented pork as a control, and drawing a standard curve of the cholesterol. The determination method refers to colorimetric method in determination of cholesterol in food safety national standard (GB 5009.128-2016). The regression equation of the linear relation of the established cholesterol standard curve (shown in figure 6) is that y is 2.4592x-0.0018 (R)20.9958). And calculating the cholesterol content in the sample according to a regression equation. The cholesterol contents of the pre-fermented pork and the fermented sour pork are shown in fig. 7, and then are calculated according to the following formula: the degradation efficiency (%) of cholesterol (cholesterol content of pork before fermentation-cholesterol content of fermented sour pork)/cholesterol content of pork before fermentation was calculated to obtain a degradation rate of the DC2 strain to cholesterol in pork of 77.2%.
Example 3
10 7-week-old female C57BL/6 mice (SJA laboratory animals Co., Ltd, Hunan) were randomly divided into two groups of 5, including a Pre-fermented pork group (Pre-FM) and a fermented sour pork group (Post-FM), and placed in a clean room where food and water were freely available. After 3 days of acclimation, two groups of mice were fed with pre-fermented pork (pork loin) and fermented sour pork prepared in example 2, respectively. Mice were weighed every 3 days. After 15 days, the mice were euthanized to collect blood samples. The content of Total Cholesterol (TC), low-density lipoprotein (LDL), high-density lipoprotein (HDL), Triglyceride (TG) and serum Lipopolysaccharide Binding Protein (LBP) in blood samples was measured using a biochemical automatic analyzer (DS5J0002, sinnawa medical science and technology limited, nanjing, china). All animal experiments were approved by the ethics and welfare committee of laboratory animals of the university of south central (2020sydw 0201).
Data are presented as mean ± standard deviation of three independent experiments. Differences between all groups were assessed using analysis of variance, and statistical significance of differences between groups was assessed using Tukey's multiple comparison test. Statistical analysis used SPSS software version 19.0 (IBM corporation, Armonk, NY, USA). P <0.01 and P <0.05 were significant.
As can be seen from fig. 8, the mice fed the fermented sour pork group were significantly lighter in weight than the pre-fermented pork group (fig. 8), indicating that the weight gain of the mice fed the sour pork was relatively slow.
As can be seen from FIGS. 9 to 12, the contents of serum Lipopolysaccharide Binding Protein (LBP), Total Cholesterol (TC), low-density lipoprotein (LDL) and Triglyceride (TG) in mice fed with the fermented sour pork group were significantly lower than those in the pre-fermented pork group. There was no significant difference in high density lipoprotein in mice fed pre-fermented and fermented acid pork groups (see figure 13). The dietary sour pork provided by the invention is proved to be beneficial to reducing blood fat and controlling weight.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Sequence listing
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Claims (9)
1. Lactobacillus plantarum DC2, Latin, Lactobacillus plantarum DC2, deposited in the China center for type culture Collection, accession number: china, Wuhan university, the preservation number: CCTCC NO: m2021427, date of deposit: 22/4/2021.
2. Use of lactobacillus plantarum DC2, according to claim 1, for fermenting sour pork.
3. The use of claim 2, wherein the lactobacillus plantarum DC2 is used for fermenting low-cholesterol sour pork.
4. A method for fermenting sour pork by using lactobacillus plantarum DC2 is characterized in that lactobacillus plantarum DC2 is inoculated into pork, salt accounting for 8-12 wt% of pork, corn flour accounting for 10-12 wt% of pork and pepper accounting for 3-5 wt% of pork are added, and sealed fermentation is carried out.
5. The method according to claim 4, wherein the lactobacillus plantarum DC2 has a bacterial inoculation amount of (1-32) x 106cfu/g。
6. The method of claim 4, wherein the temperature of the fermentation is 10-45 ℃.
7. The method of claim 4, wherein the pH of the fermentation is 3.6 to 9.6.
8. The method of claim 4, wherein the fermentation time is 18 to 25 days.
9. A sour pork fermented according to the method of any one of claims 4 to 8.
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