CN103610213B - The preparation method of food-grade lactobacillus acidophilus fermentation natural antiseptic agent and product - Google Patents
The preparation method of food-grade lactobacillus acidophilus fermentation natural antiseptic agent and product Download PDFInfo
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Classifications
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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
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
- A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
- A23L3/3463—Organic compounds; Microorganisms; Enzymes
- A23L3/3571—Microorganisms; Enzymes
-
- 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
- A23V2400/113—Acidophilus
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- Life Sciences & Earth Sciences (AREA)
- Microbiology (AREA)
- Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Food Preservation Except Freezing, Refrigeration, And Drying (AREA)
Abstract
The present invention relates to preparation method and product that natural antiseptic agent is produced in the fermentation of food-grade lactobacillus acidophilus, belong to technical field of food biotechnology.Tomato juice, beer, skimmed milk, pure water are become fermentation medium according to the proportions of 5 ~ 10:15 ~ 25:5 ~ 20:65 ~ 80, and 121 DEG C of sterilizing 7min, are fermentation medium.After lactobacillus acidophilus (Lactobacillus acidophilus) NX2-6 seed culture medium expands cultivation, 2 ~ 4% inoculations, 37 DEG C of quiescent culture 36h, zymotic fluid collected by centrifugation supernatant, the freeze drying of-20 DEG C of pre-freeze 24h final vacuums, obtains natural sandy gravel.This product, to Main Pathogenic Bacteria in food and spoilage organisms, all has remarkable inhibitory action.This natural antiseptic agent antimicrobial spectrum is wide, security is high, therefore can be used for the industries such as food, medicine, feed.
Description
One, technical field
The present invention relates to preparation method and the product of food-grade lactobacillus acidophilus fermentation natural antiseptic agent, belong to technical field of food biotechnology, this anticorrisive agent can suppress the harmful microorganism in food to grow effectively, extends the shelf-life of food, is applicable to food processing and preservation and freshness.
Two, background technology
Microbial infection is the key factor causing food apoilage, according to estimates, the whole world every year about 25% food due to corrupt and discard, China about has the food of 20-30% to lose in vain because of corruption every year.In order to prevent food apoilage, be used physical method (as pickled, canning, refrigeration etc.) and carry out preserved food, but there is remarkable effect with the use of anticorrisive agent to preventing food spoilage under certain condition, therefore anticorrisive agent is as one of food additives, in the food industry extensive use.
Food preservative is as the class in food additives, it is defined as and prevents food apoilage, extends the material of food storing phase, the anticorrisive agent that People's Republic of China (PRC) GB2760-2011 " food additives use sanitary standard " clear stipulaties 28 class Permissible limits use.Food preservative can be divided into chemical preservative and the large class of natural antiseptic agent two by its source, and chemical preservative is larger, effective relative to natural antiseptic agent cheaper, output, so it is still main food fresh keeping wide spectrum additive at present.But through the discovery that studies for a long period of time, chemical preservative has certain toxicity to human body, and even some can lure cancer and teratogenesis, easily cause food poisoning when exceeding standard and using.
Along with the enhancing of growth in the living standard and Consciousness of food security, people are for the safety and sanitation of food and self health status growing interest, derive from plant, microorganism and animal natural sandy gravel because it is natural, efficient, nontoxic or low toxicity and do not destroy the significantly characteristic such as the original local flavor of food, become the new lover in international food additive market.At present, in the world generally acknowledge, safety, adopt Production by Microorganism Fermentation and developed country use primary biological anticorrisive agent have: nisin (Nisin), natamycin (Natamycin) and epsilon-polylysine (ε-poly-lysine).China's food additives standard (GB2760-2011) allows microbial source natural antiseptic agent only nisin and the natamycin two kinds used.
Nisin is that FAO (Food and Agriculture Organization of the United Nation) and the World Health Organization (FAO/WHO) confirm as safety, efficient, food preservative, up to the present more than 50 national extensive uses in the whole world reliably.Nisin is a kind of little peptide be made up of 34 amino acid residues produced through streptococcus lactis fermentation, to many gram-positive bacterias, there is bacteriostatic activity, especially remarkable bacteriostatic activity is had to Main Foods spoilage organisms such as clostridium (Clostridium) and bacillus (Bacillus), but its antimicrobial spectrum is narrower, to gramnegative bacterium unrestraint effect.Natamycin comes from actinomyces Natal mould (Streptomyces natalensis), is polyene macrolides fungistat, be merely able to suppress and kill yeast, mould and other filamentous fungis, but to bacterium and virus invalid.
Although more to the research of different microorganisms source natural antiseptic agent both at home and abroad, but what be really applied in the food industry is little, this is mainly because some of them natural antiseptic agent antimicrobial spectrum is narrow, fungistatic effect is not ideal enough, anti-microbial property is unstable, especially on the high side, productive rate is low and security can not determined, at present be also difficult to substituted chemistry anticorrisive agent status.Therefore developing wide spectrum, efficient, stable, safe antibacterial material is the required of food industry as antiseptics for natural food, is also the key of antiseptics for natural food development.Current U.S. FDA have approved with fresh-keeping for food of food security microorganism fermented food level raw material, as microgard.
Inventor screens a strain lactobacillus acidophilus NX2-6 from the traditional zymotic koumiss of Xilin Hot, Inner Mongolia area herdsman, MRS culture medium can produce antibacterial material (Theantibacterial properties and strain identification of Lactobacillus acidophilus NX2-6screened from Chigee, the Milchwissenschaft65 (2) 2010 such as WUYUNDALAI).And (Wuyun Dalai etc. are studied to the fermentation medium producing antibacterial material, lactobacillus acidophilus NX2-6 liquid fermentation bacteriocinogeny culture medium and main affecting factors screening thereof, Food Science, 2009,13:187-191 Wuyun Dalai etc., the fermentation condition optimization of lactobacillus acidophilus NX2-6 bacteriocinogeny, Food Science, 2012,3:179-183).But, because synthetic media adds materials such as there being more inorganic salts, can not use as food preservative.
Three, summary of the invention
Technical problem
Goal of the invention is for chemical preservative Problems existing in food industry and the increase in demand situation to broad-spectrum high efficacy natural antiseptic agent, there is provided lactobacillus acidophilus to ferment and produce the food-grade medium preparation method of natural antiseptic agent, it is fresh-keeping that this product can be used for food antiseptic, effectively can extend the storage life of food.
Technical scheme
Be separated the anticorrosion bacterial strain NX2-6 of the koumiss sample made from Xilin Hot, Inner Mongolia area herdsman family traditional fermentation methods, through being accredited as lactobacillus acidophilus (Lactobacillus acidophilus).First through the natural antiseptic agent that above-mentioned bacterial strains NX2-6 fermentation produces, obtain by the following method:
(1) preparation of food-grade culture medium
Tomato is cleaned, pulls an oar and make tomato juice, tomato is cleaned and makes tomato juice, tomato juice 5%, beer 15%, skimmed milk 5% are added pure water respectively, or tomato juice, beer, skimmed milk, pure water are become fermentation medium according to the proportions of 5 ~ 10:15 ~ 25:5 ~ 20:65 ~ 80,121 DEG C of sterilizing 7min, are fermentation medium.
(2) bacterial strain shake-flask seed liquid preparation
By be stored in-70 DEG C of glycerine pipes bacterial classification, be inoculated in 10% degreasing milk medium, put 37 DEG C, cultivate 10 ~ 12h.
(3) preparation of zymotic fluid
Getting seed liquor is inoculated in fermentation broth with 2 ~ 4%, puts 37 DEG C, cultivates 24h ~ 36h.By fermentation culture in the centrifugal 20min of 5000rpm, get supernatant.
(4) preparation of antibacterial product
By fermented liquid supernatant liquid after-20 DEG C of pre-freeze 24h, freeze drying, obtains natural antiseptic agent.
Natural antiseptic agent prepared by described method is to Main Pathogenic Bacteria in food and spoilage organisms, comprise 9 kinds of gram-positive bacteria (staphylococcus aureuses, micrococcus luteus, bacillus pumilus, bacillus cereus, singly increase listeria spp, Yi Shi listeria spp, grignard listeria spp, Wei Shi listeria spp) and 8 kinds of Gram-negative bacteria (bacillus colis, Pseudomonas fluorescens, Salmonella paratyphi A, salmonella typhimurium, Bacterium enteritidis, salmonella typhi, Salmonella choleraesuls hog cholera subspecies, Enterobacter sakazakii) all there is significant bacteriostasis.
The application of described natural antiseptic agent can be used for food processing and the anti-corrosive fresh-keepings such as fish, meat, fowl, egg, breast, fruits and vegetables, effectively controls microorganism in food growth, loses and extend effective period of food quality to have significant application value for minimizing food spoilage.
Natural antiseptic agent, under 4 DEG C of holding conditions, adds in Chilled Meats according to mass ratio 2-8% by described natural antiseptic agent, anticorrosion for Chilled Meats.
Beneficial effect
Major advantage of the present invention is as follows:
1, the present invention provides a kind of antibacterial product Main Pathogenic Bacteria in food and spoilage organisms to broad-spectrum high efficacy bacteriostasis first, is called for short FBF(food-based fermentates).FBF freeze-dried powder is to Main Pathogenic Bacteria in food and spoilage organisms, this product is to Main Pathogenic Bacteria in food and spoilage organisms, comprise 9 kinds of gram-positive bacteria (staphylococcus aureuses, micrococcus luteus, bacillus pumilus, bacillus cereus, singly increase listeria spp, Yi Shi listeria spp, grignard listeria spp, Wei Shi listeria spp) and 8 kinds of Gram-negative bacteria (bacillus colis, Pseudomonas fluorescens, Salmonella paratyphi A, salmonella typhimurium, Bacterium enteritidis, salmonella typhi, Salmonella choleraesuls hog cholera subspecies, Enterobacter sakazakii) all there is significant bacteriostasis.Because antibacterial product provided by the invention has obvious inhibitory action to harmful bacteria multiple in food, show high-efficiency broad spectrum characteristic.Therefore, can develop and become a kind of novel antiseptics for natural food, be widely used in food processing and the anti-corrosive fresh-keepings such as fish, meat, fowl, egg, breast, fruits and vegetables, effective control microorganism in food growth, has significant application value for the loss of minimizing food spoilage and prolongation effective period of food quality.
2, lactobacillus acidophilus NX2-6 antibacterial product provided by the invention is fresh-keeping for food antiseptic, Be very effective.Chilled Meats corrosion-resistanting test result shows: add the growth that antibacterial product effectively can suppress bacterium in Chilled Meats in Chilled Meats, slow down the generation of VBN in Chilled Meats, extend cooled meat shelf period, and FBF increases with addition, Chilled Meats extended shelf-life.Under 4 DEG C of holding conditions, add 2% its shelf life and can extend 6 days compared with the control, 4% can extend 9 days, and 8% can extend more than 9 days.
Four, accompanying drawing explanation
Fig. 1 lactobacillus acidophilus NX2-6 antibacterial product is to the fungistatic effect of part indicator bacteria.
Note: A: salmonella typhimurium CICC51005; B: salmonella typhi CMCC50071; C: Salmonella choleraesuls hog cholera subspecies ATCC13312; D: grignard listeria spp CICC21670; E: Enterobacter sakazakii CICC21563; F: Wei Shi listeria spp CICC21672; G: bacillus pumilus CMCC (B) 63202; H: staphylococcus aureus CMCC (B) 26003; I: micrococcus luteus CMCC (B) 28000.
TVB-N change in Fig. 2 different preservative treatment cooling meat pie
Note: different lowercase in each row (a, b, c, d and e) and in every a line different capitalizations (A, B, C, D, E and F) show significant difference (P<0.05).
TBARS change in Fig. 3 different preservative treatment cooling meat pie
Note: different lowercase in each row (a, b, c, d and e) and in every a line different capitalizations (A, B, C, D, E and F) show significant difference (P<0.05).
A* value change in Fig. 4 different preservative treatment cooling meat pie
Note: different lowercase (a, b, c, d and e) show significant difference (P<0.05).
PH value change in Fig. 5 different preservative treatment cooling meat pie
Note: different lowercase in each row (a, b, c, d and e) and in every a line different capitalizations (A, B, C, D, E and F) show significant difference (P<0.05).
The different preservative treatment of Fig. 6 is on the impact (quantitative descriptive analysis method) of Chilled Meats sense organ
Five, detailed description of the invention
The food antiseptic bacterial strain NX2-6(Wuyun Dalai etc. adopted in the present invention, lactobacillus acidophilus NX2-6 liquid fermentation bacteriocinogeny culture medium and main affecting factors screening thereof, Food Science, 2009,13:187-191 Wuyun Dalai etc., the fermentation condition optimization of lactobacillus acidophilus NX2-6 bacteriocinogeny, Food Science, 2012,3:179-183) from the traditional zymotic koumiss sample of Xilin Hot, Inner Mongolia area herdsman, separation screening obtains, through being accredited as lactobacillus acidophilus (Lactobacillus acidophilus).
Food antiseptic bacterial strain NX2-6 generation natural antiseptic agent of the present invention obtains by the following method.Natural antiseptic agent production process is:
1. food-grade medium preparing
Tomato is cleaned, pulls an oar and make tomato juice, tomato juice 5%, beer 15%, skimmed milk 5% are added pure water respectively, or tomato juice, beer, skimmed milk, pure water are become fermentation medium according to the proportions of 5 ~ 10:15 ~ 25:5 ~ 20:65 ~ 80,121 DEG C of sterilizing 7min, are fermentation medium.With MRS and TJA culture medium for contrast.
MRS culture medium: peptone 10.0g, beef extract 10.0g, yeast extract powder 5.0g, glucose 20.0g, Tween 80 1.0g, K2HPO43H2O2.0g, sodium acetate 5.0g, MgSO47H2O200.0mg, MnSO4H2O38.0mg, trisodium citrate water and thing 5.0g, pH6.5.
TJA culture medium: tomato juice 50mL, yeast extract 5.0g, beef extract 10.0g, lactose 20.0g, glucose 2.0g, K
2hPO
42.0g, Tween-80 1.0g, sodium acetate 5.0g, agar 15.0g, distilled water 1000mL, pH7.1 ~ 7.5,121 DEG C of sterilizing 15min.
2. bacterial strain seed liquor preparation
By the strain inoculation that is stored in glycerine pipe in 10% degreasing milk medium, 37 DEG C of quiescent culture 10 ~ 12h.
3. zymotic fluid preparation
Getting seed liquor is inoculated in fermentation broth by 3 ~ 5% inoculum concentrations, 37 DEG C of quiescent culture 24 ~ 36h.By fermentation culture in the centrifugal 20min of 5000rpm, get supernatant.
4. antibacterial product preparation
By fermented liquid supernatant liquid after-20 DEG C of pre-freeze 24h, freeze drying, obtains natural antiseptic agent.
(1) antibacterial activity of raw-food material fermentate
To 3 kinds of raw-food materials, the antibacterial activity of the bacillus acidophilus fermented product of tomato juice, beer and defatted milk carries out analysis and finds, 5% tomato juice and 15% beer fermentation thing do not detect antibacterial activity, and 5% defatted milk MRS and TJA culture medium have antibacterial activity.Therefore tomato juice, beer and skimmed milk power are carried out to the fermentate of Mixed culture, antibacterial activity increases considerably (table 1).
(2) natural antiseptic agent is to the bacteriostatic test of common pathogen and spoilage organisms
1. the indicator bacteria that bacteriostatic experiment adopts is important pathogenic bacteria and spoilage organisms in food: Salmonella paratyphi A (Salmonella paratyphi-ACICC21501), salmonella typhimurium (SalmonellatyphimuriumCICC21483), Bacterium enteritidis (Salmonella enteritidisCMCC50041), salmonella typhi (Salmonella enterica subsp.Enterica CMCC50071), Salmonella choleraesuls hog cholera subspecies (Salmonella enterica subsp.Enterica Leminor et popoffATCC13312), Enterobacter sakazakii (Enterobacter sakazakii CICC21560, CICC21563), listeria innocua (Listeria innocuaCICC10417), singly increase listeria spp (ListeriamonocytogenesCICC21662), Yi Shi listeria spp (Listeria ivanoviiCICC21663), grignard listeria spp (Listeria grayiCICC21670), Wei Shi listeria spp (Listeria welshimeriCICC21672), Escherichia coli (EscherichiacoliATCC25922), staphylococcus aureus (Staphylococcus aureusATCC25923), bacillus pumilus (Bacillus pumilusCMCC (B) 63202), bacillus cereus (Bacillus cereus AS1.1846), Pseudomonas fluorescens (PseudomonasfluorescensAS1.1802), the indicator bacterias such as micrococcus luteus (Micrococcus luteusAS1.191) are this Laboratories Accession bacterial strain.
2. indicator bacteria culture medium: each indicator bacteria is applicable to culture medium in table 2.
Nutrient agar (NA): peptone 10.0g, beef extract 3.0g, sodium chloride 5.0g, distilled water 1000mL, adjusts pH to be 7.2 ~ 7.4, (add agar 18.0 ~ 20.0g and be solid medium), 121 DEG C of sterilizing 20min.
Luria-Bertani culture medium (LB): tryptone 10.0g, yeast extract 5.0g, sodium chloride 10.0g, distilled water 1000mL, pH is adjusted to be 7.2 ~ 7.4, (add agar 18.0 ~ 20.0g and be solid medium), 121 DEG C of sterilizing 20min.
Brain-heart infusion medium (BHI): buy from Beijing road and bridge technology Co., Ltd, adds agar 18.0 ~ 20.0g and is BHI solid medium.
3. bacteriostatic test:
3.1 adopt punch method primary dcreening operation basic media components: Escherichia coli and staphylococcus aureus activation are carried out Multiplying culture, and physiological saline wash-out is made bacteria suspension and added mixing in NA to obtain the dull and stereotyped bacterial concentration of indicator bacteria be 10
6~ 10
7cfu/mL, chooses agar block with after card punch punching after culture medium solidifying, adds fermentation culture centrifuged supernatant 20 μ L in indicator bacteria flat board in it, cultivates 12h for 37 DEG C, measure and record antibacterial circle diameter (table 1) after 4 DEG C of diffusion 1h.
3.2 adopt Oxford cup double layer agar method to measure the antibacterial situation of Optimal Medium supernatant: the Oxford cup placing high-temperature sterilization on indicator bacteria flat board, add 200 μ L natural antiseptic agent freeze-dried powders redissolution liquid, after putting 4 DEG C, refrigerator diffusion 1h, cultivate 16 ~ 18h at 37 DEG C, observe and record antibacterial circle diameter size.Fungistatic effect is in table 1 and Fig. 1.Bacteriostatic test result proves: lactobacillus acidophilus NX2-6 antibacterial product is to Main Pathogenic Bacteria in food and spoilage organisms, comprise 9 kinds of gram-positive bacteria (staphylococcus aureuses, micrococcus luteus, bacillus pumilus, bacillus cereus, singly increase listeria spp, Yi Shi listeria spp, grignard listeria spp, Wei Shi listeria spp) and 8 kinds of Gram-negative bacteria (bacillus colis, Pseudomonas fluorescens, Salmonella paratyphi A, salmonella typhimurium, Bacterium enteritidis, salmonella typhi, Salmonella choleraesuls hog cholera subspecies, Enterobacter sakazakii) all there is significant bacteriostasis.
Because lactobacillus acidophilus NX2-6 antibacterial product has obvious inhibitory action to multiple food-borne pathogens and spoilage organisms, show high-efficiency broad spectrum characteristic, therefore can develop and become a kind of novel antiseptics for natural food, be widely used in food processing and the anti-corrosive fresh-keepings such as fish, meat, fowl, egg, breast, fruits and vegetables, effective control microorganism in food growth, has significant application value for the loss of minimizing food decay and prolongation effective period of food quality.
Lactobacillus acidophilus NX2-6 bacterial strain belongs to the food security bacterial classification of generally acknowledging in the world.The present invention utilizes the antibacterial product microorganism belonging to genus antiseptics for natural food of its fermenting and producing in the culture medium made by raw-food material, to food and human health security high, have no side effect, can not cause the pollution to environment, alternative chemical preservative is widely used in the industries such as food, medicine, feed.
(3) natural antiseptic agent is tested the antiseptic effect of Chilled Meats
Add lactobacillus acidophilus NX2-6 antibacterial product to Chilled Meats in minced steak end, the final concentration of natural antiseptic agent in cooling meat pie is made to reach 2%, 4% and 8%, using the process added Nisin and do not add natural antiseptic agent as positive and negative control, be designated as respectively L.1, L.2, L.3, Nisin and Control.Cooling minced meat is made meat pie by every block 50g, to be sub-packed in food pallet and with preservative film sealing, preservation at 4 DEG C, micro organism quantity and physical and chemical index in inspection meat pie.
(1) change of microorganism in Chilled Meats storage
Pig Minced Steak after different preservative treatment is made minced pork cake and is positioned over polypropylene pallet and stores 15 days with placing after the sealing of polyethylene preservative film with 4 DEG C of refrigerators.Variable concentrations natural antimicrobial substance freeze-dried powder on the impact of micro organism quantity in meat pie in table 2.Experimental result shows that natural antimicrobial all microorganisms of verifying all have inhibitory action (P<0.05).In all processed group, negative control group micro organism quantity increases the fastest, is secondly nisin group and natural antimicrobial substance processed group.Natural antimicrobial substance addition is more, and in meat pie, micro organism quantity is fewer.
In all processed group, total plate count all increases with memory time and increases (P<0.05).Compared with Control group, in all processed group, total plate count is all significantly suppressed.The meat pie of Control group was at the 6th day corruption (>7log
10cFU/gmeat), the meat pie corrupt phase of nisin and the process of natural antimicrobial substance freeze-dried powder is added more than 12 days.Wherein, the sample L.3 organized also did not have corruption at 15 days.In Control group and Nisin group, coliform quantity extends along with memory time and improves, but coliform group count but reduces in FBF processed group.L.1 group kept downward trend, from initial 2.98log at first 9 days
10cFU/g drops to 2.50, to coliform quantity increase in 15 days from the 9th day.Compared with Nisin group, the coliform L.2 and L.3 in group receives obvious suppression (P<0.05), have dropped 3.79 and 4.42log respectively to when 15 days
10cFU/g.Pseudomonad quantity is starkly lower than control group in interpolation FBF group, and there was no significant difference between control group.Control group and L.1 in group pseudomonad quantity increase with memory time and improve, but L.1 in group bacterium number be starkly lower than control group.And L.3 L.2 in group, pseudomonad presents downward trend (P<0.05) always.By the 15th day, the pseudomonad number ratio Control group L.2 in group and Nisin group have dropped 4.60 and 4.18log respectively
10cFU/g.Examine less than pseudomonad in L.3 group when it should be noted that 15 days.Although along with the prolongation of memory time, in all processed group, lactic acid bacterium number is all in raising, and the lactic acid bacteria increasing degree L.3 in group is obviously little of other processed group (P<0.05).15 days time, Control group, L.1 group and L.2 in group lactic acid bacterium number there is no significant difference, but in Nisin group and L.3 group, quantity is lower.In addition, L.3 the content of lactic acid bacteria reduces 2.85 and 2.10log respectively than Control group and Nisin group in group
10cFU/g.The starting quantity of mould yeast in all processed group is 3.80 ~ 3.90log
10cFU/g, and its quantity increases along with memory time and increases.The 6th day time, in Control group and Nisin group, mould yeast quantity is respectively 7.29 and 7.07log
10cFU/g, but L.1 organizing mould yeast number is increased to 6.18log
10cFU/g.15 days time, in two control groups, mould amount of yeast is significantly higher than FBF processed group (P<0.05).
(2) change of physicochemical property in Chilled Meats storage
Measure Chilled Meats to generally include with extending putrid and deteriorated physical and chemical index memory time: TVB-N, TBARS, a*-value and pH value etc.
As shown in Figure 2, in all samples, TVB-N value all increases along with the prolongation of memory time (P<0.05), but in each FBF processed group, the amplification of TVB-N value is less.By the 6th day, in control group, TVB-N value exceeded limit standard (≤15mg/100g meat).L.1 group and L.2 in group sample exceed limit standard respectively the 9th day and 12 days.Especially, L.3 organize sample and all do not exceed limit standard in the storage period of whole 15 days.
Sample oxidation value TBARS value is weighed, and each sample group TBARS value all raises with the prolongation of memory time, sees Fig. 3.Compared with control group, in FBF processed group, TBARS value raises faster (P<0.05).But at whole memory period, TBARS value does not all exceed limit standard 1.0mgMDA/kg meat.
In each sample group, Fig. 4 is shown in a*-value (redness) change.The redness of all meat samples all reduces with the prolongation of memory time (P<0.05).Time initial, control group and the redness L.1 organized do not have difference (P>0.05), but it is higher than L.2 group and L.3 group.By the 12nd day, the redness of Nisin group was higher than Control group and L.1 group.When the 15th day, Nisin group has the highest red scale value, is L.1 group subsequently, but it is minimum L.3 to organize redness.
Under 4 DEG C of storing temperatures, in all samples group, pH value (P<0.05) all in rising trend, is shown in Fig. 5.Test the initial stage, L.1 group, L.2 group and L.3 organize sample pH value be respectively 5.62,5.37 and 5.05.The pH of FBF processed group is all less than control group, and there was no significant difference (P>0.05) between control group.At whole memory period, compared with control group, the increasing degree of FBF group pH value is all less than control group.From the 9th day, in control group, pH raised fast.15th day, Control group and Nisin group pH value rose to 7.19 and 7.11 respectively, but L.1 group, L.2 group and L.3 group sample pH value difference 6.01,5.55 and 5.15.
(3) subjective appreciation in Chilled Meats storage
The meat pie of the different disposal sensory evaluation under 4 DEG C of storage conditions is in Table.Result shows, along with the prolongation of memory time, in each processed group, Chilled Meats sense organ all significant change occurs.FBF addition is larger, and the color of meat is poorer, but better on the impact of smell and viscosity.
As shown in Figure 6, although L.2 group and L.3 group is better to the effect of Chilled Meats in control of microorganisms, viscosity and smell, along with the raising of FBF addition in meat, what it was unfavorable for meat protects look.With regard to general effect, the 15th day time, L.1 group is all obviously better than Control group and Nisin group in each sense organ.
Table 1 lactobacillus acidophilus NX2-6 fermentation different culture media produces antibacterial substance
Adopt punch method.
The antimicrobial spectrum of table 2 natural antiseptic agent
Note: antibacterial circle diameter (mm) is repeat mean+SD 3 times. adopt Odontothrips loti.
Micro organism quantity change in table 3 different preservative treatment cooling meat pie
Note: result is expressed as mean+SD.Different lowercase in each row (a, b, c, d and e) and in every a line not
Same capitalization (A, B, C, D, E and F) shows significant difference (P<0.05).ND: show not detect.
The different preservative treatment of table 4 is on the impact of Chilled Meats sensory evaluation
Note: in table, A, B, C, D, E show the significance of difference (P<0.05).
Claims (3)
1. a preparation method for natural antiseptic agent is produced in the fermentation of food-grade lactobacillus acidophilus, it is characterized by:
1) preparation of food-grade culture medium
Cleaned by tomato, pull an oar and make tomato juice, tomato juice, beer, skimmed milk, pure water are become fermentation medium according to the proportions of weight 5 ~ 10:15 ~ 25:5 ~ 20:65 ~ 80, and 121 DEG C of sterilizing 7 min, are fermentation medium;
2) lactobacillus acidophilus of fermenting produces natural antiseptic agent
Lactobacillus acidophilus (
lactobacillus acidophilus) NX2-6 seed culture medium expand cultivate after, volume ratio 2 ~ 4% is seeded on food-grade culture medium, 37 DEG C of quiescent culture 24 ~ 36h, zymotic fluid collected by centrifugation supernatant ,-20 DEG C of pre-freeze 24 h, vacuum freeze drying, obtain natural sandy gravel.
2. the natural antiseptic agent prepared of method described in claim 1.
3. the application of natural antiseptic agent described in claim 2, is characterized by: under 4 DEG C of holding conditions, adds in Chilled Meats according to mass ratio 2-8% by natural antiseptic agent, anticorrosion for Chilled Meats.
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| CN1746291A (en) * | 2005-07-20 | 2006-03-15 | 哈尔滨商业大学 | Mixed bacterium leavening agent with high-content L-lactic acid fermenting milk and preparation thereof |
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