CN102150925B - Bacteriostat prepared by fermentation of lactobacillus and preparation method thereof - Google Patents
Bacteriostat prepared by fermentation of lactobacillus and preparation method thereof Download PDFInfo
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- CN102150925B CN102150925B CN 201110003356 CN201110003356A CN102150925B CN 102150925 B CN102150925 B CN 102150925B CN 201110003356 CN201110003356 CN 201110003356 CN 201110003356 A CN201110003356 A CN 201110003356A CN 102150925 B CN102150925 B CN 102150925B
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Abstract
The invention relates to a new method for preparing a bacteriostat by fermentation of lactobacillus. The new method comprises the steps of utilizing concentrated lactalbumin powder for reconstitution so as to get reconstituted concentrated lactalbumin with the protein content of 2.3%-3.2% (w/w), adding 0.4-0.6 per mill (w/w) of FeSO4.7H2O, sterilizing through the conventional method, inoculating a lactobacillus fermenting agent for fermentation, and finally getting the bacteriostat with significant bacteriostasis function.
Description
Technical field
The invention belongs to the food fresh keeping field, be specifically related to the new method that a kind of lactobacillus-fermented prepares bacteriostatic agent.
Background technology
Along with the development of economy and society, people are increasingly high to food safety requirements, the putrid and deteriorated attention that more and more causes people of food.According to statistics, the whole world has the food of 10 %～20 % to be lost in various corruption every year approximately, wherein mainly is the corruption that microorganism is caused.Recently, the means and methods of control food spoilage is multiple, comprises the application of improvement food processing technology, technology of the package, storage condition and antiseptic and inhibiting bacteria function agent etc.Wherein, enormous function has been brought into play in the use of bacteriostatic agent in development of food industry.And the kind of bacteriostatic agent constantly increases, and use amount increases year by year, and the bacteriostasis, preservation that therefore utilizes bacteriostatic agent to carry out food remains a kind of indispensable important means.Be based on currently, we must correctly use the food bacteriostatic agent.Therefore, research food bacteriostasis, preservation method has become current important research project.
The food bacteriostatic agent can suppress or kill the putrefactive microorganisms that contains in the food, prevents the variation of food in storage with this.The present food bacteriostatic agent of China is most to be synthetic bacteriostatic agent, finds that through long term studies some synthetic bacteriostatic agents lure carcinous, teratogenesis and are prone to cause problem such as food poisoning.Some understanding are incomplete, without the application of the bacteriostatic agent of complete detection, can more or less work the mischief to human body.So searching safely, new bacteriostatic agent is imperative efficiently.Along with people's is to natural, the increasing cognition of healthy food; Now more and more pay attention to for the safety problem of food bacteriostatic agent both at home and abroad; The chemical bacteriostatic agent that can be used for the food bacteriostatic agent more and more is restricted; This has produced very big obstruction to food production, transportation, storage, presses for to research and develop out efficient natural food bacteriostatic agent.
Lactoferrin has antibacterial, the antivirus action of wide spectrum, and the growth and breeding that comprises multiple spoilage organisms such as enteropathogenic E, salmonella is had inhibitory action, as bacteriostatic agent plurality of applications is arranged all in food, medicine and feed industry.Because lactoferrin costs an arm and a leg, thereby the bacteriostatic agent of seeking new similar lactoferrin effect is essential.
Summary of the invention
In order to solve the deficiency of prior art; The present invention breaks through the thinking of original separation and Extraction lactoferrin; Utilize genuine milk acid bacteria fermentation WPC and contain the compound of iron ion, excavate the natural biological bacteriostatic agent that has close antibacterial, bactericidal effect with the genuine milk ferritin.
Another object of the present invention is to the application of said bacteriostatic agent, be used to prolong the shelf life of food.
Concrete preparation method is: utilize concentrated whey protein powder to be recovered to the recovery WPC of protein content 2.3% ~ 3.2%, and add 0.4 ‰ ~ 0.6 ‰ FeSO
47H
2O after the sterilization of employing conventional method, is cooled to 37-42 ℃, inoculation 2%-5% lactic acid bacteria fermenting agent, and 37 ℃ of fermentation 12 ~ 16h get tunning, i.e. bacteriostatic agent.This tunning has good antagonistic property, after freeze drying, processes freeze-dried powder and uses.
Said concentrated whey protein powder can be selected WPC80 (protein content of whey is 80%) for use among the present invention.
Said lactic acid bacteria fermenting agent has conventional implication among the present invention, is meant the lactic acid bacteria pure culture that is used for fermentations such as dairy products, include but not limited to Lactobacillus rhamnosus (
Laetobacillus rhamnosus), lactobacillus acidophilus (
Laetobacillus acidophilus), Lactobacillus casei (
Lactobacillus case).
The present invention confirms its antagonistic property with tunning through bacteriostatic experiment through lactobacillus-fermented iron content lactalbumin.And through the antagonistic property of antibacterial substance composition is discovered, lactobacillus-fermented has been strengthened the compatibilization of iron ion, thereby has strengthened the antagonistic property of tunning.
The specific embodiment
Employed material can obtain through following approach among the present invention:
PURE WHEY (WPC80): sky high mountain, Shanghai industrial corporation;
Lactobacillus rhamnosus grx10 (preserving number is CGMCC No:2526): China Committee for Culture Collection of Microorganisms common micro-organisms center;
Lactobacillus acidophilus (
Laetobacillus acidophilus) ATCC 4356: purchase in China Committee for Culture Collection of Microorganisms common micro-organisms center;
Lactobacillus casei (
Lactobacillus caseATCC 10639), enteropathogenic E (
E.coliACCC 01628), salmonella (
Salmonella typhiCICC 21913), Shigella (
Shigella dysenteriaeCMCC 51272), vibrio parahaemolytious (
Vibrio parahaemolyticusMCCC 1H00057), staphylococcus aureus (
Staphylococcus aureusATCC 25922), saccharomyces cerevisiae (
Saccharomyces cerevisiaeCICC 31123), the alternate Mucor (
Mucor alternansCGMCC 3.3418): purchase Institute of Micro-biology in the Chinese Academy of Sciences.
Percent concentration related among the present invention all refers to concentration expressed in percentage by weight (w/w) like no specified otherwise.
1, the preparation of bacteriostatic agent
Utilizing concentrated whey protein powder (WPC80) to be recovered to protein content is 2.3% ~ 3.2% recovery WPC, and adds 0.4 ‰ ~ 0.6 ‰ FeSO
47H
2O after the sterilization of employing conventional method, is cooled to about 40 ℃; The inoculating lactic acid bacterium leavening agent is (as inoculating 3% Lactobacillus rhamnosus grx10; Culture presevation number: CGMCC No:2526), 37 ℃ of fermentation 12 ~ 16h process freeze-dried powder with this tunning through freeze drying and use.
2, the antagonistic property of bacteriostatic agent
The enteropathogenic E ACCC 01628 that activation is good, salmonella CICC 21913, Shigella CMCC 51272, vibrio parahaemolytious MCCC 1H00057, staphylococcus aureus ATCC 25922, saccharomyces cerevisiae CICC 31123, alternate Mucor CGMCC 3.3418
,In the corresponding liquid culture medium, add the bacteriostatic agent freeze-dried powder of 1.25% the present invention preparation with 3% inoculated and cultured simultaneously, 37 ℃ of Mixed culture are through measuring OD
580nmAnd with do not add bacteriostatic agent, fermentation WPC (adopt identical lactalbumin reconstituted milk, do not add molysite) and lactoferrin and make comparisons with the fermenting process of preparing acquisition under the same conditions of same lactic acid bacteria.
Table 1 bacteriostatic agent is to the bacteriostasis of different spoilage organisms
| ? | Bacteriostatic agent | Lactoferrin | The fermentation WPC |
| Enteropathogenic E ACCC 01628 | ++ | + | + |
| Salmonella CICC 21913 | +++ | + | ++ |
| Shigella CMCC 51272 | +++ | ++ | + |
| Vibrio parahaemolytious MCCC 1H00057 | +++ | ++ | + |
| Staphylococcus aureus ATCC 25922 | +++ | + | ++ |
| Saccharomyces cerevisiae CICC 31123 | +++ | + | ++ |
| Alternate Mucor CGMCC 3.3418 | ++ | +++ | - |
Annotate: " +++" represent that fungistatic effect is remarkable; " ++ " expression fungistatic effect is general; A little less than "+" expression fungistatic effect; The no fungistatic effect of "-" expression.
Can find out that by table 1 in fermentation WPC, lactoferrin and the rejection characteristic of three kinds of materials of bacteriostatic agent to seven kinds of microorganisms, bacteriostatic agent is to enteropathogenic E, salmonella, vibrio parahaemolytious, Shigella
,The inhibitory action of staphylococcus aureus and saccharomyces cerevisiae is bigger than lactoferrin, fermentation WPC inhibitory action; Lactoferrin is stronger with fermentation WPC inhibitory action than bacteriostatic agent to the inhibitory action of Mucor.Show that the bacteriostatic agent that the present invention makes has stronger inhibitory action to the common spoilage organisms of major part.
3, the heat endurance of bacteriostatic agent
The bacteriostatic agent that the present invention makes is handled 5min, 10min, 15min, 20min, 30min at 70 ℃ respectively; Handle 5min, 15min, 30min for 80 ℃; Handle 5min, 10min for 100 ℃; Handle 5min for 121 ℃, 10min is right with the measured in solution before and after handling
E.coliThe antagonistic property of SDZC07.Find that antibacterial circle diameter is 16.50mm before the bacteriostatic agent heat treatment, handle 15min or 80 ℃ down through 70 ℃ and handle the 5min antibacterial circle diameter down basically about 15.8mm, not remarkable to spoilage organisms fungistatic effect difference.Show that 15min under 70 ℃, handled by bacteriostatic agent or 80 ℃ of antagonistic property property of handling 5min down are stable.
4, the pH of bacteriostatic agent stability
The bacteriostatic agent that the present invention is made is respectively 2.0,3.0,4.0,5.0,6.0,7.0,8.0,9.0,10.0 with HCl and its pH of NaOH adjusting, and the measured in solution before and after handling is right
E.coliThe antagonistic property of SDZC07.Find the increase of bacteriostatic agent along with pH, antibacterial circle diameter is elder generation and increases the trend that afterwards reduces, and when pH8.0, reaches maximum, is 15.80mm; And pH is 6.0,7.0,8.0, and 9.0 o'clock, antibacterial circle diameter maintained in the scope of 14.80mm ~ 15.80mm.Show bacteriostatic agent in pH4.0 ~ 9.0 scopes, stable to the fungistatic effect of spoilage organisms.
The preparation of embodiment 1. bacteriostatic agent freeze-dried powders
Concentrated whey protein powder (WPC80) is recovered to 2.8% reconstituted milk albumin with deionized water, adds 0.5 ‰ FeSO
47H
2O after 85 ℃/30s sterilization, is cooled to 37-42 ℃, and inoculation 3% Lactobacillus rhamnosus grx10 leavening at 37 ℃ of bottom fermentation 16h, after freeze drying, can make and has remarkable bacteria resistance function and get the bacteriostatic agent freeze-dried powder.
Embodiment 2.
Concentrated whey protein powder is recovered to 2.3% reconstituted milk albumin with deionized water, adds 0.4 ‰ FeSO
47H
2O after 85 ℃/30s sterilization, is cooled to 37-42 ℃, and the lactobacillus acidophilus ATCC 4356 of inoculation 2% at 37 ℃ of bottom fermentation 16h, gets tunning, makes bacteriostatic agent.
Embodiment 3
Concentrated whey protein powder (WPC80) is recovered to 3.2% reconstituted milk albumin with deionized water, adds 0.6 ‰ FeSO
47H
2O after 85 ℃/30s sterilization, is cooled to 37-42 ℃, and inoculation 5% Lactobacillus casei at 37 ℃ of bottom fermentation 16h, after freeze drying, can make and has remarkable bacteria resistance function and get the bacteriostatic agent freeze-dried powder.
The application of embodiment 4. bacteriostatic agents in yogurt
In yogurt, add 0.3% bacteriostatic agent freeze-dried powder; Preserve in 30 days and compare with the yogurt that does not add bacteriostatic agent; Find to add the yogurt and the yogurt that does not add bacteriostatic agent of bacteriostatic agent; Viable lactic acid bacteria quantity does not have significant change, but the yogurt quality of adding bacteriostatic agent meets the requirements, and the yogurt to 15 that does not add bacteriostatic agent of the present invention day has mould and yeast growth.
The application of embodiment 5. bacteriostatic agents in meat products
Show condition 30% is adopted in the making of common fermentation sausage, lean meat 70%, salt 2.5 %, sugar 3%; Liquor 1%, ginger 0.1%, green onion 0.1%, white green pepper powder 0.3%; Monosodium glutamate 0.3%, five-spice powder 0.05%, lactic acid bacteria 4% is added 0.3% bacteriostatic agent freeze-dried powder on this basis; At 4 ℃ of condition bottom fermentation 24h, Exposure to Sunlight was put into the indoor maturation that hangs about 3 days after the can, was finished product after 2 weeks.Find to add more un-added the comparing of sausage of bacteriostatic agent, not only to not influence of product special flavour, quality meets the requirements, and the shelf-life can prolong more than one times.
Claims (3)
1. the bacteriostatic agent of a lactobacillus-fermented is characterized in that it being by making through following method: concentrated whey protein powder is recovered to the recovery WPC that protein content is 2.3% ~ 3.2% (w/w), and adds the FeSO of 0.4 ‰ ~ 0.6 ‰ (w/w)
47H
2O carries out sterilization, is cooled to 37-42 ℃, inoculation 2%-5% (w/w) lactic acid bacteria fermenting agent, and 37 ℃ of fermentation 12 ~ 16h get tunning, i.e. bacteriostatic agent; Said lactic acid bacteria fermenting agent be Lactobacillus rhamnosus (
Laetobacillus rhamnosus), lactobacillus acidophilus (
Laetobacillus acidophilus) or Lactobacillus casei (
Lactobacillus case).
2. the preparation method of the bacteriostatic agent of a lactobacillus-fermented is characterized in that, is concentrated whey protein powder is recovered to the recovery WPC that protein content is 2.3% ~ 3.2% (w/w), and adds the FeSO of 0.4 ‰ ~ 0.6 ‰ (w/w)
47H
2O carries out sterilization, is cooled to 37-42 ℃, inoculation 2%-5% (w/w) lactic acid bacteria fermenting agent, and 37 ℃ of fermentation 12 ~ 16h get tunning, i.e. bacteriostatic agent; Wherein said lactic acid bacteria fermenting agent be Lactobacillus rhamnosus (
Laetobacillus rhamnosus), lactobacillus acidophilus (
Laetobacillus acidophilus) or Lactobacillus casei (
Lactobacillus case).
3. preparation method according to claim 2 is characterized in that processing freeze-dried powder with after the tunning sterilization through freeze drying.
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| CN103421715B (en) * | 2013-08-03 | 2015-06-10 | 青岛蔚蓝生物集团有限公司 | Lactobacillus rhamnosus and application thereof |
| CN105853465B (en) * | 2016-03-25 | 2019-06-25 | 浙江大学 | A method of inhibit vibrio parahaemolytious in mussel body to breed by lactic acid bacteria biological antagonism |
| CN109430368A (en) * | 2018-12-14 | 2019-03-08 | 马山县山山农业科技有限公司 | Joint lactic acid bacteria antibacterial liquid is eluted to carry out the diversification method of beef carcase disinfection |
| CN112210507B (en) * | 2020-08-10 | 2022-09-06 | 江苏微康生物科技有限公司 | Shigella-antagonistic lactobacillus rhamnosus LRa05, screening method and application thereof |
| CN114246246B (en) * | 2021-12-29 | 2023-08-18 | 湖南营养树生物科技有限公司 | Method for desensitizing isolated whey protein containing lactoferrin |
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Effective date of registration: 20190703 Address after: 226000 Bayi Industrial Park, Tianshenggang Town Street, Gangzha District, Nantong City, Jiangsu Province Patentee after: Jiangsu Haote Food Technology Co., Ltd. Address before: No. 88, South Road, University of Yangzhou, Jiangsu, Jiangsu Patentee before: Yangzhou University |
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