CN117603860A - Lactobacillus harbini and application thereof in fermentation of porridge - Google Patents
Lactobacillus harbini and application thereof in fermentation of porridge Download PDFInfo
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- 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
- 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
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/065—Microorganisms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23L7/00—Cereal-derived products; Malt products; Preparation or treatment thereof
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- C12R2001/00—Microorganisms ; Processes using microorganisms
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- C12R2001/225—Lactobacillus
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- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
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Abstract
The invention discloses a Haabine lactobacillus strain and application thereof in sour porridge fermentation, belonging to the technical field of microorganisms, wherein the preservation number of the Haabine lactobacillus strain (Schleiferilactobacillus harbinensis) GXUN74706 is CGMCC No.28712. The strain is separated from Guangxi traditional fermented sour porridge, has higher activity and better growth, and has good acid production capacity and antibacterial effect; the inoculation of the strain for fermenting the sour porridge can reach the index level of the natural fermented sour porridge in a short time. The strain can be used as an alternative bacterium for acid porridge fermentation, improves the success rate of acid porridge fermentation, and promotes the industrial production of local acid porridge.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to a lactobacillus harbine and application thereof in fermentation of porridge.
Background
In the long food culture circles in China, the food culture circles with local features are formed all over the country due to the influence of natural conditions, and the sour porridge is food which is prepared by fermenting with lactic acid bacteria and has very special taste and certain nutritional value in Guangxi, shanxi and inner Mongolian western regions, and is popular among local people. The sour gruel is a food which is prepared by taking grains such as millet, rice, millet, glutinous rice and the like and water as raw materials and naturally fermenting by utilizing microorganisms in the environment. The preparation method effectively retains probiotics such as lactobacillus, gives rich nutritional ingredients and unique flavor, and has good taste.
Lactic acid bacteria are almost existing in all traditional fermented foods, and the fermented starch raw materials can generate various organic acids, so that the pH value of the foods is reduced, and the high-concentration acid inhibits the growth of bacteria and mold, so that the shelf life and the safety of the foods are improved. Thus, related studies on cereal fermented foods have been one of the hot spots of fermented food research. At present, the fermentation of the sour porridge product is mainly small-scale home fermentation, and the problems of low fermentation efficiency, uneven quality of the fermented product and the like exist. The acid-producing bacteria are separated out efficiently, and the acid porridge fermentation strain is screened, so that the method has great promotion effect on the development of acid porridge products and the development of Guangxi special economy.
Disclosure of Invention
The invention aims to provide a lactobacillus harbine and application thereof in acid porridge fermentation, so as to solve the problems in the prior art, and the lactobacillus harbine GXUN74706 can be used as an alternative bacterium for acid porridge fermentation, so that the success rate of acid porridge fermentation is improved, and the industrial production of local acid porridge is promoted.
In order to achieve the above object, the present invention provides the following solutions:
the invention provides a Halbine lactobacillus (Schleiferilactobacillus harbinensis) GXUN74706 which is preserved in the China general microbiological culture Collection center of China Committee for culture Collection of microorganisms for 10 months and 23 days in 2023, wherein the preservation address is North Chen Xway No. 1, 3 in the Korean region of Beijing, and the preservation number is CGMCC No.28712.
The invention also provides a food starter, which comprises the strain, bacterial liquid or fermentation liquid of the Harbin lactobacillus GXUN74706.
The invention also provides application of the lactobacillus harbine GXUN74706 in preparing a food starter.
The invention also provides an application of the lactobacillus harbine GXUN74706 in fermentation of sour porridge.
Further, the lactobacillus harbine GXUN74706 was inoculated to cereal for fermentation.
Further, the cereal comprises rice, millet and/or polished glutinous rice.
Further, the lactobacillus harbine GXUN74706 is inoculated in the form of a fermentation broth or a bacterial suspension.
Further, the inoculum size of the lactobacillus harbine GXUN74706 is 10-15% by volume percent.
The invention also provides an application of the lactobacillus harbine GXUN74706 in preparing porridge-like food.
The invention discloses the following technical effects:
the invention provides a lactobacillus harbine strain, wherein lactobacillus harbine (Schleiferilactobacillus harbinensis) GXUN74706 is separated from Guangxi traditional fermented sour porridge, has higher activity and better growth, has good acid production capacity and antibacterial effect, can be used for fermenting sour porridge, is inoculated with the strain for continuous fermentation for 96 hours, and can obtain sour porridge with pH of 3.60, acidity value of 2.58 DEG T, total protein content of 385.33 mug/mL and total amino acid content of 166.67 mug/mL, and the index level of the naturally fermented sour porridge can be reached in a short time. The strain is derived from fermented food, is used as probiotics, and can play a role in inhibiting pathogenic bacteria in food storage; the method can be applied to the sour porridge fermentation industry to play important biological functions of improving food quality and nutrition, promoting digestion and absorption, enhancing immunity and the like; as an alternative bacterium for the fermentation of the sour porridge, the success rate of the fermentation of the sour porridge can be improved, and the industrialized production of the Guangxi local sour porridge is promoted.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a 16S rDNA phylogenetic tree of Lactobacillus harbini GXUN 74706;
FIG. 2 is a gram-stain microscopic image of Lactobacillus harbini GXUN 74706;
FIG. 3 is a diagram showing the result of scanning electron microscope observation of Lactobacillus harbini GXUN 74706; left scale = 2 μm; right scale = 1 μm;
FIG. 4 is a growth curve of Lactobacillus harbini GXUN 74706;
FIG. 5 is a graph showing the acidogenic effects of Lactobacillus harbini GXUN 74706;
fig. 6 is a graph showing the bacteriostatic effect of lactobacillus harbine GXUN 74706;
FIG. 7 is a graph showing pH change in the continuous fermentation of a porridge of Lactobacillus harbini GXUN74706 fermented with acid for 96 hours;
FIG. 8 is a graph showing titratable acid changes of a halbumin lactobacillus GXUN74706 fermented porridge for 96 hours in continuous fermentation;
FIG. 9 is a bar graph of total amino acid change for 96h of continuous fermentation of a porridge of Lactobacillus harbini GXUN74706 fermentation;
FIG. 10 is a bar graph showing the total protein change of the continuous fermentation of the porridge of the fermented porridge of Halbumin Lactobacillus GXUN74706 for 96 hours.
Detailed Description
Various exemplary embodiments of the invention will now be described in detail, which should not be considered as limiting the invention, but rather as more detailed descriptions of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. In addition, for numerical ranges in this disclosure, it is understood that each intermediate value between the upper and lower limits of the ranges is also specifically disclosed. Every smaller range between any stated value or stated range, and any other stated value or intermediate value within the stated range, is also encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the invention described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from consideration of the specification of the present invention. The specification and examples of the present invention are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are intended to be inclusive and mean an inclusion, but not limited to.
The invention provides a lactobacillus harbine isolated from Guangxi traditional fermented porridge, which is named as lactobacillus harbine GXUN74706, classified and named as Schleiferilactobacillus harbinensis, and has a preservation unit: the collection number of the Chinese microorganism strain collection management committee is CGMCC No.28712, and the Halbumin lactobacillus isolated from the Guangxi traditional fermented food acid porridge is easy to culture, has faster growth, stronger acid production capacity and better antibacterial effect on food pathogenic bacteria, and can be used as acid porridge fermentation bacteria to be applied to the acid porridge fermentation industry. The lactobacillus harbine separated by the invention is used as a single strain for fermentation of the sour porridge, so that indexes such as pH, titratable acidity, total protein content, total amino acid content and the like of the sour porridge reach or even are superior to the level of the sour porridge by natural fermentation. The strain can be preliminarily determined to be an alternative strain for fermentation of the porridge.
The following describes embodiments of the present invention in detail.
Example 1
In this example, a strain was isolated from Guangxi traditional fermented sour porridge and was named GXUN74706. The method is used for carrying out 16S rDNA sequencing to construct a 16S rDNA phylogenetic tree, morphological and physiological biochemical identification analysis.
The physiological and biochemical results of the strain are shown in Table 1, and the strain can utilize various sugar alcohols such as glucose, maltose and the like.
TABLE 1 physiological and biochemical identification Table
Remarks: "+" is utilized and "-" is not utilized.
As shown in FIG. 1, the strain was subjected to 16S rDNA sequencing to construct a 16S rDNA phylogenetic tree, and the result shows that the strain is in the same branch as that of Lactobacillus harbini.
As shown in fig. 2 and 3, the microscopic examination result and the scanning electron microscope result prove that the strain provided by the embodiment is lactobacillus harbinensis.
The strain is identified as lactobacillus harbinensis by molecular biology, systematic evolution analysis, morphological analysis and physiological and biochemical characteristic analysis, and is named as Schleiferilactobacillus harbinensis GXUN74706. The Harbin lactobacillus GXUN74706 is preserved in the China general microbiological culture Collection center (China Committee for culture Collection of microorganisms) at the date of 10 and 23 of 2023, and has a detailed address of No. 1, no. 3, and a preservation number of CGMCC No.28712.
The results show that the lactobacillus harbine GXUN74706 has better activity, can hydrolyze main sugar starch in the porridge to convert into glucose, maltose and the like, and can utilize the metabolism of the strain to produce lactic acid, thereby having the potential of prolonging the shelf life of porridge products.
Example 2
The growth curve, acid production performance and antibacterial performance of the Harbin lactobacillus GXUN74706 are detected in the embodiment.
Growth curve for the strain, at CaCO 3 Acid production ability of MRS solid culture medium and antibacterial performance of the MRS solid culture medium on food pathogenic bacteria (escherichia coli, tubercle bacillus and staphylococcus aureus) are analyzed.
As shown in FIG. 4, the strain is drawn into a growth curve, and the result shows that the strain grows well, has a long logarithmic phase and reaches a stationary phase OD (OD) after 20 hours 600 The value reached 1.50.
As shown in FIG. 5, the strain was tested for CaCO 3 Transparent circle size of MRS solid culture medium, and the result shows that the strain has good acid production performance and can be used for CaCO 3 The size of transparent circle produced by MRS solid culture medium is 40mm.
As shown in FIG. 6, the size of the bacteriostasis circle of the fermentation liquor and the supernatant of the strain on three food-borne pathogens is detected, and the result shows that the strain has good bacteriostasis effect on the three food-borne pathogens (escherichia coli, staphylococcus aureus and tubercle bacillus), the diameter of the bacteriostasis transparent circle of the fermentation liquor is 13.0mm, 15.0mm and 15.5mm in sequence, and the diameter of the bacteriostasis transparent circle of the supernatant of the strain is 10.1mm, 17.5mm and 15.5mm in sequence.
The strain has good activity and CaCO (CaCO) after analysis of growth curve, acid production performance and antibacterial effect 3 Transparent ring of MRS solid culture mediumThe size reaches 40mm, the fermentation liquor and the supernatant have good growth state and acid production capacity, the antibacterial circle sizes of the fermentation liquor and the supernatant on three food pathogenic bacteria (escherichia coli, staphylococcus aureus and tubercle bacillus) are respectively 13.0mm, 15.0mm, 15.5mm, 10.1mm, 17.5mm and 15.5mm, the fermentation liquor and the supernatant have good effects of inhibiting the food pathogenic bacteria in the acid porridge, and the bacteria can be primarily determined to be applied to fermenting the acid porridge.
The Halbine lactobacillus GXUN74706 has good growth state and is characterized by CaCO 3 The MRS solid culture medium has good acid production capability and good antibacterial effect on three food pathogenic bacteria (escherichia coli, staphylococcus aureus and tubercle bacillus), and according to the prior research, the MRS solid culture medium is nontoxic and harmless and can be used for fermentation of acid porridge.
Example 3
The embodiment provides a preparation method of fermented sour porridge, which adopts lactobacillus harbine GXUN74706 screened in embodiment 1 for fermentation to obtain the fermented sour porridge, and specifically comprises the following steps:
step 1, culturing lactobacillus harbine GXUN74706 to obtain fermentation liquor; the culture method comprises the following steps: streaking Lactobacillus harbini GXUN74706 on MRS solid culture medium, culturing at 37deg.C for 48 hr to obtain single colony, selecting single colony, inoculating on MRS broth culture medium, and shake culturing at 37deg.C and 180r/min for 24 hr to obtain OD 600 1.0 fermentation broth.
Step 2, after washing rice, according to the volume ratio of water to rice of 5:1 is put into a fermentation bottle for high-pressure steam sterilization (105 ℃ C., 5 min) to prepare the rice culture medium.
Step 3, inoculating and fermenting the fermentation liquor with 12.5% (V/V) inoculum size, and carrying out centrifugal resuspension twice before inoculation to obtain bacterial suspension, wherein the bacterial suspension is specifically as follows: firstly, 3500r/min fermentation liquor is centrifuged for 10min, the supernatant is discarded, physiological saline is added into the bacterial precipitate to be resuspended to the original volume, and then the solution is centrifuged, and an equal volume of 2% glucose solution is added into the precipitate to be resuspended, thus obtaining bacterial suspension.
And step 4, pouring the bacterial suspension into a rice culture medium, uniformly stirring, and placing the rice culture medium into an incubator for continuous fermentation for 96 hours to obtain the fermented sour porridge.
Comparative example 1
Naturally fermenting rice to obtain fermented sour porridge, which specifically comprises the following steps: after washing rice, the volume ratio of the rice to water is 5:1 is put into a fermentation bottle to prepare a rice culture medium, and after being added with a 2% glucose solution with the same volume as that of the embodiment 3 and uniformly stirred, the rice culture medium is put into an incubator to be continuously fermented for 96 hours, thus obtaining the natural fermented sour porridge.
Effect example 1
1. Detection of pH and titratable acids during fermentation
The pH and titratable acid tests were performed on the porridge samples of example 3 and comparative example 1, which were continuously fermented for 96 hours, and the results are shown in FIGS. 7 and 8.
The results of FIG. 7 show that example 3 and comparative example 1 both showed a decrease in pH after 96h fermentation, but the pH of the inoculated fermented porridge was decreased faster than that of the natural fermentation to 3.6.
The results in FIG. 8 show that both example 3 and comparative example 1 showed an upward trend in titratable acidity values after 96h fermentation, but the acidity value of the inoculated fermented porridge increased faster than that of the natural fermentation to 2.58℃T.
By analyzing the pH and titratable acid change in the fermentation process, compared with natural fermentation, the pH of the porridge with the added Haabine lactobacillus GXUN74706 is faster after 96 hours of fermentation, the acidity value of the porridge is faster to rise, and the porridge is stabilized at pH 3.6 and acidity value of 2.58 DEG T after fermentation is finished, which is superior to the fermentation rate of the porridge obtained by natural fermentation.
2. Detection of total protein content and total amino acid content in fermentation process
The total protein content and total amino acid content of the porridge samples continuously fermented for 96 hours in example 3 and comparative example 1 were measured, and the results are shown in fig. 9 and 10.
The results of FIG. 9 show that the total protein content of both example 3 and comparative example 1 was varied to different degrees after 96 hours of fermentation, wherein the total protein content of the inoculated fermented sour porridge showed a gradual increase trend and the total protein content was higher than that of the naturally fermented sour porridge at each stage of fermentation, and the content was stabilized at 385.33 μg/mL after the completion of fermentation.
The results of FIG. 10 show that the total amino acid content of both example 3 and comparative example 1 changed to different degrees after 96 hours of fermentation, wherein the total amino acid content of the inoculated fermented sour porridge showed a tendency of decreasing before increasing and the total amino acid content was higher than that of the naturally fermented sour porridge at the initial stage and the later stage of fermentation, and the content was stabilized at 166.67. Mu. Mol/mL after the completion of fermentation.
By analyzing the total protein content and the total amino acid content in the fermentation process, compared with natural fermentation, the sour porridge added with the Haabine lactobacillus GXUN74706 has the advantages that the total protein content shows an increasing trend after 96 hours of fermentation, the total amino acid content shows a decreasing trend before increasing trend, the total protein content is 385.33 mug/mL after the fermentation is finished, and the total amino acid content is 166.67 mug/mL higher than that of the sour porridge subjected to natural fermentation.
In summary, the invention provides a strain of Harbin lactobacillus, wherein the Harbin lactobacillus (Schleiferilactobacillus harbinensis) GXUN74706 is separated from Guangxi traditional fermented sour porridge, has good activity and good growth, has good acid production capacity, has good antibacterial effect on three food pathogens (such as escherichia coli, tubercle bacillus and staphylococcus aureus), can be used for fermenting sour porridge, and can obtain sour porridge with pH of 3.60, acidity value of 2.58 DEG T, total protein content of 385.33 mu g/mL and total amino acid content of 166.67 mu mol/mL after the sour porridge is fermented for 96 hours by inoculating the Harbin lactobacillus GXUN74706, so that the fermentation success rate of the product is high and the safety of the sour porridge product is improved.
The above embodiments are only illustrative of the preferred embodiments of the present invention and are not intended to limit the scope of the present invention, and various modifications and improvements made by those skilled in the art to the technical solutions of the present invention should fall within the protection scope defined by the claims of the present invention without departing from the design spirit of the present invention.
Claims (9)
1. The lactobacillus harbinensis (Schleiferilactobacillus harbinensis) GXUN74706 is characterized in that the lactobacillus harbinensis is preserved in the China general microbiological culture Collection center (China general microbiological culture Collection center) for 10 and 23 days in 2023, and the preservation address is North Chen Xway No. 1 and 3 in the Korean region of Beijing city, and the preservation number is CGMCC No.28712.
2. A food starter culture comprising the strain, the bacterial liquid or the fermentation liquid of Lactobacillus harbini GXUN74706 according to claim 1.
3. Use of lactobacillus harbine GXUN74706 as claimed in claim 1 in the preparation of a food starter.
4. Use of lactobacillus harbine GXUN74706 according to claim 1 for fermentation of sour porridge.
5. The use according to claim 4, wherein the lactobacillus harbine GXUN74706 is inoculated to cereal for fermentation.
6. The use according to claim 5, wherein the cereal comprises rice, millet and/or polished glutinous rice.
7. The use according to claim 5, wherein the lactobacillus harbine GXUN74706 is inoculated in the form of a fermentation broth or a bacterial suspension.
8. The use according to claim 5, characterized in that the inoculum size of lactobacillus harbine GXUN74706 is 10-15% in volume percent.
9. Use of lactobacillus harbine GXUN74706 according to claim 1 for the preparation of a porridge.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040005677A1 (en) * | 2002-07-05 | 2004-01-08 | Eddington John M. | Novel lactobacillus strains and use thereof in fermentation process for L-lactic acid production |
| CN110358714A (en) * | 2019-08-01 | 2019-10-22 | 浙江一鸣食品股份有限公司 | A kind of lactic bacteria composition of high yield folic acid |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040005677A1 (en) * | 2002-07-05 | 2004-01-08 | Eddington John M. | Novel lactobacillus strains and use thereof in fermentation process for L-lactic acid production |
| CN110358714A (en) * | 2019-08-01 | 2019-10-22 | 浙江一鸣食品股份有限公司 | A kind of lactic bacteria composition of high yield folic acid |
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| 杨士章;张焕新;施帅;陈玉勇;: "板鸭中优势乳酸菌的分离、筛选及鉴定", 食品科学, no. 07, 1 April 2010 (2010-04-01) * |
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