CN115322943A - Method for separating lactic acid bacteria from sour honey - Google Patents
Method for separating lactic acid bacteria from sour honey Download PDFInfo
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- CN115322943A CN115322943A CN202211156743.9A CN202211156743A CN115322943A CN 115322943 A CN115322943 A CN 115322943A CN 202211156743 A CN202211156743 A CN 202211156743A CN 115322943 A CN115322943 A CN 115322943A
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/36—Adaptation or attenuation of cells
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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
- C12N1/02—Separating microorganisms from their culture media
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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
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- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
- C12R2001/25—Lactobacillus plantarum
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Abstract
The invention discloses a method for separating lactic acid bacteria from sour honey, which comprises the following steps: s1, diluting acid honey to obtain a bacterial liquid; s2, inoculating the bacterial liquid obtained in the step S1 into a lactic acid bacteria liquid culture medium, and performing multiple rounds of domestication culture to obtain a bacterial liquid to be separated, wherein the domestication culture comprises the following steps: performing shake culture at 34-37 deg.C; s3, diluting the bacterial liquid to be separated obtained in the step S2, and coating the diluted bacterial liquid to be separated on a substrate containing CaCO 3 The lactic acid bacteria are separated from the solid medium and cultured in an inverted manner. The invention successfully extracts the lactic acid bacteria from the sour honey by a special domestication method, and the bacterial strain is beneficial bacteria and can be widely applied to food and human bodies.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to a method for separating lactic acid bacteria from sour honey.
Background
Sour bees belong to the genus of stingless bees, and are named because the sweet and sour honey produced by the produced honey is sour, the sour honey is natural honey brewed and processed by collecting pollen and nectar from the sour bees in the species of the stingless bees, and the sour bees preferentially collect plant nectar with high medicinal value generally, so that the sour honey has high health-care value and has the effects of resolving food stagnation, promoting digestion, delaying senescence, detoxifying, calming nerves, beautifying, resisting wrinkles and the like.
The raw honey of sour honey contains various microorganisms, mainly including yeast, lactic acid bacteria, bacillus, monascus and the like, wherein lactobacillus is the most commonly used probiotic microorganism group, is gram-positive bacteria (G +), does not produce spores, has a chain or club shape in cell morphology, is mostly facultative anaerobe, has lactic acid as a main fermentation product, and also has a few byproducts, such as potassium acid, acetic acid, succinic acid, CO 2 And the like. The lactobacillus is commonly used in the fermentation of dairy products, meat products, pickled vegetables and the like, is an important component of human and other animal respiratory tract, gastrointestinal tract and genitourinary tract microbial communities, and also has the effective effects of regulating micro-ecological balance, enhancing intestinal barrier function, regulating immune response and the like. The traditional strain separation method has very large workload and slow speed.
Therefore, it is necessary and very significant to develop a method for rapidly extracting lactic acid bacteria from sour honey.
Disclosure of Invention
The invention aims to provide a method for separating lactic acid bacteria from sour honey, so as to realize the successful separation of lactic acid bacteria from sour honey, and the method has the advantages of high efficiency and high accuracy.
The invention is realized by the following technical scheme:
a method for separating lactic acid bacteria from sour honey comprises the following steps:
s1, diluting acid honey to obtain a bacterial liquid;
s2, inoculating the bacterial liquid obtained in the step S1 into a lactic acid bacteria liquid culture medium, and performing multiple rounds of domestication culture to obtain a bacterial liquid to be separated, wherein the domestication culture comprises the following steps: performing shake culture at 34-37 deg.C;
s3, diluting the bacterial liquid to be separated obtained in the step S2, and coating the diluted bacterial liquid to be separated on a substrate containing CaCO 3 The lactic acid bacteria are separated from the solid medium and cultured in an inverted manner.
In the conventional method for separating lactic acid bacteria, after the step S1 (dilution), the mixed bacteria solution is directly used for separating lactic acid bacteria by using a plate, which is very inefficient.
The lactic acid bacteria in the sour honey have high concentration content, and a single colony is difficult to obtain by direct separation, so that the single colony can be obtained after separation by performing dilution treatment in the step S1.
If the diluted acid honey is directly subjected to the inverted culture of the step S3, the acid honey p H is between 2 and 4, has high acidity, and is directly coated on the product containing CaCO 3 The lactic acid bacteria can directly hydrolyze CaCO on a solid culture medium 3 It is difficult to directly separate lactobacillus from sour honey through step S3.
The applicant found through experiments that:
the diluted acid honey is firstly subjected to multiple rounds of domestication culture in the step S2 of the invention, and then the inverted culture in the step S3 is carried out, so that the lactic acid bacteria can be successfully separated from the acid honey, the acidity of the bacterial liquid after the multiple rounds of domestication culture in the step S2 of the invention is fully reduced, and the separation effect of the lactic acid bacteria is better.
In conclusion, the invention uses the special culture medium MRS for lactobacillus to carry out multiple rounds of domestication culture (8-12 rounds), so that the lactobacillus in the mixed bacteria liquid becomes the dominant bacteria, and the separation speed and the separation accuracy are greatly improved.
Further, in step S1, dilution is performed with physiological saline.
Further, in step S1, the specific dilution process of the sour honey is as follows:
placing the acid honey into 0.9% normal saline for dilution and culture for 24h, wherein the weight ratio of the acid honey to the normal saline is 1: 1g of acid honey sample is diluted and cultured in 100mL of 0.9% physiological saline for 24h.
Further, in step S2, the rotation speed of shaking culture is 60-80rpm.
Further, in step S2, the acclimatization culture is carried out for 8 to 12 rounds, and the time of each round is 24 hours.
Further, the pH of the lactic acid bacteria culture medium is 6.2-6.4.
Further, the lactobacillus liquid culture medium comprises the following components in percentage by mass:
Further, in step S3, the bacterial liquid to be separated is diluted 8-10 times with physiological saline.
Further, in step S3, the lactic acid bacteria liquid medium contains CaCO 3 And agar.
Further, caCO 3 And agar in the lactobacillus liquid culture medium in a mass ratio of 2% and 1.5%, respectively.
The hardness of the culture medium is suitable for the growth of the lactic acid bacteria by reasonably controlling the dosage of the agar.
Compared with the prior art, the invention has the following advantages and beneficial effects:
1. according to the invention, in the process of extracting and separating the lactic acid bacteria, a special domestication method is reasonably designed according to the characteristics of the acid honey, the diluted acid honey is domesticated and cultured, and then inverted and cultured, so that the lactic acid bacteria can be successfully separated from the acid honey.
2. The strain separated by the method is beneficial bacterium and can be widely applied to food and human bodies.
Drawings
The accompanying drawings, which are included to provide a further understanding of the embodiments of the invention and are incorporated in and constitute a part of this application, illustrate embodiment(s) of the invention and together with the description serve to explain the principles of the invention. In the drawings:
FIG. 1 is a colony morphology of the isolated strain of the present invention producing calcium lysocircles on a solid plate;
FIG. 2 is the colony morphology of the isolated strain of the present invention on MRS solid medium;
FIG. 3 is a gram-stained microscopic photograph of isolated bacteria of the present invention;
FIG. 4 shows the shape of the isolated strain of the present invention under a scanning electron microscope;
FIG. 5 is a graph of the effect of different salt concentrations on the growth of isolates;
FIG. 6 is a graph of the effect of different initial pH values on the growth of isolates;
FIG. 7 is a graph of the effect of different temperatures on the growth of isolates;
FIG. 8 is a graph showing the acid and gas evolution test of glucose.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail below with reference to examples and accompanying drawings, and the exemplary embodiments and descriptions thereof are only used for explaining the present invention and are not meant to limit the present invention.
Example 1:
a method for separating lactic acid bacteria from sour honey comprises the following steps:
s101, filling 1g of sour honey into a sterilized triangular flask, adding 100mL of 0.9% normal saline for dilution, and fully and uniformly mixing to obtain a bacterial liquid.
S102, inoculating the bacterial liquid obtained in the step S101 into a lactic acid bacteria liquid culture Medium (MRS), performing acclimation culture for 24 hours at the temperature of 34-37 ℃ and the rpm of 60-80, and performing acclimation culture for 10 times to obtain a bacterial liquid to be separated;
the lactobacillus liquid culture medium consists of the following components:
S103, diluting the bacterial liquid to be separated obtained in the step S102 by 8-10 times by using sterile physiological saline, and coating the bacterial liquid to be separated containing CaCO 3 The amount of the inoculated lactobacillus solution was 0.2 ml, the inoculated solution was uniformly spread on the surface of a plate using a sterile coating bar, and the plate was cultured in an inverted state at 37 ℃ for 48 hours to obtain a separation plate producing a caltrop, as shown in FIG. 1. Performing gram staining and microscopic examination on the single bacterial colony which generates the calcium soluble ring on the obtained separation plate; and selecting the single bacterial colony with the microscopic observation result according with the characteristics of the lactic acid bacteria for scanning electron microscope observation.
Containing CaCO 3 The formula of the solid culture medium for separating the lactic acid bacteria is as follows:
The lactic acid bacteria isolated in this example were identified as follows:
1) And observation of colony and cell morphology
The identification method comprises the following steps: respectively streaking the screened lactobacillus strains on an MRS solid culture medium, culturing for 48h at 37 ℃, and observing the color, shape, size and other characteristics of colonies; meanwhile, single bacterial colony is picked by using an inoculating loop for gram staining, and the staining condition and the shape of the thallus are observed under a microscope.
Bacteria with consistent shapes are separated and observed from the sour honey, and the shapes of the bacteria colonies on an MRS culture medium are wet and smooth, white or milky white, the edges are neat, and the diameters of the bacteria colonies are 1-2mm. Selecting the bacterial colonies for continuous separation and purification to obtain a primary screening bacterial strain for preservation, wherein the bacterial strain morphology is shown in figure 2; the gram staining result of the strain shows that the strain is a gram-positive strain, and the result is shown in figure 3, wherein the strain is bluish purple, rod-shaped and has two blunt ends.
2) Physiological and biochemical test
Performing physiological and biochemical identification on the screened lactobacillus strains, wherein the physiological tests comprise the influences of different salt concentrations, different initial pH values and different temperatures on the growth of the separated strains; the biochemical tests comprise a catalase test, acid production tests of different carbon sources (comprising fructose, galactose, glucose, lactose, maltose, mannose, xylose, sucrose and soluble starch), acid and gas production tests of glucose and nitrite degradation capability tests.
a. Effect of different salt concentrations on the growth of isolates
The identification method comprises the following steps: inoculating the activated bacterial suspension into MRS liquid culture medium with NaCl concentration of 0%, 2%, 4%, 6%, 8% and 10% according to 1% (V/V), performing anaerobic culture at 37 ℃ for 24h, measuring OD value of the bacterial solution at 600nm, and drawing a curve by taking NaCl concentration as abscissa and OD600 as ordinate.
FIG. 5 shows the growth conditions of 7 isolates (S1, S2, S3, S4, S5, S6 and S7) obtained by the method of this example under different salinity, from which it can be seen that the growth of 7 strains of bacteria is gradually inhibited as the salt concentration increases, or cannot tolerate very high salt concentration, and the growth is sensitive to the salinity in the culture medium, wherein the salt tolerance of the strain S1 is relatively good, and the salinity of more than 6% seriously affects the growth of the bacteria. The light absorption values of 7 isolates continuously decrease with the increase of salinity, the inhibition trends are approximately similar, and basically no growth occurs when the salinity is more than or equal to 8 percent.
b. Effect of different initial pH values on isolate growth
The identification method comprises the following steps: adjusting the pH value of the MRS liquid culture medium with 1mol/L HCl and 1mol/L NaOH to be 1, 2, 3, 4, 5, 6, 7 and 8 respectively. Inoculating the activated bacterial suspension into the culture medium according to the proportion of 1% (V/V), culturing for 24h under the anaerobic condition at 37 ℃, and measuring the OD value of the bacterial liquid under 600 nm. pH value as abscissa, OD 600 Is a vertical coordinate, is plottedAnd (5) making a curve.
FIG. 6 shows the growth of 7 isolates (S1, S2, S3, S4, S5, S6 and S7) obtained by the method of this example at different initial pH values. As can be seen from FIG. 6, the medium was neither too acidic nor too alkaline to promote the growth of the isolate, and the OD of the bacterial suspension was the greatest when the initial pH of the medium was 6 to 7. Therefore, the optimal initial pH value of the 7 isolates is 6-7; almost no strain grows on a medium with a pH below 3; at pH 4, the isolate still had a certain amount of growth; at pH 8, the OD values of 7 isolates decreased significantly, and the test results showed that the strains had a more favorable pH of around 7.
c. Effect of different temperatures on the growth of isolates
The identification method comprises the following steps: inoculating the activated bacterial suspension into MRS liquid culture medium according to 1% (V/V), respectively culturing at 15 deg.C, 20 deg.C, 25 deg.C, 30 deg.C, 35 deg.C, 40 deg.C, and 45 deg.C under anaerobic condition for 24 hr, and measuring OD value of bacterial liquid at 600 nm. pH value as abscissa, OD 600 The graph is plotted as ordinate.
FIG. 7 shows the growth of 7 isolates (S1, S2, S3, S4, S5, S6 and S7) obtained by the method of this example at different temperatures. As seen from the figure, the optimum growth temperature of the S5 strain was 35 ℃ and the optimum growth temperatures of the remaining isolates were 30 ℃. The growth amounts of the S1 and S6 strains at 25 ℃ are very low, and the other strains can also grow. The 7 isolates showed little growth below 20 ℃ and above 45 ℃.
d. Catalase assay
The identification method comprises the following steps: inoculating the isolated strain on a suitable slant, which is usually a PYG agar slant, and culturing at 37 ℃ for 24 hours, wherein the PYG medium is prepared by adding 1.0g of glucose into a PY basic medium; the formula of the PY culture medium comprises 0.5g of peptone, 1.0g of yeast extract, 0.5g of trypticase casein, 4.0mL of salt solution and 100mL of distilled water; the component of the salt solution is anhydrous CaCl 2 0.2g,MgSO 4 .7H 2 O 0.48g,K 2 HPO 4 1.0g,KH 2 PO 4 1.0g,NaHCO 3 10.0g, naCl 2.0g, adding CaCl 2 And MgSO 4 .7H 2 Dissolving in mixed OTo 300mL of distilled water was added 500mL of water, and the other salts were slowly added with stirring. Stirring was continued until all dissolved, 200mL of distilled water was added, and after mixing, the mixture was stored at 4 ℃.
A loop of the culture grown on a slant of PYG agar was applied to a clean slide and a drop of 3% -15% H2O2 was added to the slide, which was positive if any bubbles were formed and negative if no bubbles were formed.
All of the 7 isolates obtained by the method of this example were negative without producing air bubbles.
e. Carbohydrate fermentation acidogenesis test
The identification method comprises the following steps: PY basic culture medium is adopted, various sugars, alcohols and certain glycoside carbohydrates are respectively added into the PY basic culture medium, and the fermentation acid production condition is tested. After sterilization at 115 ℃, the inoculum was cultured for 48h at 30 ℃. The configuration requirements are shown in table 1:
TABLE 1 carbohydrate Medium
For the test bacteria using the PY-based carbohydrate medium described above, BTB-MR reagent can be used for the detection of the acid production result. The difference in color developed by the reagent may indicate the degree of acid production.
The formula of the reagent is as follows: bromothymol blue (BTB) 0.2g, potassium-based red (MR) 0.1g,95% ethanol 300mL, distilled water 200mL.
The indicated range of color development is shown in table 2:
TABLE 2
The results of identifying the 7 isolates obtained by the method of this example are shown in Table 3:
TABLE 3 acid production test on different carbon sources
In Table 3, "+" is positive, "-" is negative, and "+" is high in acid yield; as can be seen from Table 3: the 7 isolates can utilize the carbon source tested at present and produce acid, and the difference is not large.
f. Acid and gas evolution test from glucose
The identification method comprises the following steps: culture medium (1L)
30g of glucose and 0.5mL of Tween 80 were added to the PY basal medium, and 6g of agar was added to prepare a soft agar column. Subpackaging the test tubes with the height of 4-5 cm. To facilitate the observation of acid production, 1.4mL of potassium bromphenol violet with a concentration of 1.6g/100mL can be added to the medium as an indicator. Sterilizing at 112 deg.C for 20 min.
Inoculation and culture
A large amount of fresh and strong-activity strains are used for puncture inoculation and are cultured for 48 hours at 37 ℃.
The results of identifying the 7 isolates obtained by the method of this example are shown in FIG. 8: as can be seen from FIG. 8, no bubbles were produced in all 7 tubes, i.e., no gas was produced in 7 isolates, which was preliminarily determined to be homolactic ferments; yellowing of the indicator in the medium indicates acid production, and the acid production of both S3 and S4 bacteria is greater than that of the other strains.
g. Nitrite degradation test
The identification method comprises the following steps: the residual nitrite content of 7 isolates isolated was measured after culturing in MRS liquid medium containing 125. Mu.g/mL of nitrite at 37 ℃ for 48 hours, and the results are shown in Table 2. The determination of the nitrite content adopts naphthyl ethylenediamine hydrochloride spectrophotometry in GB/T5009.33-2016 (determination of nitrite and nitrate in food).
The results of identifying the 7 isolates obtained by the method of this example are shown in Table 4:
TABLE 4 nitrite degrading ability of different strains
As can be seen from Table 4, the degradation ability of the 7 isolates to nitrite was high, and both exceeded 99.5%, and the final pH of the culture broth was around 4.
In conclusion of the physiological and biochemical test results, the isolated strain can be preliminarily identified as lactobacillus according to Bergey's Manual of identification of bacteria and ' Classification identification and Experimental methods of lactic acid bacteria '.
3) And the micro-nano morphology on the surface of the lactobacillus isolate is observed by using a Nigri SU8220 cold Field Emission Scanning Electron Microscope (FESEM), the sample is frozen and dried, then the sample is fixed on a sample table by using a conductive adhesive, a gold coating is sprayed on the surface of the sample by using an ion sputtering instrument, and the micro-nano morphology on the surface of the sample is represented by using the FESEM.
As shown in FIG. 4, FESEM is used for representing the micro-nano morphology of the surface of a sample, clearly shows that bacteria are rod-shaped, two ends are blunt and round, and the micro-nano morphology of Lactobacillus plantarum (Lactobacillus plantarum) is met.
And (3) identifying the isolated strain as Lactobacillus plantarum (Lactobacillus plantarum) by combining the morphological and physiological biochemical analysis and the micro-nano morphology result on the surface of the Lactobacillus isolated strain, and successfully extracting the Lactobacillus plantarum by the method.
The above-mentioned embodiments, objects, technical solutions and advantages of the present invention are further described in detail, it should be understood that the above-mentioned embodiments are only examples of the present invention, and are not intended to limit the scope of the present invention, and any modifications, equivalent substitutions, improvements and the like made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (10)
1. A method for separating lactic acid bacteria from sour honey is characterized by comprising the following steps:
s1, diluting acid honey to obtain a bacterial liquid;
s2, inoculating the bacterial liquid obtained in the step S1 into a lactic acid bacteria liquid culture medium, and performing multiple rounds of domestication culture to obtain a bacterial liquid to be separated, wherein the domestication culture comprises the following steps: performing shake culture at 34-37 deg.C;
s3, diluting the bacterial liquid to be separated obtained in the step S2, and coating the diluted bacterial liquid to be separated on a substrate containing CaCO 3 The lactic acid bacteria are separated from the solid medium and cultured in an inverted manner.
2. The method for separating lactic acid bacteria from sour honey according to claim 1, wherein in step S1, dilution with physiological saline is performed.
3. The method for separating lactic acid bacteria from sour honey according to claim 2, wherein in step S1, the specific dilution process of sour honey is as follows:
placing the sour honey into 0.9% normal saline for dilution and culture for 24h, wherein the weight ratio of the sour honey to the normal saline is 1.
4. The method for separating lactic acid bacteria from sour honey according to claim 1, wherein in step S2, the rotation speed of shaking culture is 60-80rpm.
5. The method for separating lactic acid bacteria from sour honey according to claim 1, wherein in step S2, the acclimatization culture is performed for 8-12 rounds, and the time of each round is 24h.
6. The method for separating lactic acid bacteria from sour honey of claim 1, wherein the pH of the lactic acid bacteria culture medium is 6.2-6.4.
7. The method for separating lactic acid bacteria from sour honey according to claim 1, wherein the lactic acid bacteria liquid medium consists of the following components in percentage by mass:
peptone 1%, beef extract 1%, yeast powder 0.5%, dipotassium hydrogen phosphate 0.2%, diammonium hydrogen citrate 0.2%, sodium acetate 0.5%, glucose2％，Tween-80 0.1％，MgSO 4 ·7H 2 O 0.058g，MnSO 4 ·4H 2 0.025g of O and the balance of water.
8. The method for separating lactic acid bacteria from sour honey according to claim 1, wherein in step S3, the bacterial liquid to be separated is diluted 8-10 times with normal saline.
9. The method for separating lactic acid bacteria from sour honey of claim 1, wherein in step S3, the lactic acid bacteria liquid medium contains CaCO 3 And agar.
10. The method for separating lactic acid bacteria from sour honey of claim 9, wherein CaCO 3 And agar in the lactobacillus liquid culture medium in a mass ratio of 2% and 1.5%, respectively.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109182165A (en) * | 2018-08-21 | 2019-01-11 | 云南农业大学 | One plant of Lactobacillus helveticus and its application during bee raising |
| CN111979222A (en) * | 2020-09-03 | 2020-11-24 | 青海雪峰牦牛乳业有限责任公司 | Screening method of lactobacillus paracasei with high acid yield |
| CN112852676A (en) * | 2021-03-04 | 2021-05-28 | 江苏永盛生物科技有限公司 | Method for separating, screening, detecting and identifying bacteriocin-producing strain |
| CN113774003A (en) * | 2021-11-11 | 2021-12-10 | 北京挑战农业科技有限公司 | Lactobacillus buchneri and application thereof in preparation of low-moisture fermented feed |
-
2022
- 2022-09-22 CN CN202211156743.9A patent/CN115322943A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109182165A (en) * | 2018-08-21 | 2019-01-11 | 云南农业大学 | One plant of Lactobacillus helveticus and its application during bee raising |
| CN111979222A (en) * | 2020-09-03 | 2020-11-24 | 青海雪峰牦牛乳业有限责任公司 | Screening method of lactobacillus paracasei with high acid yield |
| CN112852676A (en) * | 2021-03-04 | 2021-05-28 | 江苏永盛生物科技有限公司 | Method for separating, screening, detecting and identifying bacteriocin-producing strain |
| CN113774003A (en) * | 2021-11-11 | 2021-12-10 | 北京挑战农业科技有限公司 | Lactobacillus buchneri and application thereof in preparation of low-moisture fermented feed |
Non-Patent Citations (4)
| Title |
|---|
| MOHD NIZAM LANI ET AL.: "MICROBIOLOGICAL QUALITY AND pH CHANGES OF HONEY PRODUCED BY STINGLESS BEES, Heterotrigona itama AND Geniotrigona thoracica STORED AT AMBIENT TEMPERATURE" * |
| SYARIFFAH NURATIQAH SYED YAACOB ET AL.: "Identification of Lactobacillus spp. And Fructobacillus spp. isolated from fresh Heterotrigona itama honey and their antagonistic activities against clinical pathogenic bacteria" * |
| 王伟等: "酸马奶酒中乳酸菌的分离及发酵性能研究" * |
| 郭延清: "溶钙圈法对浆水中乳酸菌的筛选" * |
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