CN117413924A - Composition with antibacterial and fermentation promoting effects and application thereof - Google Patents
Composition with antibacterial and fermentation promoting effects and application thereof Download PDFInfo
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- CN117413924A CN117413924A CN202311446242.9A CN202311446242A CN117413924A CN 117413924 A CN117413924 A CN 117413924A CN 202311446242 A CN202311446242 A CN 202311446242A CN 117413924 A CN117413924 A CN 117413924A
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- 238000000855 fermentation Methods 0.000 title claims abstract description 86
- 230000004151 fermentation Effects 0.000 title claims abstract description 86
- 239000000203 mixture Substances 0.000 title claims abstract description 53
- 230000000844 anti-bacterial effect Effects 0.000 title abstract description 23
- 230000001737 promoting effect Effects 0.000 title abstract description 10
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 74
- 241000193830 Bacillus <bacterium> Species 0.000 claims abstract description 44
- 238000004519 manufacturing process Methods 0.000 claims abstract description 10
- OPGYRRGJRBEUFK-UHFFFAOYSA-L disodium;diacetate Chemical compound [Na+].[Na+].CC([O-])=O.CC([O-])=O OPGYRRGJRBEUFK-UHFFFAOYSA-L 0.000 claims abstract description 9
- 239000001632 sodium acetate Substances 0.000 claims abstract description 9
- 235000017454 sodium diacetate Nutrition 0.000 claims abstract description 9
- 229920001661 Chitosan Polymers 0.000 claims abstract description 8
- 230000000694 effects Effects 0.000 claims description 24
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- 238000000034 method Methods 0.000 claims description 11
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Abstract
The invention relates to the field of bacteriostats, and discloses a composition with bacteriostasis and fermentation promotion functions and application thereof, wherein the composition comprises the following components in parts by weight: at least one of chitosan and sodium diacetate is used in combination with bacillus culture. The composition provided by the invention has equivalent or better antibacterial effect compared with antibiotics (penicillin); the composition of the present invention has excellent inhibitory performance against not only infectious microbe but also can promote fermentation without affecting ethanol production, thereby promoting ethanol production in specific embodiments.
Description
Technical Field
The invention relates to the field of bacteriostats, in particular to a composition with bacteriostasis and fermentation promotion functions and application thereof.
Background
Along with the development of the feed industry, the development of products such as microecologics, enzyme preparations, antibacterial peptides and the like is an important measure for solving the problems of feed safety, raw material deficiency, environmental pollution and the like.
At present, the research range of the tibody product is wider, the product types are various, and the tibody product has respective advantages and disadvantages. For example, the use condition of the microecological preparation is strict, and the microecological preparation cannot be suitable for various working scenes; the simple enzyme preparation can not achieve the use effect of antibiotics and has relatively high cost; the source of the antibacterial peptide is wide, the variety is many, the molecular mass is small, the natural content is low, the separation and purification are difficult, and the chemical synthesis cost is high; some Chinese herbal medicine extracts have the problems of shortage of resources, unstable drug effect and quality, high cost and the like.
Antibacterial broad spectrum and safety become key difficulties to be solved in antibiotic substitutes.
Bacillus subtilis is used as food safety level probiotics, and the strain and fermentation culture thereof are widely used as food raw materials or food production as food additives, such as bacillus subtilis, bacillus natto and culture, antibacterial peptide, protease, lactase, xylanase, chitosanase and the like.
The researches show that bacillus is a biological control bacterium with strong antibacterial activity and wide antibacterial spectrum, which is important in the nature, and can inhibit various pathogenic bacteria such as aspergillus, candida, fusarium, staphylococcus and the like. The bacillus bacteria can generate various antibacterial active substances in the growth metabolism process, and the generation of the antibacterial active substances leads to higher antibacterial activity and wider antibacterial spectrum of the bacillus. The antibacterial substances include cell wall lyase substances (such as cellulase and chitinase, etc., which kill pathogenic bacteria by lysing cell walls) and protein antibacterial substances (such as protease and lipopeptide, etc., which kill hyphae by enlarging and deforming), etc. If the bacillus fermentation product is used as a biological antibacterial substance, the problem of antibiotic substitution is solved to a certain extent.
Disclosure of Invention
The invention aims to overcome the problems in the prior art and provide a composition with antibacterial and fermentation promoting effects and application thereof.
To achieve the above object, the present invention provides, in a first aspect, a composition having both bacteriostatic and pro-fermentation effects, the composition comprising: at least one of chitosan and sodium diacetate is used in combination with bacillus culture.
In a second aspect, the invention provides a method for fermentative ethanol production, the method comprising: in the presence of the above composition, the saccharide-containing feedstock is subjected to ethanol fermentation.
In a third aspect the present invention provides the use of a composition as described above in fermentation and/or food bacteriostasis.
Through the technical scheme, the antibacterial effect of the composition provided by the invention is equivalent to or better than that of antibiotics (penicillin); the composition of the invention has excellent inhibition performance on mixed bacteria, does not influence ethanol production, and can promote ethanol production in specific embodiments; in addition, the bacillus culture-containing composition provided by the invention is friendly to animals and environment relative to antibiotics, is nontoxic and harmful, can be suitable for the fields of foods, alcohol, cultivation, cosmetics, medicines and the like, and has a wide application range.
Drawings
FIG. 1 is a graph showing acidity data of samples obtained by fermentation for 66 hours using a mixture of a liquefied mash with serious factory contamination and chemical starch as raw materials, and penicillin pairs prepared in examples and comparative examples;
FIG. 2 is a graph of the alcohol content of a sample obtained in the examples and comparative examples with penicillin using a liquefied mash and chemical starch, which are seriously contaminated in a factory, as raw materials after fermentation for 66 hours;
FIG. 3 is a graph showing the chromatographic residual sugar data of samples obtained in examples and comparative examples, wherein the compositions and penicillin pairs are prepared by using liquefied mash and chemical starch with serious factory pollution as raw materials and fermenting for 66 hours;
FIG. 4 is a graph showing the organic acid data of the compositions prepared in examples and comparative examples and penicillin pairs obtained from samples obtained after 66 hours of fermentation using highly contaminated liquefied mash from a plant and chemical starch as raw materials;
FIG. 5 is a graph of ethanol/glycerol data for samples obtained from the compositions of the examples and comparative examples and penicillin pairs after fermentation for 66 hours using highly contaminated liquefied mash from the plant and chemical starch as raw materials.
Detailed Description
The endpoints and any values of the ranges disclosed herein are not limited to the precise range or value, and are understood to encompass values approaching those ranges or values. For numerical ranges, one or more new numerical ranges may be found between the endpoints of each range, between the endpoint of each range and the individual point value, and between the individual point value, in combination with each other, and are to be considered as specifically disclosed herein.
In a first aspect, the present invention provides a composition having both bacteriostatic and pro-fermentation effects, said composition comprising: at least one of chitosan and sodium diacetate is used in combination with bacillus culture. In the present invention, the weight of the "bacillus culture" is on a dry basis.
In the invention, in order to obtain better bacteriostasis and fermentation promotion effects and facilitate efficient use and fermentation of sugar by saccharomyces cerevisiae, the composition comprises chitosan, the weight ratio of bacillus culture to chitosan is 1:0.005-0.25, and the weight ratio can be 1:0.005, 1:0.01, 1:0.015, 1:0.02, 1:0.025, 1:0.03, 1:0.035, 1:0.04, 1:0.045, 1:0.05, 1:0.055, 1:0.06, 1:0.065, 1:0.07, 1:0.075, 1:0.08, 1:0.09, 1:0.1, 1:0.15, 1:0.2, 1:0.25 or any two values of the above are formed, and the values in the range are preferably 1:0.01-0.15.
In the present invention, in order to obtain better antibacterial and fermentation promoting effects, and also to facilitate efficient use and fermentation of sugar by Saccharomyces cerevisiae, the composition includes sodium diacetate, the weight ratio of the bacillus culture to sodium diacetate is 1:0.005-0.45, and may be 1:0.005, 1:0.01, 1:0.015, 1:0.02, 1:0.025, 1:0.03, 1:0.035, 1:0.04, 1:0.045, 1:0.05, 1:0.055, 1:0.06, 1:0.065, 1:0.07, 1:0.075, 1:0.08, 1:0.09, 1:0.1, 1:0.15, 1:0.2, 1:0.25, 1:0.3, 1:0.35, 1:0.4, 1:0.45 or any two or more, and the preferred values within the ranges thereof are 1:0.01-0.25.
Preferably, the bacillus culture contains cellulase, chitinase, protease and lipopeptide; more preferably, the cellulase is present in the bacillus culture in an amount of 10-200U/g; preferably, the bacillus culture has a chitinase content of 5-50U/g; more preferably, the protease is present in the bacillus culture in an amount of 50-200U/g; further preferably, the lipopeptides are present in the bacillus culture in an amount of 5 to 30 wt.%.
In the invention, the cellulase enzyme activity is defined as NY/T912-2004.
In the invention, the chitinase activity is defined as GB/T34799-2017.
In the present invention, the proteases are acid protease, neutral protease and alkaline protease, and the protease activity=acid protease activity+neutral protease activity+alkaline protease activity.
In the present invention, the protease activity is defined as GB/T23527-2009.
In the present invention, in order to obtain better antibacterial and fermentation promoting effects, and also facilitate efficient use and fermentation of sugar by saccharomyces cerevisiae, nisin is further included in the composition, preferably, the weight ratio of bacillus culture to nisin is 1:0.005-0.45, and may be 1:0.005, 1:0.01, 1:0.015, 1:0.02, 1:0.025, 1:0.03, 1:0.035, 1:0.04, 1:0.045, 1:0.05, 1:0.055, 1:0.06, 1:0.065, 1:0.07, 1:0.075, 1:0.08, 1:0.09, 1:0.1, 1:0.15, 1:0.2, 1:0.25, 1:0.3, 1:0.35, 1:0.4, 1:0.45 or any range of values within any two ranges, and preferably, the values are 1:0.01-0.01.
In the present invention, in order to obtain better antibacterial and fermentation promoting effects, and also facilitate efficient use and fermentation of sugar by saccharomyces cerevisiae, sodium gluconate is further included in the composition, preferably, the weight ratio of bacillus culture to sodium gluconate is 1:0.01-0.3, and may be 1:0.01, 1:0.015, 1:0.02, 1:0.025, 1:0.03, 1:0.035, 1:0.04, 1:0.045, 1:0.05, 1:0.055, 1:0.06, 1:0.065, 1:0.07, 1:0.075, 1:0.08, 1:0.09, 1:0.1, 1:0.15, 1:0.2, 1:0.25, 1:0.3, or any two values within the range and the range formed by the above, and preferably, 1:0.03-0.25.
In the invention, in order to obtain better bacteriostasis and fermentation promotion effects, and simultaneously facilitate efficient use and fermentation of sugar by saccharomyces cerevisiae, sodium dehydroacetate is further included in the composition, preferably, the weight ratio of bacillus culture to sodium dehydroacetate is 1:0.02-20, and can be 1:0.02, 1:0.05, 1:0.1, 1:0.5, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:12, 1:14, 1:16, 1:18, 1:20 or any two values of the above ranges and ranges, and values in the ranges are preferably 1:1-10.
In the invention, in order to obtain better antibacterial and fermentation promotion effects, and simultaneously, the high-efficiency utilization and fermentation of the saccharomyces cerevisiae on sugar are more facilitated, and the composition also comprises calcium propionate; preferably, the weight ratio of the bacillus culture to calcium propionate is 1:0.005-1, which may be 1:0.005, 1:0.01, 1:0.05, 1:0.1, 1:0.15, 1:0.2, 1:0.25, 1:0.3, 1:0.35, 1:0.4, 1:0.45, 0.5, 1:0.6, 1:0.7, 1:0.8, 1:0.9, 1:1 or any two values above, preferably 1:0.01-0.5.
In the invention, in order to obtain better antibacterial and fermentation promoting effects, and simultaneously, the efficient utilization and fermentation of the saccharomyces cerevisiae on sugar are more facilitated, and the bacillus culture is bacillus subtilis @Bacillus subtilis) Is a culture of (a); preferably, the bacillus subtilis is at least one of collection numbers CGMCC1.821, ATCC6051, ATCC23857, ATCC21332, ATCC11774 and ATCC13952, wherein CGMCC1.821 is purchased at CGMCC, and ATCC6051, ATCC23857, ATCC21332, ATCC11774 and ATCC13952 are purchased at ATCC。
In the invention, in order to obtain better antibacterial and fermentation promotion effects and simultaneously more facilitate the efficient utilization and fermentation of sugar by saccharomyces cerevisiae, the preparation method of the bacillus culture comprises the following steps: inoculating bacillus into a fermentation medium for fermentation; preferably, the fermentation conditions include: the time is 2-4 days, and the temperature is 30-40 ℃.
In the invention, in order to make the bacillus culture and other components of the invention cooperate to obtain better bacteriostasis and fermentation promotion effects, the fermentation medium comprises the following components: 30-100 g/L saccharide, 10-50 g/L inorganic nitrogen source, 0.5-3 g/L, KH organic nitrogen source 2 PO 4 0.1-1 g/L、Na 2 HPO 4 0.1-0.2 g/L、CaCl 2 0.002-0.01 g/L、MnSO 4 0.0015-0.009 g/L、FeSO 4 0.001-0.006 g/L, wherein the saccharide is at least one of glucose, sucrose and starch; the inorganic nitrogen source may be ammonium salt, preferably ammonium sulfate, and the organic nitrogen source may be at least one of yeast extract powder, peptone, fish meal, corn steep liquor powder and soybean cake powder, and the Na 2 HPO 4 、 MnSO 4 And FeSO 4 Can be provided with crystal water in actual use, such as Na 2 HPO 4 ·12H 2 O、MnSO 4 ·H 2 O and FeSO 4 ·7H 2 O。
In the invention, in order to make the bacillus culture and other components of the invention cooperate to obtain better bacteriostasis and fermentation promotion effects, the initial pH of the fermentation medium is 6.5-7.5 and can be 6.5, 6.6, 6.7, 6.8, 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5 or any two values above or a value in a range.
In the present invention, the method further comprises drying the supernatant of the fermentation product-removed cells in order to obtain a final culture of bacillus. The supernatant liquid with the thalli removed is obtained by filtering a fermentation product.
In the invention, in order to ensure that the obtained bacillus culture has better bacteriostasis and fermentation promotion effects with other components of the invention, and is more beneficial to the efficient utilization and fermentation of sugar by saccharomyces cerevisiae, the fermentation medium also comprises a fermentation additive; preferably, the fermentation additive is present in an amount of 0.01 to 20 wt%; more preferably, the fermentation additive comprises at least one of a silicon-based antifoaming agent, an amino acid or derivative thereof, an oligopeptide, a phospholipid, a glycolipid, and a fatty acid or derivative thereof.
In the invention, in order to ensure that the obtained bacillus culture has better bacteriostasis and fermentation promotion effects with other components of the invention, and is more beneficial to the efficient utilization and fermentation of sugar by saccharomyces cerevisiae, the fermentation medium also comprises isopropyl thiogalactoside; more preferably, the concentration of the isopropyl thiogalactoside is 0.5-1.5mM, and can be; further preferably, the isopropylthiogalactoside is contacted with the fermentation medium at 2-6 hours of cultivation.
In a second aspect, the invention provides a method for fermentative ethanol production, the method comprising: in the presence of the above composition, the saccharide-containing feedstock is subjected to ethanol fermentation. Preferably, the composition is present in the ethanol fermentation system in an amount of 2-8 mg/L, which may be 2 mg/L, 3 mg/L, 4mg/L, 5 mg/L, 6 mg/L, 7 mg/L, 8 mg/L, or any two of the above values.
In the present invention, the sugar-containing feedstock is not limited, and is a common yeast in the art that can ferment to ethanol using sugar therein, preferably a liquefied mash and/or a sugar solution.
In a third aspect the present invention provides the use of a composition as described above in fermentation and/or food bacteriostasis.
Preferably, the antibacterial agent is at least one of the fields of food, alcohol, cultivation, cosmetics and medicine.
The present invention will be described in detail by examples. In the following examples, the preparation of bacillus cultures: bacillus subtilis @Bacillus subtilis) Inoculating CGMCC1.821 to LB liquid medium, culturing at 37deg.C and 200rpm for 16 hr to obtain seed liquid, inoculating into fermentation medium shake flask at a ratio of 5% (based on fermentation medium) by volume, and culturing at 37deg.C,1 mM IPTG is added when the culture is carried out for 4 hours under the condition of 200rpm, the culture is continued for 2 days, the sterile supernatant is spray-dried after the fermentation product is filtered to obtain a powder product, namely the bacillus culture, wherein the cellulase is 100U/g, the chitinase is 25U/g, the protease is 100U/g, and the lipopeptide is 10 weight percent, and the testing method of each component is as follows:
the cellulase activity determination method comprises the following steps: NY/T912-2004;
chitinase activity assay: GB/T34799-2017;
protease activity determination method: GB/T23527-2009;
lipopeptide assay method: the 1g bacillus culture was dissolved in water and mixed well, filtered through a 0.22 μm filter and analyzed by HPLC. The mobile phase analyzed by HPLC is methanol and water in a ratio of 85/15, the flow rate is 1 mL/min, the chromatographic column is a C18-ODS reversed phase chromatographic column, the column temperature is 40 ℃, and the detection wavelength is 205 nm by an ultraviolet detector.
The composition of the fermentation medium used was: glucose 50 g/L, ammonium sulfate 30g/L, yeast extract 1.5 g/L, KH 2 PO 4 0.5 g/L、Na 2 HPO 4 ·12H 2 O 0.4g/L、CaCl 2 0.006 g/L、MnSO 4 ·H 2 O 0.006 g/L、FeSO 4 ·7H 2 O 0.006 g/L，pH 7；
Glucose and ammonium sulfate were purchased from national pharmaceutical group chemicals limited;
yeast extract FM888 was purchased from Angel Yeast Co., ltd;
both Angel super brewing high activity dry yeast and acid protease are purchased from Angel Yeast Co., ltd, and saccharifying enzyme is purchased from Shandong Bioengineering Co., ltd;
urea was purchased from the company cymbidium guansu practice limited;
nisin is a commercial product of Zhejiang Yinuo biotechnology Co., ltd, food grade;
sodium gluconate is a commercial product of Shandong Baisheng biotechnology Co., ltd, and is food grade;
the chitosan oligosaccharide is a commercial product of Jiangsu Kotsumami bioengineering Co., ltd, and is food grade;
the sodium dehydroacetate is a commercial product of Shandong san Jose Biotechnology Co., ltd, and is food grade;
sodium diacetate is a commercial product of Qingdao Hovesen Biotechnology Co., ltd, food grade;
the calcium propionate is a commercial product of Henan Gaobao biotechnology Co., ltd, and is food grade;
the antibiotic product (penicillin) is a commercial product of Shandong Noribu Biotech Co., ltd, food grade.
Examples and comparative examples
The components are added in sequence according to the content of each component in the table 1 and are mixed uniformly.
TABLE 1
Note that: the units of the values in the tables are% by weight
Test case
In the test example, fermentation experiments were carried out on liquefied mash (liquefied mash produced by not performing sterilization treatment in the ethanol production process) with serious actual pollution of factories as a raw material:
1. cultivation of yeast
Step 1, weighing 8g of dry yeast in a 500mL conical flask, adding the dry yeast into 200mL of activated water, and activating the dry yeast in an incubator at 35 ℃ for 20 minutes to obtain an activated liquid;
step 2, using severely contaminated liquefied mash (pH 4.2) for yeast expansion: filling each 270g of liquefied mash into 500mL conical flask, and adding 2.5mL of activating solution;
step 3, adding 2.7mg of the corresponding bacteriostat (the composition obtained in the above examples and comparative examples and penicillin, and blank control are not added), 0.007g of acid protease, 0.014g of saccharifying enzyme and 0.27g of urea into the yeast, and culturing at 30 ℃ under ventilation for 10 hours at 45rpm to obtain the yeast.
2. Fermentation
Step 1, 60g of severely polluted liquefied mash raw material (pH 4.2) and 60g of severely polluted 30 wt% chemical starch solution are taken into a 500mL conical flask, 80g of yeast obtained by culturing in step 1 is added, and at the moment, the concentration of bacteriostat in fermentation liquor is respectively as follows: the control group was 4mg/L penicillin product and the experimental group was 4mg/L compositions prepared in examples and comparative examples, respectively.
Step 2, adding 0.076g of saccharifying enzyme into each bottle, sealing a liquid sealing pipe, uniformly mixing at a rotating speed of 100rpm, and fermenting at a temperature of 32 ℃ for 66 hours; three replicates were tested in each set.
And 3, after fermentation is completed, determining acidity by titration with a standard sodium hydroxide solution and determining alcohol content with an alcohol content meter, and calculating an average value of experimental data of 3 parallel samples, wherein the acidity result is shown in figure 1, and the alcohol content result is shown in figure 2.
Step 4, at the same time, the sugar content, organic acid, glycerol, ethanol, etc. in the mature mash was measured by a liquid phase column Aminex HPX-87H (available from Agilent corporation, model 300X 7.8 mm), and the results obtained by calculating the average value of each set of 3 parallel sample experimental data are shown in Table 2.
TABLE 2
The general acidity is used to characterize the bacteria contamination of the fermentation process, while the alcohol content is used to characterize whether the fermentation is better. As can be seen from FIG. 1, the acidity of the groups of examples 1-3 were 7.52, 7.31 and 7.47, respectively, which were lower than the acidity of the penicillin group of 7.55 and the acidity of the bacteriostatic agent-free group of 10.05, while the acidity of the groups of examples 4-8 were slightly higher than the penicillin group but lower than the bacteriostatic agent-free group; the results of the alcohol content analysis in FIG. 2 show that the alcohol content of the groups of examples 1 to 3 are 13.23%, 13.43% and 13.20%, respectively, which is higher than or equal to the alcohol content of the penicillin group and 11.81% of the alcohol content of the group without bacteriostat, while the alcohol content of the groups of examples 4 to 8 is slightly lower than that of the penicillin group and the group without bacteriostat.
To ensure accuracy of the results, liquid phase detection of the matured mash was performed simultaneously (Table 2). In the general fermentation process, the chromatographic residual total sugar and the chromatographic residual reducing sugar are used for representing whether the fermentation is thorough or not, and the higher the chromatographic residual total sugar and the chromatographic residual reducing sugar are, the less thorough the fermentation is. Total chromatographic sugars = DP4 (maltotetraose and above, oligosaccharides below decaose) +dp3 (maltotriose) +dp2 (maltose) +glucose+fructose, residual chromatographic reducing sugars = dp3+glucose+fructose. As can be seen from FIG. 3, the total and reduced sugars of the chromatograms of the groups of examples 1 to 3 were the lowest, the total sugar of the chromatograms was 13.91g/L, 13.63g/L and 13.93g/L, respectively, and the reduced sugar of the chromatograms was 3.72g/L, 3.59g/L and 3.86g/L, respectively, which were lower than the values of the penicillin group (total sugar of the chromatograms 14.30g/L, reduced sugar of the chromatograms 4.07 g/L) and the bacteriostatic agent-free group (total sugar of the chromatograms 21.51g/L and reduced sugar of the chromatograms 7.44g/L, respectively). Examples 4-8 were slightly higher in both total and reduced chromatograms than the penicillin and bacteriostatic groups.
The chromatographic organic acid is used for representing the fermentation pollution degree, and the larger the chromatographic organic acid value is, the more serious the fermentation pollution is, namely the weaker the inhibition of the bacteriostat to the mixed bacteria is. Chromatographic organic acid = succinic acid + lactic acid + acetic acid. As can be seen from FIG. 4, the minimum amount of organic acids was 2.92g/L, 2.79g/L, and 2.90g/L for the chromatograms of the groups of examples 1-3, respectively, which were lower than 3.15g/L for the penicillin group and 4.74g/L for the bacteriostatic agent-free group. Examples 4-8 were slightly higher on chromatographic organic acids than the penicillin group and the bacteriostatic agent-free group.
The ethanol/glycerol characterizes the fermentation effect, and the higher the ethanol/glycerol value is, the higher the fermentation utilization rate is. As can be seen in FIG. 5, the ethanol/glycerol values for examples 1-3 were highest, 9.70, 9.85 and 9.58, respectively, above 9.46 for the penicillin group and 7.92 for the bacteriostatic agent-free group. The ethanol/glycerol values for examples 4-8 were 8.64, 9.01, 8.89, 8.47 and 8.27, respectively, lower than the penicillin and bacteriostatic agent-free groups.
3. Fermentation experiments of sterile liquefied mash
Fermentation experiments were carried out using sterile liquefied mash (liquefied mash produced by sterilizing in ethanol production) as a raw material, and steps 1 and 2 were the same.
After fermentation, acidity and alcohol content were determined by titration with standard sodium hydroxide solution and alcohol content by alcohol content meter, and the results of acidity and alcohol content are shown in tables 3 and 4 after the average value of the experimental data of each group of 3 parallel samples was calculated.
As can be seen from tables 3 and 4, the compositions in examples and comparative examples have a promoting effect on fermentation effect, and the larger the proportion of bacillus subtilis fermented liquid, the more remarkable the promoting effect.
TABLE 3 Table 3
TABLE 4 Table 4
The results show that compared with penicillin, the composition of the examples 1-3 achieves better antibacterial effect, and is more beneficial to the efficient utilization and fermentation of sugar by the super wine yeast; the compositions of examples 4-8 and comparative examples 1-4 containing the Bacillus broth spray-dried powder were disadvantageous for fermentation of the super wine yeast due to poor inhibitory effect on the miscellaneous bacteria as compared with penicillin.
The preferred embodiments of the present invention have been described in detail above, but the present invention is not limited thereto. Within the scope of the technical idea of the invention, a number of simple variants of the technical solution of the invention are possible, including combinations of the individual technical features in any other suitable way, which simple variants and combinations should likewise be regarded as being disclosed by the invention, all falling within the scope of protection of the invention.
Claims (10)
1. A composition having both bacteriostatic and pro-yeast effects, said composition comprising: at least one of chitosan and sodium diacetate is used in combination with bacillus culture.
2. The composition of claim 1, wherein the chitosan is included in the composition, and the weight ratio of the bacillus culture to the chitosan is 1:0.005-0.25;
and/or, the composition comprises the sodium diacetate, and the weight ratio of the bacillus culture to the sodium diacetate is 1:0.005-0.45;
and/or the bacillus culture contains cellulase, chitinase, protease and lipopeptide.
3. The composition of claim 1 or 2, wherein nisin is further included in the composition, the weight ratio of the bacillus culture to nisin being 1:0.005-0.45;
and/or, the composition further comprises sodium gluconate, and the weight ratio of the bacillus culture to the sodium gluconate is 1:0.01-0.3.
4. The composition of claim 1 or 2, wherein the bacillus culture is bacillus subtilis @Bacillus subtilis) Is a culture of (a);
and/or, the composition further comprises sodium dehydroacetate, and the weight ratio of the bacillus culture to the sodium dehydroacetate is 1:0.02-20;
and/or, the composition further comprises calcium propionate, and the weight ratio of the bacillus culture to the calcium propionate is 1:0.005-1.
5. The composition of claim 4, wherein the bacillus subtilis is at least one of collection numbers CGMCC1.821, ATCC6051, ATCC23857, ATCC21332, ATCC11774 and ATCC 13952.
6. The composition of claim 1 or 2, wherein the bacillus culture is prepared by a method comprising: inoculating bacillus into a fermentation medium for fermentation.
7. The composition of claim 6, wherein the fermentation medium comprises: 30-100 g/L saccharide, 10-50 g/L inorganic nitrogen source, 0.5-3 g/L, KH organic nitrogen source 2 PO 4 0.1-1 g/L、Na 2 HPO 4 0.1-0.2 g/L、CaCl 2 0.002-0.01 g/L、MnSO 4 0.0015-0.009 g/L、FeSO 4 0.001-0.006 g/L, wherein the saccharide is at least one of glucose, sucrose and starch;
and/or, the fermentation conditions include: the time is 2-4 days, and the temperature is 30-40 ℃;
and/or the initial pH of the fermentation medium is 6.5-7.5;
and/or, the preparation method of the bacillus culture further comprises the following steps: and (3) drying the supernatant of the fermentation product with the thalli removed.
8. A method for fermentative ethanol production, the method comprising: ethanol fermentation of a saccharide-containing feedstock in the presence of a composition according to any one of claims 1-7.
9. The method of claim 8, wherein the composition is present in the ethanol fermentation system in an amount of 2-8 mg/L.
10. Use of a composition according to any one of claims 1 to 7 for fermentation and/or bacteriostasis.
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