CN106665636A - Method for inhibiting contaminative microorganism during alcoholic fermentation by adopting ethyl lauroyl arginate - Google Patents
Method for inhibiting contaminative microorganism during alcoholic fermentation by adopting ethyl lauroyl arginate Download PDFInfo
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- CN106665636A CN106665636A CN201510748675.9A CN201510748675A CN106665636A CN 106665636 A CN106665636 A CN 106665636A CN 201510748675 A CN201510748675 A CN 201510748675A CN 106665636 A CN106665636 A CN 106665636A
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- ethyl ester
- fermentation
- arginine ethyl
- ethyl lauroyl
- lauroyl arginate
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N47/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom not being member of a ring and having no bond to a carbon or hydrogen atom, e.g. derivatives of carbonic acid
- A01N47/40—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom not being member of a ring and having no bond to a carbon or hydrogen atom, e.g. derivatives of carbonic acid the carbon atom having a double or triple bond to nitrogen, e.g. cyanates, cyanamides
- A01N47/42—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom not being member of a ring and having no bond to a carbon or hydrogen atom, e.g. derivatives of carbonic acid the carbon atom having a double or triple bond to nitrogen, e.g. cyanates, cyanamides containing —N=CX2 groups, e.g. isothiourea
- A01N47/44—Guanidine; Derivatives thereof
Abstract
The invention discloses a method for inhibiting a contaminative microorganism during alcoholic fermentation by adopting ethyl lauroyl arginate. The invention relates to a method for inhibiting a contaminative microorganism in an alcoholic fermentation process. The method consists of the following parts that (1), used antibacterial substances are the ethyl lauroyl arginate and (2) and other salt compounds of the ethyl lauroyl arginate, and these compounds comprise the hydrochloride, the lactate, the citrate, the sulfate, the phosphate, the nitrate, the ascorbate and the acetate of the ethyl lauroyl arginate, other tallate and the like; (3), the using concentration of the ethyl lauroyl arginate or the salt compounds of the ethyl lauroyl arginate is 10PPM to 80PPM; (4), an applied fermentation process is the alcoholic fermentation or rum fermentation carried out by adopting raw materials of a cereal, sugarcane, beet, a plant fiber, a potato, a sweet potato and the like.
Description
Technical field
The present invention relates to a kind of method for suppressing contaminating microorganisms in alcoholic fermentation process.The method is refered in particular to using lauroyl arginine ethyl ester（English name：lauramide arginine
ethyl ester）Suppress S. cervisiae（Saccharomyces
cerevisiae）The microorganism polluted during fermenting and producing alcohol, Rum, yellow rice wine.
Background technology
Saccharomyces cerevisiae is adopted with the crops such as corn, beet, sugarcane, wheat fermenting raw materials production alcohol in the general inclined application in the whole world, its production process includes crushing, boiling, saccharification, fermentation, the distillation of raw material.The microorganism that sweat is adopted is the saccharomyces cerevisiae Pseudomonas in saccharomycete（Saccharomyces
cerevisiae).At 29 ± 1 °C, pH value is controlled 4.2 to 4.7, and sweat is frequently with batch fermentation or continuously ferments, and the pollution of bacterium is highly susceptible in fermentation for fermentation temperature control.The pollution of bacterium can not only make zymotic fluid souring, can also reduce the yield of alcohol.The microorganism of pollution includes bacillus subtilis（Bacillus subtilis）, bacillus cereus（Bacillus cereus）Deng.
Factory suppresses the pollution of bacterium using addition antibiotic.These antibiotic mainly have coban（Monensin ）With Virginia rhzomorph（Virginiamycin）, sometimes can also use penicillin.
Rum is Jing fermentation by saccharomyces cerevisiae, distillation, ageing and a kind of Spirit for producing, alcohol content 45% or so with cane milling syrup out or molasses as raw material.At 32 ± 1 °C, alcohol content generally reaches 8.0% to 10.0% to fermentation time to temperature control in sweat generally at 6 to 8 hours, fermentation termination, and cane sugar content drops to less than 2.0%.Fermentation ends are separated saccharomycete from zymotic fluid using the method for centrifugal sedimentation.Isolated zymotic fluid enters distillation process.The yeast isolated, adjusts pH2 ~ 2.5 to preserve, as the bacterial classification of lower batch fermentation using dilute acid soln.Contaminating microorganisms can in a large number produce lactic acid, show the adverse consequences such as pH declines, alcohol output is reduced, product taste changes.The bacterium of pollution mainly has lactobacillus fermenti（Lactobacillus
fermentum）, Lactobacillus plantarum（Lactobacillus plantarum）, Lactobacillus casei（Lactobacillus casei）, bacillus subtilis（Bacillus subtilis）, Leuconostoc mesenteroides（Leuconostoc
mesenteroides）, wax-like acetic acid bacteria（Bacillus cereus）.Factory adopts coban（Monensin）Or Virginia rhzomorph（Virginamicin）The content of control bacterium, the residual of antibiotic can produce potential impact to the health of people.
In sum, finding a kind of safely and effectively material is used to suppress the pollution of bacterium in alcoholic fermentation or Rum sweat to have actual meaning.
Lauroyl arginine ethyl ester is a kind of cationic surfactant that adopts arginine, ethanol, laurate for raw material obtained from a few step chemical reactions.Its CAS is encoded to 60372-77-2, and INS is encoded to 243.
Lauroyl arginine ethyl ester has broad spectrum antibiotic activity by destroying the cell membrane of microorganism.Salt can be formed with hydrochloric acid, lactic acid, citric acid, sulfuric acid, phosphoric acid, ascorbic acid, aliphatic acid etc..It is a kind of white powder, the solubility 24% or so in water at normal temperature, 54 °C or so of fusing point.Its chemical stability and antibacterial activity can be kept in the range of pH3 ~ 7.2005, FDA regarded as a kind of GRAS materials, and research on maximum utilized quantity is 200ppm.In addition, Europe approval may apply in cosmetics and in food as antibacterial material.
Lauroyl arginine ethyl ester is decomposed into aliphatic acid, ethanol and amino acid in people, animal, natural environment, is a kind of environmentally friendly material.
The content of the invention
Instant invention overcomes suppressing the deficiency of polluted bacteria method in alcoholic fermentation process using antibiotic.Material used is lauroyl arginine ethyl ester or its salt, and these salts include its hydrochloride, lactate, citrate, sulfate, phosphate, nitrate, ascorbate, acetate, and other soaps.The concentration of lauroyl arginine ethyl ester or its salt is 10PPM to 80PPM.The alcoholic fermentation process that sweat is predominantly carried out using cereal, sucrose, beet, potato, cellulose etc. as raw material.The present invention is using lauroyl arginine ethyl ester to saccharomyces cerevisiae and bacterium minimum inhibitory concentration（MIC）Difference, find suitable concn and the growth of bacterium and breeding can be made a significant impact through research, and the impact to growth and the breeding of saccharomyces cerevisiae is smaller, reaches the value-added purpose of suppression polluted bacteria.
Description of the drawings.
Fig. 1 is different lauroyl arginine ethyl ester concentration to saccharomyces cerevisiae（Saccharomyces
Cerevisiae CICC1023) growth impact.
Fig. 2 is different lauroyl arginine ethyl ester concentration to Lactobacillus casei（Lactobacillus
Paracasei CICC20241) growth impact.
Fig. 3 is different lauroyl arginine ethyl ester concentration to bacillus subtilis（Bacillus
Subtilis CICC10033) growth impact.
Fig. 4 is different lauroyl arginine ethyl ester concentration to Lactobacillus plantarum (Bacterial Strain
Code CICC20263) growth impact.
Fig. 5 is the impact of saccharomycete activity of the lauroyl arginine ethyl ester to adopting in alcoholic fermentation process.
Fig. 6 is impact of the lauroyl arginine ethyl ester to polluting lactic acid bacteria in alcoholic fermentation process.
Fig. 7 is impact of the lauroyl arginine ethyl ester to alcohol output in alcoholic fermentation process
Detailed description of the invention and citing.
Lauroyl arginine ethyl ester is a kind of cationic surfactant, and the material in mainly acting on the cell membrane of microorganism so as to cause cytoplasm is lost in and makes microorganism dead, and various microorganisms are different to the sensitivity of lauroyl arginine ethyl ester.Microorganism is concentration dependent to the sensitivity of lauroyl arginine ethyl ester, that is to say, that relevant with the content of the quantity of microorganism and lauroyl arginine ethyl ester.
The bacterial concentration polluted in Rum sweat is usually up to 106- 108CFU/ml, the bacterium of pollution mainly has lactobacillus fermenti（Lactobacillus fermentum）, Lactobacillus plantarum（Lactobacillus plantarum）, Lactobacillus casei（Lactobacillus casei）, bacillus subtilis（Bacillus subtilis）, Leuconostoc mesenteroides（Leuconostoc
mesenteroides）With wax-like acetic acid bacteria（Bacillus cereus）.
Factory controls the content of bacterium using antibiotic, if bacterial content reaches 105– 106CFU/ml, it is believed that acceptable, can reach 10 when few4CFU/ml。
The microorganism that present invention research is used comes from Chinese industrial Culture Collection（CICC）, following table is listed studies the microbial name and its bacterium numbering for adopting.Experiment have studied growth characteristics and microorganism concn of this several microorganism in different time by the lauroyl arginine ethyl ester of addition variable concentrations using certain proportioning fermentation medium.
Represent that micro organism quantity adopts visible spectrophotometer mensuration absorbance（OD values）.
Table 1, research microbial name and its bacterium numbering.
Lauroyl arginine ethyl ester is 60PPM to the Mlc for selecting saccharomyces cerevisiae, and the impact as 50PPM is not notable.Compare with saccharomyces cerevisiae, selected several lactic acid bacterias are more sensitive to lauroyl arginine ethyl ester, generally in below 20PPM, its growth is just significantly suppressed.Suitable lauroyl arginine ethyl ester concentration is found in this experiment, it is ensured that the impact to saccharomyces cerevisiae is slight, and the impact to lactobacter growth is significant, and the purpose of lactic acid bacteria inhibiting growth is reached by controlling usage amount.
The Medium Proportion for below adopting for experiment：
Lactic acid bacteria slant medium（g/L）
Sucrose
10.0
Vegetable protein
5.0
Yeast extract
8.0
Potassium dihydrogen phosphate
10.0
Agar
18.0
It is 7.4 to adjust pH with 20% potassium hydroxide solution.
Lactobacillus-fermented culture medium（g/L）
Sucrose
10.0
Yeast extract
8.0
Sodium phosphate（Na2HPO4·12H20）
22.4
Potassium dihydrogen phosphate
1.4
Magnesium sulfate（MgSO4·7H2O）
0.2
It is 7.4 to adjust pH with 20% potassium hydroxide solution.
Saccharomycete slant medium（g/L）
Glucose
20.0
Yeast extract
10.0
Peptone
20.0
Agar
18.0
It is 7.4 to adjust pH with 20% potassium hydroxide solution.
Yeast fermentation medium（g/L）
Sucrose
160.0
Brown sugar
30.0
Yeast extract
10.0
Potassium dihydrogen phosphate
5.0
Sodium chloride
2.0
It is 7.4 to adjust pH with 20% potassium hydroxide solution.
Experimentation.
The activation of bacterial classification：The glycerol tube thaw at RT containing bacterial classification of low-temperature preservation is taken, with oese bacteria suspension is dipped, be applied on slant medium well prepared in advance, be placed in quiescent culture 16h in 31 ± 0.5 DEG C of constant incubator.
The preparation of bacteria suspension：The bacterium colony grown on slant medium is all shoveled out, is mixed with Sterile Salines of the 50mL containing 0.2% NaCl, shaken up standby.
The Amplification Culture of thalline：The bacteria suspension 0.5mL for preparing is drawn with 1mL liquid-transfering guns, in being inoculated into the preprepared 250mL triangular flasks containing 50mL seed culture mediums, 31 ± 0.5 DEG C of constant incubator quiescent culture is placed in.
The measure of different lauroyl arginine ethyl ester concentration conditions hypothallus growth curves：Again respectively the lauroyl arginine ethyl ester of addition 10,20,30,40,50,60PPM presses some bottles of cultural method Amplification Culture in [0032] in original fermentation medium, it is placed in 31 ± 0.5 DEG C of constant incubator quiescent culture, one bottle is taken out at regular intervals, nutrient solution OD values are determined, the curve that the OD values of nutrient solution change with incubation time is made.
Nutrient solution OD values（Absorbance）Measure：Using 723PC type visible spectrophotometers, with the nonvaccinated culture medium through sterilizing as blank, the OD values of its nutrient solution under 600nm wavelength are determined（Absorbance）.
Experimental result.
Figure one, by different time OD values（Absorbance）Change, show different lauroyl arginine ethyl ester concentration to saccharomyces cerevisiae（Saccharomyces cerevisiae
CICC1023) the impact of growth.Find out from figure one and there is large effect to saccharomycetic growth when of that month osmanthus acyl arginine ethyl ester concentration reaches 60PPM, the impact in 50PPM is slight.
Figure two, by different time OD values（Absorbance）Change, show different lauroyl arginine ethyl ester concentration to Lactobacillus casei（Lactobacillus
Paracasei CICC20241) growth impact.Experiment proves that the growth of Lactobacillus casei reaches and completely suppresses when lauroyl arginine ethyl ester concentration reaches 40PPM.
Figure three, by different time OD values（Absorbance）Change, show different lauroyl arginine ethyl ester concentration to bacillus subtilis（Bacillus subtilisCICC10033) growth impact.Experiment proves that the growth of bacillus subtilis reaches and completely suppresses when lauroyl arginine ethyl ester concentration reaches 10PPM.
Figure four, by different time OD values（Absorbance）Change, show different lauroyl arginine ethyl ester concentration to Lactobacillus plantarum (Bacterial
Strain CICC20263) growth impact.Figure four proves that lauroyl arginine ethyl ester is 20PPM to the full inhibition concentrations of Lactobacillus plantarum.
Productive experiment.
The purpose of this experiment is to assess impact of the lauroyl arginine ethyl ester to contaminating microorganisms, saccharomyces cerevisiae and alcohol output in Rum sweat.This experiment is completed in Rum workshop, and experimental program already has accounted for a complete production cycle.
Experimental program：
Control group --- without any antibacterial substance.Lauroyl arginine ethyl ester group --- the lauroyl arginine ethyl ester of addition 55PPM.Normal group --- the addition normal productive process antibacterial substance for using and the concentration for normally using.
Saccharomycetic process and preparation：Saccharomycete is processed according to normal productive process using dilute acid soln, and requires to be added in fermentation tank according to normal process.The antibacterial substance for using is added separately in fermentation tank according to the amount of design.
Fermentation tank feed supplement and ferment control：Once, feed supplement six times altogether, supplementary raw material is the molasses aqueous solution, and it is 10 that Jing determines the quantity of contaminating microorganisms for fermentation tank feed supplement per hour6CFU/ml, with Typical Representative meaning.Whole fermentation time continues 9 hours, and when carbon dioxide is no longer produced, sucrose concentration drops below 1%（W/W）When fermentation stop.
Sampling：Fermentation ends, fermentation broth agitation is uniform, sampling.By sample centrifugation, lower sediment thing is used for microbiological analysis, and upper strata is for alcohol content analysis.
Analysis：Analysis project totally three, respectively yeast activity, polluted bacteria are counted and alcohol output.
Experimental result：
Using workshop yeast activity assay method, it is 81.5% to determine initial yeast activity.Figure five shows the control of the saccharomycete activity in three experimental group zymotic fluids of fermentation termination, it was demonstrated that the lauroyl arginine ethyl ester of 55PPM has slight impact to saccharomycetic activity.
Figure six shows the impact that addition lauroyl arginine ethyl ester grows to polluted bacteria, it is illustrated that prove to be substantially less than two other experimental group using the concentration of contaminating microorganisms after lauroyl arginine ethyl ester.
Figure seven shows impact of the addition lauroyl arginine ethyl ester to alcohol output.
Prove in plant experiment, lauroyl arginine ethyl ester has obvious inhibitory action to the bacterium polluted in alcoholic fermentation.It is slight to saccharomycetic growth effect.Alcohol output reduces by 0.6% compared with other inhibitor for using, but using LAE cost less than the inhibitor that is currently in use, Integrated comparative will not reduce factory account.
In Rum fermentation is applied to, the reduction of contaminating microorganisms has obvious facilitation to the mouthfeel for improving product.
Comprehensive narration has been done in the main points of the present invention, citing, application, it will be understood that present disclosure is not only limited to these citings, and without prejudice to present disclosure, the especially experimental program of content mentioned above other modifications can be carried out.
Claims (5)
1. the present invention relates to be applied to the growth and breeding of contaminating microorganisms in alcoholic fermentation, Rum, yellow wine fermentation suppression sweat using antibacterial material lauroyl arginine ethyl ester.
2., according to claim 1, chemical substance used is other salt compounds of lauroyl arginine ethyl ester and this material.
3. these salt compounds include its hydrochloride, lactate, citrate, sulfate, phosphate, nitrate, ascorbate, acetate and other soaps etc..
4., according to claim 1, application includes the alcoholic fermentation carried out using raw materials such as cereal, sugarcane, beet, string, potato, sweet potatoes, Rum fermentation.
5., according to claim 2, the concentration of these materials is 10ppm to 80ppm.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108992437A (en) * | 2017-11-03 | 2018-12-14 | 华东师范大学 | Purposes of the lauroyl arginine ethyl ester as antibacterial agent for animals |
| CN113546055A (en) * | 2018-06-22 | 2021-10-26 | 华东师范大学 | Novel pharmaceutical coated articles, method of manufacture and uses thereof |
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| CN1049687C (en) * | 1992-01-27 | 2000-02-23 | 国家造酒集团联盟 | The use of polyether ionophore antibiotics to control bacterial growth in alcohol fermentation |
| CN102088858A (en) * | 2008-07-02 | 2011-06-08 | 米雷特实验室股份公司 | Use of cationic surfactants as sporicidal agents |
| CN103773810A (en) * | 2012-10-18 | 2014-05-07 | 中粮集团有限公司 | Method for producing alcohol by fermentation |
| CN104364383A (en) * | 2012-06-15 | 2015-02-18 | 纳幕尔杜邦公司 | Use of virginiamycin for bacterial contamination control in fermentations using zymomonas mobilis |
-
2015
- 2015-11-07 CN CN201510748675.9A patent/CN106665636A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1049687C (en) * | 1992-01-27 | 2000-02-23 | 国家造酒集团联盟 | The use of polyether ionophore antibiotics to control bacterial growth in alcohol fermentation |
| CN102088858A (en) * | 2008-07-02 | 2011-06-08 | 米雷特实验室股份公司 | Use of cationic surfactants as sporicidal agents |
| CN104364383A (en) * | 2012-06-15 | 2015-02-18 | 纳幕尔杜邦公司 | Use of virginiamycin for bacterial contamination control in fermentations using zymomonas mobilis |
| CN103773810A (en) * | 2012-10-18 | 2014-05-07 | 中粮集团有限公司 | Method for producing alcohol by fermentation |
Non-Patent Citations (1)
| Title |
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| 张赛赛等: "一种新型食品防腐剂――L-月桂酰胺精氨酸盐酸盐乙醇酯", 《食品工业科技》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108992437A (en) * | 2017-11-03 | 2018-12-14 | 华东师范大学 | Purposes of the lauroyl arginine ethyl ester as antibacterial agent for animals |
| CN108992437B (en) * | 2017-11-03 | 2021-10-12 | 华东师范大学 | Use of lauroyl arginine ethyl ester as veterinary antibacterial agent |
| CN113546055A (en) * | 2018-06-22 | 2021-10-26 | 华东师范大学 | Novel pharmaceutical coated articles, method of manufacture and uses thereof |
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