CN117264800A - Lactobacillus plantarum for reducing content of fusel in yellow wine and application thereof - Google Patents
Lactobacillus plantarum for reducing content of fusel in yellow wine and application thereof Download PDFInfo
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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
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
- C12G3/00—Preparation of other alcoholic beverages
- C12G3/02—Preparation of other alcoholic beverages by fermentation
- C12G3/021—Preparation of other alcoholic beverages by fermentation of botanical family Poaceae, e.g. wheat, millet, sorghum, barley, rye, or corn
- C12G3/022—Preparation of other alcoholic beverages by fermentation of botanical family Poaceae, e.g. wheat, millet, sorghum, barley, rye, or corn of botanical genus Oryza, e.g. rice
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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
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- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- 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 belongs to the technical field of microorganisms, and relates to lactobacillus plantarum for reducing the content of fusel in yellow wine and application thereof. Lactobacillus plantarum (Lactobacillus plantarum) AR1018 is preserved in China general microbiological culture collection center (CGMCC) with the preservation number of 25626 in 2022, 8 and 31; the strain has better capability of resisting ethanol stress and acid stress, has better utilization capability of sucrose, also has better antibacterial activity, and can reduce the total amount of fusel in fermentation liquor by 37.31 percent by co-culture fermentation with yeast. The method is characterized in that the method is used as an auxiliary starter in the process of brewing the yellow wine, the total amount of higher alcohols in the yellow wine can be obviously reduced, 0.5% -2.0% of inoculation amount is added in the fermentation process, and after the stages of pre-fermentation at 28 ℃, post-fermentation at 20 ℃, squeezing, clarification and wine frying, the obtained yellow wine has bright color and luster, mellow taste, and the content of fusel is reduced by 8.72% -33.36%, so that the flavor quality and drinking comfort of the yellow wine are effectively improved.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to lactobacillus plantarum for reducing the content of fusel in yellow wine and application thereof.
Background
The yellow wine is a fermented wine brewed by taking rice grains as raw materials and adding saccharification fermenting agents such as yeast, wheat starter and the like, and has the advantages of rich nutrition, unique flavor, moderate alcohol degree and mild alcohol property, and meets the requirements of consumers in the current big health age on low alcohol degree, nutrition and the like. Yellow wine is a natural fermented wine with long history, is extremely rich in names in China and even the world, and is called as 'three great old wines in the world' together with western grape wine and beer. Yellow wine is taken as a special wine seed in China, and competitors in the international market are relatively few, so that the yellow wine has great export potential. However, consumer acceptance of yellow wine products remains low with market share, which is closely linked to the poor drinking comfort of yellow wine, which is "easy to get on the head, easy to get drunk".
The fusel is taken as an important flavor component generated in the yellow brewing process, is a generic name of monovalent alcohols with more than two carbon chain frameworks, is mainly generated by metabolism of brewing microorganisms in the fermentation process, and plays an important role in aroma and flavor development of the yellow wine. However, when the content of the miscellaneous alcohol in the yellow wine is too high, the miscellaneous flavor can be brought to the yellow wine to influence the flavor quality, and symptoms such as thirst, headache and the like are easy to generate after drinking, so that the yellow wine has poor drinking comfort. The miscellaneous alcohol produced in the process of brewing the yellow wine mainly comprises n-propanol, isobutanol, isoamyl alcohol, phenethyl alcohol and the like, wherein the content of the isoamyl alcohol is the highest, and tens of times of the content of the n-propanol and the isobutanol is the main reason for leading to the 'top' of the yellow wine. Therefore, reducing the content of isoamyl alcohol in the yellow wine has important significance for controlling the content of miscellaneous alcohol in the yellow wine and improving the quality and the drinking comfort of the yellow wine.
Disclosure of Invention
In order to solve the problem of poor drinking comfort of the traditional yellow wine that the yellow wine is easy to get on the head and drunk, the invention provides lactobacillus plantarum for reducing the content of fusel in the yellow wine and application thereof.
The aim of the invention can be achieved by the following technical scheme:
the invention firstly provides lactobacillus plantarum (Lactobacillus plantarum) for reducing the content of fusel in yellow wine, which is marked as AR1018 strain and is preserved in China general microbiological culture collection center (CGMCC) with the preservation number of No.25626 and the address: the institute of microorganisms of national academy of sciences of China, no. 1, no. 3, north Chen West Lu, the Korean region of Beijing.
The invention further provides application of the lactobacillus plantarum for reducing the content of the fusel in the yellow wine, wherein the lactobacillus plantarum for reducing the content of the fusel in the yellow wine is used as an auxiliary fermenting agent for brewing in the process of brewing the yellow wine, so that the flavor of the yellow wine can be obviously improved, and the content of the fusel in the yellow wine, especially the isoamyl alcohol in the yellow wine, can be obviously reduced.
In one embodiment of the invention, the lactobacillus plantarum which reduces the content of the fusel in the yellow wine is added in an inoculation amount of 0.5-2.0% as an auxiliary fermentation agent in the process of brewing the yellow wine, and the lactobacillus plantarum and the yeast are fermented together.
In one embodiment of the invention, the processes of pre-fermentation at 28 ℃ and post-fermentation at 20 ℃, squeezing, clarifying and decocting are adopted in the process of brewing the yellow wine.
In one embodiment of the invention, the specific process conditions of the lactobacillus plantarum for reducing the content of the fusel in the yellow wine as an auxiliary starter in the brewing process of the yellow wine are as follows:
(1) Adding 10% (v/v) wheat starter into cooled rice, inoculating 9% (v/v) yeast, standing at 28deg.C, and performing main fermentation;
(2) Inoculating 1% (v/v) lactobacillus plantarum fermentation liquor into the fermentation liquor within 2-9 days of fermentation, stirring uniformly, and standing for fermentation;
(3) After the main fermentation starts for 5 days, the temperature is reduced to 20 ℃ for standing fermentation for 10 days, and stirring is carried out once a day;
(4) After 15 days of main fermentation, filtering and squeezing fermented mash, clarifying the obtained filtrate overnight, blending and decocting to obtain yellow wine.
The invention also provides a preservation method of the lactobacillus plantarum for reducing the content of the fusel in the yellow wine, which comprises the steps of adding 20% glycerol into the bacterial liquid of the lactobacillus plantarum for reducing the content of the fusel in the yellow wine, uniformly mixing, and placing in a refrigerator at the temperature of minus 80 ℃ for 2-3 years.
The invention also provides an activating method of the lactobacillus plantarum for reducing the content of the fusel in the yellow wine, the glycerol freezing tube is taken out during the activating process, the glycerol freezing tube is melted under the ice bath condition, the bacterial liquid is inoculated into a MRS culture dish prepared in advance by using a sterile inoculating loop, the bacteria liquid is placed into an anaerobic incubator for growth, and complete bacterial colony grows in the culture dish for about 40 hours, so that the bacteria can be passaged.
Compared with the prior art, the invention has the following advantages:
the strain provided by the invention has better capability of resisting ethanol stress and acid stress, has better utilization capability on sucrose, also has better antibacterial activity, and can reduce the total amount of fusel in fermentation liquor by 37.31% by co-culture fermentation with yeast. The yellow wine is taken as an auxiliary starter in the brewing process of the yellow wine, the total amount of higher alcohols in the yellow wine can be obviously reduced, 0.5% -2.0% of inoculation amount is added in the fermentation process, and after the stages of pre-fermentation at 28 ℃, post-fermentation at 20 ℃, squeezing, clarification and wine frying, the obtained yellow wine has bright color and luster, mellow taste, the content of hetero alcohol is reduced by 8.72% -33.36%, especially the content of isoamyl alcohol in the yellow wine can be reduced by 8.07% -38.48%, and the flavor quality and the drinking comfort level of the yellow wine are effectively improved.
Drawings
FIG. 1 extreme tolerance of lactic acid bacteria under different concentrations of ethanol stress (5% -15%);
FIG. 2 extreme tolerance of lactic acid bacteria under different acid stresses (pH 4.0-3.3);
fig. 3 analysis of sucrose utilization characteristics of lactic acid bacteria ("" represents significant differences in growth of lactic acid bacteria in sucrose environments);
FIG. 4 comparison of antibacterial activity of lactic acid bacteria (a: lactobacillus plantarum AR1018 antibacterial zone; b: lactic acid bacteria without antibacterial zone);
FIG. 5 content of fusel by co-fermentation of different lactic acid bacteria with yeast.
Detailed Description
The invention will now be described in detail with reference to the drawings and specific examples.
Example 1
Isolation, culture, performance screening and preservation of lactobacillus plantarum AR 1018.
1. Purpose of experiment
The embodiment provides an operation flow for separating lactic acid bacteria capable of improving the flavor and quality of yellow rice wine from natural fermentation food and a bacteria identification and preservation method.
2. Experimental equipment
2.1 laboratory apparatus
ML104 type electronic balance (Mettler Toledo, metrele-tolidol instruments Shanghai Co., ltd.), PL2002 type electronic balance (Mettler Toledo, metrele-tolidol instruments Shanghai Co., ltd.), HIRAYAMA AUTOCLAVE (Japanese autoclave), SW-CJ-2D type ultra clean bench (Suzhou purification equipments Co., ltd.), 3-18K type centrifuge (sigma product), BUGBOX type anaerobic workstation (RUSKINN product), enzyme-labeled instrument (MD company), gas chromatograph-mass spectrometer (Siemet Fed.), XSP-BM-2C type biological optical microscope (Shanghai Aimer optical instruments Co., ltd.), and other usual appliances (conical flask, beaker, measuring cylinder, freezing tube, inoculating loop, alcohol lamp, culture dish and liquid-transferring device etc.)
2.2 major reagents
Beef extract powder (Beijing land bridge technologies Co., ltd., lot number 160901), casein peptone (Beijing land bridge technologies Co., ltd., lot number 160801), yeast extract powder (Beijing land bridge technologies Co., ltd., lot number 160815), glucose (General-Reagent, lot number 160817), sodium acetate trihydrate (Shanghai source leaf Biotechnology Co., lot number 161013), dipotassium phosphate (Shanghai source leaf Biotechnology Co., lot number 160720), magnesium sulfate heptahydrate (Shanghai source leaf Biotechnology Co., lot number 170205), manganese sulfate monohydrate (Shanghai run chemical Co., lot number 160208), tween 80 (General-Reagent, lot number 160509), glycerol (General-Reagent, lot number 170310)
3. Experiment contents and method
3.1 preparation of MRS Medium
The formulation of MRS medium per liter is as follows: 10g of beef extract powder, 10g of casein peptone, 5g of yeast extract powder, 20g of glucose, 8.3g of sodium acetate trihydrate, 2g of dipotassium hydrogen phosphate, 0.58g of magnesium sulfate heptahydrate, 0.25g of manganese sulfate monohydrate, 1mL of tween-80, and sterilizing at 115 ℃ for 20min by using deionized water to fix the volume to 1L.
3.2 isolation of seed
Collecting yellow wine fermentation liquor, diluting by a concentration gradient dilution method, coating 100uL of the dilution liquor on an MRS culture medium plate, carrying out anaerobic culture at 37 ℃ for 40-56h, respectively picking colonies with different characters to a new MRS culture medium plate according to the shape, the size, the color and the like of the colonies, and carrying out streaking separation and purification to obtain separated strains; carrying out gram staining and catalase contact test on the obtained isolated strain, and selecting strains with gram staining positive and catalase contact negative according to the characteristics of lactic acid bacteria, such as gram staining positive and catalase free; extracting the DNA of the obtained strain, using a general primer 27F/1492R (27F: 5 '-AGAGTTTGATCCTGGGCTCAG-3', 1490 2R:5 '-GGTTACCTTGTTACGACTT-3') of a lactobacillus 16srDNA fragment, carrying out PCR amplification, and then sending to a biological engineering (Shanghai) stock company for sequencing, and carrying out homologous comparison with a gene sequence in an NCBI gene library; and (5) identifying to obtain lactobacillus strains.
3.3 Strain screening
3.3.1 analysis of the tolerance of lactic acid bacteria under ethanol stress
Absolute ethyl alcohol filtered by a sterile 0.22 mu m organic phase filter membrane is added into an MRS culture medium after high-pressure steam sterilization, and ethanol stress culture mediums with the ethanol content of 5%, 10% and 15% are respectively prepared, so that the continuous change of ethanol stress in the yellow wine fermentation process is simulated. After lactic acid bacteria are selected for activation, single colony is selected and cultured in 1mL MRS culture medium at 37 ℃ overnight, then inoculated in 10mL MRS culture medium at 37 ℃ for expansion culture for 12 hours, and the optical density value (OD) of the bacterial suspension at 600nm 600 ) Is adjusted to about 1.0 (+ -0.05) as a lactic acid bacteria seed liquid. Then respectively inoculating into 10mL of ethanol stress culture medium and common MRS culture medium (the inoculum size is 5%) and then placing inCulturing in a 37 deg.C incubator for 24 hr, and measuring OD of bacterial suspension by using enzyme-labeled instrument after culturing 600 The results are shown in FIG. 1.
3.3.2 analysis of tolerance to lactic acid bacteria under acid stress
Acid stress culture medium with pH of 4.0 is added into MRS culture medium, and hydrochloric acid solution is added on the basis to prepare acid stress culture medium with pH of 3.6 and 3.3, and high-pressure steam sterilization is carried out, so as to simulate the continuous change of acid stress in the yellow wine fermentation process. After lactic acid bacteria are selected for activation, single colony is selected and cultured in 1mL MRS culture medium at 37 ℃ overnight, then inoculated in 10mL MRS culture medium at 37 ℃ for expansion culture for 12 hours, and the optical density value (OD) of the bacterial suspension at 600nm 600 ) Is adjusted to about 1.0 (+ -0.05) as a lactic acid bacteria seed liquid. Inoculating lactobacillus seed solution into 10mL of acid stress culture medium and common MRS culture medium (inoculum size is 5%), culturing in 37 deg.C incubator for 24 hr, and measuring OD of bacterial suspension by enzyme-labeled instrument 600 The results are shown in FIG. 2.
3.3.3 analysis of sucrose utilization Performance of lactic acid bacteria
Glucose in the MRS medium was replaced with sucrose, sucrose was used as the sole glycogen, and sucrose MRS medium with a sucrose content of 20g/L was prepared. After lactic acid bacteria are selected for activation, single colony is selected and cultured in 1mL MRS culture medium at 37 ℃ overnight, then inoculated in 10mL MRS culture medium at 37 ℃ for expansion culture for 12 hours, and the optical density value (OD) of the bacterial suspension at 600nm 600 ) Is adjusted to about 1.0 (+ -0.05) as a lactic acid bacteria seed liquid. Inoculating seed solution into 10mL sucrose MRS culture medium and common MRS culture medium (with inoculum size of 5%) after high pressure steam sterilization, culturing in 37 deg.C incubator for 24 hr, and measuring OD of bacterial suspension by enzyme-labeled instrument 600 The results are shown in FIG. 3.
Wherein, lactic acid bacteria which have better capability of resisting ethanol stress and acid stress and better capability of utilizing sucrose, namely lactobacillus plantarum (Lactobacillus plantarum) AR1018, are selected and preserved in China general microbiological culture collection center (CGMCC) NO.25626 in 8-31 of 2022.
3.3.4 analysis of antibacterial Properties of lactic acid bacteria
The antibacterial activity of the lactic acid bacteria is determined by an oxford cup method. After lactic acid bacteria are selected for activation, single colony is selected and cultured in 1mL MRS culture medium at 37 ℃ overnight, then inoculated in 10mL MRS culture medium at 37 ℃ for expansion culture for 12 hours, and the optical density value (OD) of the bacterial suspension at 600nm 600 ) Is regulated to about 1.0 (+ -0.05) as lactobacillus seed liquid for standby. 100. Mu.L of an E.coli strain solution cultured overnight at 37℃was plated on a solid LB medium plate. After the belt plates are dried, the sterile oxford cups are gently and uniformly distributed to be inserted into a culture medium for fixation, 200 mu L of lactobacillus seed solution is added into the oxford cups, and the mixture is subjected to forward culture in a 37 ℃ incubator for 36 hours. After the cultivation is finished, the oxford cup is taken down, and the diameter of the inhibition zone is measured by using a digital vernier caliper, and the result is shown in figure 4. The strain AR1018 also shows better antibacterial activity.
3.3.5 Co-cultivation fermentation of lactic acid bacteria and Yeast
After lactic acid bacteria are selected for activation, single colony is selected and cultured in 1mL MRS culture medium at 37 ℃ overnight, then inoculated in 10mL MRS culture medium at 37 ℃ for expansion culture for 12 hours, and the optical density value (OD) of the bacterial suspension at 600nm 600 ) Is adjusted to about 1.0 (+ -0.05) as a lactic acid bacteria seed liquid. Yeast F23 was removed from the tube, spread on YPD solid medium, incubated at 28℃for 2d, and single colony was picked for 3 generations. F23 single colonies were picked up and grown overnight at 28℃in 10mL liquid YPD medium (10 g/L yeast extract, 20g/L plant (soybean) peptone, 20g/L glucose) to give an Optical Density (OD) at 600nm 600 ) Is adjusted to about 1.0 (+ -0.05) as a yeast seed solution. 15mL of yeast seed solution and 7.5mL of lactic acid bacteria seed solution (volume ratio is 2:1) are simultaneously inoculated into a 500mL shaking flask containing 150mL of wort culture medium (6%brix), a negative control of pure yeast culture is arranged, and the bacterial suspension is taken out for qualitative and quantitative analysis on the content of fusel in the fermentation broth by adopting a gas chromatography-mass spectrometer (GC-MS) at 48h (28 ℃ C., 150 rpm) of shaking culture.
3.3.6 analysis of the ability of lactic acid bacteria to reduce the content of fusel by Co-fermentation with Yeast
Sample preparation: 2g of sodium chloride, 5mL of yeast-lactic acid bacteria fermentation broth and 10. Mu.L of 2-octanol (dissolved in ethanol) with a concentration of 200. Mu.g/mL were added to a 20mL headspace sample bottle and capped and sealed.
HS-SPME extraction conditions: the aged SPME extraction head is penetrated into a bottle, the extraction head is exposed outside, extracted for 30min at 50 ℃, taken out and rapidly inserted into a GC sample inlet for sample injection analysis.
GC conditions: the chromatographic column is DB-WAX; column temperature (42 ℃ C. For 3 minutes followed by a rise to 120 ℃ C. At a rate of 6 ℃ C./min, and finally a rise to 230 ℃ C. At 8 ℃ C./min for 10 min); the temperature of the sample inlet is 250 ℃; adopting a non-split mode, wherein the spacer purging rate is 5mL/min; the carrier gas was helium at a flow rate of 1.0mL/min.
MS conditions: EI is used as an ion source, the ionization voltage is 70eV, the temperature of the ion source is 260 ℃, the temperature of a transmission line is 250 ℃, and the acquisition mass number range is 30-400 amu.
The fusel substances are subjected to Xcalibur software carried by an instrument, qualitative determination is completed by comparing mass spectrum libraries such as NIST, wiley and the like, quantitative calculation is completed according to the content of an internal standard, and the result of the fusel content reduction capability of the fermentation of different lactic acid bacteria and the yeast is shown in figure 5.
3.4 preservation and activation of seed
Adding 20% glycerol into the bacterial liquid, mixing, and storing in a refrigerator at-80deg.C for 2-3 years. And (3) taking out the glycerol cryopreservation tube during activation, melting under ice bath conditions, inoculating the bacterial liquid into a prepared MRS culture dish by using a sterile inoculating loop, placing the culture dish into an anaerobic incubator for growth, and growing complete bacterial colonies in the culture dish for about 40 hours, thus carrying out passage.
Example 2 Effect of Lactobacillus plantarum AR1018 on the content of fusel in yellow wine
In the embodiment, lactobacillus plantarum AR1018 is taken as an auxiliary fermenting agent to cooperatively participate in the yellow wine brewing, the influence of residual reducing sugar, amino acid nitrogen, ethanol and the like in the fermentation process on fermentation parameters in the yellow wine brewing process is analyzed, the content of volatile flavor substances in the yellow wine after the fermentation is finished is measured, and the influence of lactobacillus plantarum AR1018 on the content of fusel in the yellow wine is evaluated.
2.1 yellow wine brewing control experiment
(1) Cleaning commercially available semen oryzae Sativae, soaking in water for 24 hr, steaming under normal pressure for 25min until the rice is soft and not rotten, and rapidly taking out without white core and spreading for cooling;
(2) Adding cooled cooked rice into sterilized container, adding 10% (v/v) wheat starter, inoculating 10% (v/v) yeast, standing at 28deg.C, and performing primary fermentation;
(3) After the main fermentation starts for 5 days, the temperature is reduced to 20 ℃ for standing fermentation for 10 days, and stirring is carried out once a day;
(4) After 15 days of main fermentation, filtering and squeezing fermented mash, clarifying the obtained filtrate overnight, blending and decocting to obtain the final product.
The product obtained in this example had an alcohol content of 19.5% vol, a residual reducing sugar content of 3.63g/L, an amino acid nitrogen content of 1.87g/L, a fusel content of 13.31g/L, and an isoamyl alcohol content of 6.94g/L, and had a typical flavor of yellow wine.
2.2 Lactobacillus plantarum AR1018 synergistic yellow wine brewing experiment
(1) Cleaning commercially available semen oryzae Sativae, soaking in water for 24 hr, steaming under normal pressure for 25min until the rice is soft and not rotten, and rapidly taking out without white core and spreading for cooling;
(2) After lactobacillus plantarum AR1018 was activated, single colonies were picked up and cultured overnight at 37℃in 1mL of MRS medium, then inoculated in 10mL of MRS medium and expanded at 37℃for 12 hours, and the Optical Density (OD) of the bacterial suspension at 600nm was measured 600 ) Is regulated to about 1.0 (+ -0.05) as an accessory starter of lactobacillus plantarum AR1018 for standby.
(3) Adding cooled rice into the sterilized container, adding 10% (v/v) wheat starter, inoculating 8% (v/v) yeast and 2% (v/v) lactobacillus plantarum fermentation liquid, standing at 28deg.C, and performing main fermentation;
(4) After the main fermentation starts for 5 days, the temperature is reduced to 20 ℃ for standing fermentation for 10 days, and stirring is carried out once a day;
(5) After 15 days of main fermentation, filtering and squeezing fermented mash, clarifying the obtained filtrate overnight, blending and decocting to obtain the final product.
The alcohol content of the product obtained in the embodiment is 17.3%vol, the content of residual reducing sugar is 4.01g/L, the content of amino acid nitrogen is 2.12g/L, the content of fusel is reduced by 21.35%, the content of isoamyl alcohol is reduced by 26.07%, besides the typical flavor of yellow wine, the mellow flavor is obviously weakened, and the fragrance of flowers and fruits is stronger.
2.3 Lactobacillus plantarum AR1018 synergistic yellow wine brewing experiment
(1) Cleaning commercially available semen oryzae Sativae, soaking in water for 24 hr, steaming under normal pressure for 25min until the rice is soft and not rotten, and rapidly taking out without white core and spreading for cooling;
(2) After lactobacillus plantarum AR1018 was activated, single colonies were picked up and cultured overnight at 37℃in 1mL of MRS medium, then inoculated in 10mL of MRS medium and expanded at 37℃for 12 hours, and the Optical Density (OD) of the bacterial suspension at 600nm was measured 600 ) Is regulated to about 1.0 (+ -0.05) as an accessory starter of lactobacillus plantarum AR1018 for standby.
(3) Adding cooled rice into the sterilized container, adding 10% (v/v) wheat starter, inoculating 9.5% (v/v) yeast, standing at 28deg.C, and performing primary fermentation;
(4) Inoculating lactobacillus plantarum fermentation liquor with the concentration of 0.5% (v/v) into the fermentation liquor within 2-9 days of fermentation, stirring uniformly, and standing for fermentation;
(5) After the main fermentation starts for 5 days, the temperature is reduced to 20 ℃ for standing fermentation for 10 days, and stirring is carried out once a day;
(6) After 15 days of main fermentation, filtering and squeezing fermented mash, clarifying the obtained filtrate overnight, blending and decocting to obtain the final product.
The product obtained in this example had an alcohol content of 20.1% vol, a residual reducing sugar content of 1.52g/L, an amino acid nitrogen content of 2.38g/L, a reduction in the amount of fusel alcohol of 8.72%, and an isoamyl alcohol content of 18.48%, with the exception of the typical flavor of yellow wine, which was slightly reduced in body odor.
The previous description of the embodiments is provided to facilitate a person of ordinary skill in the art in order to make and use the present invention. It will be apparent to those skilled in the art that various modifications can be readily made to these embodiments and the generic principles described herein may be applied to other embodiments without the use of the inventive faculty. Therefore, the present invention is not limited to the above-described embodiments, and those skilled in the art, based on the present disclosure, should make improvements and modifications without departing from the scope of the present invention.
Claims (10)
1. The lactobacillus plantarum for reducing the content of fusel in yellow wine is characterized in that the lactobacillus plantarum is marked as AR1018 strain and is preserved in China general microbiological culture collection center (CGMCC) No.25626 in the 8 th month 31 of 2022.
2. The lactobacillus plantarum for reducing the content of fusel in yellow wine according to claim 1, wherein the lactobacillus plantarum has the capability of resisting ethanol stress and acid stress, has the capability of utilizing sucrose and also has antibacterial activity.
3. The use of the lactobacillus plantarum for reducing the content of the fusel in the yellow wine according to claim 1 or 2, which is characterized in that the lactobacillus plantarum for reducing the content of the fusel in the yellow wine is used as an auxiliary fermenting agent for brewing in the process of brewing the yellow wine.
4. The use of lactobacillus plantarum for reducing the content of fusel in yellow wine according to claim 3, wherein the lactobacillus plantarum for reducing the content of fusel in yellow wine is used for reducing the content of fusel in yellow wine when being used as an auxiliary starter in the brewing process of yellow wine.
5. The use of lactobacillus plantarum for reducing the content of fusel in yellow wine according to claim 4, wherein the lactobacillus plantarum for reducing the content of fusel in yellow wine is used for reducing the content of isoamyl alcohol in yellow wine when being used as an auxiliary starter in the brewing process of yellow wine.
6. Use of lactobacillus plantarum for reducing the content of fusel in yellow wine according to claim 3, wherein the lactobacillus plantarum for reducing the content of fusel in yellow wine is added in an inoculation amount of 0.5% -2.0% as an auxiliary starter in the process of brewing yellow wine, and is fermented together with yeast.
7. The use of lactobacillus plantarum for reducing the content of fusel in yellow wine according to claim 6, wherein the process of pre-fermentation at 28 ℃, post-fermentation at 20 ℃, squeezing, clarifying and decocting is adopted in the process of brewing yellow wine.
8. The use of lactobacillus plantarum for reducing the content of fusel in yellow wine according to claim 3, wherein the specific process conditions of the lactobacillus plantarum for reducing the content of fusel in yellow wine as an auxiliary starter in the brewing process of yellow wine are as follows:
(1) Adding 10% (v/v) wheat starter into cooled rice, inoculating 9% (v/v) yeast, standing at 28deg.C, and performing main fermentation;
(2) Inoculating 1% (v/v) lactobacillus plantarum fermentation liquor into the fermentation liquor within 2-9 days of fermentation, stirring uniformly, and standing for fermentation;
(3) After the main fermentation starts for 5 days, the temperature is reduced to 20 ℃ for standing fermentation for 10 days, and stirring is carried out once a day;
(4) After 15 days of main fermentation, filtering and squeezing fermented mash, clarifying the obtained filtrate overnight, blending and decocting to obtain yellow wine.
9. The method for preserving the lactobacillus plantarum for reducing the content of the fusel in the yellow wine according to claim 1 or 2, which is characterized in that glycerin with the final concentration of 20% is added into the bacterial liquid of the lactobacillus plantarum for reducing the content of the fusel in the yellow wine, and the lactobacillus plantarum can be preserved for 2-3 years after being uniformly mixed in a refrigerator with the temperature of-80 ℃.
10. The method for activating the lactobacillus plantarum for reducing the content of the fusel in the yellow wine, which is characterized in that a glycerol cryopreservation tube is taken out during activation, melted under ice bath conditions, bacterial liquid is inoculated into a prepared MRS culture dish by using a sterile inoculating loop, the culture dish is placed into an anaerobic incubator for growth, and after complete bacterial colonies grow in the culture dish, the culture dish can be passaged.
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