CN104673703A - Bacillus capable of simultaneously promoting saccharomyces cerevisiae to produce ethyl alcohol and flavor substances and application of bacillus - Google Patents
Bacillus capable of simultaneously promoting saccharomyces cerevisiae to produce ethyl alcohol and flavor substances and application of bacillus Download PDFInfo
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Abstract
The invention relates to bacillus capable of simultaneously promoting saccharomyces cerevisiae to produce ethyl alcohol and flavor substances, and an application of the bacillus, and belongs to the technical field of bioengineering. The bacillus provided by the invention comprises bacillus licheniformis CGMCC NO.3963, bacillus cereus CCTCC NO:M 2013369, bacillus lentus CCTCC NO:M 2013370 and bacillus methylotrophicus CCTCC NO: 2013371. After the bacillus and the saccharomyces cerevisiae are subjected to mixed culture, the yield of ethyl alcohol can be improved by over 20%; meanwhile, metabolism aldehydes, alcohols, esters, acids, and other flavor substances of the saccharomyces cerevisiae are improved; the bacillus is applied to production of fermented wine; the wine yield can be improved; and meanwhile, the flavor and the quality of the fermented wine are significantly improved.
Description
Technical field
The present invention relates to and can promote that yeast saccharomyces cerevisiae produces genus bacillus and the application thereof of alcohol and flavour substances simultaneously, belong to technical field of bioengineering.
Background technology
Brewing wine industry is the advantage national tradition production of China, is also the important component part of foodstuffs industry.Total output value in 2013 reaches more than 6,000 hundred million yuan, accounts for foodstuffs industry total value and is about 6-7%.Therefore the development of brewing wine industry all has great importance for China's food service industry even development of national economy.
China white wine is the Typical Representative of brewing wine.But compared with international brewing wine manufacturing, there are the following problems: first, and production efficiency is on the low side, and namely yeast of white wine is low.In the solid-state wine unstrained spirits of white wine, ethanol maximum concentration is only about 7%, and the highest liquor ratio of raw material is 50% (little Qu Qingxiang) only, and minimum only about 35% (giving off a strong fragrance), far below theoretical the yield of liquor (65%); Secondly, liquor flavor metabolism is uncontrollable, and important flavour substances anabolism is more weak, causes quality percentage on the low side, and the annual high quality liquor produced only accounts for 5 ~ 10% of national white wine output at present.Such as, quality percentage only 6% (the representative inside data of enterprise) of fen-flavor type white spirit, cannot meet the consumers demand of the people to quality products.Therefore, the inevitable requirement that the yield of liquor of white wine and quality percentage are liquor industry Sustainable developments is improved.
Yeast saccharomyces cerevisiae is the important microbe of liquor production, provide not only alcohol, and provides important flavour substances, and the alcohol metabolism and the local flavor metabolism that therefore improve yeast saccharomyces cerevisiae are the important contents that liquor fermentation needs regulation and control.But because liquor production belongs to open type spontaneous fermentation, Wine brewing yeast strain and metabolism thereof are all difficult to control, although have employed certain method in liquor production, all produce little effect.Such as, the yeast that metabolic activity is stronger is added in liquor production, but it is high to meet alcohol vigor, the yeast that can meet local flavor metabolic activity high is again less, and a lot of yeast is all difficult to the severe environment adapting to liquor fermentation, and, the yeast saccharomyces cerevisiae of strengthening does not quantitatively have advantage, the yeast saccharomyces cerevisiae in native country can not be replaced, therefore, directly strengthen the mode effect of high-quality yeast saccharomyces cerevisiae and not obvious.Therefore, if can the metabolism of yeast saccharomyces cerevisiae be a kind of more efficiently approach in direct regulation and control actual production environment.But be different from controlling fermentation, the spontaneous fermentation of liquor production determines the difficulty of its metabolic regulation.In addition, in current liquor production, the yield of liquor and local flavor promote and are difficult to can meet simultaneously, and when general the yield of liquor improves, local flavor then can decline, and when local flavor improves, the yield of liquor then can decline.Except above difficulty, be also difficult to improve yeast-alcohol and local flavor metabolism simultaneously.
Present invention finds 4 bacillus, can the alcohol metabolism of Effective Regulation yeast saccharomyces cerevisiae and local flavor metabolic capacity, because above-mentioned bacterial strains comes from brewed spirit, therefore brewed spirit environment is suitable for, be applied in liquor production, simultaneously effectively to improve alcohol and the local flavor metabolic capacity of yeast saccharomyces cerevisiae in production system, finally can improve the yield of liquor and the quality of white wine simultaneously, solve the difficulty that the yield of liquor that exists in current liquor production and flavor quality are difficult to improve simultaneously.Meet the requirement of liquor industry Sustainable development.Meanwhile, the present invention can be applied to other fermentation food field, for promoting that the efficient of fermentation food and fine quality production have important practice significance.
Summary of the invention
The invention provides the genus bacillus of improving yeast saccharomyces cerevisiae metabolism alcohol and flavour substances, and be applied in liquor production the method improving yeast of white wine and quality.
First object of the present invention is to provide 4 strains can promote that yeast saccharomyces cerevisiae produces the genus bacillus of alcohol and flavour substances simultaneously.
Described genus bacillus is bacillus licheniformis Bacillus licheniformis CGMCC NO.3963; Bacillus cereus Bacillus cereus BC-1, i.e. B.cereus CCTCC NO:M 2013369; Bacillus lentus Bacillus lentus BL-1, i.e. B.lentus CCTCC NO:M 2013370; Methylotrophic bacillus methylotrophicus BM-1, i.e. B.methylotrophicusCCTCC NO:M 2013371.
Described bacillus licheniformis B.licheniformis CGMCC NO.3963, be preserved in China General Microbiological culture presevation administrative center on June 28th, 2010, deposit number is CGMCC NO.3963; Described bacillus cereus B.cereus CCTCC NO:M 2013369, be preserved in China typical culture collection center on August 8th, 2013, deposit number is CCTCC NO:M 2013369; Described bacillus lentus B.lentus CCTCC NO:M 2013370, be preserved in China typical culture collection center on August 8th, 2013, deposit number is CCTCC NO:M 2013370; Described Methylotrophic genus bacillus B.methylotrophicus CCTCC NO:M 2013371, be preserved in China typical culture collection center on August 8th, 2013, deposit number is CCTCC NO:M 2013371.
Described flavour substances comprises: the acetaldehyde of aldehydes, 2 methyl propanal, the 3-tertiary butyl-4-methoxyphenol, the 2-methylpropanol of alcohols, 3-methyl butanol, phenyl aldehyde, phenylacetic aldehyde, Phenylacetic acid ethylester, Phenylethyl ethanoate, caproic acid phenethyl ester, the phenylethyl isobutyrate of the fragrance same clan, 2 of ketone and furans, 3-Dihydrobenzofuranes, acids 2 Methylpropionic acid, caproic acid, sad, capric acid, and farnesol, β-geraniol.
Described bacillus licheniformis B.licheniformis CGMCC NO.3963, is characterized in that, by itself and yeast saccharomyces cerevisiae mixed fermentation, can improve yeast saccharomyces cerevisiae alcohol metabolism ability; Increase the ability of yeast saccharomyces cerevisiae metabolism flavour substances simultaneously, comprise acetaldehyde, the 3-tertiary butyl-4-methoxyphenol, 3-methyl butanol, phenyl aldehyde, phenylacetic aldehyde, phenylethyl alcohol, Phenylethyl ethanoate, caproic acid phenethyl ester, phenylethyl isobutyrate, 2,3-Dihydrobenzofuranes, 2 Methylpropionic acid, caproic acid, sad, capric acid, trans-nerolidol, farnesol, Tetramethylpyrazine.
Described bacillus cereus B.cereus CCTCC NO:M 2013369, is characterized in that, by itself and yeast saccharomyces cerevisiae mixed fermentation, can improve yeast saccharomyces cerevisiae alcohol metabolism ability; Increase the ability of yeast saccharomyces cerevisiae metabolism flavour substances simultaneously, comprise acetaldehyde, 2 methyl propanal, the 3-tertiary butyl-4-methoxyphenol, 2,4-DI-tert-butylphenol compounds, 2-methylpropanol, 3-methyl butanol, phenyl aldehyde, phenylacetic aldehyde, phenylethyl alcohol, Phenylacetic acid ethylester, Phenylethyl ethanoate, caproic acid phenethyl ester, phenylethyl isobutyrate, ethyl octylate, ethyl decylate, 3-hydroxy-2-butanone, 2,3-Dihydrobenzofuranes, 2 Methylpropionic acid, caproic acid, sad, capric acid, trans-nerolidol, farnesol, β-geraniol, dorinone.
Described bacillus lentus B.lentus CCTCC NO:M 2013370, is characterized in that, by itself and yeast saccharomyces cerevisiae mixed fermentation, can improve yeast saccharomyces cerevisiae alcohol metabolism ability; Increase the ability of yeast saccharomyces cerevisiae metabolism flavour substances simultaneously, comprise acetaldehyde, methyl catechol, 4-vinyl guaiacol, the 3-tertiary butyl-4-methoxyphenol, 2,4-DI-tert-butylphenol compounds, 2-methylpropanol, 3-methyl butanol, phenyl aldehyde, phenylethyl alcohol, Phenylacetic acid ethylester, Phenylethyl ethanoate, caproic acid phenethyl ester, phenylethyl isobutyrate, ethyl decylate, 3-hydroxy-2-butanone, 2,3-Dihydrobenzofuranes, 2 Methylpropionic acid, caproic acid, sad, capric acid, trans-nerolidol, farnesol, β-geraniol.
Described Methylotrophic genus bacillus B.Methylotrophicus CCTCC NO:M 2013371, is characterized in that, by itself and yeast saccharomyces cerevisiae mixed fermentation, can improve yeast saccharomyces cerevisiae alcohol metabolism ability; Increase the ability of yeast saccharomyces cerevisiae metabolism flavour substances simultaneously, comprise acetaldehyde, methyl catechol, 4-vinyl guaiacol, 2-methylpropanol, 2-methyl-4-amylene-1-ol, phenyl aldehyde, phenylacetic aldehyde, Phenylacetic acid ethylester, Phenylethyl ethanoate, phenylethyl isobutyrate, ethyl octylate, ethyl decylate, 3-hydroxy-2-butanone, 2,3-Dihydrobenzofuranes, 2 Methylpropionic acid, caproic acid, sad, capric acid, farnesol, dorinone.
4 described bacillus all have following microbial characteristic: Gram-positive, amphimicrobian, hydrolyzable casein, gelatin and starch; Can directly grow in Chinese sorghum solid medium, its feature is not need to carry out saccharification process to Chinese sorghum solid medium, directly can utilize sorghum starch and Growth and metabolism; Can tolerate 60 DEG C of high growth temperatures, NaCl and KCl of tolerance 15 ~ 18%, can grow in pH3.8 ~ 4, can tolerate the ethanol of 10 ~ 12%.Above feature can survive in brewed spirit complex environment, and forms promoter action to yeast saccharomyces cerevisiae metabolism.Meanwhile, due to this bacterium for a long time in brewed spirit environment with yeast saccharomyces cerevisiae mutual survival, evolution can with the mutual effect of producing Yeast looks, and promote the Growth and metabolism of yeast saccharomyces cerevisiae.
Second object of the present invention is to provide a kind of method simultaneously promoting yeast saccharomyces cerevisiae product alcohol and flavour substances, is that yeast saccharomyces cerevisiae and following at least one genus bacillus are carried out mixed culture: bacillus licheniformis B.licheniformis CGMCC NO.3963, bacillus cereus B.cereus CCTCC NO:M 2013369, bacillus lentus B.lentus CCTCC NO:M 2013370, Methylotrophic genus bacillus B.methylotrophicus CCTCC NO:M 2013371.
Described yeast saccharomyces cerevisiae is any one yeast saccharomyces cerevisiae.
Described method is specifically: by yeast saccharomyces cerevisiae and genus bacillus seed liquor, is that 0.001:1000 to 1000:0.001 is inoculated in substratum according to thalline number ratio after inoculation, in 25 DEG C to 40 DEG C, 0rpm to 200rpm cultivates 16h to 72h.
Described method, to substratum no requirement (NR).In one embodiment of the invention, described substratum is Chinese sorghum liquid nutrient medium.
3rd object of the present invention is to provide the application of described genus bacillus in wine brewing.
Described application is applied to by genus bacillus during brewing wine is produced to improve brewing wine the yield of liquor and quality.
Described application is by genus bacillus liquid microbial inoculum or solid fungicide, and after mixing with wine brewing distiller's yeast kind or fermentation raw material, ferment, the yield of liquor and the quality of brewing wine all significantly improve.
The strengthening application that described application is described genus bacillus in one embodiment of the invention in brewing wine is produced, adopts the mode of liquid bacterial agent or solid fungicide to be inoculated in production system, improves the yield of liquor and product special flavour substance classes and content.
Described liquid bacterial agent production method, in one embodiment of the invention, be that the seed liquor of activation is inoculated in the liquid culture medium of sterilizing by the inoculum size (i.e. every 100g liquid culture medium inoculation seed liquor 1-5g) of 1 ~ 5% (w/w), cultivate 24h ~ 72h for 25 ~ 40 DEG C.
Described liquid culture medium, in one embodiment of the invention, for Chinese sorghum liquid nutrient medium, making method is: 20 ~ 200g Chinese sorghum sample after crushed, adds the water of mL/g meter 1 ~ 4 times, boiling 1-5h, in the pasty state, after cooling, add the saccharifying enzyme of 10 ~ 50 units/g, keep 2 ~ 10h in 40 ~ 100 DEG C, filtration, centrifugal gained filtrate, pol is 10 ~ 15
obx.
The production method of described solid fungicide, in one embodiment of the invention, by in the seed liquor of activation inoculation (first order seed) Chinese sorghum solid medium, (inoculum size is 1 ~ 5%, w/w), solid state fermentation 32 ~ 72h in 25 ~ 40 DEG C of environment, makes solid-state bacteria preparation.Chinese sorghum solid medium production method: Chinese sorghum is pulverized, after adding water in the ratio of 1:1 ~ 1:2, boiling 30-60min.
Genus bacillus of the present invention is brewageed screening environment from Chinese Maotai-flavor liquor and obtains, because Maotai-flavor liquor brewages features such as having high temperature, peracid, high alcohol concn, impart above genus bacillus and be different from conventional genus bacillus, there is the feature of unique withstand high temperatures, peracid and high alcohol concn, can adapt to brewing wine severe brewage environment, and promote the metabolism of yeast saccharomyces cerevisiae.Simultaneously, in mixed fermentation, the raising of yeast flavor metabolism is not by the material of genus bacillus own metabolism, first after mixed fermentation, the growth of genus bacillus receives certain suppression, the metabolic substd overwhelming majority that yeast improves simultaneously is not that genus bacillus institute energy metabolism produces, therefore, described genus bacillus mixes the metabolic capacity that can improve yeast saccharomyces cerevisiae with yeast.
Present invention finds 4 bacillus, effectively can improve alcohol and the local flavor metabolic capacity of yeast saccharomyces cerevisiae in production system simultaneously, final the yield of liquor and the quality simultaneously improving white wine, solve the difficulty that the yield of liquor that exists in current liquor production and flavor quality are difficult to improve simultaneously, meet the requirement of liquor industry Sustainable development.
Biomaterial preservation
Bacillus licheniformis B.licheniformis CGMCC NO.3963, be preserved in China General Microbiological culture presevation administrative center on June 28th, 2010, deposit number is CGMCC NO.3963;
Bacillus cereus Bacillus cereus BC-1, be preserved in China typical culture collection center on August 8th, 2013, preservation address is Wuhan, China Wuhan University, and deposit number is CCTCC NO:M 2013369;
Bacillus lentus Bacillus lentus BL-1, be preserved in China typical culture collection center on August 8th, 2013, preservation address is Wuhan, China Wuhan University, and deposit number is CCTCC NO:M 2013370;
Methylotrophic bacillus methylotrophicus BM-1, be preserved in China typical culture collection center on August 8th, 2013, preservation address is Wuhan, China Wuhan University, and deposit number is CCTCC NO:M 2013371.
Accompanying drawing explanation
Fig. 1: the growth of yeast saccharomyces cerevisiae after single culture and mixed culture, A, yeast saccharomyces cerevisiae,, singly cultivates; ●, add bacillus licheniformis; △, adds bacillus cereus; ▼, adds bacillus lentus; ◇, adds Methylotrophic genus bacillus.
Embodiment
Flavour substances detects: use headspace solid-phase microextraction technology (HS-SPME) and gas chromatography-mass spectrum (GC-MS) method to analyze volatile products, get 8mL sample, put into the headspace sampling bottle that 3g NaCl is housed, adding 10 μ L concentration is 42.60mgL
-14-methyl-2-amylalcohol be interior mark.By ml headspace bottle in 50 DEG C of constant temperature extracting 45min.GC-MS analysis is carried out after having extracted.
The mixed culture of embodiment 1 yeast saccharomyces cerevisiae and genus bacillus
Yeast saccharomyces cerevisiae seed liquor: picking one ring S. cervisiae mud is inoculated in YEPD liquid nutrient medium, 30 DEG C, and 200rpm cultivates 16h.Genus bacillus seed liquor: picking one ring genus bacillus bacterium mud is inoculated in LB liquid nutrient medium, 37 DEG C, and 200rpm cultivates 16h.Preparation Chinese sorghum diffusion juice, inoculation yeast saccharomyces cerevisiae seed liquor and genus bacillus seed liquor, both are 1 × 10 by final concentration
6cfu/mL.At 30 DEG C, 200rpm cultivates.Sample the method be coated with by flat board and measure cell concentration; Cultivate 24 ~ 48h, bacterium liquid, in the centrifugal 10min of 8000rpm, measures alcohol concn and flavour substances composition and content.Yeast saccharomyces cerevisiae cell concentration over time curve is shown in Fig. 1, and alcohol concn change is in table 1, and flavor compound changes in table 2 to table 3.
By measuring genus bacillus and saccharomycetic cell concentration in fermenting process, find that the growth of genus bacillus can reduce, the growth of yeast saccharomyces cerevisiae changes not quite compared with single culture.
As can be seen from Table 1, after genus bacillus mixed culture, the producing and ethanol ability of yeast saccharomyces cerevisiae improves 17%-40.2%.
As can be seen from table 2 to table 3, after genus bacillus and yeast mixed culture, what flavour substances content was all improved comprises: described flavour substances comprises: the acetaldehyde of aldehydes, 2 methyl propanal, the 3-tertiary butyl-4-methoxyphenol, the 2-methylpropanol of alcohols, 3-methyl butanol, phenyl aldehyde, phenylacetic aldehyde, Phenylacetic acid ethylester, Phenylethyl ethanoate, caproic acid phenethyl ester, the phenylethyl isobutyrate of the fragrance same clan, 2 of ketone and furans, 3-Dihydrobenzofuranes, acids 2 Methylpropionic acid, caproic acid, sad, capric acid, and farnesol, β-geraniol.
Experiment prove, genus bacillus be do not produce or pole low yield alcohol, acid, ester.Due to genus bacillus growth reduction, yeast growth during mixed culture, and the kind of flavour substances and concentration thereof can improve, indicate the metabolic capacity that not genus bacillus self has high-yield ethanol and important alcohol, acid, ester class flavour substances, but genus bacillus facilitates the metabolic capacity of yeast saccharomyces cerevisiae.
Table 1 adds different genus bacillus yeast saccharomyces cerevisiae and produces alcohol concn (g/l)
Table 2 adds the change (μ g/L) of aldehydes, alcohols and aromatic series flavour substances after different genus bacillus
Table 3 adds the change (μ g/L) of ester class, acids and other flavour substancess after different genus bacillus
Embodiment 2 genus bacillus is applied to during brewing wine is produced improves brewing wine the yield of liquor and quality
Genus bacillus CCTCC NO:M 2013369 and CCTCC NO:M 2013370 of the present invention is made liquid bacterial agent or solid fungicide respectively, ferments after mixing with wine brewing distiller's yeast kind or fermentation raw material.
Described liquid bacterial agent production method: the seed liquor of activation is inoculated in the liquid culture medium of sterilizing by the inoculum size (i.e. every 100g liquid culture medium inoculation seed liquor 1-5g) of 1 ~ 5% (w/w), cultivates 24h ~ 72h for 25 ~ 40 DEG C.
Described liquid culture medium is Chinese sorghum extract substratum, making method is: 20 ~ 200g Chinese sorghum sample after crushed, add the water of mL/g meter 1 ~ 4 times, boiling 1-5h, in the pasty state, after cooling, add the saccharifying enzyme of 10 ~ 50 units/g, keep 2 ~ 10h in 40 ~ 100 DEG C, filtration, centrifugal gained filtrate, pol is 10 ~ 15 ° of Bx.
The production method of described solid fungicide is that solid state fermentation 32 ~ 72h in 25 ~ 40 DEG C of environment, makes solid-state bacteria preparation by the seed liquor of activation inoculation (first order seed) Chinese sorghum solid medium (inoculum size is 1 ~ 5%, w/w).
Chinese sorghum solid medium production method: Chinese sorghum is pulverized, after adding water in the ratio of 1:1 ~ 1:2, boiling 30-60min.
In Maotai-flavor liquor is produced, apply this two kinds of genus bacillus respectively, carry out sensory evaluation to the white wine that application is produced, result is as shown in table 4, can find out, after application genus bacillus, quality of white spirit is improved.Wherein, according to genus bacillus and yeast flavor metabolic characteristics, the ability of the acid of genus bacillus product, alcohol, ester is very low, and the sweet-smelling improved in white wine, sugariness, sour are mainly alcohols thing and the acid contribution of yeast metabolism, puts the Ester contribution that fragrant intensity is mainly yeast metabolism generation.Result shows, in white wine the important local flavor such as acid, alcohol, ester be have genus bacillus to promote yeast saccharomyces cerevisiae metabolism produces.
Table 4 key application bacteria combination produces Liquor Tasting result
Comment wine personnel: national white wine judging panel 1; Provincial white wine judging panel 5; Brewery judging panel 2.
Point system: to make number one weighting 3 points by sequence, second weighting 2 points, the 3rd weighting 1 point.
1 ~ 4 round going out wine of producing at Maotai-flavor liquor has carried out the application of genus bacillus, produce white wine quality and to be all improved and the yield of liquor is improved, result is as shown in table 5.
The yield of liquor (%) that genus bacillus bacterium produces white wine applied by table 5
The mixed culture of embodiment 3 yeast saccharomyces cerevisiae and genus bacillus
Yeast saccharomyces cerevisiae and B.licheniformis CGMCC NO.3963, B.lentus CCTCC NO:M 2013370 are made seed liquor respectively.After mixing in the ratio of bacterium number 1:1 in B.licheniformis CGMCC3963, B.lentus CCTCC NO:M 2013370, be inoculated in together with yeast saccharomyces cerevisiae in YPD substratum, after making inoculation, the bacterium number of yeast and genus bacillus is respectively 10
4individual/mL, 10
10individual/mL, with 30 DEG C of quiescent culture 72h.Found that, compared with single culture, the substances content such as acetaldehyde, 2 methyl propanal, the 3-tertiary butyl-4-methoxyphenol, 2-methylpropanol, 3-methyl butanol, phenyl aldehyde, phenylacetic aldehyde, Phenylacetic acid ethylester, Phenylethyl ethanoate, caproic acid phenethyl ester, phenylethyl isobutyrate, 2,3-Dihydrobenzofuranes, 2 Methylpropionic acid, caproic acid, sad, capric acid, farnesol, β-geraniol has rising to a certain degree.
The mixed culture of embodiment 4 yeast saccharomyces cerevisiae and genus bacillus
Yeast saccharomyces cerevisiae and B.cereus CCTCC NO:M 2013369, B.lentus CCTCC NO:M 2013370, B.methylotrophicus CCTCC NO:M 2013371 are made seed liquor respectively.After mixing in the ratio of bacterium number 1:2:2 in B.cereus CCTCC NO:M 2013369, B.lentus CCTCC NO:M 2013370, B.methylotrophicus CCTCC NO:M 2013371, be inoculated in together with yeast saccharomyces cerevisiae in substratum, after making inoculation, the bacterium number of yeast and genus bacillus is respectively 10
8individual/mL, 10
3individual/mL, cultivates 16h with 40 DEG C of 100rpm.Found that, compared with single culture, the substances content such as acetaldehyde, 2 methyl propanal, the 3-tertiary butyl-4-methoxyphenol, 2-methylpropanol, 3-methyl butanol, phenyl aldehyde, phenylacetic aldehyde, Phenylacetic acid ethylester, Phenylethyl ethanoate, caproic acid phenethyl ester, phenylethyl isobutyrate, 2,3-Dihydrobenzofuranes, 2 Methylpropionic acid, caproic acid, sad, capric acid, farnesol, β-geraniol has rising to a certain degree.Meanwhile, the liquor output rate of yeast also improves 22.3%.
Although the present invention with preferred embodiment openly as above; but it is also not used to limit the present invention, any person skilled in the art, without departing from the spirit and scope of the present invention; all can do various changes and modification, what therefore protection scope of the present invention should define with claims is as the criterion.
Claims (10)
1. can promote that yeast saccharomyces cerevisiae produces the genus bacillus of alcohol and flavour substances simultaneously, it is characterized in that, described genus bacillus be following any one: (1) bacillus cereus Bacillus cereus CCTCC NO:M 2013369, be preserved in China typical culture collection center on August 8th, 2013, deposit number is CCTCC NO:M 2013369; (2) bacillus lentus Bacillus lentus CCTCC NO:M 2013370, be preserved in China typical culture collection center on August 8th, 2013, deposit number is CCTCC NO:M 2013370; (3) Methylotrophic bacillus methylotrophicus CCTCC NO:M 2013371, be preserved in China typical culture collection center on August 8th, 2013, deposit number is CCTCC NO:M 2013371.
2. genus bacillus according to claim 1, it is characterized in that, described flavour substances comprises: the acetaldehyde of aldehydes, 2 methyl propanal, the 3-tertiary butyl-4-methoxyphenol, the 2-methylpropanol of alcohols, 3-methyl butanol, phenyl aldehyde, phenylacetic aldehyde, Phenylacetic acid ethylester, Phenylethyl ethanoate, caproic acid phenethyl ester, the phenylethyl isobutyrate of the fragrance same clan, 2,3-Dihydrobenzofuranes of ketone and furans, acids 2 Methylpropionic acid, caproic acid, sad, capric acid, and farnesol, β-geraniol.
3. genus bacillus according to claim 1, is characterized in that, when described genus bacillus and yeast saccharomyces cerevisiae mixed fermentation, can improve the ability that yeast saccharomyces cerevisiae produces alcohol ability and increases yeast saccharomyces cerevisiae metabolism flavour substances; The flavour substances that described bacillus cereus CCTCC NO:M 2013369 can improve comprises acetaldehyde, 2 methyl propanal, the 3-tertiary butyl-4-methoxyphenol, 2,4-DI-tert-butylphenol compounds, 2-methylpropanol, 3-methyl butanol, phenyl aldehyde, phenylacetic aldehyde, phenylethyl alcohol, Phenylacetic acid ethylester, Phenylethyl ethanoate, caproic acid phenethyl ester, phenylethyl isobutyrate, ethyl octylate, ethyl decylate, 3-hydroxy-2-butanone, 2,3-Dihydrobenzofuranes, 2 Methylpropionic acid, caproic acid, sad, capric acid, trans-nerolidol, farnesol, β-geraniol, dorinone; The flavour substances that described bacillus lentus CCTCC NO:M 2013370 can improve comprises acetaldehyde, methyl catechol, 4-vinyl guaiacol, the 3-tertiary butyl-4-methoxyphenol, 2,4-DI-tert-butylphenol compounds, 2-methylpropanol, 3-methyl butanol, phenyl aldehyde, phenylethyl alcohol, Phenylacetic acid ethylester, Phenylethyl ethanoate, caproic acid phenethyl ester, phenylethyl isobutyrate, ethyl decylate, 3-hydroxy-2-butanone, 2,3-Dihydrobenzofuranes, 2 Methylpropionic acid, caproic acid, sad, capric acid, trans-nerolidol, farnesol, β-geraniol; The flavour substances that described Methylotrophic genus bacillus CCTCC NO:M 2013371 can improve comprises acetaldehyde, methyl catechol, 4-vinyl guaiacol, 2-methylpropanol, 2-methyl-4-amylene-1-ol, phenyl aldehyde, phenylacetic aldehyde, Phenylacetic acid ethylester, Phenylethyl ethanoate, phenylethyl isobutyrate, ethyl octylate, ethyl decylate, 3-hydroxy-2-butanone, 2,3-Dihydrobenzofuranes, 2 Methylpropionic acid, caproic acid, sad, capric acid, farnesol, dorinone.
4. one kind promotes that yeast saccharomyces cerevisiae produces the method for alcohol and flavour substances simultaneously, it is characterized in that, be that yeast saccharomyces cerevisiae and following any one or multiple genus bacillus are carried out mixed culture: B.licheniformis CGMCC NO.3963, B.cereus CCTCC NO:M 2013369, B.lentus CCTCC NO:M 2013370, B.methylotrophicus CCTCC NO:M2013371.
5. method according to claim 4, it is characterized in that, described method is specifically: by yeast saccharomyces cerevisiae and genus bacillus seed liquor, be that 0.001:1000 to 1000:0.001 is inoculated in substratum according to thalline number ratio after inoculation, in 25 DEG C to 40 DEG C, 0rpm to 200rpm cultivates 16h to 72h.
6. method according to claim 5, is characterized in that, described substratum is Chinese sorghum liquid nutrient medium.
7. the application of genus bacillus described in claim 1 in food.
8. the application of genus bacillus described in claim 1 in wine brewing.
9. application according to claim 8, is characterized in that, described application is applied to by genus bacillus during brewing wine is produced to improve brewing wine the yield of liquor and quality; Described application is by genus bacillus liquid microbial inoculum or solid fungicide, after mixing, ferments with wine brewing distiller's yeast kind or fermentation raw material.
10. application according to claim 8 or claim 9, it is characterized in that, described liquid bacterial agent production method: seed liquor is inoculated in the liquid culture medium of sterilizing by the inoculum size of every 100g liquid culture medium inoculation seed liquor 1-5g, cultivate 24h ~ 72h for 25 ~ 40 DEG C, the making method of described liquid culture medium is: 20 ~ 200g Chinese sorghum after crushed, add the water of 1 ~ 4 times of quality, boiling 1-5h, in the pasty state, add the saccharifying enzyme of 10 ~ 50U by every g Chinese sorghum after cooling, keep 2 ~ 10h in 40 ~ 100 DEG C, filtration, centrifugal gained filtrate, pol is 10 ~ 15 ° of Bx; The production method of described solid fungicide: by the inoculum size inoculation Chinese sorghum liquid nutrient medium of seed liquor by every 100g solid medium inoculation seed liquor 1-5g, solid state fermentation 32 ~ 72h in 25 ~ 40 DEG C of environment, makes solid-state microbial inoculum.
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| CN201410747485.0A CN104673703B (en) | 2014-12-09 | 2014-12-09 | Bacillus and its application of saccharomyces cerevisiae production alcohol and flavor substance can be promoted simultaneously |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105838751A (en) * | 2016-03-17 | 2016-08-10 | 齐鲁工业大学 | Method for improving output of tetramethylpyrazine produced through fermentation by Bacillus |
| CN106929438A (en) * | 2016-11-18 | 2017-07-07 | 天津科技大学 | One plant height produces the saccharomyces cerevisiae and its construction method of Tetramethylpyrazine |
| CN110923163A (en) * | 2019-11-22 | 2020-03-27 | 郑州轻工业大学 | Bacillus methylotrophicus and application thereof |
| CN113151245A (en) * | 2021-05-28 | 2021-07-23 | 湖北工业大学 | Flavor compound microbial inoculum for reducing rice wine fusel oil and application thereof |
| CN117413924A (en) * | 2023-11-02 | 2024-01-19 | 北京衍微科技有限公司 | Composition with antibacterial and fermentation promoting effects and application thereof |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109988720B (en) * | 2019-01-28 | 2020-09-04 | 佛山市海天(高明)调味食品有限公司 | Yeast ZB412 and application thereof |
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| CN111235067B (en) * | 2020-03-16 | 2021-10-01 | 河南仰韶酒业有限公司 | White spirit enhanced yeast for making hard liquor, preparation method and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102268397A (en) * | 2011-07-25 | 2011-12-07 | 江南大学 | Bacillus licheniformis capable of highly producing specific flavor, and application thereof in Chinese liquor brewing |
| CN102766576A (en) * | 2012-07-19 | 2012-11-07 | 江南大学 | Brewing function oriented microbe combination method and application of combination bacterium in liquor-making industry |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101503654A (en) * | 2009-01-21 | 2009-08-12 | 江苏双沟酒业股份有限公司 | Aromatic Qu liquor pit mud and preparation thereof |
| CN102140427B (en) * | 2010-09-29 | 2012-10-03 | 湖北白云边酒业股份有限公司 | Method for preparing intensified daqu applied to nongjiang-flavor Chinese spirits |
| CN102277326B (en) * | 2011-08-13 | 2012-11-21 | 天津工业生物技术研究所 | Microbial mixed colony capable of enhancing flavor of brewed white liquor |
| CN102382739A (en) * | 2011-10-05 | 2012-03-21 | 古贝春集团有限公司 | Aroma mixed-steaming mixed-combustion small cellar-flowing process |
| CN102337193B (en) * | 2011-10-18 | 2012-10-10 | 李家民 | Method for preparing starter for improving content of health flavor components of white spirit |
| CN103013764A (en) * | 2013-01-06 | 2013-04-03 | 四川郎酒集团有限责任公司 | Preparation method of ultrahigh-temperature yeast for making hard liquor |
-
2014
- 2014-12-09 CN CN201710164338.4A patent/CN106754580B/en active Active
- 2014-12-09 CN CN201710164340.1A patent/CN106754581B/en active Active
- 2014-12-09 CN CN201410747485.0A patent/CN104673703B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102268397A (en) * | 2011-07-25 | 2011-12-07 | 江南大学 | Bacillus licheniformis capable of highly producing specific flavor, and application thereof in Chinese liquor brewing |
| CN102766576A (en) * | 2012-07-19 | 2012-11-07 | 江南大学 | Brewing function oriented microbe combination method and application of combination bacterium in liquor-making industry |
Non-Patent Citations (2)
| Title |
|---|
| 孟醒等: "酿酒酵母与地衣芽孢杆菌相互作用及基于蛋白组学的作用机制分析", 《微生物学通报》 * |
| 王海燕等: "清香型小曲白酒中微生物组成及功能微生物的分析", 《酿酒科技》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105838751A (en) * | 2016-03-17 | 2016-08-10 | 齐鲁工业大学 | Method for improving output of tetramethylpyrazine produced through fermentation by Bacillus |
| CN106929438A (en) * | 2016-11-18 | 2017-07-07 | 天津科技大学 | One plant height produces the saccharomyces cerevisiae and its construction method of Tetramethylpyrazine |
| CN110923163A (en) * | 2019-11-22 | 2020-03-27 | 郑州轻工业大学 | Bacillus methylotrophicus and application thereof |
| CN110923163B (en) * | 2019-11-22 | 2021-06-25 | 郑州轻工业大学 | Bacillus methylotrophicus and application thereof |
| CN113151245A (en) * | 2021-05-28 | 2021-07-23 | 湖北工业大学 | Flavor compound microbial inoculum for reducing rice wine fusel oil and application thereof |
| CN113151245B (en) * | 2021-05-28 | 2022-06-03 | 湖北工业大学 | Flavor compound microbial inoculum for reducing rice wine fusel oil and application thereof |
| CN117413924A (en) * | 2023-11-02 | 2024-01-19 | 北京衍微科技有限公司 | Composition with antibacterial and fermentation promoting effects and application thereof |
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| CN106754581B (en) | 2019-11-26 |
| CN106754580A (en) | 2017-05-31 |
| CN106754581A (en) | 2017-05-31 |
| CN106754580B (en) | 2019-11-26 |
| CN104673703B (en) | 2018-01-16 |
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