CN103451060A - Sugarcane fruit wine and brewing method thereof - Google Patents
Sugarcane fruit wine and brewing method thereof Download PDFInfo
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Abstract
The invention discloses a sugarcane fruit wine and a brewing method thereof. The brewing method comprises the following steps of obtaining sugarcane juice by squeezing cleaned sugarcane, adjusting the pH value to 3.5-4.5; adding pectinase to process; adopting macroporous resin to process, and then adjusting the sugar content of the sugarcane juice by using cane sugar; sterilizing and cooling to serve as a fermentation medium; firstly inoculating an ester-producing yeast culture solution in the fermentation medium; still fermenting for 12-24 hours at 25-28 DEG C, and then inoculating in a brewing yeast culture solution; continuing to still ferment for 168-240 hours at 10-14 DEG C; finally, centrifugally separating fermented mash, and removing a yeast body to obtain supernatant of the fermented mash; adding immobilized ester-producing yeast to the supernatant of the fermented mash; ageing for 240-360 DEG C at 6-10 DEG C; finally, centrifugally separating the aged fermented mash, filtering the supernatant liquor by an ultrafiltration membrane, so as to obtain the sugarcane fruit wine which is clear, rich in fragrance, soft in taste, and coordinated in wine flavor and fruit flavor.
Description
Technical field
The present invention relates to a kind of fruit wine, particularly a kind of sugarcane fruit wine and brew method thereof, belong to technology and brewing technology.
Background technology
Sugar cane juice flavor is sweet cold in nature, has clearing heat and promoting fluid, nourishing and the effect such as moisturizes.Sugar cane juice has peculiar flavour, is rich in the sugars such as sucrose, fructose, glucose, also contains the trace element of the needed by human such as iron, calcium, phosphorus, manganese, zinc, both can, directly as nutritional drink, can be used as again the brewing materials of fruit wine, fruit vinegar etc.With other fruit raw materials, compare, sugarcane has the advantages such as wide material sources, rich amount cheapness, unique flavor, sugar abundance, is more suitable for fruit wine, fruit vinegar etc. and brewages.
The product of brewageing that to utilize sugarcane be raw material mainly contains alcohol fuel, Rum etc.Aspect the sugar cane juice fermentation fuel ethanol, fermentation technique is only pursued alcohol getting rate, and avoids as far as possible the generation of flavour substances.Rum belongs to liquor, mainly with cane molasses, it is brewing materials, its fermentation technique there are certain requirements the generation of flavour substances, and whole brewing process has mixed fermentation (yeast, bacterium etc.), leavening temperature high (30-33 ℃), fermentation period short (36-48h), long characteristics such as (2-10) of oak barrel during aging time.The domestic document that has some to utilize sugar cane juice brew sugarcane white wine (liquor), sugar cane juice and other fruit juice mixing making fruit wine, the fermentation technique major embodiment of these documents is fermented bacterium single (only for yeast saccharomyces cerevisiae), leavening temperature is higher, ageing is default or not enough, and the product of institute's brew has much room for improvement aspect flavouring essence quality.Chinese patent CN102628013 discloses a kind of method of utilizing pure sugar cane juice to produce scent type sugarcane fruit wine, but, Shortcomings part still, major embodiment is: (1) fermented bacterium only adopts single yeast saccharomyces cerevisiae, fruit wine ester Studies of The Aromatic Substances produces not enough, need to add oak product and promote fruit wine fragrance, easily cover fruital; (2) the fermenting process temperature is controlled higher (16-22 ℃), the easy volatilization loss of fruital that sugarcane is intrinsic.
Summary of the invention
Primary and foremost purpose of the present invention is that the shortcoming that overcomes prior art, with not enough, provides a kind of brew method of sugarcane fruit wine.
Another object of the present invention is to provide the sugarcane obtained by above-mentioned brew method fruit wine.
Purpose of the present invention is achieved through the following technical solutions: a kind of brew method of sugarcane fruit wine comprises following steps:
(1) pre-treatment of sugarcane: sugarcane is through cleaning, squeezing, solid-liquid separation, obtain natural sugarcane juice, its pH value is adjusted to 3.5-4.5, add polygalacturonase to carry out enzymolysis processing, then adopt the sugar cane juice after macroporous resin is processed pectinase enzymatic hydrolysis to be processed, then adjust the sugar degree of sugar cane juice with sucrose, through sterilizing, cooling, as fermention medium;
(2) fermentation of sugarcane fruit wine: first access the ester-producing yeast nutrient solution in the fermention medium obtained in step (1), the 5%-10% that the access volume is the fermention medium volume, static fermentation 12-24h under 25-28 ℃; Then access the yeast saccharomyces cerevisiae nutrient solution, the 1%-5% that the access volume is the fermention medium volume, continue static fermentation 168-240h in 10-14 ℃; Finally, fermentation liquid is carried out to centrifugation, remove the yeast thalline, obtain the fermentation liquid supernatant; Add the immobilization ester-producing yeast again in the fermentation liquid supernatant, in 6-10 ℃ of ageing 240-360h;
(3) separation of sugarcane fruit wine: the fermentation liquid after ageing is through centrifugation, and the immobilization ester-producing yeast in precipitation is back to the ageing of next batch, and upper pure mellow wine liquid is again through ultrafiltration membrance filter, the sugarcane fruit wine that can clarify.
In order to realize better the present invention, the pH value described in step (1) is preferably uses acetic acid to be regulated;
The consumption of the polygalacturonase described in step (1) is preferably in every liter of natural sugarcane juice and adds the 2000-4000U polygalacturonase; More preferably in every liter of natural sugarcane juice, add the 3000-4000U polygalacturonase;
The condition optimization of the enzymolysis processing described in step (1) is 40-50 ℃ of reaction 60-90min;
Macroporous resin described in step (1) can be selected XDA-5, S-8, AB-8, X-5 etc., and is not limited to mentioned resin model; These resins method (resin manufacturer provides) according to the rules carry out pre-treatment and regeneration; Macroporous resin is for adsorbing polyphenols and partial pigment;
The concrete steps that sugar cane juice after employing macroporous resin described in step (1) is processed pectinase enzymatic hydrolysis is processed are as follows: macroporous resin column on the sugar cane juice after pectinase enzymatic hydrolysis is processed, upper column flow rate is 1-2BV/h, collects resin column leak point effluent liquid before;
Sugar degree described in step (1) is preferably 180-200g/L;
Sterilizing described in step (1) adopts the conventional heating sterilization method, and condition generally can be selected 115-121 ℃ of sterilizing 10-25min;
Cell concentration in ester-producing yeast nutrient solution described in step (2) is preferably (1.0-1.5) * 10
8individual/ml;
One or both in preferably abnormal Brunswick Durham yeast (Wickerhamomyces anomalus) CGMCC2.470 of described ester-producing yeast and abnormal Brunswick Durham yeast (Wickerhamomyces anomalus) CGMCC2.297;
Described ester-producing yeast nutrient solution is that ester-producing yeast is inoculated in the substratum that is applicable to the cultivation ester-producing yeast and cultivates and obtain to logarithmic phase or stationary phase; Preferably prepare by the following method: in the malt juice liquid medium that the pH value that ester-producing yeast is inoculated into to sterilizing is 4.5-5.0, under 25-28 ℃, the condition of 150-200r/min, cultivate and obtain;
Cell concentration in yeast saccharomyces cerevisiae nutrient solution described in step (2) is preferably (1.2-1.8) * 10
8individual/ml;
A kind of in described yeast saccharomyces cerevisiae preferably saccharomyces cerevisiae (Saccharomyces cerevisiae) CICC1596, yeast saccharomyces cerevisiae (Saccharomyces cerevisiae) CICC1215, yeast saccharomyces cerevisiae (Saccharomyces cerevisiae) CGMCC2.614 and yeast saccharomyces cerevisiae (Saccharomyces cerevisiae) CGMCC2.1190 or at least two kinds;
Described yeast saccharomyces cerevisiae nutrient solution is that yeast saccharomyces cerevisiae is inoculated in the substratum that is applicable to the cultivation yeast saccharomyces cerevisiae and cultivates and obtain to logarithmic phase or stationary phase; Preferably prepare by the following method: in the malt juice liquid medium that the pH value that yeast saccharomyces cerevisiae is inoculated into to sterilizing is 4.5-5.0, under 28-30 ℃, the condition of 150-200r/min, cultivate and obtain;
The condition optimization of the centrifugation described in step (2) and step (3) is the centrifugal 20-30min of 4000-5000rpm;
Immobilization ester-producing yeast described in step (2) can adopt conventional process for fixation to prepare, preferably obtain as follows: ester-producing yeast suspension and sodium alginate soln are mixed, obtain mixed solution, then mixed solution is splashed in calcium chloride solution, calcification, the gel beads obtained is the immobilization ester-producing yeast; More preferably obtain as follows: by cell concentration, be (2.0-3.0) * 10
8the sodium alginate soln that the ester-producing yeast suspension of individual/ml and concentration are 30g/L 1:8 by volume mixes, and obtains mixed solution; Then mixed solution is splashed into the isopyknic 30g/L calcium chloride solution of blended liquid phase in, calcification 4h, the gel beads of formation is the immobilization ester-producing yeast;
The consumption of the immobilization ester-producing yeast described in step (2) is preferably by every liter of fermentation liquid supernatant access immobilization ester-producing yeast 10-30g;
The preferred molecular weight cut-off of ultra-filtration membrane described in step (3) is no more than the ultra-filtration membrane of 150kD; Be preferably the ultra-filtration membrane that molecular weight cut-off is 100-150kD;
A kind of sugarcane fruit wine, made by above-mentioned brew method; Its ethanol content is 8.0%(v/v) more than, total ester content is more than 3.5g/L, residual reducing sugar content is below 3.0g/L; Being preferably its ethanol content is 8.1-9.6%(v/v), total ester content is 3.5-4.2g/L.
The principle of the invention is: adopt polygalacturonase to carry out enzymolysis processing, pectin in the degradable sugar cane juice, be conducive to the sugarcane clarification of juice; Adopt macroporous resin to carry out adsorption treatment, can remove polyphenol substance and the partial pigment of sugar cane juice, can reduce the oxidizing brown stain of sugar cane juice, be conducive to improve the non-biostability of sugarcane fruit wine product; Sugar cane juice, through fermentation by saccharomyces cerevisiae, can be converted into alcohol by fermentable sugar; Sugar cane juice multiparity ester yeast fermentation, can generate Ester, is conducive to improve the fragrance of fruit wine; Through the ageing of immobilization ester-producing yeast, can further improve the flavor quality of fruit wine again.The present invention is integrated pectinase enzymatic hydrolysis technology, macroporous adsorption resin technology, mixed bacterium low temperature fermentation technology, immobilized cell ageing technology etc., solve the haze problem of sugarcane fruit wine and improved non-biostability, solved the aroma substance Generating Problems and improved the flavouring effect, solved aroma substance and lost problem and improve the fragrance effect that keeps.
The present invention, with respect to prior art, has following advantage and beneficial effect:
(1) to select sugar cane juice be brewing materials in the present invention, with respect to existing fruit wine raw material, has the advantages such as wide material sources, unique flavor, and the novel approach that utilizes of a sugarcane resource both can be provided, and can increase again the fruit wine of novel sapor.By the method for the invention brew, the giving off a strong fragrance of fruit wine, aroma and fruital are coordinated, and mouthfeel is soft, suitable popular crowd.
(2) the present invention adopts polygalacturonase to carry out enzymolysis processing to natural sugarcane juice, adopts macroporous resin to carry out adsorption treatment to polyphenol and the partial pigment of natural sugarcane juice, is conducive to the sugarcane clarification of juice, and the non-biostability of sugarcane fruit wine product is improved.
(3) the present invention adopts ester-producing yeast and yeast saccharomyces cerevisiae to carry out mixed fermentation at lord ferment period, and adopt the immobilization ester-producing yeast to carry out ageing to wine liquid, both guaranteed the suitable wine degree of fruit wine, there is again the effect of producing the ester flavouring, the fruit wine flavouring essence quality is improved, thereby has made up the shortcoming of fragrance deficiency in liquid brewing technique.
(4) the present invention adopts low temperature fermentation technique, and for fruit wine mesophilic digestion technique, the fruit wine mouthfeel is softer; Simultaneously, low temperature fermentation technique has the effect that keeps fragrance, and the volatile quantity of aroma substance under cold condition that original aroma substance of sugar cane juice and fermentation generate all significantly reduces.
Embodiment
Below in conjunction with embodiment, the present invention is described in further detail, but embodiments of the present invention are not limited to this.
Embodiment 1
(1) pre-treatment of sugarcane
Sugarcane, through cleaning, squeezing, solid-liquid separation, obtains natural sugarcane juice.Get 0.6L natural sugarcane juice (reducing sugar content is 116g/L), with acetic acid, its pH value is adjusted to 3.5, add 0.12g pectase preparation (Jining and U.S. biotechnology company limited, product specification 20000U/g), 50 ℃ of enzymolysis 90min of constant temperature; Sugar cane juice after enzymolysis enters the XDA-5 macroporous adsorptive resins with the flow velocity of 1.0BV/h, collects the effluent liquid of removing polyphenol substance and partial pigment; Then in effluent liquid, add 44.5g sucrose and 30mL water, make the volume of effluent liquid and sugar degree be respectively 0.6L and 180g/L; Then, the 2000mL triangular flask of packing into, in 121 ℃ of sterilizing 10min, cooling rear standby.
(2) fermentation of sugarcane fruit wine
Yeast enlarged culturing flow process is: inclined-plane → shake-flask culture.
The shake-flask culture base: wort (12 °) 300mL, regulate pH4.5 with acetic acid, be distributed into 3 500mL triangular flasks, in 121 ℃ of sterilizing 15min, cooling rear standby.
Abnormal Brunswick Durham yeast (Wickerhamomyces anomalus) CGMCC2.470(is purchased to Chinese common micro-organisms culture presevation administrative center) 2 shake-flask culture bases of slant strains access, each triangular flask access 1 ring bacterial classification, be placed under 25 ℃, the condition of 150r/min and cultivate 20h, can obtain ripe yeast shake-flask culture liquid, yeast count is 1.02 * 10
8individual/mL.
By the wherein abnormal Brunswick Durham yeast culture liquid centrifugation (4000rpm, 30min) of 1 bottle of maturation, collect thalline, with after sterilized water washing 2 times, get the part thalline and be mixed with the 25mL bacteria suspension (cell concentration is about 3.0 * 10
8individual/mL), with 200mL sodium alginate soln (concentration is 30g/L), mix, the constant speed of then take splashes into 225mL calcium chloride solution (concentration is 30g/L), calcification 4h, through centrifugation (4000rpm, 30min), obtain gel beads, be the immobilization ester-producing yeast.
Yeast saccharomyces cerevisiae CICC1596(is purchased to Chinese industrial microbial strains preservation administrative center) slant strains access other 1 shake-flask culture base, inoculum size is 1 ring bacterial classification, be placed under 28 ℃, the condition of 150r/min and cultivate 16h, can obtain ripe yeast shake-flask culture liquid, yeast count is 1.24 * 10
8individual/mL.
By the fermention medium of the abnormal Brunswick Durham yeast culture liquid of 60mL maturation access 0.6L, in 25 ℃ of static fermentation 24h, then access the yeast saccharomyces cerevisiae nutrient solution of 6mL maturation, the static fermentation 240h of continuation under 14 ℃; Then, fermentation liquid is placed under the condition of 4000rpm, 30min and carries out centrifugation, collect supernatant liquor 620mL, another 2000mL triangular flask of packing into, access 18.6g immobilization ester-producing yeast, be placed in 6 ℃ of lower ageing 360h.
(3) separation of sugarcane fruit wine
Karusen after ageing is carried out under the condition of 4000rpm, 30min to centrifugation, collect clear liquid, adopt again the ultrafiltration membrance filter system that the molecular weight that dams is 150kD to carry out 1 ultrafiltration, can obtain sugarcane fruit wine 0.54L aseptic, yellow clarification, the ethanol content of sugarcane fruit wine is 8.1%(v/v), total ester content (measuring according to GB/T10345-2007) is 4.2g/L, and reducing sugar content is 2.9g/L.This sugarcane fruit wine gives off a strong fragrance, and aroma and fruital (sugarcane delicate fragrance) are coordinated, and mouthfeel is soft.
Embodiment 2
(1) pre-treatment of sugarcane
Sugarcane, through cleaning, squeezing, solid-liquid separation, obtains natural sugarcane juice.Get 1.8L natural sugarcane juice (reducing sugar content is 122g/L), with acetic acid, its pH value is adjusted to 4.5, add 0.36g pectase preparation (Jining and U.S. biotechnology company limited, product specification 20000U/g), 40 ℃ of enzymolysis 90min of constant temperature; Sugar cane juice after enzymolysis enters the S-8 resin column with the flow velocity of 2.0BV/h, collects the effluent liquid of removing polyphenol substance and partial pigment; Then in effluent liquid, add 170g sucrose and 80mL water, make the volume of effluent liquid and sugar degree be respectively 1.8L and 200g/L; Then, the 5000mL triangular flask of packing into, in 115 ℃ of sterilizing 25min, cooling rear standby.
(2) fermentation of sugarcane fruit wine
Yeast enlarged culturing flow process is: inclined-plane → shake-flask culture.
The shake-flask culture base: wort (12 °) 300mL, regulate pH5.0 with acetic acid, be distributed into 3 500mL triangular flasks, in 121 ℃ of sterilizing 15min, cooling rear standby.
Abnormal Brunswick Durham yeast CGMCC2.297(is purchased to Chinese common micro-organisms culture presevation administrative center) 2 shake-flask culture bases of slant strains access, each triangular flask access 1 ring bacterial classification, be placed under 28 ℃, the condition of 200r/min and cultivate 20h, can obtain ripe yeast shake-flask culture liquid, yeast count is 1.48 * 10
8individual/mL.
The Hansenula anomala medium centrifugal of 1 bottle of maturation is wherein separated to (5000rpm, 20min), collect thalline, with after sterilized water washing 2 times, get the part thalline and be mixed with the 30mL bacteria suspension (cell concentration is about 2.0 * 10
8individual/mL), with 240mL sodium alginate soln (concentration is 30g/L), mix, the constant speed of then take splashes into 270mL calcium chloride solution (concentration is 30g/L), calcification 4h, through centrifugation (5000rpm, 20min), obtain gel beads, be the immobilization ester-producing yeast.
Yeast saccharomyces cerevisiae CICC1215(is purchased to Chinese industrial microbial strains preservation administrative center) slant strains access other 1 shake-flask culture base, inoculum size is 1 ring bacterial classification, be placed under 30 ℃, the condition of 200r/min and cultivate 18h, can obtain ripe yeast shake-flask culture liquid, yeast count is 1.77 * 10
8individual/mL.
By the fermention medium of the Hansenula anomala nutrient solution of 90mL maturation access 1.8L, in 28 ℃ of static fermentation 12h, then access the yeast saccharomyces cerevisiae nutrient solution of 90mL maturation, the static fermentation 216h of continuation under 12 ℃; Then, karusen is placed under the condition of 5000rpm, 20min and carries out centrifugation, collect supernatant liquor 1.86L, another 5000mL triangular flask of packing into, access 18.6g immobilization ester-producing yeast, be placed in 10 ℃ of lower ageing 240h.
(3) separation of sugarcane fruit wine
Karusen after ageing is carried out under the condition of 5000rpm, 20min to centrifugation, collect clear liquid, adopt again the ultrafiltration membrance filter system that the molecular weight that dams is 100kD to carry out 1 ultrafiltration, can obtain sugarcane fruit wine 1.60L aseptic, yellow clarification, the ethanol content of sugarcane fruit wine is 9.6%(v/v), total ester content (measuring according to GB/T10345-2007) is 3.5g/L, and reducing sugar content is 2.2g/L.This sugarcane fruit wine gives off a strong fragrance, and aroma and fruital are coordinated, and mouthfeel is soft.
Embodiment 3
(1) pre-treatment of sugarcane
Sugarcane, through cleaning, squeezing, solid-liquid separation, obtains natural sugarcane juice.Get 7.0L natural sugarcane juice (reducing sugar content is 128g/L), with acetic acid, its pH value is adjusted to 4.0, add 1.05g pectase preparation (Jining and U.S. biotechnology company limited, product specification 20000U/g), 45 ℃ of enzymolysis 60min of constant temperature; Sugar cane juice after enzymolysis enters the AB-8 resin column with the flow velocity of 1.5BV/h, collects the effluent liquid of removing polyphenol substance and partial pigment; Then in effluent liquid, add 506g sucrose and 190mL water, make the volume of effluent liquid and sugar degree be respectively 7.0L and 190g/L; Then, the 10L fermentor tank of packing into, in 121 ℃ of sterilizing 15min, cooling rear standby.
(2) fermentation of sugarcane fruit wine
Yeast enlarged culturing flow process is: inclined-plane → shake-flask culture.
The shake-flask culture base: wort (12 °) 1.2L, regulate pH4.8 with acetic acid, be distributed into 6 1000mL triangular flasks, in 121 ℃ of sterilizing 15min, cooling rear standby.
Abnormal Brunswick Durham yeast (Wickerhamomyces anomalus) CGMCC2.470(is purchased to Chinese common micro-organisms culture presevation administrative center) 4 shake-flask culture bases of slant strains access, each triangular flask access 1 ring bacterial classification, be placed under 28 ℃, the condition of 200r/min and cultivate 22h, can obtain ripe yeast shake-flask culture liquid, yeast count is 1.36 * 10
8individual/mL.
The Hansenula anomala medium centrifugal of 2 bottles of maturations is wherein separated to (4500rpm, 20min), collect thalline, with after sterilized water washing 2 times, get the part thalline and be mixed with the 100mL bacteria suspension (cell concentration is about 3.0 * 10
8individual/mL), with 800mL sodium alginate soln (concentration is 30g/L), mix, the constant speed of then take splashes into 900mL calcium chloride solution (concentration is 30g/L), calcification 4h, through centrifugation (4500rpm, 20min), obtain gel beads, be the immobilization ester-producing yeast.
Yeast saccharomyces cerevisiae CGMCC2.614(is purchased to Chinese common micro-organisms culture presevation administrative center) slant strains access other 2 shake-flask culture bases, each triangular flask access 1 ring bacterial classification, be placed under 20 ℃, the condition of 150r/min and cultivate 20h, can obtain ripe yeast shake-flask culture liquid, yeast count is 1.75 * 10
8individual/mL.
By the fermention medium of the Hansenula anomala nutrient solution of 350mL maturation access 7.0L, in 26 ℃ of static fermentation 18h, then access the yeast saccharomyces cerevisiae nutrient solution of 350mL maturation, in 14 ℃ of static fermentation 168h of continuation; Then, karusen is placed under the condition of 4500rpm, 20min and carries out centrifugation, collect clear liquid 7.2L, another 10L fermentor tank of packing into, access 144g immobilization ester-producing yeast, be placed in 8 ℃ of lower ageing 288h.
(3) separation of sugarcane fruit wine
Karusen after ageing is carried out under the condition of 4500rpm, 20min to centrifugation, collect clear liquid, adopt again the ultrafiltration membrance filter system that the molecular weight that dams is 150kD to carry out 1 ultrafiltration, can obtain sugarcane fruit wine 6.2L aseptic, yellow clarification, the ethanol content of sugarcane fruit wine is 8.6%(v/v), total ester content (measuring according to GB/T10345-2007) is 3.6g/L, and reducing sugar content is 2.8g/L.This sugarcane fruit wine gives off a strong fragrance, and aroma and fruital are coordinated, and mouthfeel is soft.
Embodiment 4
(1) pre-treatment of sugarcane
Sugarcane, through cleaning, squeezing, solid-liquid separation, obtains natural sugarcane juice.Get 3.0L natural sugarcane juice (reducing sugar content is 120g/L), with acetic acid, its pH value is adjusted to 4.0, add 0.45g pectase preparation (20000U/g), 45 ℃ of enzymolysis 75min of constant temperature; Sugar cane juice after enzymolysis enters the X-5 resin column with the flow velocity of 1.5BV/h, collects the effluent liquid of removing polyphenol substance and partial pigment; Then in effluent liquid, add 204.5g sucrose and 120mL water, make the volume of effluent liquid and sugar degree be respectively 3.0L and 180g/L; Then, the 5L fermentor tank of packing into, in 121 ℃ of sterilizing 10min, cooling rear standby.
(2) fermentation of sugarcane fruit wine
Yeast enlarged culturing flow process is: inclined-plane → shake-flask culture.
The shake-flask culture base: wort (12 °) 0.8L, regulate pH5.0 with acetic acid, be distributed into 4 1000mL triangular flasks, in 121 ℃ of sterilizing 15min, cooling rear standby.
Abnormal Brunswick Durham yeast CGMCC2.297(is purchased to Chinese common micro-organisms culture presevation administrative center) 3 shake-flask culture bases of slant strains access, each triangular flask access 1 ring bacterial classification, be placed under 25 ℃, the condition of 200r/min and cultivate 20h, can obtain ripe yeast shake-flask culture liquid, yeast count is 1.25 * 10
8individual/mL.
The Hansenula anomala medium centrifugal of 1 bottle of maturation is wherein separated to (4500rpm, 20min), collect thalline, with after sterilized water washing 2 times, get the part thalline and be mixed with the 60mL bacteria suspension (cell concentration is about 2.5 * 10
8individual/mL), with 480mL sodium alginate soln (concentration is 30g/L), mix, the constant speed of then take splashes into the calcium chloride solution (concentration is 30g/L) of 540mL, calcification 4h, through centrifugation (4500rpm, 20min), obtain gel beads, be the immobilization ester-producing yeast.
Yeast saccharomyces cerevisiae CGMCC2.1190(is purchased to Chinese common micro-organisms culture presevation administrative center) slant strains access other 1 shake-flask culture base, inoculum size is 1 ring bacterial classification, be placed under 30 ℃, the condition of 200r/min and cultivate 16h, can obtain ripe yeast shake-flask culture liquid, yeast count is 1.62 * 10
8individual/mL.
By the fermention medium of the Hansenula anomala nutrient solution of 225mL maturation access 3.0L, static fermentation 18h under 25 ℃, then access the yeast saccharomyces cerevisiae nutrient solution of 75mL maturation, the static fermentation 240h of continuation under 10 ℃; Then, karusen is placed under the condition of 4500rpm, 20min and carries out centrifugation, collect clear liquid 3.0L, another 5L fermentor tank of packing into, access 60g immobilization ester-producing yeast, be placed in 6 ℃ of lower ageing 336h.
(3) separation of sugarcane fruit wine
Karusen after ageing is carried out under the condition of 4500rpm, 20min to centrifugation, collect clear liquid, adopt again the ultrafiltration membrance filter system that the molecular weight that dams is 150kD to carry out 1 ultrafiltration, can obtain sugarcane fruit wine 2.6L aseptic, yellow clarification, the ethanol content of sugarcane fruit wine is 8.2%(v/v), total ester content (measuring according to GB/T10345-2007) is 4.0g/L, and reducing sugar content is 2.8g/L.This sugarcane fruit wine gives off a strong fragrance, and aroma and fruital are coordinated, and mouthfeel is soft.
Comparative Examples 1
(1) pre-treatment of sugarcane
Sugarcane, through cleaning, squeezing, solid-liquid separation, obtains natural sugarcane juice.Get 0.6L natural sugarcane juice (reducing sugar content is 116g/L), with acetic acid, its pH value is adjusted to 3.5, add 0.12g pectase preparation (Jining and U.S. biotechnology company limited, product specification 20000U/g), 50 ℃ of enzymolysis 90min of constant temperature; Sugar cane juice after enzymolysis enters the XDA-5 macroporous adsorptive resins with the flow velocity of 1.0BV/h, collects the effluent liquid of removing polyphenol substance and partial pigment; Then in effluent liquid, add 44.5g sucrose and 30mL water, make the volume of effluent liquid and sugar degree be respectively 0.6L and 180g/L; Then, the 2000mL triangular flask of packing into, in 121 ℃ of sterilizing 10min, cooling rear standby.
(2) fermentation of sugarcane fruit wine
Yeast enlarged culturing flow process is: inclined-plane → shake-flask culture.
The shake-flask culture base: wort (12 °) 100mL, regulate pH4.5 with acetic acid, be distributed into 1 500mL triangular flask, in 121 ℃ of sterilizing 15min, cooling rear standby.
Yeast saccharomyces cerevisiae CICC1596(is purchased to Chinese industrial microbial strains preservation administrative center) slant strains access other 1 shake-flask culture base, inoculum size is 1 ring bacterial classification, be placed under 28 ℃, the condition of 150r/min and cultivate 16h, can obtain ripe yeast shake-flask culture liquid, yeast count is 1.24 * 10
8individual/mL.
By the fermention medium of the yeast saccharomyces cerevisiae nutrient solution of 30mL maturation access 0.6L, under 28 ℃, static fermentation 60h(wine degree no longer raises).
(3) separation of sugarcane fruit wine
Fermentation liquid is carried out under the condition of 4000rpm, 30min to centrifugation, collect clear liquid, adopt again the ultrafiltration membrance filter system that the molecular weight that dams is 150kD to carry out 1 ultrafiltration, can obtain sugarcane fruit wine 0.52L aseptic, yellow clarification, the ethanol content of sugarcane fruit wine is 10.9%(v/v), total ester content (measuring according to GB/T10345-2007) is 0.4g/L, and reducing sugar content is 2.0g/L.This sugarcane fruit wine fragrance extremely a little less than, sugarcane delicate fragrance is lost, entrance is thin, soft sense deviation.
Comparative Examples 2
(1) pre-treatment of sugarcane
Sugarcane, through cleaning, squeezing, solid-liquid separation, obtains natural sugarcane juice.Get 0.6L natural sugarcane juice (reducing sugar content is 116g/L), with acetic acid, its pH value is adjusted to 3.5, add 0.12g pectase preparation (Jining and U.S. biotechnology company limited, product specification 20000U/g), 50 ℃ of enzymolysis 90min of constant temperature; Sugar cane juice after enzymolysis enters the XDA-5 macroporous adsorptive resins with the flow velocity of 1.0BV/h, collects the effluent liquid of removing polyphenol substance and partial pigment; Then in effluent liquid, add 44.5g sucrose and 30mL water, make the volume of effluent liquid and sugar degree be respectively 0.6L and 180g/L; Then, the 2000mL triangular flask of packing into, in 121 ℃ of sterilizing 10min, cooling rear standby.
(2) fermentation of sugarcane fruit wine
Yeast enlarged culturing flow process is: inclined-plane → shake-flask culture.
The shake-flask culture base: wort (12 °) 200mL, regulate pH4.5 with acetic acid, be distributed into 2 500mL triangular flasks, in 121 ℃ of sterilizing 15min, cooling rear standby.
Abnormal Brunswick Durham yeast (Wickerhamomyces anomalus) CGMCC2.470(is purchased to Chinese common micro-organisms culture presevation administrative center) 1 shake-flask culture base of slant strains access, each triangular flask access 1 ring bacterial classification, be placed under 25 ℃, the condition of 150r/min and cultivate 20h, can obtain ripe yeast shake-flask culture liquid, yeast count is 1.02 * 10
8individual/mL.
To cultivate ripe abnormal Brunswick Durham yeast culture liquid centrifugation (4000rpm, 30min), collect thalline, and with after sterilized water washing 2 times, get the part thalline and be mixed with the 25mL bacteria suspension (cell concentration is about 3.0 * 10
8individual/mL), with 200mL sodium alginate soln (concentration is 30g/L), mix, the constant speed of then take splashes into 225mL calcium chloride solution (concentration is 30g/L), calcification 4h, through centrifugation (4000rpm, 30min), obtain gel beads, be the immobilization ester-producing yeast.
Yeast saccharomyces cerevisiae CICC1596(is purchased to Chinese industrial microbial strains preservation administrative center) slant strains access other 1 shake-flask culture base, inoculum size is 1 ring bacterial classification, be placed under 28 ℃, the condition of 150r/min and cultivate 16h, can obtain ripe yeast shake-flask culture liquid, yeast count is 1.24 * 10
8individual/mL.
By the fermention medium of the yeast saccharomyces cerevisiae nutrient solution of 6mL maturation access 0.6L, under 22 ℃, static fermentation 120h(wine degree no longer raises); Then, fermentation liquid is placed under the condition of 4000rpm, 30min and carries out centrifugation, collect supernatant liquor 560mL, another 2000mL triangular flask of packing into, access 16.8g immobilization ester-producing yeast, be placed in 10 ℃ of lower ageing 240h.
(3) separation of sugarcane fruit wine
Karusen after ageing is carried out under the condition of 4000rpm, 30min to centrifugation, collect clear liquid, adopt again the ultrafiltration membrance filter system that the molecular weight that dams is 150kD to carry out 1 ultrafiltration, can obtain sugarcane fruit wine 0.48L aseptic, yellow clarification, the ethanol content of sugarcane fruit wine is 10.1%(v/v), total ester content (measuring according to GB/T10345-2007) is 1.5g/L, and reducing sugar content is 2.0g/L.This sugarcane fruit wine fragrance is faint, and sugarcane delicate fragrance is lost, and entrance is thin, soft sense deviation.
Comparative Examples 3
(1) pre-treatment of sugarcane
Sugarcane, through cleaning, squeezing, solid-liquid separation, obtains natural sugarcane juice.Get 0.6L natural sugarcane juice (reducing sugar content is 116g/L), with acetic acid, its pH value is adjusted to 3.5, add 0.12g pectase preparation (Jining and U.S. biotechnology company limited, product specification 20000U/g), 50 ℃ of enzymolysis 90min of constant temperature; Sugar cane juice after enzymolysis enters the XDA-5 macroporous adsorptive resins with the flow velocity of 1.0BV/h, collects the effluent liquid of removing polyphenol substance and partial pigment; Then in effluent liquid, add 56.5g sucrose and 30mL water, make the volume of effluent liquid and sugar degree be respectively 0.6L and 200g/L; Then, the 2000mL triangular flask of packing into, in 121 ℃ of sterilizing 10min, cooling rear standby.
(2) fermentation of sugarcane fruit wine
Yeast enlarged culturing flow process is: inclined-plane → shake-flask culture.
The shake-flask culture base: wort (12 °) 200mL, regulate pH4.5 with acetic acid, be distributed into 2 500mL triangular flasks, in 121 ℃ of sterilizing 15min, cooling rear standby.
Abnormal Brunswick Durham yeast (Wickerhamomyces anomalus) CGMCC2.470(is purchased to Chinese common micro-organisms culture presevation administrative center) 1 shake-flask culture base of slant strains access, each triangular flask access 1 ring bacterial classification, be placed under 25 ℃, the condition of 150r/min and cultivate 20h, can obtain ripe yeast shake-flask culture liquid, yeast count is 1.02 * 10
8individual/mL.
Yeast saccharomyces cerevisiae CICC1596(is purchased to Chinese industrial microbial strains preservation administrative center) slant strains access other 1 shake-flask culture base, inoculum size is 1 ring bacterial classification, be placed under 28 ℃, the condition of 150r/min and cultivate 16h, can obtain ripe yeast shake-flask culture liquid, yeast count is 1.24 * 10
8individual/mL.
By the fermention medium of the abnormal Brunswick Durham yeast culture liquid of 30mL maturation access 0.6L, in 25 ℃ of static fermentation 12h, then access the yeast saccharomyces cerevisiae nutrient solution of 30mL maturation, the static fermentation 240h of continuation under 14 ℃.
(3) separation of sugarcane fruit wine
Fermentation liquid is placed under the condition of 4000rpm, 30min and carries out centrifugation, collect supernatant liquor, adopt again the ultrafiltration membrance filter system that the molecular weight that dams is 150kD to carry out 1 ultrafiltration, can obtain sugarcane fruit wine 0.53L aseptic, yellow clarification, the ethanol content of sugarcane fruit wine is 10.8%(v/v), total ester content (measuring according to GB/T10345-2007) is 2.2g/L, and reducing sugar content is 2.4g/L.This sugarcane fruit wine fragrance a little less than, it is thin that entrance shows slightly.
Above-described embodiment is preferably embodiment of the present invention; but embodiments of the present invention are not restricted to the described embodiments; other any do not deviate from change, the modification done under spirit of the present invention and principle, substitutes, combination, simplify; all should be equivalent substitute mode, within being included in protection scope of the present invention.
Claims (10)
1. the brew method of a sugarcane fruit wine is characterized in that comprising following steps:
(1) pre-treatment of sugarcane: sugarcane is through cleaning, squeezing, solid-liquid separation, obtain natural sugarcane juice, its pH value is adjusted to 3.5-4.5, add polygalacturonase to carry out enzymolysis processing, then adopt the sugar cane juice after macroporous resin is processed pectinase enzymatic hydrolysis to be processed, then adjust the sugar degree of sugar cane juice with sucrose, through sterilizing, cooling, as fermention medium;
(2) fermentation of sugarcane fruit wine: first access the ester-producing yeast nutrient solution in the fermention medium obtained in step (1), the 5%-10% that the access volume is the fermention medium volume, static fermentation 12-24h under 25-28 ℃; Then access the yeast saccharomyces cerevisiae nutrient solution, the 1%-5% that the access volume is the fermention medium volume, continue static fermentation 168-240h in 10-14 ℃; Finally, fermentation liquid is carried out to centrifugation, remove the yeast thalline, obtain the fermentation liquid supernatant; Add the immobilization ester-producing yeast again in the fermentation liquid supernatant, in 6-10 ℃ of ageing 240-360h;
(3) separation of sugarcane fruit wine: the fermentation liquid after ageing is through centrifugation, and the immobilization ester-producing yeast in precipitation is back to the ageing of next batch, and upper pure mellow wine liquid, again through ultrafiltration membrance filter, obtains the sugarcane fruit wine of clarification.
2. the brew method of sugarcane fruit wine according to claim 1 is characterized in that:
The consumption of the polygalacturonase described in step (1) is to add the 2000-4000U polygalacturonase in every liter of natural sugarcane juice;
The condition of the enzymolysis processing described in step (1) is 40-50 ℃ of reaction 60-90min.
3. the brew method of sugarcane fruit wine according to claim 1, it is characterized in that: the concrete steps that the sugar cane juice after the employing macroporous resin described in step (1) is processed pectinase enzymatic hydrolysis is processed are as follows: macroporous resin column on the sugar cane juice after pectinase enzymatic hydrolysis is processed, upper column flow rate is 1-2BV/h, collects resin column leak point effluent liquid before.
4. the brew method of sugarcane fruit wine according to claim 1, it is characterized in that: the sugar degree described in step (1) is 180-200g/L.
5. the brew method of sugarcane fruit wine according to claim 1 is characterized in that:
Cell concentration in ester-producing yeast nutrient solution described in step (2) is (1.0-1.5) * 10
8individual/ml;
Cell concentration in yeast saccharomyces cerevisiae nutrient solution described in step (2) is (1.2-1.8) * 10
8individual/ml.
6. the brew method of sugarcane fruit wine according to claim 5 is characterized in that:
Described ester-producing yeast is one or both in abnormal Brunswick Durham yeast (Wickerhamomyces anomalus) CGMCC2.470 and abnormal Brunswick Durham yeast (Wickerhamomyces anomalus) CGMCC2.297;
Described yeast saccharomyces cerevisiae is a kind of in yeast saccharomyces cerevisiae (Saccharomyces cerevisiae) CICC1596, yeast saccharomyces cerevisiae (Saccharomyces cerevisiae) CICC1215, yeast saccharomyces cerevisiae (Saccharomyces cerevisiae) CGMCC2.614 and yeast saccharomyces cerevisiae (Saccharomyces cerevisiae) CGMCC2.1190 or at least two kinds.
7. the brew method of sugarcane fruit wine according to claim 1 is characterized in that:
Immobilization ester-producing yeast described in step (2) obtains as follows: ester-producing yeast suspension and sodium alginate soln mixed, obtains mixed solution, then mixed solution is splashed in calcium chloride solution, and calcification, the gel beads obtained is the immobilization ester-producing yeast;
The consumption of the immobilization ester-producing yeast described in step (2) is for accessing immobilization ester-producing yeast 10-30g by every liter of fermentation liquid supernatant.
8. the brew method of sugarcane fruit wine according to claim 7, it is characterized in that: described immobilization ester-producing yeast obtains as follows: by cell concentration, be (2.0-3.0) * 10
8the sodium alginate soln that the ester-producing yeast suspension of individual/ml and concentration are 30g/L 1:8 by volume mixes, and obtains mixed solution; Then mixed solution is splashed into the isopyknic 30g/L calcium chloride solution of blended liquid phase in, calcification 4h, the gel beads of formation is the immobilization ester-producing yeast.
9. the brew method of sugarcane fruit wine according to claim 1 is characterized in that:
PH value described in step (1) is for being used acetic acid to be regulated;
Macroporous resin described in step (1) is XDA-5, S-8, AB-8 or X-5;
The condition of the sterilizing described in step (1) is 115-121 ℃ of sterilizing 10-25min;
The condition of the centrifugation described in step (2) and step (3) is the centrifugal 20-30min of 4000-5000rpm;
Ultra-filtration membrane described in step (3) is the ultra-filtration membrane that molecular weight cut-off is no more than 150kD.
10. a sugarcane fruit wine, made by the described brew method of claim 1～9 any one.
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104152327A (en) * | 2014-08-30 | 2014-11-19 | 枣庄新华酒业有限公司 | Pure grain containing sugar cane juice brewing process |
| CN105543032A (en) * | 2016-01-25 | 2016-05-04 | 运城学院 | Method for brewing perry through mixed fermentation |
| CN106635832A (en) * | 2016-12-07 | 2017-05-10 | 昆明理工大学 | Wickerhamomyces anomalus and application thereof |
| CN107460081A (en) * | 2017-09-21 | 2017-12-12 | 广西叶茂食品有限责任公司 | A kind of method that Sugarcane fruit wine is prepared using nanofiltration sugar-cane juice |
| CN109880716A (en) * | 2018-09-28 | 2019-06-14 | 江苏荣泽食品有限公司 | A kind of strawberry compound fruit wine and its brewing method |
| CN111518651A (en) * | 2020-04-13 | 2020-08-11 | 浙江工商大学 | Production process of yellow wine with ester-fragrance yeast reactor |
| CN111575135A (en) * | 2020-06-16 | 2020-08-25 | 山东理工大学 | Preparation method of sugarcane juice fruit wine with low alcoholic strength |
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|---|---|---|---|---|
| CN104152327A (en) * | 2014-08-30 | 2014-11-19 | 枣庄新华酒业有限公司 | Pure grain containing sugar cane juice brewing process |
| CN105543032A (en) * | 2016-01-25 | 2016-05-04 | 运城学院 | Method for brewing perry through mixed fermentation |
| CN106635832A (en) * | 2016-12-07 | 2017-05-10 | 昆明理工大学 | Wickerhamomyces anomalus and application thereof |
| CN106635832B (en) * | 2016-12-07 | 2019-07-16 | 昆明理工大学 | One plant of abnormal Brunswick Durham saccharomycete and its application |
| CN107460081A (en) * | 2017-09-21 | 2017-12-12 | 广西叶茂食品有限责任公司 | A kind of method that Sugarcane fruit wine is prepared using nanofiltration sugar-cane juice |
| CN109880716A (en) * | 2018-09-28 | 2019-06-14 | 江苏荣泽食品有限公司 | A kind of strawberry compound fruit wine and its brewing method |
| CN111518651A (en) * | 2020-04-13 | 2020-08-11 | 浙江工商大学 | Production process of yellow wine with ester-fragrance yeast reactor |
| CN111575135A (en) * | 2020-06-16 | 2020-08-25 | 山东理工大学 | Preparation method of sugarcane juice fruit wine with low alcoholic strength |
| CN114410489A (en) * | 2021-12-31 | 2022-04-29 | 云南大学 | Abnormal yeast Weikehan yeast CAP5 strain and application thereof |
| CN114410489B (en) * | 2021-12-31 | 2023-06-23 | 云南大学 | Wilkham yeast CAP5 strain with abnormal condition and application thereof |
| CN116694419A (en) * | 2023-06-08 | 2023-09-05 | 广西大学 | Sweet sugarcane fruit wine produced at low temperature and without sulfur in whole process and brewing method |
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