CN113416658B - Hansenula sordida and application thereof in yellow peach fruit wine - Google Patents
Hansenula sordida and application thereof in yellow peach fruit wine Download PDFInfo
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- CN113416658B CN113416658B CN202110592294.1A CN202110592294A CN113416658B CN 113416658 B CN113416658 B CN 113416658B CN 202110592294 A CN202110592294 A CN 202110592294A CN 113416658 B CN113416658 B CN 113416658B
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
- A23L5/27—Removal of unwanted matter, e.g. deodorisation or detoxification by chemical treatment, by adsorption or by absorption
- A23L5/276—Treatment with inorganic compounds
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/30—Physical treatment, e.g. electrical or magnetic means, wave energy or irradiation
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- 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
- C12G3/024—Preparation of other alcoholic beverages by fermentation of fruits other than botanical genus Vitis
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- 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
- C12G3/026—Preparation of other alcoholic beverages by fermentation with health-improving ingredients, e.g. flavonoids, flavones, polyphenols or polysaccharides, added before or during the fermentation stage; with flavouring ingredients added before or during the fermentation stage
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- C12H1/00—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages
- C12H1/02—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages combined with removal of precipitate or added materials, e.g. adsorption material
- C12H1/04—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages combined with removal of precipitate or added materials, e.g. adsorption material with the aid of ion-exchange material or inert clarification material, e.g. adsorption material
- C12H1/0408—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages combined with removal of precipitate or added materials, e.g. adsorption material with the aid of ion-exchange material or inert clarification material, e.g. adsorption material with the aid of inorganic added material
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- C12H1/00—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages
- C12H1/02—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages combined with removal of precipitate or added materials, e.g. adsorption material
- C12H1/06—Precipitation by physical means, e.g. by irradiation, vibrations
- C12H1/063—Separation by filtration
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- C12H1/00—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages
- C12H1/12—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages without precipitation
- C12H1/14—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages without precipitation with non-precipitating compounds, e.g. sulfiting; Sequestration, e.g. with chelate-producing compounds
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- C12H1/00—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages
- C12H1/12—Pasteurisation, sterilisation, preservation, purification, clarification, or ageing of alcoholic beverages without precipitation
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- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
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Abstract
The invention discloses a Hanseng yeast with spores and application thereof in yellow peach fruit wine. The Hanseniaspora pseudomonad is Hanseniaspora pseudolliermondii HT9, and the preservation number of the Hanseniaspora pseudolliermondii in China center for type culture Collection is CCTCC No: m2021444. The Hansenula over spore can be applied to processing of yellow peach fruit wine, and comprises the following steps: juicing raw materials, protecting color and performing enzymolysis, adjusting components, activating and inoculating yeast, fermenting, stopping fermentation, adding gum and filtering. When being applied, the Hanseng yeast with spores has the advantages of increasing the complexity of the aroma of the yellow peach fruit wine, improving the richness of the aroma of the yellow peach fruit wine, improving the quality of the yellow peach fruit wine and the like.
Description
Technical Field
The invention relates to the technical field of food processing, in particular to Hansheng yeast with spores and application thereof in yellow peach fruit wine.
Background
Yellow peach is a special fruit in Hunan province and is planted in large quantities in places such as Taoism, Huaishi and Chenzhou. The yellow peach is rich in various nutrient substances and trace elements, has strong fragrance, is delicious and tasty, and is deeply favored by consumers. The yield and market price of the yellow peaches are high, play an important role in assisting farmers in poverty and richness shedding and are a key supporting industry of our province. Yellow peaches are generally mature and centralized on the market in 7-8 months per year, are not stored at normal temperature, have less than 2 months in fresh eating period, have low fresh-selling commodity fruit rate of the yellow peaches, have less than 10 percent of deep processing proportion, and are non-commodity fruits which do not reach the standard in weight, appearance and the like, are treated by fruit farmers at low price or directly discarded, so that great waste and environmental pollution are caused, and the processing of the yellow peaches becomes a necessary link for improving the industrial value of the yellow peaches. The current yellow peach processed products mainly comprise canned yellow peaches and dried yellow peaches. The yellow peach fruit wine is an ideal edible way and deep processing mode for yellow peaches, can reserve the original nutrient components in the yellow peaches to the maximum extent, and can meet the diversified demands of the fruit wine market. The development of the yellow peach fruit wine not only can obviously improve the economic value of the yellow peach and guarantee the income of fruit growers, but also can meet the increasing health requirements of consumers.
The quality of the yeast performance not only has great influence on the taste and flavor of the fruit wine, but also is important for the formation of the special color and style of the fruit wine. At present, most of the fruit wine production in China adopts active dry yeast powder brewed by wine imported from abroad, because of differences of fruit varieties, cultivation environments, process conditions and the like, introduced strains cannot completely show good quality of the wine, products lack typicality, and meanwhile, the abundance of introduced foreign microbial strains can influence the richness and diversity of local resources. The distribution of wild saccharomyces cerevisiae in different production areas and different fruits has certain regularity. Therefore, the exploration and development of new yeast capable of improving the mouthfeel and flavor of the fruit wine are urgently needed.
Disclosure of Invention
The invention aims to overcome the defects of the prior art, provides the Hansheng yeast with spores, correspondingly provides the application of the Hansheng yeast with spores in the processing of yellow peach fruit wine, and has the advantages of increasing the complexity of the aroma of the yellow peach fruit wine, improving the richness of the aroma, improving the quality of the yellow peach fruit wine and the like.
In order to solve the technical problems, the invention adopts the following technical scheme.
A Hanseniaspora pseudomonad, wherein the Hanseniaspora pseudollifermonii is HT9, and the preservation number of the Hanseniaspora pseudollifermonii in the China center for type culture collection is CCTCC No: m2021444.
As a general technical concept, the invention also provides an application of the Hanseng yeast with spores in the processing of yellow peach fruit wine.
The above application, preferably, comprises the following steps:
s1, juicing the raw materials, protecting color and performing enzymolysis: cleaning yellow peaches, reducing pesticide residues, removing cores and pulping to obtain yellow peach juice, taking the mass of the yellow peach juice as a reference, adding 0.7-1% of D-sodium erythorbate, 0.5-1% of vitamin C and 2-2.5% of citric acid by mass percent to protect the color, continuously adding 0.1-0.12% of cellulase and 0.1-0.15% of pectinase to carry out enzymolysis, and centrifuging to obtain the yellow peach juice;
s2, component adjustment: adding cane sugar and a fermentation aid into the yellow peach juice obtained in the step S1, wherein the addition amount of the cane sugar is 8-12% of the mass of the yellow peach juice, and the addition amount of the fermentation aid is 0.2-0.3 g/L of the yellow peach juice, so as to obtain the yellow peach juice containing cane sugar and the fermentation aid;
s3, yeast activation inoculation: the Hanseng sporulata yeast is cultured and activated by adopting an YPD solid culture medium and an YPD liquid culture medium in sequence, the Saccharomyces cerevisiae is activated by adopting water and fruit juice in sequence, the activated Hanseng sporulata yeast is inoculated into the yellow peach juice containing cane sugar and fermentation auxiliary agents obtained in the step S2, and after 2-3 days of fermentation, the activated Saccharomyces cerevisiae is inoculated;
s4, fermentation: carrying out ultrasonic auxiliary fermentation on the inoculated yellow peach juice, and then carrying out main fermentation;
s5, terminating fermentation: when the alcoholic strength of the yellow peach fermentation liquor reaches 10-11 degrees, adding sulfur dioxide, treating at the low temperature of-2-3 ℃, performing ultrasonic treatment, separating and pouring to obtain yellow peach wine base;
s6, glue adding and filtering: and (3) adding glue to the yellow peach wine base by using bentonite, filtering and filling to obtain the yellow peach fruit wine.
In the above application, preferably, in step S1, the pesticide residue reduction is performed by using an ozone composite high-voltage pulse electric field to perform pesticide residue degradation, that is, the cleaned yellow peaches are subjected to high-voltage pulse electric field treatment while being subjected to ozone cleaning treatment, the concentration of ozone is 5mg/L to 15mg/L, the time of the ozone cleaning treatment is 15min to 30min, and the conditions of the high-voltage pulse electric field treatment are as follows: the electric field intensity is 2 kV/cm-3 kV/cm, the pulse time is 50 mus-150 mus, and the number of pulses is 50-100.
In the above application, preferably, in step S3, the specific process of yeast activation inoculation is as follows: firstly culturing Hanseng sporulata in an YPD solid culture medium for 2-3 days, then culturing in an YPD liquid culture medium at 26-28 ℃ for 18-24 hours to obtain a zymogen liquid, centrifuging the zymogen liquid at 6-8 ℃ for 3-5 minutes at the rotating speed of 5000-6000 rpm/min, collecting thalli, then cleaning and re-suspending the thalli by adopting sterile water for 2-3 times, adjusting the bacteria liquidAt a concentration of 106-7cfu/mL to obtain activated Hanseng yeast with spore, and refrigerating and storing at 4-6 ℃.
In the above application, preferably, in step S3, the activated hansenula sporulata yeast is inoculated according to a proportion of 1% to 2%, the activated saccharomyces cerevisiae is inoculated according to a proportion of 2% to 5%, and the saccharomyces cerevisiae is saccharomyces cerevisiae D47.
In the above application, preferably, in step S4 and step S4, the specific process of the ultrasonic-assisted fermentation is as follows: and (3) intermittently treating the yellow peach juice inoculated in the step (S3) by adopting low-frequency ultrasonic, wherein the ultrasonic power is 100-150W, the ultrasonic frequency is 10-15 kHz, the ultrasonic treatment is stopped for 6S every 0.5S, the circulation is stopped after the continuous treatment is carried out for 5-8 h, the yellow peach juice enters main fermentation, the yellow peach juice is fermented for 4-7 days at the temperature of 18-20 ℃, and the yellow peach juice is relatched for 2-3 times during the fermentation period.
In the above application, preferably, in step S5, the addition amount of sulfur dioxide is 30mg/L to 50mg/L, the time of the low-temperature treatment is 5 days to 7 days, the power of the ultrasonic treatment is 300W to 500W, the frequency of the ultrasonic treatment is 40kHz to 50kHz, and the time of the ultrasonic treatment is 30min to 40 min.
In the above application, preferably, in step S1, the color protection temperature is 35 to 40 ℃, the color protection time is 1 to 1.5 hours, the enzymolysis temperature is 40 to 45 ℃, the enzymolysis time is 2 to 3 hours, and the centrifugation is performed by a butterfly centrifugation method.
In the above application, preferably, in step S6, the bentonite is added in an amount of 50g/L to 60g/L, the filtration is performed by using a cross-flow membrane, the cross-flow membrane is a disc cross-flow membrane, and the filtration flow rate is 10m/S to 15 m/S.
The invention relates to a spore-containing Hansheng yeast HT9 obtained by screening and identifying the surfaces of yellow peach fruits in the yellow peach producing area of Mayan yellow peach in Hunan province, wherein the strain is separated from natural fermented mash of Mayan yellow peach in Hunan province. The Hansenula maxima HT9 of the present invention is mixed with commercial active dry yeast for fermentation. The ultrasonic-assisted fermentation is adopted, so that the rapid propagation of yeast is facilitated, the fermentation starting speed is high, the utilization rate of substrate glucose is high, and the generation of higher alcohol is reduced. Fermenting to preset alcohol content with small amount of SO2And (6) stopping the fermentation of the yellow peach wine by low-temperature ultrasonic treatment, and filling after glue feeding and cross-flow membrane filtration to obtain the yellow peach fruit wine.
Hanseniaspora (H.) is one of the leading wild Saccharomyces occurring in the early stage of natural fermentation of fruit wine, and is mainly selected from Hanseniaspora vinifera (H.vinae), Hanseniaspora viticola (H.uvarum), Hanseniaspora cactus (H.opuntiae), Hansenula montmorifolium (H.guilliermondii) and Hansenula osmansoni (H.osmophila). Compared with saccharomyces cerevisiae, the ethanol production capacity of h. fermentation is lower, which plays an important role in increasing volatile compounds of wine. Therefore, the invention provides the excellent Hanseng yeast with spores for the fermentation of the yellow peach fruit wine, and has important significance for improving the color, the aroma, the sense and the style of the yellow peach fruit wine.
Compared with the prior art, the invention has the advantages that:
(1) the invention provides a spore Hanseng yeast which can be applied to brewing yellow peach fruit wine and has the advantages of increasing the complexity of the aroma of the yellow peach fruit wine, improving the richness of the aroma, improving the quality of the yellow peach fruit wine and the like.
(2) The application of the Hanseng sporozoite yeast in the yellow peach fruit wine utilizes the Hanseng sporozoite yeast HT9 to be mixed with the saccharomyces cerevisiae for fermentation, can obviously improve the content of ester aroma substances such as ethyl acetate, phenethylacetate and the like, and thus effectively increases the aroma in the yellow peach fruit wine. The content of ester substances in the wine after mixed fermentation reaches 38.37mg/L, which is 11.54mg/L higher than that of the wine fermented by only using saccharomyces cerevisiae D47; the ethyl acetate and the ethyl butyrate are obviously increased, the ethyl acetate is increased from 18.6mg/L to 21.6mg/L, the ethyl acetate has pleasant flower and fruit fragrance and pineapple fragrance, the threshold value is only 3.6 mu g/L, and the contribution to the fruit fragrance of the fruit wine is large.
(3) The invention adopts the ozone composite high-voltage pulse electric field to degrade the pesticide residues, can effectively reduce the use concentration of ozone, simultaneously solves the problems that the pesticide residues are not completely reduced by the ozone and the degradation products with more toxicity are generated due to incomplete degradation at present, realizes the quick and effective degradation of the pesticide residues and ensures the food safety. Ozone is a strong oxidant, but is rarely applied in actual water treatment, mainly because of high economic cost, and how to reduce the ozone consumption and improve the utilization rate of ozone is a difficult point. The ozone composite high-voltage pulse electric field can generate a synergistic effect, the pulse discharge improves the solubility of ozone in water, the high-voltage pulse discharge and ozone oxidation fully utilize the liquid-electric cavitation effect generated by the high-voltage pulse discharge, the periodic expansion and contraction of bubbles between electrodes generate a large amount of fine bubbles, the contact area is increased to promote the ozone to enter a liquid phase or react at a gas-liquid interface in a large amount, the utilization efficiency of ozone oxidation is improved, and therefore, the ozone composite high-voltage pulse electric field has a good degradation effect on pesticide residues.
(4) The invention adopts low-frequency ultrasonic wave for auxiliary fermentation after yeast inoculation, can accelerate the utilization of yeast to substrate glucose by low-frequency ultrasonic wave, accelerates reproductive metabolism, quickly increases yeast value and iterates, and increases CO2The generation of the alcohol is reduced, and meanwhile, the influence of other flavor substances such as esters and acids is small; on the other hand, the content of pectin in the yellow peaches is high, the content of fusel alcohol such as 2-methyl-1-propanol, 2-methyl-1-butanol and 3-methyl-1-butanol after fermentation is generally high, and free radicals generated by low-frequency ultrasonic cavitation react with tartaric acid, ethanol, ferrous iron, copper ions, mannitol and the like, so that the generation of high-grade alcohol is reduced, and the improvement of the quality and the flavor of the fruit wine is facilitated.
(5) According to the invention, a small amount of sulfur dioxide, freezing and ultrasonic treatment are adopted to terminate fermentation after the main fermentation is finished, so that the stability of the quality of the yellow peach fruit wine in the later period is ensured. Because the yellow peach fruit wine has low alcohol content and high residual sugar content, microorganisms in the product are still in an active period, and the stability in the later period is difficult to ensure, the freezing treatment after fermentation can better inhibit the continuous activity of the microorganisms such as residual saccharomycetes and the like, is favorable for accelerating the sedimentation of pulp suspended substances in the wine, and increases the stability of the pulp suspended substances; the low-temperature treatment has small influence on flavor components and overall aroma quality of the wine body, is beneficial to keeping fresh and mellow taste and improving the sensory quality of the yellow peach fruit wine; meanwhile, the cavity effect and the high-energy micro bubbles generated by the ultrasonic treatment of 40 kHz-50 kHz can enable the wine body to be in a transient high-temperature and high-pressure state, accelerate the breaking of microbial cells such as yeast and the like, and better ensure the microbial stability of the product.
A Hanseniaspora yeast is Hanseniaspora pseudoquillermonii HT9, and the preservation number of the Hanseniaspora pseudoquillermonii in China center for type culture Collection is CCTCC No: m2021444, the preservation unit address is located at Wuhan university in China, and the preservation date is 2021, 4 months and 25 days.
The Hansenula sporogenes of the invention has the colony morphology characteristics that: the strain is streaked and inoculated on a YPD (glucose 2%, yeast powder 1%, peptone 2% and agar 2%) culture medium, and cultured for 2d at 28 ℃, the colony is circular, the center point is slightly convex, the color is milky, the edge color is slightly light, the color is similar to a concentric circle, the surface is smooth, the diameter of the colony is 3-4 mm, the cell morphology is spindle-shaped, and the spore is produced by budding and reproduction. Culturing in malt extract agar culture medium and YPD agar culture medium at 28 deg.C for 3 days, and continuously transferring generation culture, wherein the culture characteristics and morphological characteristics have no obvious change, and the biological properties of the strain are basically stable.
The Hansenula sporogenes provided by the invention is subjected to 18S region rDNA sequencing, and the sequencing result is shown as SEQ NO: 1 is shown.
Drawings
FIG. 1 is a colony morphology map of Hansenula sporogenes of example 1 of the present invention.
Detailed Description
The invention is further described below with reference to the drawings and specific preferred embodiments of the description, without thereby limiting the scope of protection of the invention. The materials and equipment used in the following examples are commercially available.
Example 1:
the Hanseniaspora pseudochinensis (Hanseniaspora pseudoquillermonii) is Hanseniaspora pseudoquillermonii HT9, and the preservation number of the Hanseniaspora pseudoquillermonii in the China center for type culture Collection is CCTCC No: m2021444, the address of the preservation unit is located in the preservation center of Wuhan university, China, and the preservation date is 2021, 4 months and 25 days.
Separation, purification and identification of Hansenula persica HT 9:
and (3) taking 10g of soil and the surface of the yellow peach fruit respectively in 90mL of YPD liquid culture medium, carrying out shaking culture for 24h, taking supernatant liquid for gradient dilution, coating the supernatant liquid on the YPD liquid culture medium, and carrying out inverted culture for 2-3 d. Juicing the yellow peaches, putting the juiced yellow peaches into a sterile triangular flask, fermenting, coating the juiced yellow peaches on a YPD culture medium in a gradient dilution mode, and culturing for 2-3 days. The culture temperature was 28 ℃. And selecting colonies with typical yeast characteristics, carrying out plate streaking, purifying, inoculating to YPD slant culture for 12-24 h, and storing in a refrigerator at 4 ℃. And (3) subjecting the separated yeasts to primary screening (TTC color development method), secondary screening (Du's tube method), tertiary screening (sugar fermentation test) and quaternary screening (yellow peach fruit wine fermentation method), and finally obtaining a flavor-enhanced yeast HT9 which is identified as Hansheng yeasts with spores.
The strain is subjected to morphological observation and 18SrDNA amplification sequencing, and all identification results are comprehensively analyzed. The specific identification results are as follows:
morphological observation results: when the bacterial colony is cultured in YPD medium at 28 deg.c for 2 days, the colony is circular, slightly raised in the center, milky white, slightly light in the edge, concentric and smooth in surface. The diameter of the bacterial colony is 3 mm-4 mm, the bacterial colony is spindle-shaped when observed by a microscope, and the bacterial colony germinates and produces ascospores. FIG. 1 is a colony morphology map of Hansenula sporulata HT9 of the present invention. The strain is cultured in wort agar culture medium and YPD agar culture medium at 28 deg.c for 3 days separately, and through successive passage and generation culture, the strain has no obvious change in culture characteristic and morphological characteristic and stable biological characteristic.
18SrDNA amplification sequencing analysis and identification method and result: identification and sequencing by Shanghai Bioengineering Co., Ltd; the sequencing results were aligned with existing sequences in the NCBI database and confirmed to be a Hanseniaspora pseudoquillermonii (Hanseniaspora pseudoquillermonii). The sequence of the obtained 18SrDNA is shown as SEQ NO: 1 is shown.
Example 2:
the application of the Hansenula over spore yeast in the processing of yellow peach fruit wine adopts the Hansenula over spore yeast of the embodiment 1, and comprises the following steps:
s1, juicing the raw materials, protecting color and performing enzymolysis: after cleaning the yellow peaches, adopting a high-concentration ozone composite high-voltage pulse electric field to carry out pesticide residue degradation, namely, carrying out high-voltage pulse electric field treatment on the cleaned yellow peaches while carrying out ozone cleaning treatment in an ozone pool, wherein the concentration of ozone in the ozone pool is 10mg/L, the cleaning treatment time is 30min, and the conditions of the high-voltage pulse electric field treatment are as follows: the method comprises the steps of treating the yellow peach juice with the electric field intensity of 2kV/cm, the pulse time of 100 microseconds and the pulse number of 80 microseconds, removing cores and pulping to obtain the yellow peach juice, adding 0.8% of D-sodium erythorbate (namely accounting for 0.8% of the mass of the yellow peach juice), 0.5% of vitamin C and 2.3% of citric acid by mass percentage based on the mass of the yellow peach juice, protecting the color for 1 hour at 35 ℃, continuously adding 0.12% of cellulase and 0.1% of pectinase, carrying out enzymolysis for 2 hours at 45 ℃, and carrying out butterfly centrifugation to obtain the yellow peach juice.
S2, component adjustment: and (4) adding cane sugar and a fermentation aid into the yellow peach juice obtained in the step (S1), wherein the mass of the cane sugar accounts for 8% of the mass of the yellow peach juice, and the addition amount of the fermentation aid is 0.2g/L of the yellow peach juice, so that the yellow peach juice containing the cane sugar and the fermentation aid is obtained.
S3, yeast activation inoculation: culturing Hansenula sporogenes HT9 in YPD solid culture medium for 2-3 days, culturing in YPD liquid culture medium at 28 deg.C for 24 hr to obtain zymocyte liquid, centrifuging at 6-8 deg.C and 5000rpm/min for 5min, collecting thallus, washing with sterile water for 2 times, re-suspending with sterile water to obtain activated seed to be inoculated, and regulating the concentration of bacteria liquid to 106-7cfu/mL, the refrigeration temperature is 4-6 ℃; activating Hanseng yeast HT9 with spores, inoculating according to the proportion of 1% -2%, fermenting for 2-3 days, inoculating activated Saccharomyces cerevisiae D47 (produced by France Raman company), wherein the inoculation proportion is 2-5%, and the activated Saccharomyces cerevisiae is activated by water and then activated by fruit juice to obtain yellow peach juice after inoculation.
S4, fermentation: adopting ultrasonic auxiliary fermentation, pumping the inoculated yellow peach juice into a container for low-frequency ultrasonic intermittent treatment, keeping the flow rate of the yellow peach juice at 0.03-0.04 fermentation volume per second, keeping the ultrasonic power at 100W and the frequency at 10kHz, stopping the ultrasonic treatment for 6s every 0.5s, stopping the circulation after continuous treatment for 6h, entering main fermentation, fermenting for 4d at 18 ℃, and pouring for 2 times during the fermentation period.
S5, terminating fermentation: when the alcoholic strength of the yellow peach fermentation liquor reaches 10 degrees, adding 30mg/L sulfur dioxide, carrying out low-temperature treatment at-2 ℃ for 5 days, then carrying out ultrasonic treatment with the power of 300W and the frequency of 40kHz for 30min, separating and pouring to obtain the yellow peach wine base.
S6, glue adding and filtering: determining the glue adding amount through a stability experiment, adding bentonite into the yellow peach wine base by adopting bentonite with the adding amount of 55g/L, then filtering by adopting a cross-flow membrane, wherein the cross-flow membrane is a disc cross-flow membrane, the flow rate is 12m/s, the loss rate after the fruit wine is filtered is only 0.4%, and filling to obtain the yellow peach fruit wine.
Comparative example 1:
a method for processing yellow peach wine by saccharomyces cerevisiae, which is basically the same as the method in the embodiment 2, and is characterized in that: in step S3, only Saccharomyces cerevisiae D47 is used for fermentation, and mixed fermentation is not used.
The comparison of the volatile aroma of the yellow peach wine prepared in example 2 and the yellow peach wine prepared in comparative example 1 is shown in table 1.
Table 1 comparison of major volatile fragrance materials for example 2 and comparative example 1
As can be seen from Table 1, the volatile aroma content of the HT9+ D47 mixed fermented yellow peach wine of example 2 is significantly higher than that of the D47 fermented yellow peach wine of comparative example 1. Wherein esters and alcohols are increased more, ethyl acetate is increased from 18.6mg/L to 21.6mg/L, the ethyl acetate has pleasant flower and fruit fragrance and pineapple fragrance, and the threshold value is only 3.6 mu g/L, so that the ethyl acetate is an important contribution to fruit fragrance of fruit wine; other esters with significantly increased content include ethyl octanoate, ethyl hexanoate, ethyl decanoate, etc., most of which produce a pleasant fruit aroma. The ethyl caprylate has fruit fragrance such as fruit, brandy wine fragrance, mushroom, coconut, cream, fat fragrance and the like, the fragrance intensity of the ethyl caprylate is second to that of the ethyl caproate in esters, and the ethyl caprate has fruit fragrance and brandy wine fragrance, so that the yellow peach wine fragrance fermented by mixing the Hanseng spore yeast HT9 and the Saccharomyces cerevisiae D47 in example 2 is more complicated and rich.
The invention utilizes the screened Hanseng yeast with spores HT9 and the saccharomyces cerevisiae to mix and ferment to brew the yellow peach wine, firstly adopts ultrasonic-assisted fermentation to accelerate yeast proliferation and utilize substrate glucose, simultaneously reduces the generation of high-grade alcohol, adopts low-temperature and ultrasonic treatment after fermentation to the preset alcohol content, stops the fermentation of the yellow peach wine in time, and then obtains the yellow peach wine through glue clarification, cross-flow membrane filtration sterilization and finally filling. The application of the strain obviously improves the content of ester and other aroma substances, increases the complexity of wine body aroma, improves the quality of yellow peach wine, has golden color, strong fruit aroma, mellow wine body and harmonious aroma, has the typical characteristics of yellow peach fruit wine, has low high alcohol content, does not make people feel headache after drinking, and has no headache.
The foregoing is merely a preferred embodiment of the invention and is not intended to limit the invention in any manner. Although the present invention has been described with reference to the preferred embodiments, it is not intended to be limited thereto. Those skilled in the art can make many possible variations and modifications to the disclosed embodiments, or equivalent modifications, without departing from the spirit and scope of the invention, using the methods and techniques disclosed above. Therefore, any simple modification, equivalent replacement, equivalent change and modification made to the above embodiments according to the technical essence of the present invention are still within the scope of the protection of the technical solution of the present invention.
Sequence listing
<110> agricultural product processing research institute of Hunan province
<120> Hansheng yeast with spores and application thereof in yellow peach fruit wine
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<170> SIPOSequenceListing 1.0
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<212> DNA
<213> Hansenula polymorpha HT9 (Hanseniaspora pseudoquilliermonodii HT 9)
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Claims (10)
1. A Hanseniaspora pseudochinensis yeast is characterized in that the Hanseniaspora pseudoquilliermonodii is a Hanseniaspora pseudochinensis HT9, and the preservation number of the Hanseniaspora pseudolliermonodii in China center for type culture Collection is CCTCC No: m2021444.
2. Use of the Hansenula sordida yeast of claim 1 in the processing of yellow peach wine.
3. Use according to claim 2, characterized in that it comprises the following steps:
s1, juicing the raw materials, protecting color and performing enzymolysis: cleaning yellow peaches, reducing pesticide residues, removing cores and pulping to obtain yellow peach juice, taking the mass of the yellow peach juice as a reference, adding 0.7-1% of D-sodium erythorbate, 0.5-1% of vitamin C and 2-2.5% of citric acid by mass percent to protect the color, continuously adding 0.1-0.12% of cellulase and 0.1-0.15% of pectinase to carry out enzymolysis, and centrifuging to obtain the yellow peach juice;
s2, component adjustment: adding cane sugar and a fermentation aid into the yellow peach juice obtained in the step S1, wherein the addition amount of the cane sugar is 8-12% of the mass of the yellow peach juice, and the addition amount of the fermentation aid is 0.2-0.3 g/L of the yellow peach juice, so as to obtain the yellow peach juice containing cane sugar and the fermentation aid;
s3, yeast activation inoculation: the Hanseng sporulata yeast is cultured and activated by adopting an YPD solid culture medium and an YPD liquid culture medium in sequence, the Saccharomyces cerevisiae is activated by adopting water and fruit juice in sequence, the activated Hanseng sporulata yeast is inoculated into the yellow peach juice containing cane sugar and fermentation auxiliary agents obtained in the step S2, and after 2-3 days of fermentation, the activated Saccharomyces cerevisiae is inoculated;
s4, fermentation: carrying out ultrasonic auxiliary fermentation on the inoculated yellow peach juice, and then carrying out main fermentation;
s5, terminating fermentation: when the alcoholic strength of the yellow peach fermentation liquor reaches 10-11 degrees, adding sulfur dioxide, treating at the low temperature of-2-3 ℃, performing ultrasonic treatment, separating and pouring to obtain yellow peach wine base;
s6, glue adding and filtering: and (3) adding glue to the yellow peach wine base by using bentonite, filtering and filling to obtain the yellow peach fruit wine.
4. The application of claim 3, wherein in step S1, the pesticide residue is degraded by using an ozone composite high-voltage pulse electric field, that is, the cleaned yellow peaches are subjected to high-voltage pulse electric field treatment simultaneously with the ozone cleaning treatment, the concentration of the ozone is 5 mg/L-15 mg/L, the time of the ozone cleaning treatment is 15 min-30 min, and the conditions of the high-voltage pulse electric field treatment are as follows: the electric field intensity is 2 kV/cm-3 kV/cm, the pulse time is 50 mus-150 mus, and the number of pulses is 50-100.
5. The use according to claim 3, wherein in step S3, the specific process of yeast activation inoculation is as follows: fermenting Hansheng with sporeCulturing a mother in an YPD solid culture medium for 2-3 days, culturing in an YPD liquid culture medium at 26-28 ℃ for 18-24 hours to obtain a zymogen liquid, centrifuging the zymogen liquid at 6-8 ℃ for 3-5 minutes at the rotating speed of 5000-6000 rpm/min, collecting thalli, cleaning and re-suspending the thalli by using sterile water for 2-3 times, and adjusting the concentration of the bacteria liquid to 106-7cfu/mL to obtain activated Hanseng yeast with spore, and refrigerating and storing at 4-6 ℃.
6. The use of claim 5, wherein in step S3, the activated Hansenula sporogenes is inoculated at a ratio of 1-2%, the activated Saccharomyces cerevisiae is inoculated at a ratio of 2-5%, and the Saccharomyces cerevisiae is Saccharomyces cerevisiae D47.
7. The use of claim 3, wherein in step S4, the specific process of the ultrasonic-assisted fermentation is as follows: and (3) intermittently treating the yellow peach juice inoculated in the step (S3) by adopting low-frequency ultrasonic, wherein the ultrasonic power is 100-150W, the ultrasonic frequency is 10-15 kHz, the ultrasonic treatment is stopped for 6S every 0.5S, the circulation is stopped after the continuous treatment is carried out for 5-8 h, the yellow peach juice enters main fermentation, the yellow peach juice is fermented for 4-7 days at the temperature of 18-20 ℃, and the yellow peach juice is relatched for 2-3 times during the fermentation period.
8. The use according to any one of claims 3 to 7, wherein in step S5, the addition amount of sulfur dioxide is 30mg/L to 50mg/L, the time of the low-temperature treatment is 5 days to 7 days, the power of the ultrasonic treatment is 300W to 500W, the frequency of the ultrasonic treatment is 40kHz to 50kHz, and the time of the ultrasonic treatment is 30min to 40 min.
9. The use according to any one of claims 3 to 7, wherein in step S1, the color protection temperature is 35 ℃ to 40 ℃, the color protection time is 1h to 1.5h, the enzymolysis temperature is 40 ℃ to 45 ℃, the enzymolysis time is 2h to 3h, and the centrifugation is performed by a butterfly centrifugation method.
10. The use according to any one of claims 3 to 7, wherein in step S6, the bentonite is added in an amount of 50g/L to 60g/L, the filtration is performed by adopting cross-flow membrane filtration, the cross-flow membrane is a disc cross-flow membrane, and the filtration flow rate is 10m/S to 15 m/S.
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| CN106916758A (en) * | 2017-05-08 | 2017-07-04 | 中国农业大学 | A kind of Hansenula yeast and its application in wine production |
| CN112553091A (en) * | 2020-12-07 | 2021-03-26 | 贵州理工学院 | Non-saccharomyces cerevisiae and fermentation method for increasing fragrance of blueberry fruit wine by using same |
| CN112625928A (en) * | 2021-01-15 | 2021-04-09 | 江南大学 | Hansenula polymorpha strain capable of increasing wine brewing aroma |
| CN112746029A (en) * | 2021-01-22 | 2021-05-04 | 西北农林科技大学 | Hansenula polymorpha strain QTX22 for producing aroma substances at high yield and application thereof |
| CN112812980A (en) * | 2021-02-19 | 2021-05-18 | 宁夏农产品质量标准与检测技术研究所(宁夏农产品质量监测中心) | Mixed fermentation process based on Hansenula polymorpha and saccharomyces cerevisiae |
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| CN106916758A (en) * | 2017-05-08 | 2017-07-04 | 中国农业大学 | A kind of Hansenula yeast and its application in wine production |
| CN112553091A (en) * | 2020-12-07 | 2021-03-26 | 贵州理工学院 | Non-saccharomyces cerevisiae and fermentation method for increasing fragrance of blueberry fruit wine by using same |
| CN112625928A (en) * | 2021-01-15 | 2021-04-09 | 江南大学 | Hansenula polymorpha strain capable of increasing wine brewing aroma |
| CN112746029A (en) * | 2021-01-22 | 2021-05-04 | 西北农林科技大学 | Hansenula polymorpha strain QTX22 for producing aroma substances at high yield and application thereof |
| CN112812980A (en) * | 2021-02-19 | 2021-05-18 | 宁夏农产品质量标准与检测技术研究所(宁夏农产品质量监测中心) | Mixed fermentation process based on Hansenula polymorpha and saccharomyces cerevisiae |
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| The Sensory Quality Improvement of Citrus Wine through Co-Fermentations with Selected Non-Saccharomyces Yeast Strains and Saccharomyces cerevisiae;Lanlan Hu等;《Microorganisms》;20020226;第8卷(第323期);1-17页 * |
| 靖州杨梅果酒发酵菌种筛选、鉴定及发酵性能研究;阳秀莲 等;《食品与机械》;20190131;第35卷(第3期);41-47页 * |
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