CN114574375B - Saccharomyces cerevisiae, starter and application thereof in preparation of fermented food - Google Patents
Saccharomyces cerevisiae, starter and application thereof in preparation of fermented food Download PDFInfo
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- CN114574375B CN114574375B CN202210216386.4A CN202210216386A CN114574375B CN 114574375 B CN114574375 B CN 114574375B CN 202210216386 A CN202210216386 A CN 202210216386A CN 114574375 B CN114574375 B CN 114574375B
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- saccharomyces cerevisiae
- starter
- fermentation
- fermented
- whiskey
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Abstract
The invention relates to the field of fermentation, and discloses saccharomyces cerevisiae, a starter and application thereof in preparation of fermented food. The preservation number of the saccharomyces cerevisiae is CGMCC No.21165. The saccharomyces cerevisiae has the advantages of high fermentation starting speed and high alcohol yield, and can rapidly and fully utilize maltose, isomaltose and maltotriose, and high yield of ethyl decanoate. The strain is particularly suitable for brewing whiskey, and the brewed whiskey has mellow taste and sweet and refreshing taste and has the special flavor of whiskey.
Description
Technical Field
The invention relates to the field of fermentation, and discloses a saccharomyces cerevisiae, a starter and a preparation method of the starter, wherein the saccharomyces cerevisiae or the application of the starter in preparation of fermented food and a brewing method of whiskey.
Background
Whiskey is distilled liquor prepared from malt and grains by saccharification, fermentation, distillation, ageing and blending. Glucose and fructose obtained by saccharification of malt and cereal raw materials easily permeate into yeast cells and can be directly absorbed and fermented by yeast, but the utilization of maltose, isomaltose and maltotriose requires the action of maltose penetrating enzyme, maltotriose penetrating enzyme, maltase and the like, and the absorption and utilization efficiency of saccharomyces cerevisiae is low and the fermentation speed is low. The fermentation time of whiskey is 2-3d, and the main basis for whiskey yeast selection is the speed and ability of sugar to alcohol.
Yeast is an important microorganism for whisky fermentation, and can be used for converting sugar into alcohol, and can also produce substances such as organic acid, esters, aldehydes, higher alcohols and the like, so that the yeast is an important source of aroma compounds. It is considered that higher fatty acid ethyl ester in whiskey is the main component of the fragrance, and that ethyl decanoate is the main fragrance component.
Therefore, the utilization rate and the utilization speed of maltose, isomaltose and maltotriose by yeasts and the yield of ethyl decanoate are important indexes for whiskey yeast breeding, which are helpful for improving the wine yield, saving grain raw materials and improving the product quality.
Disclosure of Invention
The invention aims to provide a saccharomyces cerevisiae, which is used for brewing whiskey wine, and has the advantages of high utilization rate of maltose, isomaltose and maltotriose, high fermentation speed and high alcohol production capacity; and the output of ethyl decanoate is high; the invention also provides a starter, a preparation method of the starter, application of the saccharomyces cerevisiae or the starter in preparation of fermented food, and a brewing method of whiskey.
In order to achieve the above purpose, the first aspect of the present invention provides a strain of saccharomyces cerevisiae (Saccharomyces cerevisiae), which has a preservation number of CGMCC No.21165.
In a second aspect the invention provides a starter comprising Saccharomyces cerevisiae (Saccharomyces cerevisiae) as described above.
In a third aspect, the present invention provides a process for the preparation of a starter comprising fermentation culturing a Saccharomyces cerevisiae as described above in a fermentation medium.
In a fourth aspect the present invention provides the use of a Saccharomyces cerevisiae as described above or a starter as described above in the preparation of a fermented food product.
A fifth aspect of the present invention provides a method of brewing whiskey, the method comprising: contacting the Saccharomyces cerevisiae or the starter as described above with the saccharifying liquid and fermenting to obtain fermented mash; distilling the fermented liquor for at least 2 times, taking wine in sections, selecting section fractions, adjusting the alcohol content to 50-80% by volume, and aging in a oak barrel.
The beneficial effects of the invention are as follows:
the saccharomyces cerevisiae is a safe strain which is screened from old yeast reserved in Gansu farmers and can be used for food, can be widely applied to the field of fermented products, and can also be used for preparing a starter.
The saccharomyces cerevisiae has the advantages of high utilization rate of maltose, isomaltose and maltotriose, high fermentation speed and high alcohol yield.
The brewing yeast of the invention is used for brewing whiskey, the content of ethyl decanoate in the final fermented mash is high, and the distilled yeast has the special flavor of whiskey.
The saccharomyces cerevisiae CGMCC No.21165 is used as a starter, is easy to culture and prepare, and has high concentration of the starter saccharomyces cerevisiae viable bacteria.
Additional features and advantages of the invention will be set forth in the detailed description which follows.
Preservation of organisms
The strain provided by the invention is classified and named as Saccharomyces cerevisiae Saccharomyces cerevisiae, and is preserved in China general microbiological culture Collection center (CGMCC) for 11 months in 2020, wherein the preservation number is CGMCC No.21165, and the preservation address is North Chen West Lu No. 1 and No. 3 in the Korean region of Beijing city.
Drawings
In order that the invention may be more readily understood, a more particular description of the invention will be rendered by reference to specific embodiments thereof which are illustrated in the appended drawings, in which,
FIG. 1 shows the colony morphology of Saccharomyces cerevisiae on a culture medium according to the present invention;
FIG. 2 shows the morphology of Saccharomyces cerevisiae according to the present invention as observed by a microscope.
Detailed Description
The endpoints and any values of the ranges disclosed herein are not limited to the precise range or value, and are understood to encompass values approaching those ranges or values. For numerical ranges, one or more new numerical ranges may be found between the endpoints of each range, between the endpoint of each range and the individual point value, and between the individual point value, in combination with each other, and are to be considered as specifically disclosed herein.
The first aspect of the invention provides a strain of saccharomyces cerevisiae Saccharomyces cerevisiae, and the preservation number of the saccharomyces cerevisiae is CGMCC No.21165.
The saccharomyces cerevisiae is obtained by screening old yeast reserved in Gansu farmers, and the method comprises the following steps: taking old ferment, and gradient diluting to 10 with sterile physiological saline -6 Each dilution gradient YPD plate was incubated at 28.+ -. 1 ℃ for 72h. The inoculating needle picks out the colonies with different colony forms to the YPD plate for streaking until single colony with uniform size and uniform form appears. Selecting bacterial strains with round and oval cell morphology and budding reproduction. The strain which is separated into the tentative saccharomycetes is activated for 3 generations in YPD liquid culture medium, then physiological and biochemical identification and molecular biological identification are carried out, and a plurality of aspects such as growth, fermentation performance, appearance quality characteristics, texture characteristics, fermentation flavor substances, sensory quality and the like of the saccharomycetes are researched, and after a plurality of rounds of researches and demonstration,the saccharomyces cerevisiae is finally obtained by screening a plurality of wild yeasts.
The screened Saccharomyces cerevisiae is cultured on YPD solid culture medium for 48 hours at 28+/-1 ℃, as shown in figure 1, colonies with the diameter of about 3.0mm can be grown, and the colonies are glossy yellow white dots, have regular edges and uniform texture and are easy to pick. As shown in FIG. 2, the cell morphology under the microscope of Saccharomyces cerevisiae is oval, and the diameter size is 1.5-3.0 μm.
The saccharomyces cerevisiae disclosed by the invention also has the following advantages:
(1) The sugar utilization rate is high, the maltose conversion rate can reach 98.90%, the isomaltose conversion rate can reach 99.07%, and the maltotriose conversion rate can reach 90.42%.
(2) The fermentation speed is high: fermenting at 30 ℃ for 2h, wherein the fermentation speed is high, the fermentation time is short, and the fermentation time is 38h.
(3) The alcohol yield is high: the predicted alcohol yield was 438.61LA/T and the actual alcohol yield was 382.80 LA/T.
(4) Has special flavor of whisky: the final fermentation broth ethyl decanoate yield was 20.36mg/100mL. The distilled whiskey has mellow taste and sweet and refreshing taste, and has the special flavor of whiskey.
The method for culturing the saccharomyces cerevisiae provided by the invention is not particularly required, as long as the saccharomyces cerevisiae can be proliferated, for example, the living saccharomyces cerevisiae can be inoculated into a saccharomyces cerevisiae culture medium, and the culture is carried out for 12-48 hours at a temperature of 20-40 ℃ under aerobic conditions, so that a culture solution is obtained. The Saccharomyces cerevisiae medium may be any of a variety of suitable media known in the art for culturing Saccharomyces cerevisiae, and may be, for example, molasses medium, 5-10 Bessel medium or YPD medium.
The preparation method of the molasses culture medium comprises the following steps: the molasses is diluted to 6-10 DEG Be, and 1-3g/L yeast powder and 0.2-0.8g/L ammonium sulfate are added. Then, the mixture can be prepared by autoclaving at 115 ℃ for 15min.
The wort medium may be prepared in a manner conventional in the art, for example, by comminuting barley malt according to 1:3-5, wherein the water ratio is added, the temperature is kept at 42-48 ℃ for 20-40min, the temperature is kept at 62-66 ℃ for 30-50min, and the temperature is kept at 68-72 ℃ for 15-25min. It is to be understood that the raw materials for preparing the wort medium may also include, for example, rye, corn, wheat and the like.
Wherein the YPD medium formulation preferably comprises: 5-15g/L of yeast extract, 15-25g/L of peptone and 15-25g/L of glucose. The YPD medium can be sterilized at 118-123 deg.C for 15-25min. When the medium is a solid medium, it preferably further comprises agar 15-20g/L.
The method of the present invention is not particularly limited as long as it is capable of enriching the cells from the culture solution, and may be, for example, a centrifugation and/or filtration method, and the centrifugation (for example, centrifugation at 1000-12000rpm for 5-20min in a refrigerated centrifuge to obtain a cell pellet) and the filtration conditions may be known conditions.
In a second aspect the invention provides a starter comprising Saccharomyces cerevisiae (Saccharomyces cerevisiae) as described above.
The form of the starter according to the present invention may be not particularly limited, for example, the starter may be a liquid starter, a semi-liquid starter, or a solid starter.
Preferably, the viable bacteria concentration of the Saccharomyces cerevisiae in the starter is 10 8 CFU/g or more.
The preparation method may be a preparation method conventional in the art.
In a third aspect, the present invention provides a process for the preparation of a starter comprising fermentation culturing a Saccharomyces cerevisiae as described above in a fermentation medium.
According to the present invention, the fermentation medium may be various media conventionally used in the art for culturing Saccharomyces cerevisiae, for example, various media suitable for culturing Saccharomyces cerevisiae such as YPD medium, wort medium and molasses medium as described above.
According to the invention, saccharomyces cerevisiae CGMCC No.21165 can be fermented in a fermentation medium to make the number of viable bacteria reach 10 8 The fermentation liquid obtained by CFU/mL can be used as a liquid fermentation agent.
Preferably, the conditions of the fermentation culture include: the temperature is 20-40 ℃, the pH is 2.5-6, and the time is 24-36h.
Concentrating the fermentation liquor to obtain semi-liquid leavening agent; the concentration method can be a conventional technical means in the art, so long as the concentration of the fermentation broth can be realized, for example, centrifugation, filtration and the like, so long as the activity of the saccharomyces cerevisiae is not significantly affected.
The centrifugation of the fermentation broth may be carried out according to a method conventional in the art, for example, the cell pellet may be obtained by centrifugation in a refrigerated centrifuge at 1000-12000rpm for 5-20min.
Concentrating the fermentation broth, washing with buffer solution, adding protective agent, and adjusting viable bacteria concentration to 10 10 And (3) mixing the materials uniformly above CFU/g, and drying to obtain the solid fermentation agent.
The buffer may be a buffer conventional in the art, for example, PBS buffer or physiological saline.
Such drying means include, but are not limited to, lyophilization, drying, air drying, vacuum drying, spray drying, and the like.
The protective agent may be various protective agents conventional in the art, and for example, may be at least one of skim milk powder, maltodextrin, trehalose, dextran, glycerin, and the like. The amount of the protective agent can be selected and adjusted as desired by those skilled in the art.
In a fourth aspect the present invention provides the use of a Saccharomyces cerevisiae as described above or a starter as described above in the preparation of a fermented food product.
Preferably, the fermented food is alcoholic fermented food or fermented pasta.
Preferably, the alcoholic fermented food product is whiskey.
Preferably, the fermented pasta is a fermented bread.
The raw materials and the preparation method of the fermented bread are all conventional in the art, and are not described herein.
The fermentation of whisky can be to inoculate Saccharomyces cerevisiae CGMCC No.21165 into raw materials (such as wort) to be treated according to the conventional use in the conventional production process of fermenting whisky, and ferment or survive under the temperature and pressure capable of propagating the Saccharomyces cerevisiae CGMCC No.21165. Through adding CGMCC No.21165 into the fermentation substrate, the metabolite thereof ensures that the fermented wine has a certain excellent characteristic of appearance, fragrance and the like, and improves the organoleptic quality characteristics of the product.
A fifth aspect of the present invention provides a method of brewing whiskey, the method comprising: contacting the Saccharomyces cerevisiae or the starter as described above with the saccharifying liquid and fermenting to obtain fermented mash; distilling the fermented liquor for at least 2 times, taking wine in sections, selecting section fractions, adjusting the alcohol content to 50-80% by volume, and aging in a oak barrel.
The conditions of the fermentation may be conventional conditions for whiskey fermentation culture known in the art, for example, the temperature of the fermentation may be 20-40 ℃.
The saccharification liquid may be a saccharification liquid obtained by saccharification of conventional grains in the art, and may be prepared from barley malt as a main raw material, or may be prepared by blending other grains such as corn, wheat, rye, and the like as raw materials, and may be selected by those skilled in the art according to desired flavors. The specific preparation method of whiskey may be performed according to a method conventional in the art, and will not be described herein.
The medium formulations referred to in the examples below were as follows:
YPD medium (wt%): yeast powder (1%), peptone (2%), glucose (2%), agar (2%), and heat to dissolve, and autoclaving at 121 ℃ for 15-20min.
Saccharomyces cerevisiae medium: barley malt was crushed according to 1:4, adding water, maintaining at 45deg.C for 30min, at 65deg.C for 40min, at 70deg.C for 20min, filtering, adding agar (2%), and autoclaving at 115deg.C for 15min to obtain Saccharomyces cerevisiae culture medium.
Molasses medium: the molasses is diluted to 8-10 DEG Be, 2g/L of yeast powder and 0.5g/L of ammonium sulfate are added, and the mixture is autoclaved for 15min at 115 ℃.
The control strain was beer active dry yeast purchased from Angel yeast.
The reagents and materials used were all commercially available without specific description.
Unless otherwise indicated, the operations are all conventional in the art.
Example 1
The embodiment is used for explaining the separation, purification and characteristic identification of the saccharomyces cerevisiae CGMCC No.21165.
The Saccharomyces cerevisiae CGMCC No.21165 is separated from old yeast reserved in Gansu farmers.
Taking old ferment, and gradient diluting to 10 with sterile physiological saline -6 Each dilution gradient YPD plate was incubated at 28.+ -. 1 ℃ for 72h. The inoculating needle picks out the colonies with different colony forms to the YPD plate for streaking until single colony with uniform size and uniform form appears.
Selecting bacterial strains with round and oval cell morphology and budding reproduction. The strain which is separated into the tentative saccharomycetes is activated for 3 generations in YPD liquid culture medium, physiological and biochemical identification and molecular biological identification are carried out, and a plurality of aspects such as the growth, fermentation performance, appearance quality characteristics, texture characteristics, fermentation flavor substances, sensory quality and the like of the saccharomycetes are researched, and a strain of saccharomyces cerevisiae is finally screened from a plurality of wild saccharomycetes through multiple rounds of research and demonstration.
(1) Morphological identification
The screened Saccharomyces cerevisiae is cultured on YPD solid culture medium for 48 hours at 28+/-1 ℃, as shown in figure 1, colonies with the diameter of about 3.0mm can be grown, and the colonies are glossy yellow white dots, have regular edges and uniform texture and are easy to pick. As shown in FIG. 2, the cell morphology under the microscope of Saccharomyces cerevisiae is oval, and the diameter size is 1.5-3.0 μm.
(2) Physiological and biochemical identification
The screened strains were subjected to physiological and biochemical identification using the French Mei Liai API identification system, and the identification results are shown in Table 1 below. The strain is Saccharomyces cerevisiae (Saccharomyces cerevisiae) after physiological and biochemical identification.
TABLE 1
| Project | Project | Results | Project | Project | Results | Project | Project | Results |
| 0 | None | - | XLT | Xylitol | - | CEL | Cellobiose | - |
| GLU | D-glucose | + | GAL | D-galactose | + | LAC | D-lactose | - |
| GLY | Glycerol | - | INO | Inositol (inositol) | - | MAL | D-maltose | + |
| 2KG | 2-ketogluconate | - | SOR | Sorbitol | - | SAC | D-sucrose | + |
| ARA | L-arabinose | - | MDG | alpha-methyl-D-glucose | + | TRE | Trehalose | + |
| XYL | D-xylose | - | NAG | N-acetyl-glucoside | - | MLZ | D-pine triple sugar | + |
| ADO | Adonis amurensis L | - | RAF | D-raffinose | + |
Note [ ]. In the table, "+" indicates positive biochemical reaction results, and "-" indicates negative biochemical reaction results.
(3) Molecular characterization
Cloning and sequencing the ITS1/ITS4 of the isolated strain, wherein the nucleotide sequence of the ITS1/ITS4 gene is as follows, and comparing the ITS1/ITS4 sequence of the strain with the sequence of saccharomyces cerevisiae, wherein the similarity of the ITS1/ITS4 sequence of the strain with the Saccharomyces cerevisiae sequence reaches 99.99%.
TTAAGTTTCTTTCTTGCTATTCCAAACGGTGAGAGATTTCTGTGCTTTTGTTAT AGGACAATTAAAACCGTTTCAATACAACACACTGTGGAGTTTTCATATCTTTGCAA CTTTTTCTTTGGGCATTCGAGCAATCGGGGCCCAGAGGTTAACAAACACAAACAAT TTTATCTATTCATTAAATTTTTGTCAAAAACAAGAATTTTCGTAACTGGAAATTTTA AAATATTAAAAACTTTCAACAACGGATCTCTTGGTTCTCGCATCGATGAAGAACGC AGCGAAATGCGATACGTAATGTGAATTGCAGAATTCCGTGAATCATCGAATCTTTG AACGCACATTGCGCCCCTTGGTATTCCAGGGGGCATGCCTGTTTGAGCGTCATTTC CTTCTCAAACATTCTGTTTGGTAGTGAGTGATACTCTTTGGAGTTAACTTGAAATTG CTGGCCTTTTCATTGGATGTTTTTTTTTTCCAAAGAGAGGTTTCTCTGCGTGCTTGA GGTATAATGCAAGTACGGTCGTTTTAGGTTTTACCAACTGCGGCTAATCTTTTTTAT ACTGAGCGTATTGGAACGTTATCGATAAGAAGAGAGCGTCTAGGCGAACAATGTT CTTAAAGTTGACCTC
The isolated strain is determined as Saccharomyces cerevisiae (Saccharomyces cerevisiae) to be preserved in China general microbiological culture Collection center, with preservation address: the collection number of the microbiological institute of China is CGMCC No.21165, and the collection date is 11 months 11 days in 2020.
Example 2
The embodiment is used for explaining the application research of saccharomyces cerevisiae in whisky fermentation.
(1) Preparing an activated saccharomycete liquid: inoculating the yeast strain CGMCC No.21165 of the invention into a Saccharomyces cerevisiae culture medium, and culturing at 28 ℃ to 10 8 And (3) obtaining an activated liquid by CFU/mL or more.
Brewage of whiskey: selecting whisky malt, crushing, preserving heat and saccharifying until the specific gravity of the wort is 1.045 at 65 ℃ and the ratio of raw materials to water is 1:4, and obtaining the whisky fermented wort.
Inoculating the activated yeast liquid into the whiskey fermented wort with 10% of inoculation amount, and fermenting at 32 ℃. Determination of CO Using a weighing method 2 Loss of weight as CO 2 The loss of weight within 0.1 g/day indicates the end of fermentation. After fermentation, fermenting mash is distilled for 2 times, wine is taken in sections, the section fraction is selected, and after the alcohol content is adjusted to 65%, the oak barrel is aged.
The saccharomyces cerevisiae of the invention starts fermentation after 2 hours of brewing and inoculation, and reaches the fermentation end at the 38 th fermentation time. The control strain starts fermentation after 6 hours of inoculation, and the fermentation is completed for 44 hours to reach the fermentation end point.
(2) Determination of predicted alcohol yield:
preparing malt flour: the disc spacing of the DLFU disc standard mill was adjusted to 0.7mm, and the distilled malt was crushed to prepare malt flour.
Preparation of saccharified wort: saccharified wort was prepared using the EBC recommended hot water extraction method (hot water saccharification at constant temperature of 65℃for 1 h). The wort demand was 250mL, and the wort specific gravity (initial specific gravity, OG) at 20℃was measured.
Inoculating: 1.25 (+ -0.1) g of yeast is weighed into a 100mL conical flask (proportionally reducing the amount of yeast used if the total amount of wort is less than 250 mL), 30mL of saccharified wort is transferred into a 100mL conical flask containing yeast, the resulting bacterial suspension is stirred with a sterile glass rod, then transferred into a 500mL conical flask, the small conical flask is washed three times with the appropriate amount of saccharified wort, the washing solution is incorporated into the large conical flask, and the remaining wort is poured into it together.
Fermentation: fermenting in water bath at 33 (+ -0.2) deg.C for 44 hr until the water level is equal to that of the conical flask. Or placing the mixture in a constant temperature incubator for culturing after preheating.
And (3) filtering: after fermentation, the broth was cooled to 20 ℃, filtered, and the first 50mL was returned to the re-filtration, during which time the funnel was covered with a petri dish and a 250mL graduated cylinder was used as the recipient. The specific gravity of the fermentation broth at 20℃was determined.
FE (fermentable extraction rate,%):
FE (fermentable extraction rate) = (f×se)/100
F (fermentability) = ((OG-FG)/OG) ×100×0.814)
SE (soluble extraction rate,%) = (og× 2.279)/SG Initial initiation
OG＝1000×(SG Initial initiation -1)
FG＝1000×(SG Finally (3) -1)
Wherein, OG: the initial specific gravity, reserving five decimal places; FG: the specific gravity is finished, and five decimal places are reserved; SG: the specific gravity was measured.
The predicted alcohol yield (PSY) is calculated according to the following formula:
PSY(LA/T)＝FE×6.06
wherein, FE: fermentable extraction (%); 6.06: experience coefficient.
The predicted alcohol yield of the brewing whiskey using the Saccharomyces cerevisiae of the present invention was calculated to be 438.61LA/T and the predicted alcohol yield of the control strain was calculated to be 403.69LA/T.
(3) Measurement of the actual alcohol yield:
experimental materials
High performance liquid chromatography, chromatographic column: aminex HPX-87H (300 mm. Times.7.8 mm)
Experimental method
Liquid phase conditions: column temperature: 65 ℃; mobile phase 0.005moL/L H 2 SO4 at a flow rate of 0.600 mL/min. The detector is a differential detector; the fermentation broth was filtered through a 0.45 μm pore size filter, and added to a sample bottle with a sample volume of 20. Mu.L.
The experimental results are as follows: the actual alcohol yield of brewing whiskey using the Saccharomyces cerevisiae of the present invention was 382.80LA/T, and the predicted alcohol yield of the control strain was 344.40LA/T.
As can be seen from the data, the Saccharomyces cerevisiae has the advantages of high fermentation speed, short fermentation time and high alcohol yield.
Example 3
This example is a study to demonstrate the determination of the utilization of maltose, isomaltose and maltotriose by Saccharomyces cerevisiae.
(1) Experimental materials
High performance liquid chromatography, chromatographic column: aminex HPX-87H (300 mm. Times.7.8 mm)
(2) Experimental method
Liquid phase conditions: column temperature: 65 ℃; mobile phase 0.005moL/L H 2 SO4 at a flow rate of 0.600 mL/min. The detector is a differential detector; the sample loading was 20. Mu.L.
The experimental results are as follows:
in the whisky brewed wort, the maltose content is 207.54mg/mL, the isomaltose content is 54.64mg/mL and the maltotriose content is 11.06mg/mL.
After fermentation, the content of maltose in the fermented mash of the saccharomyces cerevisiae is 2.28 mg/mL, the content of isomaltose is 0.51mg/mL, the content of maltotriose is 1.06mg/mL, namely the maltose conversion rate is 98.90%, the isomaltose conversion rate is 99.07%, and the maltotriose conversion rate is 90.42%.
Whereas the control strain had a maltose conversion of 90.60%, an isomaltose conversion of 89.00% and a maltotriose conversion of 80.02%. The saccharomyces cerevisiae disclosed by the invention has higher utilization rate of maltose, isomaltose and maltotriose.
Example 4
This example is used to demonstrate the ability of Saccharomyces cerevisiae to produce ethyl decanoate
(1) Experimental materials
An ultrasonic cleaner and a headspace solid-phase microextraction automatic sample injection system; SPME fiber: carboxen/DVB/PDMS,50/30,2cm; column DB-WAX 57CB column (50 m ×0.25mm,0.2 μm, agilent Co., U.S.) or equivalent column.
(2) Analytical procedure
Sample preparation: accurately transferring 5mL to 20mL of prepared fermented mash into a screw head space sample injection bottle. Accurately adding 150 mu L of trichloropropane (10 mg/L) internal standard solution, shaking uniformly and measuring.
B, sample measurement:
reference chromatographic conditions:
a, headspace solid-phase microextraction extraction conditions: and (3) taking a prepared headspace test sample, placing the prepared headspace test sample in a headspace solid-phase microextraction automatic sampling system, and vibrating at a constant temperature of 60 ℃ for 40 minutes to balance the concentration of the headspace flavor substances. SPME-fiber is inserted, extracted for 30min, and then inserted into the sample inlet for 3min.
b gas chromatograph-mass spectrometer parameters: sample inlet: the temperature is 250 ℃, the flow rate is 1mL/min, and the flow is not divided into a flow distribution mode.
c, temperature programming conditions:
d mass spectrometry conditions: the ion source is electron bombardment source (EI), the temperature of the ion source is set to 230 ℃, the temperature of the quadrupole is set to 150 ℃, the electron energy is 70ev, the interface temperature is set to 280 ℃, and the scanning range of the mass spectrum is 30-500Da.
C, quality control: the blank signal was measured 2 times using a blank headspace bottle beginning with each measurement. Monthly n-alkane samples were used to determine if the method retention time was normal. The content of the internal standard peak area in the sample is observed, for example, the deviation is more than 50 percent, and the state of the instrument is required to be checked or the extraction fiber is required to be replaced.
(3) Result calculation
The type of flavor chemical in the sample is characterized by means of NIST spectrogram library retrieval. And the relative concentration was calculated from the ratio of the compound peak area to the internal standard peak area.
Wherein:
x-the relative content of flavor compounds in the sample in mg/L;
vi-the volume of the internal standard added to the sample in mL;
vs-volume of sample solution added to headspace bottle, unit mL;
ci-concentration of internal standard solution in mg/L;
ii-internal standard peak area intensity;
is-flavor compound peak area intensity.
(4) Detection result
According to the determination, the brewing whiskey of the saccharomyces cerevisiae disclosed by the invention has the ethyl decanoate content of 20.36mg/100mL, and the control strain is 8.00mg/100mL. As can be seen by comparison, the special flavor of the whisky can be obtained when the whisky is brewed by the saccharomyces cerevisiae. In addition, the whiskey wine brewed by the saccharomyces cerevisiae has mellow taste, is sweet and refreshing, and has better flavor.
Example 5
This example is used to illustrate the study of the use of Saccharomyces cerevisiae in fermented bread.
Fermented bread was prepared using the Saccharomyces cerevisiae and the control strain of the present invention, respectively, according to the following procedure.
The steps are as follows: weighing ingredients, stirring dough to form a film, standing, cutting into blocks, rounding, standing, exhausting and shaping, proofing and baking.
The tissue state of the bread was observed and the toast texture was measured. After cooling the baked bread for 1 hour, it was cut into toast slices 15mm thick.
The prepared bread was evaluated according to the bread sensory evaluation table of table 2, and the bread sensory evaluation scores are shown in table 3.
TABLE 2
TABLE 3 Table 3
| Strain | CGMCC No.21165 | Control strain |
| Color | 8 | 8 |
| Morphology of the product | 8 | 7 |
| Flavor of | 9 | 8 |
| Mouthfeel of the product | 9 | 8 |
| Total score | 34 | 31 |
From table 3, it can be seen that by performing sensory scoring on the bread fermented by different leaven agents, different saccharomyces cerevisiae affects the experimental result to a great extent under the same fermentation condition, wherein the bread fermented by CGMCC No.21165 has full and complete appearance, smooth and unbroken surface, proper cavity size, softness and palatability, elasticity, no sticking to teeth, and composite aroma of baked flavor of bread, grains and whiskey.
The preferred embodiments of the present invention have been described in detail above, but the present invention is not limited thereto. Within the scope of the technical idea of the invention, a number of simple variants of the technical solution of the invention are possible, including combinations of the individual technical features in any other suitable way, which simple variants and combinations should likewise be regarded as being disclosed by the invention, all falling within the scope of protection of the invention.
SEQUENCE LISTING
<110> middle grain group Co., ltd
COFCO NUTRITION AND HEALTH RESEARCH INSTITUTE Co.,Ltd.
<120> Saccharomyces cerevisiae, starter and their use in the preparation of fermented foods
<130> 72853
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 632
<212> DNA
<213> Saccharomyces cerevisiae
<400> 1
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tctaggcgaa caatgttctt aaagttgacc tc 632
Claims (10)
1. A strain of Saccharomyces cerevisiae (Saccharomyces cerevisiae) is characterized in that the preservation number of the Saccharomyces cerevisiae is CGMCC No.21165.
2. A starter culture comprising the saccharomyces cerevisiae of claim 1.
3. The starter according to claim 2, wherein the viable bacteria concentration of the saccharomyces cerevisiae in the starter is 10 8 CFU/g or more.
4. A starter according to claim 2 or 3, wherein the starter is a liquid starter, a semi-liquid starter or a solid starter.
5. A process for producing a starter, which comprises culturing the Saccharomyces cerevisiae of claim 1 in a fermentation medium.
6. The method of claim 5, wherein the fermentation culture conditions comprise: the temperature is 20-40 ℃, the pH is 2.5-6, and the time is 24-36h.
7. Use of the saccharomyces cerevisiae of claim 1 or the starter of any of claims 2-4 in the preparation of fermented food.
8. The use according to claim 7, wherein the fermented food is an alcoholic fermented food or a fermented pasta.
9. A method of brewing whisky, the method comprising: contacting the saccharomyces cerevisiae of claim 1 and/or the starter of any of claims 2-4 with a saccharification liquid and fermenting to obtain a fermented mash; distilling the fermented liquor for at least 2 times, taking wine in sections, selecting section fractions, adjusting the alcohol content to 50-80% by volume, and aging in a oak barrel.
10. The method of claim 9, wherein the fermentation temperature is 20-40 ℃.
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