LU501542B1 - Yeast tolerant to high sugar and low ph, and application thereof - Google Patents
Yeast tolerant to high sugar and low ph, and application thereof Download PDFInfo
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- LU501542B1 LU501542B1 LU501542A LU501542A LU501542B1 LU 501542 B1 LU501542 B1 LU 501542B1 LU 501542 A LU501542 A LU 501542A LU 501542 A LU501542 A LU 501542A LU 501542 B1 LU501542 B1 LU 501542B1
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- 240000004808 Saccharomyces cerevisiae Species 0.000 title claims abstract description 71
- 238000000855 fermentation Methods 0.000 claims abstract description 76
- 230000004151 fermentation Effects 0.000 claims abstract description 69
- 238000009923 sugaring Methods 0.000 claims abstract description 60
- 235000013399 edible fruits Nutrition 0.000 claims abstract description 54
- 241001092040 Crataegus Species 0.000 claims abstract description 53
- 235000014493 Crataegus Nutrition 0.000 claims abstract description 28
- 235000009917 Crataegus X brevipes Nutrition 0.000 claims abstract description 27
- 235000013204 Crataegus X haemacarpa Nutrition 0.000 claims abstract description 27
- 235000009685 Crataegus X maligna Nutrition 0.000 claims abstract description 27
- 235000009444 Crataegus X rubrocarnea Nutrition 0.000 claims abstract description 27
- 235000009486 Crataegus bullatus Nutrition 0.000 claims abstract description 27
- 235000017181 Crataegus chrysocarpa Nutrition 0.000 claims abstract description 27
- 235000009682 Crataegus limnophila Nutrition 0.000 claims abstract description 27
- 235000004423 Crataegus monogyna Nutrition 0.000 claims abstract description 27
- 235000002313 Crataegus paludosa Nutrition 0.000 claims abstract description 27
- 235000009840 Crataegus x incaedua Nutrition 0.000 claims abstract description 27
- 238000000034 method Methods 0.000 claims abstract description 15
- 239000000463 material Substances 0.000 claims abstract description 12
- 238000009629 microbiological culture Methods 0.000 claims abstract description 4
- 229930006000 Sucrose Natural products 0.000 claims description 21
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 21
- 239000005720 sucrose Substances 0.000 claims description 21
- 239000001888 Peptone Substances 0.000 claims description 13
- 108010080698 Peptones Proteins 0.000 claims description 13
- 235000019319 peptone Nutrition 0.000 claims description 13
- 238000010438 heat treatment Methods 0.000 claims description 12
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 10
- 239000008103 glucose Substances 0.000 claims description 10
- 238000011081 inoculation Methods 0.000 claims description 9
- 238000009835 boiling Methods 0.000 claims description 8
- 238000001816 cooling Methods 0.000 claims description 8
- 238000012258 culturing Methods 0.000 claims description 4
- 230000000813 microbial effect Effects 0.000 abstract description 9
- 235000014663 Kluyveromyces fragilis Nutrition 0.000 abstract description 6
- 244000253911 Saccharomyces fragilis Species 0.000 abstract description 6
- 235000018368 Saccharomyces fragilis Nutrition 0.000 abstract description 6
- 229940031154 kluyveromyces marxianus Drugs 0.000 abstract description 6
- 244000005700 microbiome Species 0.000 abstract description 3
- 240000000171 Crataegus monogyna Species 0.000 abstract 2
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 55
- 239000000796 flavoring agent Substances 0.000 description 13
- 235000019634 flavors Nutrition 0.000 description 13
- 230000012010 growth Effects 0.000 description 8
- 238000004519 manufacturing process Methods 0.000 description 7
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- 235000012055 fruits and vegetables Nutrition 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 238000011161 development Methods 0.000 description 3
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- 239000011780 sodium chloride Substances 0.000 description 3
- 238000009395 breeding Methods 0.000 description 2
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- 239000002994 raw material Substances 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 235000004876 Physalis pruinosa Nutrition 0.000 description 1
- 240000006285 Physalis pubescens Species 0.000 description 1
- 235000009230 Physalis pubescens Nutrition 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000003712 anti-aging effect Effects 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
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- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000002860 competitive effect Effects 0.000 description 1
- 238000005034 decoration Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 235000013569 fruit product Nutrition 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 238000001000 micrograph Methods 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 235000019629 palatability Nutrition 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 239000006041 probiotic Substances 0.000 description 1
- 235000018291 probiotics Nutrition 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000001953 sensory effect Effects 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
- C12N1/165—Yeast isolates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B7/00—Preservation or chemical ripening of fruit or vegetables
- A23B7/02—Dehydrating; Subsequent reconstitution
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B7/00—Preservation or chemical ripening of fruit or vegetables
- A23B7/08—Preserving with sugars
- A23B7/085—Preserving with sugars in a solution of sugar
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B7/00—Preservation or chemical ripening of fruit or vegetables
- A23B7/10—Preserving with acids; Acid fermentation
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23G—COCOA; COCOA PRODUCTS, e.g. CHOCOLATE; SUBSTITUTES FOR COCOA OR COCOA PRODUCTS; CONFECTIONERY; CHEWING GUM; ICE-CREAM; PREPARATION THEREOF
- A23G3/00—Sweetmeats; Confectionery; Marzipan; Coated or filled products
- A23G3/34—Sweetmeats, confectionery or marzipan; Processes for the preparation thereof
- A23G3/36—Sweetmeats, confectionery or marzipan; Processes for the preparation thereof characterised by the composition containing organic or inorganic compounds
- A23G3/364—Sweetmeats, confectionery or marzipan; Processes for the preparation thereof characterised by the composition containing organic or inorganic compounds containing microorganisms or enzymes; containing paramedical or dietetical agents, e.g. vitamins
- A23G3/366—Sweetmeats, confectionery or marzipan; Processes for the preparation thereof characterised by the composition containing organic or inorganic compounds containing microorganisms or enzymes; containing paramedical or dietetical agents, e.g. vitamins containing microorganisms, enzymes
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23G—COCOA; COCOA PRODUCTS, e.g. CHOCOLATE; SUBSTITUTES FOR COCOA OR COCOA PRODUCTS; CONFECTIONERY; CHEWING GUM; ICE-CREAM; PREPARATION THEREOF
- A23G3/00—Sweetmeats; Confectionery; Marzipan; Coated or filled products
- A23G3/34—Sweetmeats, confectionery or marzipan; Processes for the preparation thereof
- A23G3/36—Sweetmeats, confectionery or marzipan; Processes for the preparation thereof characterised by the composition containing organic or inorganic compounds
- A23G3/48—Sweetmeats, confectionery or marzipan; Processes for the preparation thereof characterised by the composition containing organic or inorganic compounds containing plants or parts thereof, e.g. fruits, seeds, extracts
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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Abstract
The present disclosure relates to the technical field of microorganism, and in particular to yeast tolerant to high sugar and low pH, and an application thereof. The yeast provided in the present disclosure, which is tolerant to high sugar and low pH, was preserved at China General Microbiological Culture Collection Center on June 10, 2021 with a number of CGMCC No.22681. Application of the yeast tolerant to high sugar and low pH to fermented preserved hawthorn fruit, and the producing method for fermented preserved fruit are important scope of protection of the present disclosure. Firstly, yeast was fermented to obtain a seed solution; then, stoned hawthorns were added to a sugaring liquor, which was heated, boiled and cooled; then the seed liquor was used to ferment processed material; the fermentation was continued, and hawthorns were roasted, thereby obtaining finished product of fermented preserved hawthorn fruit. For the first time, the present disclosure discloses CGMCC No.22681 Kluyveromyces marxianus, which is tolerant to high sugar and low pH, and application thereof. The CGMCC No.22681 Kluyveromyces marxianus provided in the present disclosure enables microbial fermentation to be performed under the condition of high sugar content and a low pH, and thus has excellent industrial application value.
Description
Specification ————— e054? YEAST TOLERANT TO HIGH SUGAR AND LOW PH, AND APPLICATION
THEREOF Technical Field The present disclosure relates to the technical field of microorganism, and in particular to yeast tolerant to high sugar and low pH, and an application thereof. Background Preserved fruit is a slightly transparent product with fruits and vegetables as raw materials, is made by means of sugaring, drying and other technologies, and is free of icing sugar on surface. Preserved fruit can be eaten directly, or be added to other foods as decoration or flavor improvement, and is widely received by consumers. At present, traditional preserved fruit dominates the market, but it has problems such as single flavor, poor color, transparency and fullness, and short shelf life or the like. Microbial fermentation is a biotechnology that helps to preserve food, to enhance the nutritional value and to improve the sensory characteristics. Research on microbial fermentation of fruits and vegetables has attracted more and more attention, wherein the fermented products have enhanced nutritional value and palatability, and also have special fermentation flavors and nutrients, such as organic acids, vitamins, Amino acids and antioxidant substances, and thus have the functions of maintaining the balance of intestinal flora, promoting digestion and absorption of human body, anti-oxidation, anti-aging, improving human immunity, and preventing cardiovascular diseases and so on.
1
Specification EEE ————————— 5 0,0 The introduction of microbial fermentation into the production of traditional preserved fruit can preserve the nutrients of fruits and vegetables, prolong the supply period of fruits and vegetables, and adjust the demands of low and peak seasons; moreover, due to the unique effect of microbial fermentation, product flavors are improved, and industrial problems such as single flavor, excessive microorganisms and so on of traditional preserved fruit can be solved. At the same time, the health value of preserved fruits is improved, and the development prospect of product market is broadened, so microbial fermentation will definitely become the research hotspot and development direction of the preserved fruit industry.
Current research on the fermented preserved fruit mainly focuses on the means of low-sugar liquid fermentation or solid fermentation, wherein the production process is complicated, and the product taste and flavor are not uniform. Therefore, there 1s still a lack of competitive fermented preserved fruit products on the market. It has become a breakthrough direction in this field as for how to realize microbial fermentation while performing high-sugar sugaring of preserved fruit. Therefore, the breeding of fermented probiotics with excellent performance, high-sugar tolerance and low pH 1s still one of the important projects to solve the application issue of fermented preserved fruit, and is also of great value in practice and academic research.
In an article written by Ying WANG et al, i.e. Development of Fermented Preserved Fruit with Physalis Pubescens, it is disclosed that, when the pH of 2
Specification ————— e054? fermentation is in a range from 3.89 to 3.75, the preserved fruit is glossy, the appearance is relatively full, the surface is smooth and elastic, and the sweetness is suitable. In an article written by Shaoning CHENG et al, i.e. Research on Growth Conditions of Alcohol-Fermentation Yeast for Waste Sugar Solution from Production of Superior Preserved Fruit, it is disclosed that the optimal growth condition for yeast is a pH of 3; there is rare literature on fermentation at lower pH and higher sugaring. In addition, sugaring fermentation is not used in the above production process of preserved fruit. The Baume degree is higher for sugaring fermentation, but the yeast is less tolerant to high sugar and low pH conditions.
As a result, it is desirable to find a strain tolerant to high sugar and low pH conditions, so as to perform microbial fermentation during high-sugar sugaring of preserved fruit.
Summary of the Invention In order to solve the above technical problem, the present disclosure provides yeast tolerant to high sugar and low pH, wherein the yeast was preserved at China General Microbiological Culture Collection Center on June 10, 2021 with a number of CGMCC No.22681.
Application of the afore-mentioned yeast tolerant to high sugar and low pH to fermented preserved hawthorn fruit 1s also the scope of protection of the present disclosure.
The present disclosure further claims a producing method for fermented 3
Specification 542 preserved fruit, wherein the most prominent characteristic of such producing method is that the CGMCC No.22681 yeast is used for fermenting the preserved hawthorn fruit.
Specifically, a producing method for fermented preserved fruit, including: (1) fermenting CGMCC No.22681 yeast inoculating, fermenting and culturing CGMCC No.22681 yeast in a seed medium; (2) stoning and adding hawthorns into a sugaring liquor, heating, boiling and cooling the sugaring liquor, and adding sucrose so as to adjust the sugar content of the sugaring liquor; (3) adding cultured seed liquor of CGMCC No.22681 yeast in step (1) to the material obtained in step (2) for continuous fermentation; and (4) roasting the fermented material, thereby obtaining yeast-fermented preserved hawthorn fruit.
Preferably, in step (1), the seed medium is peptone of 2 to 10 g/L and glucose of 30 to 200 g/L, and the fermentation culture is performed at a pH of 5.0 to 6.0 and a temperature of 28 to 37°C for 20 to 28 h.
Preferably, in step (1), the seed medium 1s peptone of 5 g/L. and glucose of 50 to 200 g/L with a fermentation culture at a pH of 5.0 to 6.0; and the fermentation culture is performed at a pH of 5.5 and a temperature of 32°C for 24 h.
Preferably, in step (2), stoning and adding hawthorns into a sugaring liquor 4
Specification ————————————————————————————————————5{) | 5 40 with a sugar content of 42 to 48%Brix, heating, boiling and cooling the sugaring liquor, and adding sucrose so as to adjust the sugar content of the sugaring liquor.
Preferably, in step (2), stoning and adding hawthorns into a sugaring liquor with a sugar content of 42 to 48%Brix, heating the sugaring liquor to 92 to 98°C and maintaining the temperature for 10 to 20 min, heating and boiling the sugaring liquor for 10 to 20 min, naturally cooling the sugaring liquor and immersing the hawthorns for 20 to 28 h, and then adding sucrose so as to adjust the sugar content of the sugaring liquor to 48 to 52%.
Preferably, in step (3), an inoculation quantity of the seed liquor is 10%, and the continuous fermentation is to be performed at a temperature of 30 to 34°C for 45 to 50 h; and in step (4), roasting the fermented materials at a temperature of 55 to 65°C for 6 to 10 h, thereby obtaining preserved hawthorn fruit fermented by yeast.
Preferably, the producing method for fermented preserved fruit includes: (1) fermenting CGMCC No.22681 yeast inoculating, fermenting and culturing CGMCC No.22681 yeast in a seed medium; the seed medium is peptone of 5 g/L and glucose of 50 to 200 g/L with a fermentation culture at a pH of 5.0 to 6.0; and the fermentation culture is performed at a pH of 5.5 and a temperature of 32°C for 24 h; (2) stoning and adding hawthorns into a sugaring liquor with a sugar content of 45%Brix, heating the sugaring liquor to 92 to 98°C and maintaining the
Specification ————— e054? temperature for 15 min, heating and boiling the sugaring liquor for 15 min, naturally cooling the sugaring liquor and immersing the hawthorns for 24 h, and then adding sucrose so as to adjust the sugar content of the sugaring liquor to 50%; (3) adding cultured seed liquor of CGMCC No.22681 yeast in step (1) to the material obtained in step (2), wherein an inoculation quantity of the seed liquor is 10%, and the fermentation is performed at a temperature of 32°C for 48 h; and (4) roasting the fermented material at a temperature of 60°C for 8 h, thereby obtaining yeast-fermented preserved hawthorn fruit.
For the first time, the present disclosure discloses CGMCC No.22681 Kluyveromyces marxianus, which is tolerant to high sugar and low pH, and application thereof. The CGMCC No.22681 Kluyveromyces marxianus provided in the present disclosure enables microbial fermentation to be performed under the condition of high sugar content and a low pH, and thus has excellent industrial application value. Ordinary yeast cannot survive under the condition of high sugar content and a low pH, while the yeast of the present disclosure still retains outstanding activity. Embodiment 3 shows that thallus of CGMCC No.22681 yeast grew well under the condition of 55%Brix and a pH of 2.3.
The beneficial effects of the present disclosure are as follows: (1) compared with ordinary yeast, the CGMCC No.22681 yeast provided in the present disclosure is excellent in sugaring tolerance; ordinary yeast can no longer grow smoothly at 55%Brix, but the CGMCC No.22681 yeast provided in the present disclosure can grow well and can be fermented to produce alcohol; and 6
Specification El) | 5, {0 (2) preserved hawthorn fruit obtained after fermentation by using the method provided in the present disclosure has uniform and bright color, moderate sweetness, good toughness and a thick fermentation flavor. Description of Drawings Fig. 1 is a flat photo and a microphotograph of SL 6# strain; Fig. 2 is a polygenetic tree of SL 6# strain; Fig. 3 is a curve showing the fermentation process of CGMCC No.22681 yeast; Fig. 4 shows growing data of CGMCC No.22681 yeast thallus under different sugar contents and pHs; Fig. 5 shows contrast photos of fermentation processes of ordinary Saccharomyces cerevisiae and CGMCC No.22681 yeast; and Fig. 6 shows photos of preserved hawthorn fruit fermented by CGMCC No.22681 yeast. Embodiments For better understanding of the present disclosure by one skilled in the art, the present disclosure will be further explained with reference to embodiments.
Producer of alcohol-fermentation active dry yeast: Angel Yeast Co., Ltd; other raw materials used in the present disclosure, such as sucrose, peptone, sodium chloride and so on, are all conventional experimental materials, and are ordinary market products.
With reference to the following embodiments, the present disclosure will be 7
Specification El) | 5, {0 further explained. It should be understood that the embodiments below are only used for explaining the present disclosure, rather than for limiting the scope of the present disclosure.
The yeast of the present disclosure was preserved at China General Microbiological Culture Collection Center (CGMCC) on June 10, 2021, wherein the address is Institute of Microbiology, Chinese Academy of Sciences at NO.1-3 Beichen West Road, Chaoyang District, Beijing, and the preservation number is CGMCC No.22681.
Embodiment 1 Breeding of SL 6# strain Strains of alcohol-fermentation yeast preserved in the laboratory were activated in an activation medium (sucrose of 20 g/L, peptone of 5 g/L, sodium chloride of 0.5 g/L, and a pH of 6.0), then the yeast suspension was irradiated under UV lamp of 30 W for respectively 10 to 120 s, the yeast solution diluted with sterile water was spread onto a screening medium (sucrose of 500 g/L, peptone of g/L, sodium chloride of 0.5 g/L, a pH of 6.0, and agar of 20 g/L) at a temperature of 30°C for 2 to 3 days. The obtained culture with the largest colony was transferred to a new solid plate, which was streaked until single colony appeared. After secondary screening, a strain of pure culture was finally obtained, wherein the number is SL 6#.
8
Specification eel) | 5 £0 Embodiment 2 Identification of SL 6# 18SrDNA identification was performed for the SL 6# obtained in Embodiment 1, an evolutionary tree was built by using part of sequence obtained from NCBI blast, and SL 6# was determined to be Kluyveromyces marxianus from the evolutionary tree built by MP method.
As shown in Fig. 2, the strain obtained in the present disclosure is indeed Kluyveromyces marxianus yeast.
Embodiment 3 Fermentation of CGMCC No.22681 yeast Batches of fermentation were performed for CGMCC No.22681 yeast obtained in Embodiment 2. The fermentation medium was as follows: 300 g of hawthorns was added to 1 L of sucrose water solution with a sugar content of 40%Brix and then was steamed for 30 min, and sucrose was added to adjust the sugar content to 55%Brix with a natural pH (pH 2.3); the inoculation quantity was 10%, the fermentation temperature was 32°C and the fermentation lasted for 48 h. The fermentation liquor was taken for the detection of thallus growth and alcohol content, and Fig. 3 shows the curve for thallus growth, fermentation and alcohol production. As shown in Fig. 3, CGMCC No0.22681 yeast can still maintain a relatively high level of thallus growth, fermentation, metabolism and alcohol 9
Specification El) | 5, {0 production when the sugar content is 55% and the pH is 2.3, thallus growth can be stabilized after fermentation of 24 h, and the alcohol content can be greater than 12% only after fermentation of 36 h.
While other components of the fermentation medium maintained unchanged, different sugar contents were respectively adjusted so as to test thallus growth of CGMCC No.22681 yeast under different sugar contents; while other components of the fermentation medium maintained unchanged, different pHs of the fermentation liquor were respectively adjusted so as to test thallus growth of CGMCC No.22681 yeast under different pHs. Fermentation liquors were taken after fermentation for 24 h so as to respectively test the thallus OD600, please refer to Fig. 4 for the results. As shown in Fig. 4, CGMCC No.22681 yeast can grow well as long as the sugar content is lower than 60%, can still be tolerant when the sugar content is 80%, and the optimal sugar content is 30%; CGMCC No.22681 yeast can grow well when pH is in a range from 2.5 to 8.5, can be tolerant when the pH is as low as 1.5, and the optimal pH is 6.5.
Embodiment 4 Preserved hawthorn fruit fermented by CGMCC No.22681 yeast Hawthorns were stoned and added into a sugaring liquor, which was made of sucrose, with a sugar content of 45%Brix, the sugaring liquor was heated to 95°C and the temperature was maintained for 15 min, the sugaring liquor was heated to boil for 15 min, the sugaring liquor was naturally cooled and the hawthorns were
Specification eel) | 5 £0 immersed for 24 h, and then sucrose was added so that the sugar content of the sugaring liquor was adjusted to 50%Brix; then cultured seed liquor (which is cultured under the following conditions: peptone of 5 g/L, glucose of 50 g/L, a pH of 5.5 and a temperature of 33°C for 24 h) of CGMCC No.22681 yeast was added, wherein an inoculation quantity of the seed liquor was 10%, and the fermentation was carried on at a temperature of 33°C for 48 h; then hawthorns were fished out and were roasted in an environment of 60°C for 8 h, thereby obtaining yeast-fermented preserved hawthorn fruit with a thick fermentation flavor.
Embodiment 5 Preserved hawthorn fruit fermented by CGMCC No.22681 yeast Hawthorns were stoned and added into a sugaring liquor, which was made of sucrose, with a sugar content of 45%Brix, the sugaring liquor was heated to 92°C and the temperature was maintained for 17 min, the sugaring liquor was heated to boil for 17 min, the sugaring liquor was naturally cooled and the hawthorns were immersed for 24 h, and then sucrose was added so that the sugar content of the sugaring liquor was adjusted to 50%Brix; then cultured seed liquor (which is cultured under the following conditions: peptone of 8 g/L, glucose of 50 g/L, a pH of 5.8 and a temperature of 32°C for 24 h) of CGMCC No.22681 yeast was added, wherein an inoculation quantity of the seed liquor was 10%, and the fermentation was carried on at a temperature of 32°C for 45 h; then hawthorns were fished out and were roasted in an environment of 55°C for 8 h, thereby obtaining 11
Specification eel) | 5 £0 yeast-fermented preserved hawthorn fruit with a thick fermentation flavor.
Embodiment 6 Preserved hawthorn fruit fermented by CGMCC No.22681 yeast Hawthorns were stoned and added into a sugaring liquor, which was made of sucrose, with a sugar content of 45%Brix, the sugaring liquor was heated to 95°C and the temperature was maintained for 15 min, the sugaring liquor was heated to boil for 15 min, the sugaring liquor was naturally cooled and the hawthorns were immersed for 24 h, and then sucrose was added so that the sugar content of the sugaring liquor was adjusted to 50%Brix; then cultured seed liquor (which is cultured under the following conditions: peptone of 10 g/L, glucose of 80 g/L, a pH of 6.0 and a temperature of 37°C for 24 h) of CGMCC No.22681 yeast was added, wherein an inoculation quantity of the seed liquor was 10%, and the fermentation was carried on at a temperature of 37°C for 48 h; then hawthorns were fished out and were roasted in an environment of 60°C for 8 h, thereby obtaining yeast-fermented preserved hawthorn fruit with a thick fermentation flavor.
Embodiment 7 Preserved hawthorn fruit fermented by CGMCC No.22681 yeast Hawthorns were stoned and added into a sugaring liquor, which was made of sucrose, with a sugar content of 45%Brix, the sugaring liquor was heated to 95°C 12
Specification El) | 5, {0 and the temperature was maintained for 15 min, the sugaring liquor was heated to boil for 15 min, the sugaring liquor was naturally cooled and the hawthorns were immersed for 24 h, and then sucrose was added so that the sugar content of the sugaring liquor was adjusted to 50%Brix; then cultured seed liquor (which is cultured under the following conditions: peptone of 5 g/L, glucose of 50 g/L, a pH of 5.0 and a temperature of 35°C for 22 h) of CGMCC No0.22681 yeast was added, wherein an inoculation quantity of the seed liquor was 10%, and the fermentation was carried on at a temperature of 35°C for 46 h with a fermentation pH of 2.3; then hawthorns were fished out and were roasted in an environment of 55°C for 8 h, thereby obtaining yeast-fermented preserved hawthorn fruit with a thick fermentation flavor.
As for the advantages of CGMCC No.22681 yeast provided by the present disclosure in fermenting preserved hawthorn fruit, the following tests were performed by the inventor: under the same fermentation conditions, ordinary alcohol-fermentation active dry yeast purchased on market was inoculated, the fermentation was continued at 35°C for 46 h, and there found no obvious bubbling or other fermentation phenomenon; the fermentation was continued for 96 h, and there found neither obvious fermentation flavor, nor obvious bubbling and bacterial precipitation, please refer to Fig. 5 for specific fermentation situation; as shown in Fig. 5, ordinary alcohol-fermentation active dry yeast has no fermentation effects on preserved hawthorn fruit, indicating that ordinary alcohol-fermentation active 13
Specification 801542 dry yeast is not tolerant to the environment having a sugar content as high as 50%Brix and a pH of 2.3.
Hawthorns fermented by ordinary yeast were roasted under the same conditions so as to obtain preserved hawthorn fruit, but there was no fermentation flavor, and the taste was the same as ordinary preserved hawthorn fruit. It is thus determined that ordinary alcohol-fermentation active dry yeast cannot tolerate high sugar content of 50% and a low pH of 2.3 at the same time, and cannot be applied to the production of fermented preserved hawthorn fruit.
14
Claims (10)
1. Yeast tolerant to high sugar and low pH, wherein the yeast was preserved at China General Microbiological Culture Collection Center on June 10, 2021 with a number of CGMCC No.22681.
2. Application of the yeast according to claim 1, which is tolerant to high sugar and low pH, to fermented preserved hawthorn fruit.
3. A producing method for fermented preserved fruit, wherein the CGMCC No.22681 yeast is used for fermenting the preserved hawthorn fruit.
4. À producing method for fermented preserved fruit, including: (1) fermenting CGMCC No.22681 yeast inoculating, fermenting and culturing CGMCC No.22681 yeast in a seed medium; (2) stoning and adding hawthorns into a sugaring liquor, heating, boiling and cooling the sugaring liquor, and adding sucrose so as to adjust the sugar content of the sugaring liquor; (3) adding cultured seed liquor of CGMCC No.22681 yeast in step (1) to the material obtained in step (2) for continuous fermentation; and (4) roasting the fermented material, thereby obtaining yeast-fermented preserved hawthorn fruit.
5. The producing method for fermented preserved fruit according to claim 4, wherein: in step (1), the seed medium is peptone of 2 to 10 g/L. and glucose of 30 to 200 g/L, and the fermentation culture 1s performed at a pH of 5.0 to 6.0 and a 1
Claims 801542 temperature of 28 to 37°C for 20 to 28 h.
6. The producing method for fermented preserved fruit according to claim 4, wherein: in step (1), the seed medium is peptone of 5 g/L and glucose of 50 to 200 g/L with a fermentation culture at a pH of 5.0 to 6.0; and the fermentation culture 1s performed at a pH of 5.5 and a temperature of 32°C for 24 h.
7. The producing method for fermented preserved fruit according to claim 4, wherein: in step (2), stoning and adding hawthorns into a sugaring liquor with a sugar content of 42 to 48%Brix, heating, boiling and cooling the sugaring liquor, and adding sucrose so as to adjust the sugar content of the sugaring liquor.
8. The producing method for fermented preserved fruit according to claim 4, wherein: in step (2), stoning and adding hawthorns into a sugaring liquor with a sugar content of 42 to 48%Brix, heating the sugaring liquor to 92 to 98°C and maintaining the temperature for 10 to 20 min, heating and boiling the sugaring liquor for 10 to 20 min, naturally cooling the sugaring liquor and immersing the hawthorns for 20 to 28 h, and then adding sucrose so as to adjust the sugar content of the sugaring liquor to 48 to 52%.
9. The producing method for fermented preserved fruit according to claim 4, wherein: in step (3), an inoculation quantity of the seed liquor is 10%, and the continuous fermentation is to be performed at a temperature of 30 to 34°C for 45 to 50 h; and in step (4), roasting the fermented materials at a temperature of 55 to 65°C for 6 to 10 h, thereby obtaining preserved hawthorn fruit fermented by yeast.
2
Claims ‘N
10. The producing method for fermented preserved fruit according to claim 4, including: (1) fermenting CGMCC No.22681 yeast inoculating, fermenting and culturing CGMCC No.22681 yeast in a seed medium; the seed medium is peptone of 5 g/L and glucose of 50 to 200 g/L with a fermentation culture at a pH of 5.0 to 6.0; and the fermentation culture is performed at a pH of 5.5 and a temperature of 32°C for 24 h; (2) stoning and adding hawthorns into a sugaring liquor with a sugar content of 45%Brix, heating the sugaring liquor to 92 to 98°C and maintaining the temperature for 15 min, heating and boiling the sugaring liquor for 15 min, naturally cooling the sugaring liquor and immersing the hawthorns for 24 h, and then adding sucrose so as to adjust the sugar content of the sugaring liquor to 50%; (3) adding cultured seed liquor of CGMCC No.22681 yeast in step (1) to the material obtained in step (2), wherein an inoculation quantity of the seed liquor is 10%, and the fermentation is performed at a temperature of 32°C for 48 h; and (4) roasting the fermented material at a temperature of 60°C for 8 h, thereby obtaining yeast-fermented preserved hawthorn fruit.
3
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