CN113151040A - Aroma-enhancing type direct-vat-set starter, and preparation method and application thereof - Google Patents
Aroma-enhancing type direct-vat-set starter, and preparation method and application thereof Download PDFInfo
- Publication number
- CN113151040A CN113151040A CN202110146326.5A CN202110146326A CN113151040A CN 113151040 A CN113151040 A CN 113151040A CN 202110146326 A CN202110146326 A CN 202110146326A CN 113151040 A CN113151040 A CN 113151040A
- Authority
- CN
- China
- Prior art keywords
- freeze
- lactobacillus plantarum
- dried powder
- orientalis
- preparation
- Prior art date
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- Granted
Links
- 239000007858 starting material Substances 0.000 title claims abstract description 50
- 238000002360 preparation method Methods 0.000 title claims abstract description 44
- 240000006024 Lactobacillus plantarum Species 0.000 claims abstract description 77
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims abstract description 77
- 229940072205 lactobacillus plantarum Drugs 0.000 claims abstract description 77
- 239000000843 powder Substances 0.000 claims abstract description 64
- 238000000034 method Methods 0.000 claims abstract description 17
- 241000235645 Pichia kudriavzevii Species 0.000 claims abstract description 15
- 238000009629 microbiological culture Methods 0.000 claims abstract description 5
- 238000004321 preservation Methods 0.000 claims abstract description 3
- 238000013329 compounding Methods 0.000 claims abstract 2
- 241001052560 Thallis Species 0.000 claims description 46
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- 240000006677 Vicia faba Species 0.000 claims description 33
- 235000002098 Vicia faba var. major Nutrition 0.000 claims description 33
- 238000000855 fermentation Methods 0.000 claims description 31
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 20
- 239000007787 solid Substances 0.000 claims description 17
- 229920001817 Agar Polymers 0.000 claims description 14
- 239000008272 agar Substances 0.000 claims description 14
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 13
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims description 13
- 239000003223 protective agent Substances 0.000 claims description 13
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 claims description 12
- 239000001888 Peptone Substances 0.000 claims description 12
- 108010080698 Peptones Proteins 0.000 claims description 12
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 claims description 12
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 claims description 12
- 238000004108 freeze drying Methods 0.000 claims description 12
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- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 11
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- 239000011814 protection agent Substances 0.000 claims 1
- 150000002148 esters Chemical class 0.000 abstract description 48
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- FKRCODPIKNYEAC-UHFFFAOYSA-N ethyl propionate Chemical compound CCOC(=O)CC FKRCODPIKNYEAC-UHFFFAOYSA-N 0.000 abstract description 24
- MDHYEMXUFSJLGV-UHFFFAOYSA-N phenethyl acetate Chemical compound CC(=O)OCCC1=CC=CC=C1 MDHYEMXUFSJLGV-UHFFFAOYSA-N 0.000 abstract description 24
- YYZUSRORWSJGET-UHFFFAOYSA-N octanoic acid ethyl ester Natural products CCCCCCCC(=O)OCC YYZUSRORWSJGET-UHFFFAOYSA-N 0.000 abstract description 17
- HNAGHMKIPMKKBB-UHFFFAOYSA-N 1-benzylpyrrolidine-3-carboxamide Chemical compound C1C(C(=O)N)CCN1CC1=CC=CC=C1 HNAGHMKIPMKKBB-UHFFFAOYSA-N 0.000 abstract description 12
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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
- C12N1/20—Bacteria; Culture media therefor
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C19/00—Cheese; Cheese preparations; Making thereof
- A23C19/06—Treating cheese curd after whey separation; Products obtained thereby
- A23C19/061—Addition of, or treatment with, microorganisms
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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
- A23L17/00—Food-from-the-sea products; Fish products; Fish meal; Fish-egg substitutes; Preparation or treatment thereof
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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
- A23L17/00—Food-from-the-sea products; Fish products; Fish meal; Fish-egg substitutes; Preparation or treatment thereof
- A23L17/65—Addition of, or treatment with, microorganisms or enzymes
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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
- A23L19/00—Products from fruits or vegetables; Preparation or treatment thereof
- A23L19/20—Products from fruits or vegetables; Preparation or treatment thereof by pickling, e.g. sauerkraut or pickles
-
- 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
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/065—Microorganisms
-
- 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
- C12N1/04—Preserving or maintaining viable microorganisms
-
- C—CHEMISTRY; METALLURGY
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Abstract
The invention provides an aroma-enhancing type direct vat set starter, a preparation method and application thereof, and belongs to the technical field of fermented food processing. The aroma-increasing type direct vat set starter is formed by compounding lactobacillus plantarum freeze-dried powder and Issatchenkia orientalis freeze-dried powder according to the mass ratio of 1: 0.5-3. The lactobacillus plantarum is preserved in China general microbiological culture Collection center (CGMCC) at 03 months 12 and 2020, and the preservation number is CGMCC 21288. The aroma-enhancing direct-vat-set starter provided by the invention has the advantages of simple components, convenience in preparation and simple and convenient use method, and can obviously enhance the ester content of the product. Wherein the contents of ethyl propionate, ethyl butyrate, ethyl caproate, ethyl caprylate, ethyl caprate and phenethyl acetate are respectively increased to 1.1-8.0 times, 1.1-4.4 times, 1.0-4.0 times, 1.0-4.8 times, 1.0-7.5 times and 1.1-6.0 times.
Description
Technical Field
The invention relates to a preparation method and application of an aroma-enhancing type direct vat set starter, belonging to the technical field of fermented food processing.
Background
Fermentation is a method for preserving food by using microorganisms for thousands of years. It utilizes the metabolism of microbe, and has high nutritive value and unique flavor. In the fermentation process, the microorganisms can decompose macromolecular substances into micromolecular amino acids, increase nutrition and generate unique flavor. Micromolecular flavor peptide and free amino acid are formed in the macromolecular hydrolysis process, the amino acid can be directly used as a precursor of flavor substances, and aldehyde, acid, alcohol, ester and other aromatic components are formed through microbial catalytic deamination, decarboxylation, transamination and cracking of amino acid side chains.
Traditional fermented foods such as cheese and thick broad-bean sauce mostly belong to solid fermentation, and different from the existing developed liquid fermentation, the traditional fermented foods have the defects of non-uniform system, steric hindrance in reaction and the like in the fermentation process, the fermentation process is often uncontrollable, and the quality of the fermented products is different. Most of the existing processing modes are workshop type natural fermentation, and are greatly influenced by environmental factors. This causes problems such as unstable production quality, low production efficiency, and difficulty in standardized control. The breeding of the strain with specific flavor and the development of the direct vat set starter which is convenient and easy to use can greatly improve the quality and stability of the product, and is the preferred scheme for standardizing the production process and realizing standardized production.
Esters are important flavour substances formed by microorganisms during fermentation. Due to the low odor threshold, the ester compounds contribute pleasant fruity flavor to the overall flavor under proper concentration, and are important flavor substances in fermented foods. For example, ethyl propionate has pineapple flavor, ethyl butyrate has fruit and pineapple flavor, ethyl hexanoate has fruit flavor, ethyl octanoate has brandy-like flavor and has sweet taste, etc., ethyl decanoate has coconut flavor, phenethylacetate has honey-like and rose-like flavor, etc. Most of the research is focused on increasing the total ester content or increasing the content of single esters, especially ethyl acetate. However, there is no good starter which can simultaneously increase the contents of various esters.
Disclosure of Invention
Based on the prior art, the invention aims to solve the problem that the prior high-quality leaven which can simultaneously improve the content of various esters is lacked, and specifically breeds good microorganisms with high ester production capacity, and prepares the microorganisms into a direct vat set leaven for fermenting food.
The invention provides an aroma-enhancing type direct vat set, a preparation method and application thereof, and the technical scheme is as follows:
the aroma-increasing type direct-vat-set starter is compounded by lactobacillus plantarum freeze-dried powder and issatchenkia orientalis freeze-dried powder according to the mass ratio of 1 (0.5-3), and the total viable count of the aroma-increasing type direct-vat-set starter is (10)10～1012) CFU/g protectant solids; the lactobacillus plantarum freeze-dried powder comprises lactobacillus plantarum and lactobacillus plantarum freeze-protecting agent, wherein the lactobacillus plantarum freeze-dried powder comprises lactobacillus plantarum and lactobacillus plantarum freeze-protecting agentThe Lactobacillus plantarum is CGMCC21288, which is preserved in China general microbiological culture Collection Center (CCM) at 03 months 12 and 2020, has the address of No. 3 Xilu-1 of the Chaoyang district in Beijing, and the preservation number of CGMCC21288, and is classified and named as Lactobacillus plantarum; the Issatchenkia orientalis freeze-dried powder comprises Issatchenkia orientalis and Issatchenkia orientalis freeze-protecting agent, wherein the Issatchenkia orientalis is CICC 31431 and is characterized by being high-temperature-resistant aroma-producing yeast.
Preferably, the preparation method of the flavoring type direct vat set starter comprises the following steps:
s1, preparation of Lactobacillus plantarum thallus and Saiyi orientalis thallus:
when the lactobacillus plantarum thalli is prepared, the preparation method comprises the following steps: activating Lactobacillus plantarum strain with MRS agar plate, and performing MRS broth one-stage culture to obtain OD600nm1.0-10.0, washing the thalli for 1-2 times by using sterile physiological saline, centrifuging, collecting, and suspending by using a proper amount of sterile water for later use;
when preparing the thalli of the sayittuy orientalis, the preparation method comprises the following steps: activating the Saccharomyces cerevisiae strain with YPD agar plate, performing YPD broth primary culture to obtain OD600nm1.0-10.0, washing the thalli for 1-2 times by using sterile normal saline, centrifuging, collecting, and suspending by using a proper amount of sterile water for later use;
s2, preparing the lactobacillus plantarum thallus and the say yeast oriental thallus by the step S1, and preparing lactobacillus plantarum freeze-dried powder and the say yeast oriental freeze-dried powder:
when the lactobacillus plantarum freeze-dried powder is prepared, the preparation method comprises the following steps: mixing the lactobacillus plantarum thallus obtained in the step S1 with a lactobacillus plantarum cryoprotectant, and freeze-drying to obtain a strain with a strain concentration of (10)10～1012) CFU/g protectant solids;
when preparing the freeze-dried powder of the saccharomyces orientalis, the preparation method comprises the following steps: the sayia orientalis thallus obtained in the step of S1 is mixed with the freezing protective agent of the sayia orientalis, and then is frozen and dried to prepare the strain concentrationIs (10)10～1012) CFU/g protectant solids;
s3, preparing a flavoring type direct vat set starter: and (3) mixing the lactobacillus plantarum freeze-dried powder obtained in the step S2 and the sayia orientalis freeze-dried powder according to the mass ratio of 1 (0.5-3).
Preferably, the lactobacillus plantarum cryoprotectant comprises 10-15% (w/w) peptone, 20-30% (w/w) inulin, 20-30% (w/w) trehalose and 5-10% (w/w) sea grape polysaccharide; the Saccharum orientalis cryoprotectant comprises 25-35% (w/w) peptone, 20-30% (w/w) skim milk powder, 10-20% (w/w) trehalose and 5-10% (w/w) sea grape polysaccharide.
Preferably, the flavoring type direct vat set starter can be applied to the preparation of fermented foods, especially the fermentation of solid fermented foods, such as fermented salted fish (i.e. sour fish), fermented salted pepper, cheese, thick broad-bean sauce, etc.
Preferably, the flavoring type direct vat set starter is added in the fermentation preparation of the acid salted fish, the acid salted pepper, the cheese and the broad bean paste in an amount of (10)6～109) CFU/g material.
Compared with the prior art, the invention has the following beneficial effects:
the aroma-enhancing direct vat set starter can be used in solid fermented foods, particularly in the fermentation process of acid salted fish, acid salted hot pepper, cheese, thick broad-bean sauce and the like, the generated ester substances have aromatic odor, ethyl propionate has pineapple fragrance, ethyl butyrate has fruit fragrance and pineapple fragrance, ethyl caproate has fruit fragrance, ethyl caprylate has fragrance similar to brandy and sweet taste and the like, ethyl caprate has coconut fragrance, and phenylethyl acetate has honey-like and rose-like fragrance, so that the flavors of the fermented foods are enhanced.
The aroma-enhancing type direct-vat-set starter is simple and convenient to use, the shortcoming of unstable production of the ester flavor of a fermentation product is overcome by fermenting with the aroma-enhancing type direct-vat-set starter, and the purpose of remarkably enhancing the content of the ester substances in the product is achieved. Wherein the contents of ethyl propionate, ethyl butyrate, ethyl caproate, ethyl caprylate, ethyl caprate and phenethyl acetate are respectively increased to 1.1-8.0 times, 1.1-4.4 times, 1.0-4.0 times, 1.0-4.8 times, 1.0-7.5 times and 1.1-6.0 times.
The aroma-enhancing type direct vat set starter of the invention also has the advantages of simple components, convenient preparation and simple and convenient use method.
Detailed Description
The following examples are given to further illustrate the technical solution of the present invention to help understanding the patent, but the embodiments of the present invention are not limited by the examples, and the scope of the present invention is determined by the claims.
MRS agar medium, MRS broth medium, YPD agar medium and YPD broth medium used in examples 1 to 4 were purchased from Qingdao Haibo Biotech Ltd.
The Saiymyces orientalis CICC 31431, the Saiymyces orientalis CICC 1344, the Lactobacillus plantarum CICC 22696, the Lactobacillus plantarum CICC 20022, the Staphylococcus xylosus CICC 22943, the Saccharomyces cerevisiae CICC 1229, the Pediococcus acidilactici CICC 24367, the Saccharomyces cerevisiae CICC 1049 and the Saiymyces orientalis CICC 1926 used in the embodiments 1 to 4 are purchased from the China center for culture Collection of industrial microorganisms; aspergillus oryzae CGMCC 3.2792 was purchased from China center for general microbiological culture Collection; lactobacillus curvatus GDMCC 1.725 was purchased from the Guangdong province collection of microorganisms. Lactobacillus plantarum CGMCC21288, a lactic acid bacterium with ester-producing property screened previously, is preserved in China general microbiological culture Collection center (China Committee for culture Collection of microorganisms) 03 d 12.2020, and is addressed to Beijing City, Chaoyang district, Beichen West Lu No. 1 Hospital No. 3.
Example 1
A preparation method of a flavor-enhancing direct vat set starter and an application thereof in sour fish comprise the following steps:
s1, preparing lactobacillus plantarum thalli and sayia orientalis thalli:
when lactobacillus plantarum CGMCC21288 is prepared, the preparation method of the lactobacillus plantarum is as follows: strains stored at-80 ℃ were picked and streaked on MRS agar medium. The streaked plates were incubated for 36h in a 37 ℃ incubator. Single colonies on the plates were picked into MRS broth, shake-cultured at 37 ℃ and 200rpm for 16h, and then inoculated at 10% inoculum size into MRS broth, shake-cultured at 37 ℃ and 160rpm for 16 h. And centrifuging to collect thalli, washing the thalli for 2 times by using sterile normal saline, centrifuging again to collect thalli, and suspending by using a proper amount of sterile water for later use.
When the Issatchenkia orientalis CICC 31431 is prepared, the preparation method of the thalli is as follows: the strain stored at-80 ℃ was picked and streaked on YPD agar medium. The streaked plates were incubated in a 28 ℃ incubator for 48 h. Single colonies on the plates were picked into YPD broth, shake-cultured at 28 ℃ and 100rpm for 24h, and then inoculated at 5% inoculum size into YPD broth, and shake-cultured at 28 ℃ and 160rpm for 24 h. And centrifuging to collect thalli, washing the thalli for 1 time by using sterile normal saline, centrifuging again to collect thalli, and suspending by using a proper amount of sterile water for later use.
S2, preparing lactobacillus plantarum freeze-dried powder and east sayiyeast freeze-dried powder:
when the lactobacillus plantarum freeze-dried powder is prepared, the preparation method comprises the following steps: the lactobacillus plantarum thallus obtained in the step S1 adopts 15% (w/w) peptone, 30% (w/w) inulin, 30% (w/w) trehalose and 5% (w/w) vitis amurensis polysaccharide as cryoprotectants, the cryoprotectants are cooled to a certain degree after being sterilized (121 ℃, 15min), are uniformly mixed with the lactobacillus plantarum thallus, are pre-frozen in a glass plate at the temperature of minus 80 ℃ in an ultra-low temperature refrigerator for 24h, and are then put into a freeze dryer for freeze drying: vacuum degree of 1Pa, and cold trap at-83 deg.C for 70 hr. The survival rate of freeze-dried bacterial powder is up to 90%, and the prepared product contains 10 viable bacteria12CFU/g protectant solids;
when preparing freeze-dried thallus powder of the sayia orientalis, the preparation method comprises the following steps: the sayia orientalis thallus obtained in the step of S1 adopts 35% (w/w) peptone, 30% (w/w) skimmed milk powder, 20% (w/w) trehalose and 10% (w/w) marine glucose polysaccharide as cryoprotectants, the cryoprotectants are sterilized (121 ℃, 15min), cooled to a certain degree and uniformly mixed with the sayia orientalis thallus, and the mixture is placed in a glass plate at an ultralow temperature of-80 ℃ in a refrigeratorPre-freezing for 24h, and then putting the mixture into a freeze dryer for freeze drying: vacuum degree of 1Pa, and cold trap at-83 deg.C for 70 hr. The freeze dried powder has survival rate up to 89%, and contains viable bacteria of 9 × 1011CFU/g protectant solids.
S3, preparing various groups of leaven:
TABLE 1 sour fish fermentation group scheme
The fermented fish starter used in example 1 was prepared in accordance with the formulation shown in table 1, by mixing lyophilized bacterial powder at the ratio shown in table 1.
S4, fermenting the sour fish:
the traditional preparation method of sour fish (group 11) refers to the relation research of characteristic flavor formation and microorganisms in the fermentation process of sour fish. Doctor thesis at south of the Yangtze river. Zang jin hong is prepared. The method specifically comprises the following steps: uniformly stirring 100 parts of carp blocks, 3 parts of salt and 2 parts of white granulated sugar, pickling in a refrigerator at 4 ℃ for 48h, and drying under the conditions of constant temperature and humidity (50 ℃, 65% humidity) for 3 h. During the process, 100 parts of corn flour added with 3 parts of salt and 2 parts of white granulated sugar are fried to be cooked and then are put to room temperature, 100 parts of dried fish fillets and 25 parts of fried corn flour are mixed evenly and are put into a 5L glass jar in a mode of one layer of fish fillets and one layer of corn flour, and the mixture is covered with water and sealed and then is put into an incubator at 25 ℃ for fermentation.
The method for making the sour fish in the groups 1-10 refers to the group 11, and specifically comprises the following steps: uniformly stirring 100 parts of carp blocks, 3 parts of salt and 2 parts of white granulated sugar, pickling in a refrigerator at 4 ℃ for 48h, and drying under the conditions of constant temperature and humidity (50 ℃, 65% humidity) for 3 h. During the period, 100 parts of corn flour added with 3 parts of salt and 2 parts of white granulated sugar is fried to be cooked and then is put to room temperature, and a direct vat set starter (shown in table 1) is added, wherein the total number of viable bacteria of the direct vat set starter is 107CFU/g fish, mixing dried fish fillet 100 parts and parched corn flour 25 parts, loading into 5L glass jar with fish piece layer by corn flour layer, covering with water seal, and placing in 25 deg.C incubatorFermenting for 23 days.
The flavor substances in the sour fish are analyzed by GC-MS 7890B-7010B (Agilent company, USA), volatile organic compounds in a sample are extracted by Solid Phase Microextraction (SPME), and the contents of 6 main ester substances (ethyl propionate, ethyl butyrate, ethyl hexanoate, ethyl octanoate, ethyl decanoate and phenethylacetate) in the sour fish are measured in an MRM mode. The chromatographic conditions were as follows, capillary chromatography column: HP-5MS (30 m.times.250. mu.m.times.0.25 μm); gradient program: at 30 deg.C for 5 min; heating to 50 deg.C at 3 deg.C/min and maintaining for 3 min; raising the temperature to 150 ℃ at the speed of 5 ℃/min, and raising the temperature to 250 ℃ at the speed of 20 ℃/min and keeping the temperature for 5 min; the carrier gas is He, the flow rate is 1.0mL/min, and a non-flow splitting mode is selected.
TABLE 2 content of esters in fermented sour fish
The results of the content of the esters in the sour fish are shown in table 2, and it is known that the total amount of the esters in group 1 (lactobacillus plantarum CGMCC21288 cell freeze-dried powder, and sayittuya orientalis cic 31431 cell freeze-dried powder mixed in a mass ratio of 1: 1) is the highest, and the contents of ethyl propionate, ethyl butyrate, ethyl caproate, ethyl caprylate, ethyl caprate and phenethylacetate are all higher than those in groups 2-11, and the contents of the esters are increased to about 1.2-4.7 times, 1.1-3.9 times, 1.0-3.8 times, 1.0-2.3 times, 1.0-2.2 times and 1.3-3.4 times of the control group, so that a better flavor enhancing effect is achieved. The reason for this is probably that lactobacillus plantarum CGMCC21288 and sayia orientalis cic 31431 play a synergistic role, promoting the generation of esters. The 6 esters have pleasant aroma such as pineapple aroma, apple aroma, flower aroma, cream aroma and the like, have a low threshold value, and have an important contribution to the characteristic flavor of the sour fish. Therefore, the adoption of the aroma-enhancing type direct vat set starter can obviously improve the ester content in the sour fish, thereby improving the flavor quality of the product.
Example 2
The preparation method of the ester-producing aroma-enhancing type direct vat set starter and the application of the starter in cheese comprises the following steps:
s1, preparing lactobacillus plantarum thalli and sayia orientalis thalli:
when lactobacillus plantarum CGMCC21288 is prepared, the preparation method of the lactobacillus plantarum is as follows: strains stored at-80 ℃ were picked and streaked on MRS agar medium. The streaked plates were incubated for 36h in a 37 ℃ incubator. Single colonies on the plates were picked into MRS broth, shake-cultured at 37 ℃ and 200rpm for 16h, and then inoculated at 10% inoculum size into MRS broth, shake-cultured at 37 ℃ and 160rpm for 16 h. And centrifuging to collect thalli, washing the thalli for 2 times by using sterile normal saline, centrifuging again to collect thalli, and suspending by using a proper amount of sterile water for later use.
When the Issatchenkia orientalis CICC 31431 is prepared, the preparation method of the thalli is as follows: the strain stored at-80 ℃ was picked and streaked on YPD agar medium. The streaked plates were incubated in a 28 ℃ incubator for 48 h. Single colonies on the plates were picked into YPD broth, shake-cultured at 28 ℃ and 100rpm for 24h, and then inoculated at 5% inoculum size into YPD broth, and shake-cultured at 28 ℃ and 160rpm for 24 h. And centrifuging to collect thalli, washing the thalli for 1 time by using sterile normal saline, centrifuging again to collect thalli, and suspending by using a proper amount of sterile water for later use.
S2, preparing lactobacillus plantarum freeze-dried powder and east sayiyeast freeze-dried powder:
when the lactobacillus plantarum freeze-dried powder is prepared, the preparation method comprises the following steps: the lactobacillus plantarum thallus obtained in the step S1 adopts 15% (w/w) peptone, 30% (w/w) inulin, 30% (w/w) trehalose and 5% (w/w) vitis amurensis polysaccharide as cryoprotectants, the cryoprotectants are cooled to a certain degree after being sterilized (121 ℃, 15min), are uniformly mixed with the lactobacillus plantarum thallus, are pre-frozen in a glass plate at the temperature of minus 80 ℃ in an ultra-low temperature refrigerator for 24h, and are then put into a freeze dryer for freeze drying: vacuum degree of 1Pa, and cold trap at-83 deg.C for 70 hr. The survival rate of the freeze-dried bacterial powder is as high as90% of the total amount of viable bacteria contained in the extract12CFU/gProtectant solids；
When preparing freeze-dried thallus powder of the sayia orientalis, the preparation method comprises the following steps: the sayia orientalis thallus obtained in the step of S1 adopts 35% (w/w) peptone, 30% (w/w) skimmed milk powder, 20% (w/w) trehalose and 10% (w/w) marine glucose polysaccharide as freezing protective agents, the protective agents are sterilized (121 ℃, 15min), cooled to a certain degree and uniformly mixed with the sayia orientalis thallus, the mixture is placed in a glass plate to be pre-frozen in an ultra-low temperature refrigerator at minus 80 ℃ for 24h, and then the freeze-drying is carried out in a freeze-dryer: vacuum degree of 1Pa, and cold trap at-83 deg.C for 70 hr. The freeze dried powder has survival rate up to 89%, and the viable count is 9 × 1011CFU/gProtectant solids。
S3, preparing various groups of leaven:
TABLE 3 cheese fermentation group protocol
The cheese starter used in example 2 was prepared in accordance with the procedure shown in table 3, by mixing the lyophilized mycelia in the proportions shown in table 3.
S4, fermented cheese:
the preparation method of traditional cheese (group 11) refers to the yak cheddar cheese making technical flow protocol. The Chinese dairy industry, 2020, 02: 77-80. And (5) making. The method specifically comprises the following steps: centrifuging fresh milk without antibiotic to remove impurities, homogenizing under 30MPa and 65 deg.C, pasteurizing at 65 deg.C for 30min, and rapidly cooling to 35 deg.C. Adding 0.10g/L leaven (DOM1 lactic acid bacteria, Italy, Coleli Co.)MilkFermenting at 35 deg.C for 30 min. Mixing chymosin (rennet, bovine pepsin, Italy) with water at a ratio of 1:403min, adding into milk after fermentation, with the addition amount of 0.05g/LMilk. And (4) after adding the rennin, stirring for 30s, and standing until the milk is coagulated. Cutting the curd into cubes of 1.5cm multiplied by 1.5cm after the curd is finished, stirring and heating to 43 ℃, stewing for 30-60 min at constant temperature with continuous stirring, stopping stewing when the PH value of the separated whey is reduced to 6.15, and discharging whey for the first time. Thereafter, whey was drained every 15min and the clot was piled up for 60 min. The curd was cut to thumb size when the whey pH was discharged to 5.45. Adding salt 3.0g/L into the cut clotMilkSalting, adding salt for three times, kneading for 5min each time to distribute salt uniformly. And putting the salted coagulum into a mold to be pressed for 12-24 hours to form cheese.
The making method of the cheese in the groups 1-10 refers to the group 11, and specifically comprises the following steps: centrifuging fresh milk without antibiotic to remove impurities, homogenizing under 30Mpa at 65 deg.C, pasteurizing at 65 deg.C for 30min, and rapidly cooling to 35 deg.C. Adding 0.10g/L of leaven (DOM1 lactic acid bacteria, Italy, Coleli Co.)MilkA direct vat set starter (shown in Table 3) was added in an amount of 10 total viable cell count9CFU/gMilkFermenting at 35 deg.C for 30 min. Mixing chymosin (rennet, bovine pepsin, Italy) and water at a ratio of 1:40 for 3min, adding into milk at an amount of 0.05g/L after fermentationMilk. And (4) after adding the rennin, stirring for 30s, and standing until the milk is coagulated. Cutting the curd into cubes with the size of 1.5cm multiplied by 1.5cm after the curd is finished, stirring and heating to 43 ℃, stewing for 30-60 min at constant temperature and continuously stirring, stopping stewing when the PH value of the separated whey is reduced to 6.15, and performing primary whey discharge. Thereafter, whey was drained every 15min and the clot was piled up for 60 min. The whey discharge pH was reduced to 5.45 and the curd was cut to thumb size. Adding salt 3.0g/L into the cut clotMilkSalting, adding salt for three times, kneading for 5min each time to distribute salt uniformly. And putting the salted coagulum into a mold to be pressed for 12-24 hours to form cheese.
The flavor substances in the cheese were analyzed by GC-MS 7890B-7010B (Agilent Co., U.S.A.), volatile organic compounds in the sample were extracted by Solid Phase Microextraction (SPME), and the contents of 6 major ester substances (ethyl propionate, ethyl butyrate, ethyl caproate, ethyl caprylate, ethyl caprate and phenylethyl acetate) in the cheese were measured in MRM mode. The chromatographic conditions were as follows, capillary chromatography column: HP-5MS (30 m.times.250. mu.m.times.0.25 μm); gradient program: at 30 deg.C for 5 min; heating to 50 deg.C at 3 deg.C/min for 3 min; raising the temperature to 150 ℃ at the speed of 5 ℃/min, and raising the temperature to 250 ℃ at the speed of 20 ℃/min and keeping the temperature for 5 min; the carrier gas is He, the flow rate is 1.0mL/min, and a non-flow splitting mode is selected.
TABLE 4 content of esters in cheese
The results of the content of the esters in the cheese are shown in table 4, and it is known that the total amount of the esters in group 1 (freeze-dried powder of lactobacillus plantarum CGMCC21288 cells, freeze-dried powder of sayia orientalis cic 31431 cells were mixed in a mass ratio of 1: 1) was the highest, and the contents of ethyl propionate, ethyl butyrate, ethyl hexanoate, ethyl octanoate, ethyl decanoate and phenethyl acetate were all higher than those in groups 2 to 11, and the contents thereof were increased to about 1.3 to 3.9 times, 1.4 to 2.7 times, 1.4 to 2.6 times, 1.3 to 1.6 times, 1.3 to 1.7 times and 1.1 to 1.9 times of the control group, thereby achieving a better flavor enhancing effect. The reason for this is probably that lactobacillus plantarum CGMCC21288 and sayia orientalis cic 31431 play a synergistic role, promoting the generation of esters. The 6 esters have pleasant aroma such as pineapple aroma, apple aroma, flower aroma, cream aroma and the like, have a low threshold value, and have an important contribution to the characteristic flavor of cheese. Therefore, the adoption of the aroma-enhancing type direct vat set starter can obviously increase the content of esters in cheese, thereby improving the flavor quality of products.
Example 3
A preparation method of ester-producing flavor-enhancing direct vat set starter and its application in acid preserved chili (salted chili) comprises the following steps:
s1, preparing lactobacillus plantarum thalli and sayia orientalis thalli:
when lactobacillus plantarum CGMCC21288 is prepared, the preparation method of the lactobacillus plantarum is as follows: strains stored at-80 ℃ were picked and streaked on MRS agar medium. The streaked plates were incubated for 36h in a 37 ℃ incubator. Single colonies on the plates were picked into MRS broth, shake-cultured at 37 ℃ and 200rpm for 16h, and then inoculated at 10% inoculum size into MRS broth, shake-cultured at 37 ℃ and 160rpm for 16 h. And centrifuging to collect thalli, washing the thalli for 2 times by using sterile normal saline, centrifuging again to collect thalli, and suspending by using a proper amount of sterile water for later use.
When preparing Issatchenkia orientalis, the thallus preparation method is as follows: the strain stored at-80 ℃ was picked and streaked on YPD agar medium. The streaked plates were incubated in a 28 ℃ incubator for 48 h. Single colonies on the plate were picked into YPD broth, shake-cultured at 28 ℃ and 100rpm for 24 hours, and then inoculated at 5% inoculum size into YPD broth, and shake-cultured at 28 ℃ and 160rpm for 24 hours. And (3) centrifuging to collect thalli, washing the thalli for 1 time by using sterile physiological saline, centrifuging again to collect thalli, and suspending the thalli by using a proper amount of sterile water for later use.
S2, preparing lactobacillus plantarum freeze-dried powder and east sayiyeast freeze-dried powder:
when the lactobacillus plantarum freeze-dried powder is prepared, the preparation method comprises the following steps: the cultured lactobacillus plantarum thallus adopts 12% (w/w) peptone, 25% (w/w) inulin, 20% (w/w) trehalose and 10% (w/w) vitis amurensis polysaccharide as freezing protective agents, the protective agents are cooled to a certain degree after being sterilized (121 ℃, 15min), are uniformly mixed with the lactobacillus plantarum thallus, are placed in a glass plate to be pre-frozen for 24h in an ultra-low temperature refrigerator at minus 80 ℃, and are then placed in a freeze dryer for freeze-drying: vacuum degree of 1Pa, and cold trap at-83 deg.C for 70 hr. The freeze dried powder has survival rate up to 90%, and the viable count is 9 × 1011CFU/gProtectant solids；
When preparing freeze-dried thallus powder of the sayia orientalis, the preparation method comprises the following steps: the cultured yeast adopts 28% (w/w) peptone, 23% (w/w) skim milk powder, 20% (w/w) trehalose, and 5% (w/w) sea grape polysaccharide as raw materialsFreezing protectant, sterilizing the protectant (121 deg.C, 15min), cooling to a certain extent, mixing with the thalli of Saiyi Oriental Saiyi yeast, pre-freezing in a glass plate at-80 deg.C for 24h, and freeze-drying in a freeze-dryer: vacuum degree of 1Pa, and cold trap at-83 deg.C for 70 hr. The freeze dried powder has a survival rate of 89%, and contains viable bacteria of 1012CFU/gProtectant solids。
S3, preparing various groups of leaven:
TABLE 5 acid salted pepper fermentation group scheme
The starter culture for acid preserved chili used in example 3 is prepared by mixing lyophilized thallus powder at the ratio shown in table 5, as shown in table 5.
S4, fermenting the salted pepper:
the conventional method for preparing salted chili with acid (group 11) optimizes the fermentation process of salted chili with response to the dough processing. Chinese brewing, 2018, 10: 106-. And (5) making. The method specifically comprises the following steps: cutting cleaned red pepper into small pieces of 5mm × 5mm, cutting ginger and garlic into small pieces of 3mm × 3mm, pulverizing long-shaped rice, sieving to obtain rice flour, pre-pickling pepper for 1 hr, mixing pre-pickled pepper with small amount of ginger and garlic, adding rice flour, stirring, placing into a fermentation tank, fixing the tank opening with rice straw, sealing with liquid, and fermenting. The fermentation process conditions are that red pepper: 1.0 of long-shaped rice flour: 1.0(w/w), 15 days of fermentation time, 30 ℃ of fermentation temperature and 20 meshes of rice flour granularity.
The method for preparing the pickled pepper in groups 1-10 refers to group 11, which specifically comprises the following steps: cutting cleaned Capsici fructus into small pieces of 5mm × 5mm, cutting rhizoma Zingiberis recens and Bulbus Allii into small pieces of 3mm × 3mm, pulverizing semen oryzae, sieving to obtain rice flour, pre-pickling Capsici fructus for 1 hr, mixing pre-pickled Capsici fructus with small amount of rhizoma Zingiberis recens and Bulbus Allii, adding rice flour, adding direct vat set starter (shown in Table 5), adding total viable count of 108CFU/gChili pepperStirring, placing into a fermentation tank, fixing the tank opening with rice straw, inverting, sealing with liquid, and fermenting. The fermentation process conditions are that red pepper: 1.0 of long-shaped rice flour: 1.0(w/w), 15d of fermentation time, 30 ℃ of fermentation temperature and 20 meshes of rice flour granularity.
The flavor substances in the pickled hot pepper of the acid pickling were analyzed by using GC-MS 7890B-7010B (Agilent company, usa), volatile organic compounds in the sample were extracted by Solid Phase Microextraction (SPME), and the contents of 6 major ester substances (ethyl propionate, ethyl butyrate, ethyl hexanoate, ethyl octanoate, ethyl decanoate, and phenethylacetate) in the pickled hot pepper of the acid pickling were measured in MRM mode. The chromatographic conditions were as follows, capillary chromatography column: HP-5MS (30 m.times.250. mu.m.times.0.25 μm); gradient program: at 30 deg.C for 5 min; heating to 50 deg.C at 3 deg.C/min and maintaining for 3 min; raising the temperature to 150 ℃ at the speed of 5 ℃/min, and raising the temperature to 250 ℃ at the speed of 20 ℃/min and keeping the temperature for 5 min; the carrier gas is He, the flow rate is 1.0mL/min, and a non-flow splitting mode is selected.
TABLE 6 content of esters in fermented acid salted pepper
The results of the content of ester substances in the pickled pepper with acid are shown in table 6, and it is known that the total amount of ester substances in group 1 (freeze-dried powder of lactobacillus plantarum CGMCC21288 cell, freeze-dried powder of sayimyces orientalis cic 31431 cell mixed in a mass ratio of 1: 1) is the highest, and the contents of ethyl propionate, ethyl butyrate, ethyl caproate, ethyl caprylate, ethyl caprate and phenethyl acetate are all higher than those of group 2-11, and the contents are increased to about 1.2-5.2 times, 1.5-4.4 times, 1.2-2.6 times, 2.0-3.6 times, 1.0-2.9 times and 1.5-6.0 times of the control group, so as to achieve better flavor enhancement effect. The reason for this is probably that lactobacillus plantarum CGMCC21288 and sayia orientalis cic 31431 play a synergistic role, promoting the generation of esters. The 6 esters have pleasant aroma such as pineapple flavor, apple flavor, flower flavor, cream flavor and the like, have low threshold value, and have important contribution to the characteristic flavor of the pickled pepper. Therefore, the flavor-enhancing direct vat set starter can obviously improve the content of esters in the pickled peppers, thereby improving the flavor quality of products.
Example 4
The preparation method of the ester-producing flavor-enhancing type direct vat set starter and the application of the starter in the broad bean paste comprises the following steps:
s1, preparing lactobacillus plantarum thalli and sayia orientalis thalli:
when lactobacillus plantarum CGMCC21288 is prepared, the preparation method of the lactobacillus plantarum is as follows: strains stored at-80 ℃ were picked and streaked on MRS agar medium. The streaked plates were incubated for 36h in a 37 ℃ incubator. Single colonies on the plates were picked into MRS broth, shake-cultured at 37 ℃ and 200rpm for 16h, and then inoculated at 10% inoculum size into MRS broth, shake-cultured at 37 ℃ and 160rpm for 16 h. And centrifuging to collect thalli, washing the thalli for 2 times by using sterile normal saline, centrifuging again to collect thalli, and suspending by using a proper amount of sterile water for later use.
When preparing Issatchenkia orientalis, the thallus preparation method is as follows: the strain stored at-80 ℃ was picked and streaked on YPD agar medium. The streaked plates were incubated in a 28 ℃ incubator for 48 h. Single colonies on the plate were picked into YPD broth, shake-cultured at 28 ℃ and 100rpm for 24 hours, and then inoculated at 5% inoculum size into YPD broth, and shake-cultured at 28 ℃ and 160rpm for 24 hours. And (3) centrifuging to collect thalli, washing the thalli for 1 time by using sterile physiological saline, centrifuging again to collect thalli, and suspending the thalli by using a proper amount of sterile water for later use.
S2, preparing lactobacillus plantarum freeze-dried powder and east sayiyeast freeze-dried powder:
when the lactobacillus plantarum freeze-dried powder is prepared, the preparation method comprises the following steps: the cultured lactobacillus plantarum thallus adopts 12% (w/w) peptone, 20% (w/w) inulin, 27% (w/w) trehalose and 7% (w/w) vitis amurensis polysaccharide as freezing protective agents, the protective agents are cooled to a certain degree after being sterilized (121 ℃, 15min), are uniformly mixed with the lactobacillus plantarum thallus, are placed in a glass plate to be pre-frozen for 24h in an ultra-low temperature refrigerator at minus 80 ℃, and are then placed in a freeze dryer for freeze-drying: vacuum degree of 1Pa, cold trap of-83 deg.CFreeze-drying for 70h under the condition. The freeze dried powder has survival rate up to 90%, and the viable count is 7 × 1011CFU/gProtectant solids；
When preparing freeze-dried thallus powder of the sayia orientalis, the preparation method comprises the following steps: the cultured yeast body adopts 30% (w/w) peptone, 30% (w/w) skim milk powder, 15% (w/w) trehalose and 8% (w/w) sea grape polysaccharide as freezing protective agents, the protective agents are cooled to a certain degree and uniformly mixed with the thalli of the east sayi yeast after being sterilized (121 ℃, 15min), the mixture is placed in a glass plate to be pre-frozen for 24h in an ultra-low temperature refrigerator with the temperature of-80 ℃, and then the freeze-drying machine is used for freeze-drying: vacuum degree of 1Pa, and cold trap at-83 deg.C for 70 hr. The freeze dried powder has survival rate up to 89%, and the viable count is 6 × 1011CFU/g Protectant solids。
S3, preparing various groups of leaven:
TABLE 7 fermentation group scheme for thick broad-bean sauce
The fermented soybean paste starter used in example 4 was prepared in accordance with the following composition scheme shown in Table 7, by mixing lyophilized bacterial powder at the ratio shown in Table 7.
S4, fermenting the broad bean paste:
the traditional preparation method of the broad bean paste (group 11) refers to the influence of different raw material pretreatment processes on the quality of the broad bean paste. Food industry science and technology 2020, 41(7): 301-. And (5) making. The method specifically comprises the following steps: spore concentration of 107Respectively inoculating the Aspergillus oryzae and Aspergillus niger suspension per mL according to the inoculation amount of 1% (v/w) on a seed koji culture medium, culturing at 30 ℃ for 72h, and turning over the koji once for 24h to prepare a seed koji for later use. Mixing broad beans and water at a ratio of 1:2(w/w), soaking for 16h, shelling, steaming at 121 deg.C for 20min, exhausting gas, and cooling. The mass ratio of the broad beans to the flour is 4:1, and the flour and the seed starter are uniformly mixed according to the inoculation amount (the mass of the seed starter accounts for the total mass of the broad beans and the flour) of 0.5 percent (the mass of the seed starter accounts for 3:1 of the mass of the broad beans and the flour) (the ratio of the aspergillus oryzae to the aspergillus niger is 3:1)Mixing, placing into a triangular flask, mixing with broad bean, wrapping with eight layers of gauze, culturing in 30 deg.C incubator for 5d, shaking for 2 times every day, and allowing broad bean to grow spores on its surface to obtain the final product. Uniformly mixing the broad bean koji with saline water with the concentration of 16% in a crock according to the mass-volume ratio of 1:1.5, placing the mixture in a constant temperature box at 45 ℃ for culturing for 30 days for maturation, and turning over the sauce once every day.
The making method of the broad bean paste in the groups 1-10 refers to the group 11, and specifically comprises the following steps: the spore concentration is 107Respectively inoculating the Aspergillus oryzae and Aspergillus niger suspension per mL according to the inoculation amount of 1% (v/w) on a seed culture medium, culturing at 30 ℃ for 72h, and turning over the seed culture once for 24h to prepare the seed culture for later use. Mixing broad beans and water at a ratio of 1:2(w/w), soaking for 16h, shelling, steaming at 121 deg.C for 20min, exhausting gas, and cooling. The mass ratio of broad bean to flour is 4:1, flour and seed koji are uniformly mixed according to the inoculation amount (the ratio of aspergillus oryzae to aspergillus niger is 3:1) of 0.5% (the seed koji accounts for the total mass of broad bean and flour), and a direct vat starter (shown in table 7) is added, wherein the total number of viable bacteria added is 106CFU/gThick broad-bean sauceThen putting into a triangular flask to be fully mixed with the broad beans, wrapping with eight layers of gauze, putting into a constant temperature incubator at 30 ℃ to be cultured for 5d, shaking up for 2 times every day, and obtaining the needed broad bean koji after the broad bean petals are full of spores. Mixing broad bean koji with 16% saline water at a mass-volume ratio of 1:1.5, placing in 45 deg.C incubator for 30d, and turning over sauce once every day.
The flavor substances in the thick broad-bean sauce were analyzed by using GC-MS 7890B-7010B (Agilent company, usa), volatile organic compounds in the sample were extracted by Solid Phase Microextraction (SPME), and the contents of 6 major ester substances (ethyl propionate, ethyl butyrate, ethyl hexanoate, ethyl octanoate, ethyl decanoate, and phenethylacetate) in the thick broad-bean sauce were measured in the MRM mode. The chromatographic conditions were as follows, capillary chromatography column: HP-5MS (30 m.times.250. mu.m.times.0.25 μm); gradient program: at 30 deg.C for 5 min; heating to 50 deg.C at 3 deg.C/min and maintaining for 3 min; raising the temperature to 150 ℃ at the speed of 5 ℃/min, and raising the temperature to 250 ℃ at the speed of 20 ℃/min and keeping the temperature for 5 min; the carrier gas is He, the flow rate is 1.0mL/min, and a non-flow splitting mode is selected.
TABLE 8 content of esters in fermented Thick broad-bean sauce
The results of the content of the esters in the thick broad-bean sauce are shown in table 8, and it is known that the total amount of the esters in group 1 (the freeze-dried powder of lactobacillus plantarum CGMCC21288 cells, the freeze-dried powder of the sayittuy orientalis CICC 31431 cells are mixed in a mass ratio of 1: 1) is the highest, and the contents of ethyl propionate, ethyl butyrate, ethyl caproate, ethyl caprylate, ethyl caprate and phenethylacetate are all higher than those in groups 2-11, and the contents of the esters are increased to about 1.1-8.0 times, 1.2-2.4 times, 1.0-4.0 times, 1.2-4.8 times, 1.1-7.5 times and 1.0-5.0 times of the control group, so that a better flavor enhancing effect is achieved. The reason for this is probably that lactobacillus plantarum CGMCC21288 and sayia orientalis cic 31431 play a synergistic role, promoting the generation of esters. The 6 esters have pleasant aroma such as pineapple aroma, apple aroma, flower aroma, cream aroma and the like, have a low threshold value, and have an important contribution to the characteristic flavor of the broad bean paste. Therefore, the adoption of the aroma-enhancing type direct vat set starter can obviously improve the content of esters in the broad bean paste, thereby improving the flavor quality of the product.
In conclusion, the ester-producing aroma-enhancing type direct vat set starter prepared by the invention has the characteristics of simple preparation, convenient use, high viable count and excellent fermentation performance. The ester-producing flavor-enhancing type direct vat set starter prepared by the invention can obviously improve the content of esters in fermented products, particularly sour fish, cheese, pickled hot pepper and thick broad-bean sauce, thereby improving the flavor quality of the fermented products.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be able to cover the technical solutions and the inventive concepts of the present invention within the technical scope of the present invention.
Claims (5)
1. AThe aroma-increasing type direct-vat-set starter is characterized by being formed by compounding lactobacillus plantarum freeze-dried powder and Issatchenkia orientalis freeze-dried powder according to the mass ratio of 1 (0.5-3), and the total viable count of the aroma-increasing type direct-vat-set starter is (10)10～1012)CFU/gProtectant solids(ii) a The Lactobacillus plantarum freeze-dried powder comprises Lactobacillus plantarum and a Lactobacillus plantarum freeze-protection agent, wherein the Lactobacillus plantarum is CGMCC21288, the Lactobacillus is preserved in China general microbiological culture Collection center (CGMCC) at 12 months and 03 days of 2020, has the preservation number of CGMCC21288 and is named as Lactobacillus plantarum (Lactobacillus plantarum) in classification; the Issatchenkia orientalis freeze-dried powder comprises Issatchenkia orientalis and Issatchenkia orientalis freeze-protecting agent, and the Issatchenkia orientalis is CICC 31431.
2. A method of preparing a flavored direct vat set starter according to claim 1, comprising the steps of:
s1, preparation of Lactobacillus plantarum thallus and Saiyi orientalis thallus:
when the lactobacillus plantarum thalli is prepared, the preparation method comprises the following steps: activating Lactobacillus plantarum strain with MRS agar plate, and performing MRS broth one-stage culture to obtain OD600nm1.0-10.0, washing the thalli for 1-2 times by using sterile normal saline, centrifuging, collecting, and suspending by using a proper amount of sterile water for later use;
when preparing the thalli of the sayittuy orientalis, the preparation method comprises the following steps: activating the Saccharomyces cerevisiae strain with YPD agar plate, performing YPD broth primary culture to obtain OD600nm1.0-10.0, washing the thalli for 1-2 times by using sterile normal saline, centrifuging, collecting, and suspending by using a proper amount of sterile water for later use;
s2, preparing the lactobacillus plantarum thallus and the say yeast oriental thallus by the step S1, and preparing lactobacillus plantarum freeze-dried powder and say yeast oriental freeze-dried powder:
when the lactobacillus plantarum freeze-dried powder is prepared, the preparation method comprises the following steps: the lactobacillus plantarum thallus obtained in the step S1 and the lactobacillus plantarum cryoprotectantMixing, freeze drying to obtain strain with concentration of 1010～1012) CFU/g protectant solids;
when preparing the freeze-dried powder of the saccharomyces orientalis, the preparation method comprises the following steps: the above-mentioned S1 step is carried out by mixing the obtained east saya thallus with the freezing protective agent of east saya, then freeze-drying to obtain the strain with the concentration of 1010～1012) CFU/g protectant solids;
s3, preparing a flavoring type direct vat set starter: and (3) mixing the lactobacillus plantarum freeze-dried powder obtained in the step S2 and the sayia orientalis freeze-dried powder according to the mass ratio of 1 (0.5-3).
3. The method for preparing a flavored direct vat set starter according to claim 2, wherein the lactobacillus plantarum cryoprotectant comprises 10-15% (w/w) peptone, 20-30% (w/w) inulin, 20-30% (w/w) trehalose, 5-10% (w/w) glucosamine polysaccharide; the freeze protectant for the sayia orientalis comprises 25-35% (w/w) peptone, 20-30% (w/w) skim milk powder, 10-20% (w/w) trehalose and 5-10% (w/w) sea grape polysaccharide.
4. The use of the flavor-enhancing direct vat set starter of claim 1 for the fermentation of pickled fish, pickled chili, cheese, thick broad-bean sauce, etc.
5. The use of the flavor-enhancing direct vat set starter according to claim 4, wherein the amount of the flavor-enhancing direct vat set starter of claim 1 added is (10) when fermenting6～109)CFU/g Material(s)。
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