CN113812600B - Method for preparing milk-flavored compound perfume base by two-step enzymolysis and combined fermentation - Google Patents

Method for preparing milk-flavored compound perfume base by two-step enzymolysis and combined fermentation Download PDF

Info

Publication number
CN113812600B
CN113812600B CN202111019651.1A CN202111019651A CN113812600B CN 113812600 B CN113812600 B CN 113812600B CN 202111019651 A CN202111019651 A CN 202111019651A CN 113812600 B CN113812600 B CN 113812600B
Authority
CN
China
Prior art keywords
milk
product
lactobacillus
flavored
flavor
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN202111019651.1A
Other languages
Chinese (zh)
Other versions
CN113812600A (en
Inventor
徐丹
彭雨露
徐学明
吴淑蒙
吴凤凤
杨哪
金亚美
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Jiangnan University
Original Assignee
Jiangnan University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Jiangnan University filed Critical Jiangnan University
Priority to CN202111019651.1A priority Critical patent/CN113812600B/en
Publication of CN113812600A publication Critical patent/CN113812600A/en
Application granted granted Critical
Publication of CN113812600B publication Critical patent/CN113812600B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • 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
    • A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
    • A23L27/20—Synthetic spices, flavouring agents or condiments
    • A23L27/24—Synthetic spices, flavouring agents or condiments prepared by fermentation
    • A23L27/25—Dairy flavours
    • 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
    • A23C13/00—Cream; Cream preparations; Making thereof
    • A23C13/12—Cream preparations
    • A23C13/16—Cream preparations containing, or treated with, microorganisms, enzymes, or antibiotics; Sour cream
    • 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
    • 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
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/123—Bulgaricus
    • 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
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/125—Casei
    • 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
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/137—Delbrueckii
    • 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
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/147—Helveticus
    • 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
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/157—Lactis
    • 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
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/165—Paracasei
    • 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
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/169—Plantarum
    • 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
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/21—Streptococcus, lactococcus
    • A23V2400/249—Thermophilus

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Health & Medical Sciences (AREA)
  • Nutrition Science (AREA)
  • Dairy Products (AREA)
  • Seasonings (AREA)

Abstract

The invention discloses a method for preparing a milk-flavored compound perfume base by two-step enzymolysis and combined fermentation, and belongs to the technical field of foods. The invention provides a method for preparing milk-flavored compound perfume base by using soy protein to replace cream and adopting two-step enzymolysis combined fermentation. The two-step enzymolysis method and the lactobacillus fermentation are reasonably combined, the function of the bacteria enzyme is more effectively exerted, the utilization rate of raw materials is improved, the aroma production rate is accelerated and the aroma intensity is improved on the premise of ensuring the characteristic flavor, and meanwhile, the aroma is finer, more natural, softer and richer.

Description

Method for preparing milk-flavored compound perfume base by two-step enzymolysis and combined fermentation
Technical Field
The invention relates to a method for preparing a milk-flavored compound perfume base by two-step enzymolysis and combined fermentation, belonging to the technical field of foods.
Background
The cream is widely popular with people due to higher flavor value, and has wide application in vegetable, dessert and baked food, but has higher cost, and the types and contents of protein source nutrition and flavor substances are relatively deficient. Therefore, the cream needs to be subjected to aroma enhancement treatment, the composition of protein source substances is enriched, the consumption cost of the cream is reduced, and simultaneously, the higher flavor requirement and the higher nutritive value are achieved.
The soybean protein is a high-quality plant protein produced by taking soybean as a raw material, and can be used as one of the substitute varieties of animal protein. The food has comprehensive nutrition, high protein and low fat, does not contain cholesterol, has rich amino acid types and content, has the effects of increasing calcium, reducing cholesterol and the like, and can improve the protein content of food and enhance the nutritional value when used in food. Meanwhile, the good functional characteristics such as emulsifying property, hydration property, oil absorption, gel property and the like can further help to improve the quality of food, and the food is widely applied to flour products, minced fillet products, dairy products and meat products.
However, the soybean protein utilization in China is still in the primary stage, the product type is single, and the resource utilization rate is low, so that the research and development of new products are required to be enhanced.
In the preparation method of the milk flavor base, the treatments such as blending, essence and the like do not accord with the current trend of food greening and health, and the obtained flavor is not soft in stimulus. The biological method uses butter as a main raw material, obtains the flavor base with 150-250 times of enhanced flavor through the actions of microorganisms and enzymes, overcomes the defects, has finer and finer mouthfeel and richer flavor, can obtain certain beneficial substances, and enhances the nutritive value to a certain extent.
The fermentation engineering technology and the enzyme engineering technology are key technologies for preparing the milk flavor base, and the existing milk flavor base is prepared by simply combining a single enzyme method, a microorganism method or a one-step enzyme method with the microorganism method, namely protease and/or lipase and the like are added simultaneously during fermentation, but the milk flavor base prepared by adopting the existing method has the defects of single deficiency of protein source nutrition and flavor substances, monotonous flavor type, long flavor production period, low flavor stimulation, easy generation of stimulating flavor, lower yield, longer time consumption and the like.
For example, zhou Meiyu and the like adopt the technical means of enzyme engineering in the paper of preparing natural cream essence by using the butter through complex enzyme enzymolysis to prepare milk flavor base, and the technical scheme achieves the effect of improving the flavor intensity, but has the defects of monotonous aroma type and easy generation of pungent flavor; as another example, wang Wei adopts the technical scheme of fermentation engineering in the paper of 'research on preparing natural milk flavor essence by lactobacillus fermentation', while improving the disadvantages of monotonous aroma type and easy generation of pungent flavor of enzyme engineering, achieves the effects of unique, soft, rich and fine aroma, but has small aroma intensity, lower yield and longer time consumption; as another example, lin Weifeng adopts a technical scheme of combining enzyme engineering and fermentation engineering in the paper of 'influence of protease and lipase on fermentation characteristics and flavor of a cream-whey system', and the like, so that the defect of a certain single method is overcome to a certain extent, but the problems that the lipase is hydrolyzed by the protease while the hydrolysis of fat generates a large amount of free fatty acid to inhibit the proteolysis and the growth of lactobacillus are solved, so that the bacterial enzyme benefit cannot be maximized, resources are wasted, the cost is increased and the like are solved; there is therefore a need for a more rational and efficient process for preparing a milk-flavored base having a fermented aroma and a dense cheese flavor.
Disclosure of Invention
In order to solve the defects of single deficiency of protein source nutrition and flavor substances, monotonous aroma type, low aroma production period long rate, unsmooth flavor stimulation, insufficient green, safe and natural effects, incapability of maximizing benefits due to antagonism generated by bacteria and enzymes in the preparation process and the like of the existing milk flavor base, and meanwhile, the defects of single variety, low processing degree, low resource utilization rate and the like of products manufactured by soy proteins (including soy protein isolate, soy protein concentrate and the like) are overcome, the invention provides the preparation method of the milk flavor composite flavor base, which not only enriches soy protein processing products and extends the industrial chain of the soy protein processing products, but also provides the green natural milk flavor composite flavor base which combines fermentation and enzymolysis more reasonably and effectively, has more controllable process degree, rapid aroma production, natural strong aroma, rich, soft and full flavor and the like, and enhances the nutritive value of the milk flavor composite flavor base to a certain extent.
The invention provides a preparation method of a milk-flavored compound perfume base, which comprises the following steps:
(1) Preparing a substrate:
mixing milk fat and vegetable protein according to the mass ratio of (1:5) - (20:1) to obtain a mixture; mixing the mixture with water according to the mass ratio of (1:0) - (1:9), preparing to obtain a substrate, and homogenizing the substrate;
(2) Fermenting and carrying out proteolysis:
the lactobacillus strain is prepared according to 1×10 3 ～1×10 8 Inoculating CFU/g inoculum size to the homogenized substrate prepared in step (1), adding proteolytic enzyme into the substrate according to the mass fraction of 0.05-1%, and culturing for 6-48 h at 28-32 ℃;
(3) Fat enzymolysis
And (3) adding lipase into the product prepared in the step (2) according to the mass fraction of 0.05-1%, and carrying out enzymolysis for 1-4 h at the temperature of 35-50 ℃ to obtain the milk-flavored compound perfume base.
In one embodiment of the present invention, step (1) is: fully mixing cream and vegetable protein according to the mass ratio of (1:5) - (20:1), adding a proper amount of water to prepare a liquid substrate with the mass concentration of 10-90%, homogenizing, sterilizing, and cooling to 25-35 ℃.
In one embodiment of the present invention, the homogenizing conditions are: 75 deg.C, 20Mpa.
In one embodiment of the present invention, in the step (2), the culture is performed at 28 to 32℃for 6 to 48 hours, and then the temperature is lowered to room temperature.
In one embodiment of the invention, in the step (3), after enzymolysis for 1-4 hours at 35-50 ℃, sterilization is carried out, and the milk-flavored compound perfume base is obtained.
In one embodiment of the present invention, the sterilization conditions are sterilization at 80-95 ℃ for 15-30min.
In one embodiment of the present invention, the cream in step (1) may be one or more of cream, butter, anhydrous cream.
In one embodiment of the present invention, the plant protein in step (1) includes one or more of soy protein, peanut protein, wheat protein and other plant-derived proteins, plant protein concentrates and derivatives.
In one embodiment of the invention, the vegetable protein is soy protein.
In one embodiment of the present invention, the bacterial species in step (2) is one or more of lactobacillus bulgaricus, lactobacillus helveticus, lactobacillus delbrueckii, streptococcus thermophilus, lactobacillus casei, lactobacillus paracasei, lactococcus lactis, lactobacillus plantarum and the like.
In one embodiment of the invention, the bacteria are purchased from North Nabiotech, inc., lactobacillus bulgaricus accession number BNCC223782, lactobacillus helveticus accession number BNCC193203, lactobacillus delbrueckii accession number BNCC168312, streptococcus thermophilus accession number BNCC340813, lactobacillus casei accession number BNCC137633, lactobacillus paracasei accession number BNCC345679, lactobacillus lactis accession number BNCC223809, and Lactobacillus plantarum accession number BNCC194165.
In one embodiment of the present invention, the proteolytic enzyme in step (2) is one or more of peptidase, flavourzyme, complex protease, neutral protease, alkaline protease and acid protease.
In one embodiment of the invention, the Peptidase is peptase R, the Flavourzyme500MG, the complex protease is Protamex, the neutral protease is Neutrase 0.5L, the alkaline protease is Alcalase 2.4L, and the acid protease is purchased from the new hawk bioengineering company in lakenan.
In one embodiment of the invention, the lipase in step (3) is a lipase and/or a phospholipase.
In one embodiment of the invention, the lipase is Palatase 20000L and the phospholipase is purchased from shandong science and technology biosciences ltd.
The invention also provides the milk-flavored compound perfume base prepared by the method.
The invention also provides a product containing the milk-flavored compound fragrance base.
In one embodiment of the invention, the product is a food or a health product.
In one embodiment of the present invention, the food comprises any one of dishes, baked goods, fermented foods, cold drinks, desserts.
In one embodiment of the invention, the health product comprises low-fat body-building powder and essence capsules.
The invention also provides application of the milk-flavored compound perfume base in preparing dishes, baked foods, fermented foods, cold drinks and desserts.
In one embodiment of the invention, the application is that the milk flavor compound perfume base is directly used as a raw material in the preparation process of dishes, baked foods, fermented foods, cold drinks and desserts or is used as a base material to be subjected to spray drying treatment to prepare milk flavor essence to be applied to various products.
Advantageous effects
(1) The milk-flavored composite flavor base prepared by the invention uses the soybean protein to partially replace cream, uses the soybean protein as a protein source auxiliary substrate, combines the plant protein with animal protein, overcomes the defect of insufficient content and variety of cream protein nutrient substances, builds a brand-new composite substrate system, accords with the development trend of green healthy foods, expands the nutrition value, flavor and taste types of the milk-flavored flavor base, overcomes the defect of high production cost of the cream and insufficient utilization of the soybean protein, simultaneously expands the soybean protein utilization channel, extends the industrial chain, fully utilizes resources and increases economic benefits.
(2) The invention adopts the method of adding protease and lactobacillus together, can degrade protein more efficiently to produce polypeptide and amino acid, further improves the nutritional value of the perfume base, and the peptide and amino acid can promote the growth of lactobacillus, produce acid and perfume, greatly shorten the fermentation time, and can be metabolized by lactobacillus to produce more nutrition, flavor substances and flavor precursor substances. Under the same conditions, when protease and lactobacillus are added simultaneously, the fermentation time can reach the number of strains which are fermented for 48 hours when only the lactobacillus is added; at the same fermentation time, the total amount of amino acids when protease and lactobacillus are added simultaneously is 53.40% higher than that when only lactobacillus is added.
(3) The invention adopts the preparation method of adding lipase after fermentation and proteolysis, which not only has the advantages of both a microbiological method and an enzymatic method, improves the nutritive value, accelerates the aroma production rate and improves the aroma intensity on the premise of ensuring the characteristic flavor, and obtains the milk-flavored compound flavor base with thick fermentation aroma and cheese flavor; and the problem that the hydrolysis of protein is inhibited by a large amount of free fatty acid generated by the hydrolysis of fat when lactobacillus, protease and lipase are added together is avoided. Therefore, the microbial enzyme can be more effectively exerted, the fat and protein of the raw materials are fully hydrolyzed, and the utilization rate of the raw materials is improved. Compared with the perfume base obtained by adding lactobacillus, protease and lipase together, the perfume base prepared by the method has the advantages that the content of 3-hydroxy-2-butanone serving as a common strong-flavor compound in butter and cheese products is increased by 333.76 percent, the content of important short-chain volatile fatty acid butyric acid and caproic acid is respectively increased by 98.52 percent and 22.42 percent, and the content of long-chain fatty acid with poor flavor is reduced.
(4) The invention reasonably combines the two-step enzymolysis method with the microbial method, so that the respective degrees of fermentation, proteolysis and fat enzymolysis are more controllable, the industrial application is facilitated, the flavor and taste types of the obtained perfume base are wider, more selectivity and applicability are provided, and the perfume base can be widely used for directly perfuming dishes, baked foods, cold drinks, desserts and snacks and also can be used as a base material of milk-flavored essence.
Drawings
Fig. 1: strain growth curves for examples and comparative examples containing lactic acid bacteria.
Detailed Description
The following description of the preferred embodiments of the present invention is provided for better illustration of the invention, and should not be construed as limiting the invention.
Bacteria referred to in the following examples were purchased from North NaBiotechnology Inc., wherein Lactobacillus bulgaricus was deposited with BNCC223782, lactobacillus helveticus was deposited with BNCC193203, lactobacillus delbrueckii was deposited with BNCC168312, streptococcus thermophilus was deposited with BNCC340813, lactobacillus casei was deposited with BNCC137633, lactobacillus paracasei was deposited with BNCC345679, lactococcus lactis was deposited with BNCC223809, and Lactobacillus plantarum was deposited with BNCC194165.
Alkaline protease Alcalase 2.4L, flavourzyme Flavourzyme500MG, composite protease Protamex, lipase Palatase 20000L were purchased from Norwesterns (China) investment Co., ltd; peptidase Peptidase R, neutral protease Neutrase 0.5L was purchased from Amersham pharmacia (Jiangsu) Inc.; acid protease was purchased from Hunan Xinhong eagle bioengineering Co., ltd; phospholipase was purchased from Shandong Techno Wenxing Biotech Co.
The soy proteins referred to in the examples below were purchased from Shandong Xiangchi Bean technology Co., ltd; cream was purchased from brome food limited; cream and anhydrous cream are purchased from constant natural commerce (Shanghai) limited.
The method for detecting the colony count of lactic acid bacteria in the following examples is as follows:
the colony number of the live lactobacillus is measured by adopting a gradient dilution plate counting method every 4 hours, 0.5g of cream sample is taken from each group to carry out 10 times of gradient dilution (10 5 ～10 7 Multiple), 100. Mu.L of diluent and normal saline (blank) were added to a plate of mMRS solid medium, and the plate was placed in anaerobic condition at 30℃for inversion culture, and colony counting was performed after 48 hours, and 3 replicates were performed at the same dilution concentration for each sample.
The method for detecting volatile flavor substances in the following examples is as follows:
weighing 5g of cream sample into a 20mL headspace vial and sealing, balancing the vial in a water bath at 60 ℃ for 20min, inserting an aged 65 mu m PDMS/DVB extraction head into the headspace of the vial and adsorbing for 40min, then taking out the extraction head with adsorbed volatile substances, inserting the extraction head into a sample injection hole of a gas chromatograph-mass spectrometer, and desorbing for 10min.
The model of the capillary chromatographic column is DB-WAX 122-7032 (30 m×0.25mm×0.25 μm), and high-purity helium is used as carrier gas with the flow rate of 1mL/min. The procedure set up for the gas chromatograph is as follows: the temperature was kept at 40℃for 2min, then raised to 100℃at a rate of 6℃per min, followed by a temperature rise to 230℃at a rate of 10℃per min, and maintained for 6min. The temperature of the sample inlet is 250 ℃, the ionization mode is EI, the temperature of the ion source is 200 ℃, the electron energy is 70eV, the emission current is 200 mu A, and the mass range is 33-495 m/z by adopting a full scanning acquisition mode. And matching and searching the obtained GC-MS spectrum peaks with Wiley Library and NIST Library data, wherein the matching degree and purity are more than 900 and are regarded as effective identification results, and the peak area is used for representing the relative content of volatile flavor substances.
The method for detecting free amino acids in the following examples is as follows:
the determination of free amino acids was performed on a High Performance Liquid Chromatography (HPLC) system equipped with an ODS Hypersil capillary column, an autosampler and a uv detector. 1g of cream sample was weighed and put into a 25mL volumetric flask, 5% (w/w) trichloroacetic acid was used for volume fixation, and the sample was placed into an ultrasonic apparatus for ultrasonic treatment at 25℃for 15min, and placed for 2h, and then centrifuged at 10000rpm for 30min, and the obtained supernatant was filtered again through a filter to obtain a final filtrate for measurement.
The detection parameters were set as follows: column temperature, 40 ℃; detection wavelength, 338nm (262 nm for proline); mobile phase A (pH 7.2) (27.6 mmol/L sodium acetate-trimethylamine-tetrahydrofuran, volume ratio 500:0.11:2.50); mobile phase B (pH 7.2): (80.9 mmol/L sodium acetate-methanol-acetonitrile, volume ratio 1:2:2). Gradient elution was performed as follows: 8% B,0min;50% B,17.0min;100% B,20.1min;0% B,24.0min, flow rate 1.0mL/min.
The free amino acid concentration was calculated from the peak area using the external standard method.
The following examples relate to the following media:
mrs liquid medium: 10.0g of peptone, 5.0g of beef extract, 5.0g of yeast extract, 10.0g of malt extract, 5.0g of glucose, 10.0g of maltose, 5.0g of fructose and K 2 HPO 4 4.0g，KH 2 PO 4 2.6g，NH 4 Cl 3.0g, L-cys HCl 0.5g, tween 80 1.0g, mgSO 4 ·7H 2 O 0.1g，MnSO 4 ·7H 2 O0.05 g, adding deionized water to a constant volume of 1000mL，pH 6.2～6.8。
mrs solid medium: 1.5% agar was added on the basis of mMRS broth.
Example 1: preparation of milk-flavored compound perfume base
A method for preparing milk-flavored compound perfume base by using soy protein to replace cream through two-step enzymolysis and combined fermentation comprises the following steps:
(1) Mixing 100g of cream and 5g of soy protein, adding 7.5g of water to prepare a liquid substrate with the mass concentration of 40%, homogenizing the prepared substrate under the conditions of 75 ℃ and 20Mpa, sterilizing for 15min at the temperature of 95 ℃ after homogenizing, and cooling the substrate to 25 ℃;
(2) Preparation of Lactobacillus suspensions
Inoculating a single colony of Lactobacillus bulgaricus into 1mL of mMRS liquid culture medium, performing anaerobic culture at 30 ℃ for 18 hours to obtain seed liquid, transferring the seed liquid into the mMRS liquid culture medium with an inoculum size of 2% (v/v), culturing for 24 hours to obtain fermentation liquor, centrifuging the fermentation liquor under 4000r/min for 10 minutes, discarding supernatant, collecting thalli, and re-suspending the thalli by using sterile water to obtain bacterial suspension;
(3) Inoculating the lactobacillus bulgaricus bacterial suspension obtained in the step (2) into the product of the step (1), wherein the inoculation amount is 1.5x10 7 CFU/g; meanwhile, 0.06g of Flavourzyme500MG is added into the product, after being cultured for 15 hours in a constant-temperature incubator at 28 ℃, the product is sterilized for 15 minutes at 95 ℃ and then cooled to room temperature;
(4) Adding 0.06g of lipase Palatase 20000L into the product prepared in the step (3), carrying out constant-temperature enzymolysis for 4 hours at 37 ℃, sterilizing for 15 minutes at 95 ℃ and obtaining the milk-flavored compound perfume base 1 after sterilization.
Example 2: preparation of milk-flavored compound perfume base
A method for preparing milk-flavored compound perfume base by using soy protein to replace cream through two-step enzymolysis and combined fermentation comprises the following steps:
(1) Thoroughly mixing 120g of cream and 50g of soy protein, adding 45.43g of water to prepare a liquid substrate with the mass concentration of 70%, homogenizing the prepared substrate under the conditions of 75 ℃ and 20Mpa, sterilizing for 17min at the temperature of 90 ℃ after homogenizing, and cooling the substrate to 30 ℃;
(2) Preparation of Lactobacillus suspensions
Respectively picking single bacterial colonies of lactococcus lactis and lactobacillus casei, respectively inoculating the single bacterial colonies into 1mL of mMRS liquid culture medium, performing anaerobic culture at 30 ℃ for 18 hours to prepare seed liquid, respectively transferring the seed liquid into the mMRS liquid culture medium according to the inoculum size of 2% (v/v), culturing for 24 hours to obtain fermentation liquor, centrifuging the fermentation liquor under 4000r/min for 10 minutes, discarding supernatant, collecting thalli, and re-suspending the thalli by using sterile water to obtain bacterial suspension;
(3) Respectively mixing the lactobacillus lactis and lactobacillus casei suspension obtained in step (2) according to 2×10 7 CFU/g and 3X 10 7 Inoculating CFU/g into the product obtained in the step (1) at the same time, adding 0.05g of composite protease Protamex and 0.05g of Peptidase Peptidase R into the product, culturing in a constant temperature incubator at 30 ℃ for 10 hours, sterilizing at 90 ℃ for 17 minutes, and cooling to room temperature;
(4) Adding 0.07g of phospholipase into the product prepared in the step (3), carrying out enzymolysis for 3.5 hours at a constant temperature of 40 ℃, sterilizing for 17 minutes at a temperature of 90 ℃, and obtaining the milk-flavored compound perfume base 2 after sterilization.
Example 3: preparation of milk-flavored compound perfume base
A method for preparing milk-flavored compound perfume base by using soy protein to replace cream through two-step enzymolysis and combined fermentation comprises the following steps:
(1) Thoroughly mixing 150g of cream and 150g of soy protein, adding 804g of water to prepare a liquid substrate with the mass concentration of 25%, homogenizing the prepared substrate under the conditions of 75 ℃ and 20Mpa, sterilizing at 85 ℃ for 20min after homogenization, and cooling the substrate to 30 ℃;
(2) Preparation of Lactobacillus suspensions
Respectively picking lactobacillus paracasei and lactobacillus plantarum, inoculating single bacterial colonies, respectively inoculating into 1mL of mMRS liquid culture medium, performing anaerobic culture at 30 ℃ for 18 hours to obtain seed liquid, respectively transferring the seed liquid into the mMRS liquid culture medium according to the inoculum size of 2% (v/v), culturing for 24 hours to obtain fermentation liquor, centrifuging the fermentation liquor at 4000r/min for 10 minutes, discarding supernatant, collecting thalli, and re-suspending the thalli by using sterile water to obtain bacterial suspension;
(3) Respectively mixing lactobacillus paracasei and lactobacillus plantarum bacterial suspensions obtained in the step (2) according to the ratio of 5 multiplied by 10 7 CFU/g and 1X 10 7 Inoculating CFU/g into the product obtained in the step (1) at the same time, adding 0.3g of acid protease and 0.25g of Peptidase Peptidase R into the product, culturing in a constant temperature incubator at 32 ℃ for 7 hours, sterilizing at 85 ℃ for 20 minutes, and cooling to room temperature;
(4) Adding 0.2g of phospholipase and 0.3g of lipase Palatase 20000L into the product prepared in the step (3), carrying out constant-temperature enzymolysis for 2h at 45 ℃, sterilizing for 20min at 85 ℃ to obtain the milk-flavored compound perfume base 3.
Example 4: preparation of milk-flavored compound perfume base
A method for preparing milk-flavored compound perfume base by using soy protein to replace cream through two-step enzymolysis and combined fermentation comprises the following steps:
(1) Mixing 200g anhydrous butter and 200g soybean protein, adding 225g water to obtain liquid substrate with mass concentration of 64%, homogenizing the prepared substrate at 75deg.C and 20Mpa, sterilizing at 80deg.C for 30min, and cooling to 32deg.C;
(2) Respectively picking single bacterial colonies of Lactobacillus delbrueckii, lactobacillus helveticus and streptococcus thermophilus, respectively inoculating the single bacterial colonies into 1mL of mMRS liquid culture medium, performing anaerobic culture at 30 ℃ for 18 hours, preparing seed liquid, respectively transferring the seed liquid into the mMRS liquid culture medium according to the inoculum size of 2% (v/v), culturing for 24 hours to obtain fermentation liquor, centrifuging the fermentation liquor under 4000r/min for 10 minutes, discarding supernatant, collecting thalli, and re-suspending the thalli by using sterile water to obtain bacterial suspension;
(3) Respectively mixing the Lactobacillus delbrueckii, lactobacillus helveticus and Streptococcus thermophilus suspensions obtained in step (2) according to 2×10 7 CFU/g、3×10 7 CFU/g、4×10 7 Inoculum size of CFU/g, inoculated simultaneously onAdding 0.5g Flavourzyme500MG, 0.3g composite protease Protamex and 0.2g alkaline protease Alcalase 2.4L into the product obtained in the step (1), culturing in a constant-temperature incubator at 32 ℃ for 6 hours, sterilizing at 80 ℃ for 30 minutes, and cooling to room temperature;
(4) Adding 0.5g of phospholipase and 0.5g of lipase Palatase 20000L into the product prepared in the step (3), carrying out enzymolysis for 1h at a constant temperature of 50 ℃, and sterilizing for 30min at a temperature of 80 ℃ to obtain the milk-flavored compound perfume base 4.
Comparative example 1:
untreated cream in example 1 was used as comparative example 1.
Comparative example 2:
the specific embodiment is the same as example 1, except that only lactobacillus bulgaricus bacterial suspension is inoculated in the step (3) and the step (4) is removed; namely, the specific steps are as follows:
step (1) and step (2) are the same as in example 1;
inoculating the lactobacillus bulgaricus bacterial suspension obtained in the step (2) into the product of the step (1), wherein the inoculation amount is 1.5x10 7 CFU/g; culturing in a constant temperature incubator at 28deg.C for 15 hr, sterilizing at 95deg.C for 15min, and cooling to room temperature to obtain milk flavor compound perfume.
Comparative example 3:
the specific embodiment is the same as example 1, except that only flavourzyme is added in step (3) and steps (2) and (4) are removed; namely, the specific steps are as follows:
step (1) is the same as in example 1;
adding 0.06g of Flavourzyme500MG into the product prepared in the step (1), culturing for 15h in a constant-temperature incubator at 28 ℃, sterilizing for 15min at 95 ℃, and cooling to room temperature to obtain the milk-flavored compound perfume base.
Comparative example 4:
the specific embodiment is the same as example 1, except that step (2) and step (3) are removed, namely, the specific steps are as follows:
step (1) is the same as in example 1;
adding 0.06g of lipase Palatase 20000L into the product prepared in the step (1), carrying out constant-temperature enzymolysis for 4 hours at 37 ℃, sterilizing for 15 minutes at 95 ℃, and obtaining the milk-flavored compound perfume base after sterilization.
Comparative example 5:
the specific embodiment is the same as in example 1, except that the step (4) is removed, that is, the milk-flavored complex flavor base is prepared according to the sequence of the steps (1) to (3) of the example.
Comparative example 6:
the specific embodiment is the same as example 1, except that only lactobacillus bulgaricus bacterial suspension is inoculated in the step (3); namely, the specific steps are as follows:
step (1) and step (2) are the same as in example 1;
(3) Inoculating the lactobacillus bulgaricus bacterial suspension obtained in the step (2) into the product of the step (1), wherein the inoculation amount is 1.5x10 7 CFU/g; culturing in a constant temperature incubator at 28deg.C for 15 hr, sterilizing at 95deg.C for 15min, and cooling to room temperature;
(4) Adding 0.06g of lipase Palatase 20000L into the product prepared in the step (3), carrying out constant-temperature enzymolysis for 4 hours at 37 ℃, sterilizing for 15 minutes at 95 ℃, and obtaining the milk-flavored compound perfume base after sterilization.
Comparative example 7:
the specific embodiment is the same as in example 1, except that only flavourzyme is added in step (3); namely, the specific steps are as follows:
step (1) is the same as in example 1;
(2) Adding 0.06g of Flavourzyme500MG into the product prepared in the step (1), culturing for 15h in a constant-temperature incubator at 28 ℃, sterilizing for 15min at 95 ℃, and cooling to room temperature;
(3) Adding 0.06g of lipase Palatase 20000L into the product prepared in the step (2), carrying out constant-temperature enzymolysis for 4 hours at 37 ℃, sterilizing for 15 minutes at 95 ℃, and obtaining the milk-flavored compound perfume base after sterilization.
Comparative example 8:
the specific embodiment is the same as in example 1, except that the flavourzyme and lipase are added in step (3), and step (4) is removed; namely, the specific steps are as follows:
step (1) and step (2) are the same as in example 1;
(3) Inoculating the lactobacillus bulgaricus bacterial suspension obtained in the step (2) into the product of the step (1), wherein the inoculation amount is 1.5x10 7 CFU/g; meanwhile, 0.06g of Flavourzyme500MG and 0.06g of lipase Palatase 20000L are added into the product, and after being cultured for 15 hours in a constant-temperature incubator at 28 ℃, the product is sterilized for 15 minutes at the temperature of 95 ℃ to obtain the milk-flavored compound perfume base.
Example 5: characterization of milk-flavored Complex fragrance Properties obtained by different preparation methods
The milk-flavored complex bases prepared in examples 1 to 4 and comparative examples 1 to 8 were subjected to the determination of colony count (characterizing fermentation time and rate of aroma production), free amino acids and volatile flavor substances.
The invention focuses on the preparation process, namely, lactobacillus and protease are added into a milk-based system (soy protein and cream) to react at the same time, then lipase is added to carry out enzymolysis, and the two-step enzymolysis combined fermentation method is used for carrying out reaction to prepare the milk-flavor compound perfume base, wherein the types of the lactobacillus, the protease and the lipase have little influence on the reaction, and the obtained milk-flavor compound perfume bases 1-4 all meet the production requirements, and have the technical effects which are slightly different from those of the embodiment 1; this example only shows the corresponding properties of the milk-flavored complex bases prepared in example 1 and comparative examples 1-8.
The specific results are shown in fig. 1, table 1 and table 2.
Table 1 results of determination of free amino acid content (mg/kg) in examples and comparative examples
Table 2 peak areas of volatile flavors for examples and comparative examples
Description of the curves of fig. 1: since comparative examples 1 to 8 and example 1 relate to 4 groups of the bacteria-adding treatment groups in total, and the proteolytic and fermentation processes of comparative example 5 and example 1 are completely identical, in the growth curves of the previous strains, example 1 and comparative example 5 are coincident, so the strain growth curves represent 3 groups of data in total, to illustrate the effect of different enzyme engineering techniques when used in combination (comparative example 8 represents simultaneous co-addition of bacteria, protease and lipase; example 1/comparative example 5 represents two-step enzymolysis combined fermentation of the present invention) on the number of strain growth (characterizing fermentation time and rate of aroma production) relative to the fermentation engineering technique alone (comparative example 2). Meanwhile, although comparative examples 1 to 8 and example 1 only involved 15 hours of fermentation time, the strain growth curve provided to 48 hours of data can provide a more extensive further reference for the longer fermentation time examples.
(1) As shown in FIG. 1, the growth rate of viable bacteria in example 1 and comparative example 8 was always significantly higher than that in comparative example 2. At 16h, the number of viable bacteria in example 1 and comparative example 8 is more than that in comparative example 2, which indicates that the joint addition of protease and lactobacillus, i.e. the combination of fermentation engineering and enzyme engineering technology, can greatly promote the proliferation of lactobacillus, further cause vigorous metabolic activity and improve the rate of fragrance production, thus greatly saving time cost.
Meanwhile, compared with comparative example 8, the living bacteria number of the example 1 is always higher, the growth speed of the early fermentation stage before 8h is obviously faster, and meanwhile, the bacterial strain number can still be kept stable in the late fermentation stage after 32h, which shows that the influence of the use of different enzyme engineering technologies in combination with the fermentation engineering technology on the bacterial strain growth number (the time consumption of characterization fermentation and the aroma production rate) is different. And when the method is applied to scale-up production/industrialization, the effect is expected to be more outstanding than that of a laboratory due to the higher total amount of the substrate.
(2) As shown in Table 1, compared with comparative example 1, the remaining seven groups can increase the amino acid content to different degrees, which shows that the microbial method and the enzymatic method can both improve the nutrition and flavor value of the milk-flavored compound perfume base, and the two-step enzymolysis method adopted by the invention has more outstanding effect on the accumulation of amino acid by combined fermentation, as the example 1 is the most superior.
Meanwhile, although the total amount of amino acids of comparative example 3 and example 1 was close due to the fact that the rapid increase in the amount of lactic acid bacteria consumed peptide and amino acid as nutrients, the use of amino acids by lactic acid bacteria was further demonstrated by the smaller amino acid content of comparative example 5 compared to comparative example 3 and example 1, but the increase in the total amount of amino acids of comparative example 5 by 53.40% compared to comparative example 2, which means that the rapid increase in the amount of lactic acid bacteria consumed amino acids while also producing amino acids and accumulated more than consumed, and the addition of protease together with lactic acid bacteria was still significantly advantageous over the addition of only bacteria. The vigorous metabolic activity of the bacteria caused by the sharp increase in the amount of lactic acid bacteria in example 1 can further generate the relevant flavor substances or flavor precursor substances, so that although the total amount of amino acids of comparative example 3 and example 1 is close, example 1 is expected to be still superior to comparative example 3 in flavor and the like, while further addition of lipase in the late stage of example 1 can obtain further accumulation of amino acids, and thus the total amount of amino acids of example 1 is slightly higher than that of comparative example 3.
(3) As shown in Table 2, in the key volatile flavor substances of the milk-flavored compound flavor base, benzaldehyde, medium and short chain fatty acids (acetic acid, butyric acid and caproic acid), 3-hydroxy-2-butanone, lactones (gamma-dodecalactone and delta-dodecalactone) and heterocycles (2-amyl furan and furfural) with outstanding contribution to aroma can respectively provide almond caramel aroma, sour cream aroma, yoghourt aroma, flower fruit aroma and sweet almond aroma. Whereas hexanal has typical beany and grassy flavors, long chain fatty acids (e.g., decanoic acid, dodecanoic acid) at concentrations above a certain level have a soapy or bitter taste, and high concentrations are generally unsuitable for food production.
Examples 1 and comparative examples 2, 5, 6, 8, acetic acid and 3-hydroxy-2-butanone achieved a range from 0 to some, demonstrating the outstanding effect of fermentation engineering techniques on both flavors.
The examples 1 and comparative examples 5 to 8 have significantly improved types and contents of volatile flavor substances, especially the contents of butyric acid, valeric acid, caproic acid, heptanoic acid, caprylic acid, nonanoic acid and the like, compared with comparative examples 1 to 4, and the combination of the enzyme engineering technique and the fermentation engineering technique has significantly advantages compared with the single action.
In particular, the content of amino acids in example 1 is similar to that in comparative example 3 (Table 1), but the volatile flavor compounds in example 1 are more abundant in variety and content, and the contents of acetic acid, butyric acid, caproic acid, 3-hydroxy-2-butanone, delta-dodecalactone, furfural and the like are far superior to those in comparative example 3, and the hexanal content is obviously reduced, so that the obvious promotion effect of the vigorous metabolic activity of the proliferated lactobacillus on the accumulation of flavor substances while the proliferated lactobacillus metabolizes amino acids as nutrients is further proved.
In general, the volatile flavor compounds of example 1 and comparative example 8 are most abundant in type and content. In contrast, example 1 compared with comparative example 8, it was found that the two-step enzymolysis combined fermentation used in the present invention, i.e., the preparation method of fermenting and proteolysis performed simultaneously first and then adding lipase (example 1) contributed more significantly to the improvement of the beneficial volatile flavor substances and the reduction of the content of the irritating bad flavor compounds than the simultaneous addition of lactic acid bacteria, protease and lipase (comparative example 8), such as 3-hydroxy-2-butanone, as a common strong odor compound in butter and cheese products, and the content of example 1 increased by 333.76% than that in comparative example 8; the amounts of important short chain volatile fatty acids butyric and caproic acids, example 1, were increased by 98.52% and 22.42% respectively, compared to comparative example 8, while the amounts of poorly flavored long chain fatty acids, example 1, were reduced compared to comparative example 8.
In conclusion, the method for preparing the milk-flavored compound perfume base by the two-step enzymolysis combined fermentation is more excellent.
While the invention has been described with reference to the preferred embodiments, it is not limited thereto, and various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.

Claims (5)

1. A method for preparing a milk-flavored compound base, which is characterized by comprising the following steps:
(1) Mixing 100g cream and 5g soy protein, adding 7.5g water to obtain 40% liquid substrate, homogenizing the prepared substrate at 75deg.C and 20Mpa, sterilizing at 95deg.C for 15min, and cooling to 25deg.C;
(2) Preparation of lactobacillus suspension:
inoculating a single colony of Lactobacillus bulgaricus into 1mL of mMRS liquid culture medium, performing anaerobic culture at 30 ℃ for 18h, preparing seed liquid, transferring the seed liquid into the mMRS liquid culture medium with an inoculum size of 2% (v/v), culturing for 24h to obtain fermentation liquor, centrifuging the fermentation liquor under 4000r/min for 10min, discarding supernatant, collecting thalli, and re-suspending the thalli with sterile water to obtain bacterial suspension;
(3) Inoculating the lactobacillus bulgaricus bacterial suspension obtained in the step (2) into the product of the step (1), wherein the inoculation amount is 1.5x10 7 CFU/g; meanwhile, 0.06g Flavourzyme500MG is added into the product, after being cultured in a constant-temperature incubator at 28 ℃ for 15h, the product is sterilized at 95 ℃ for 15min and then cooled to room temperature;
(4) Adding 0.06g lipase 20000L into the product prepared in the step (3), carrying out constant-temperature enzymolysis at 37 ℃ for 4h, sterilizing at 95 ℃ for 15min, and obtaining the milk-flavored compound perfume base.
2. The milk-flavored complex flavor base prepared by the method of claim 1.
3. A product comprising the milk-flavored complex flavor base of claim 2.
4. A product according to claim 3, wherein the product is a food or a health product.
5. The product of claim 4, wherein the food product comprises any one of a baked good, a fermented good, and a dessert.
CN202111019651.1A 2021-09-01 2021-09-01 Method for preparing milk-flavored compound perfume base by two-step enzymolysis and combined fermentation Active CN113812600B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202111019651.1A CN113812600B (en) 2021-09-01 2021-09-01 Method for preparing milk-flavored compound perfume base by two-step enzymolysis and combined fermentation

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202111019651.1A CN113812600B (en) 2021-09-01 2021-09-01 Method for preparing milk-flavored compound perfume base by two-step enzymolysis and combined fermentation

Publications (2)

Publication Number Publication Date
CN113812600A CN113812600A (en) 2021-12-21
CN113812600B true CN113812600B (en) 2023-11-28

Family

ID=78923564

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202111019651.1A Active CN113812600B (en) 2021-09-01 2021-09-01 Method for preparing milk-flavored compound perfume base by two-step enzymolysis and combined fermentation

Country Status (1)

Country Link
CN (1) CN113812600B (en)

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2280551A1 (en) * 1998-08-27 2000-02-27 Kraft Foods, Inc. Highly flavored component for use in cheese manufacture and method for producing
CN103211197A (en) * 2013-04-02 2013-07-24 华南理工大学 High-concentration fermented milky flavor base, and preparation method and application thereof
CN103238808A (en) * 2013-04-15 2013-08-14 杭州艾菲曼普香精香料有限公司 Method for preparing natural milk flavour raw material by catalyzing cream through complex enzyme preparation
CN105639584A (en) * 2016-01-27 2016-06-08 广州市臻味宜源食品配料有限公司 Method for preparing natural milk flavor essence base stock
CN105919014A (en) * 2016-04-28 2016-09-07 南昌朵美生物科技有限公司 Preparation method of soybean milk essence

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20110236545A1 (en) * 2008-12-03 2011-09-29 Solae, Llc Non-Dairy Creamers Comprising Protein Hydrolysate Compositions and Method for Producing the Non-Dairy Creamers

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2280551A1 (en) * 1998-08-27 2000-02-27 Kraft Foods, Inc. Highly flavored component for use in cheese manufacture and method for producing
CN103211197A (en) * 2013-04-02 2013-07-24 华南理工大学 High-concentration fermented milky flavor base, and preparation method and application thereof
CN103238808A (en) * 2013-04-15 2013-08-14 杭州艾菲曼普香精香料有限公司 Method for preparing natural milk flavour raw material by catalyzing cream through complex enzyme preparation
CN105639584A (en) * 2016-01-27 2016-06-08 广州市臻味宜源食品配料有限公司 Method for preparing natural milk flavor essence base stock
CN105919014A (en) * 2016-04-28 2016-09-07 南昌朵美生物科技有限公司 Preparation method of soybean milk essence

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
生物法制备稀奶油-乳清复合风味乳基的研究作;周艳;中国优秀硕士学位论文全文数据库;第1.1.1节、第1.1.2节、第3.2.1节、3.3.2.1-3.3.2.2节、第4.3.3.1节、第5.5节、第83页第2段 *
蛋白酶辅助发酵结合脂肪酶酶解法制备天然奶酪风味剂;周美玉;傅亮;赵燕楠;;中国乳品工业(04);全文 *

Also Published As

Publication number Publication date
CN113812600A (en) 2021-12-21

Similar Documents

Publication Publication Date Title
JP4823318B2 (en) Process for producing fermented soybeans with increased .GAMMA.-aminobutyric acid content.
JP4815493B2 (en) Medium composition containing fermented gochujang, brewed soy sauce stock or acid-decomposed soy sauce stock, and method for producing γ-aminobutyric acid
JP5972166B2 (en) Lactic acid-containing yeast extract
KR101106668B1 (en) The preparing method for natural GABA using serial fermentation of plant originated lactic acid bacteria with GABA-producing ability and protease-producing ability
EP1893033B1 (en) Method for preparing a dairy analogue
JPWO2007097374A1 (en) Lactic acid bacteria having the ability to produce γ-aminobutyric acid
RU2603096C2 (en) Flavour and aroma modulation by fermenting milk source for producing multiple flavours and aromas with cocktail of bacteria strains
CN111248409A (en) Low-salt thick broad-bean sauce fermentation method
JP2002300862A (en) METHOD FOR PRODUCING gamma-AMINOBUTYRIC ACID-CONTAINING NATURAL FOOD MATERIAL
CN103053950A (en) Method for fermenting soybean by virtue of lactobacillus
KR100882278B1 (en) Soy yogurt fermented with kimchi lactic acid bacteria and production method
JP2006238743A (en) Method for culturing lactic acid bacteria, lactic acid bacteria cultured product, food and drink, feed and feed additive, and pharmaceutical
CN113812600B (en) Method for preparing milk-flavored compound perfume base by two-step enzymolysis and combined fermentation
RU2593719C2 (en) Flavour modulation by bio-processing using flavour forming bacteria strains
Feng et al. Safety evaluation and application of lactic acid bacteria and yeast strains isolated from Sichuan broad bean paste
KR102243418B1 (en) Novel acetic acid bacteria and process for preparing Kombucha beverages using the same
JP3796463B2 (en) Beef extract-like natural seasoning and method for producing the same
EP1000547A1 (en) Process for producing seasoning liquors with flavor of vegetables pickled in salted rice bran paste
CN106834181A (en) A kind of Pediococcus acidilactici and its application
KR20160025090A (en) Method of manufacturing fermentation coffee
KR100624704B1 (en) Microorganism seperated from fermented soybeans
CN116286438B (en) Lactobacillus plantarum from bean curd acid pulp and application thereof
JP3530851B1 (en) New lactic acid bacteria and flavor-improving seasonings using the same
JP2008228654A (en) Fermented food doubling as culture medium, and method for producing the same
KR20030076765A (en) Manufacture of Korean rice wines(Takju and Yakju) utilizing bacteriocin produced by lactic acid bacteria.

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant