CN110447880A - A kind of preparation method of delicate flavour fermented flavoring material - Google Patents
A kind of preparation method of delicate flavour fermented flavoring material Download PDFInfo
- Publication number
- CN110447880A CN110447880A CN201910876200.6A CN201910876200A CN110447880A CN 110447880 A CN110447880 A CN 110447880A CN 201910876200 A CN201910876200 A CN 201910876200A CN 110447880 A CN110447880 A CN 110447880A
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- China
- Prior art keywords
- fermentation
- delicate flavour
- preparation
- fermented
- flavoring material
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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
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- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Chemical & Material Sciences (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Seasonings (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The present invention provides a kind of preparation method of delicate flavour fermented flavoring material, is digested using compound protease and neutral proteinase to animal protein source, obtains enzymolysis product；Then it is fermented using corynebacterium glutamicum and/or bacillus subtilis to hydrolysate, obtains tunning.Present invention process is simple, and environmental protection, at low cost, taste is mellow, and nutritive value is high, and safety is good, provides a new approach for the exploitation of fermented seasonings.
Description
Technical field
The invention belongs to field of microbial fermentation, and in particular to a kind of strain fermentation animal by production glutamic acid and nucleotide
Protein sources (such as shrimp enzymolysis liquid) obtain the fermentation liquid for being rich in glutamic acid, nucleotide, flavor peptide under the condition of culture of optimization,
The fermentation liquid is suitable for field of food, as base-material or auxiliary material for making the products such as flavors and fragrances or flavouring.
Background technique
Early in 1985, researcher had found to receive the taste receptors of delicate flavour material from human oral cavity taste bud, made delicate flavour
As the 5th class sense of taste after acid, sweet tea, hardship, salty four kinds of sense of taste.But people for delicate flavour pursuit in all age never
Stop, to meet the needs of people's life, delicate flavour flavouring comes into being, with the spy of its " it is necessary that the sense of taste remembers distinctness+life "
Point possesses vigorous demand in the industry.Delicate flavour flavouring experienced four Dai Biange.Early 20th century, Japanese chemists Ikeda chrysanthemum seedling
The sodium glutamate extracted from kelp is taught, is called first generation delicate flavour flavouring in history --- monosodium glutamate；20th century mid-term, scientist from
Inosinate is extracted in bonito stick, and guanosine hydrochlorate is obtained from yeast；Sodium glutamate is compounded with nucleosides hydrochlorate makes umami effects at double
Increase, be called second generation delicate flavour flavouring in history --- strength monosodium glutamate；The 1980s, the delicate flavour flavouring of exploitation be
Monosodium glutamate is added on the basis of the meat products such as ox, chicken, crab meat and prepares flavor type delicate flavour flavoring agent, is called the seasoning of third generation delicate flavour in history
Product --- compound monosodium glutamate；On the basis of third generation delicate flavour flavoring agent, addition amino acid, vitamin, Chinese herbal medicine etc. are at separately
Forth generation nutrition reinforced type flavouring is sent out.
To sum up, since Ikeda chrysanthemum seedling professor in more than the 100 years times of the delicate flavour ingredient sodium glutamate extracted in kelp,
Four generation flavouring are that the ticbit on dining table has played irreplaceable role.But Gourmet Powder Factory extracts paddy from fermentation liquid
It when propylhomoserin, generally separated directly from the fermentation liquid containing thallus and protein, extract glutamic acid, be converted into sodium glutamate,
The isolated monosodium glutamate of crystallization leads to the production of waste water and losing for part nutritional ingredient of a large amount of high concentrations generated in production
It loses, therefore the problems such as there is wastage of material, by-product contamination environment.At present at home and abroad in the market, third generation flavor type is fresh
The sales volume of taste flavouring substantially exceeds monosodium glutamate.Forth generation delicate flavour flavouring can supplement amino acid, small molecule necessary to human body
The nutritional ingredients such as peptide, vitamin and microelement have huge development potentiality, meanwhile, it has been found that whether any generation production
Product, their production are crude product to be obtained using the methods of microbial fermentation or enzymatic hydrolysis, then obtain through complicated separation, purification process
To purer flavor components, the compounding of last heterogeneity obtains final product, therefore production process is complicated, at high cost, together
When influenced by raw material itself there may be special smell, if yeast extract has special yeast stink, influence product
Flavor effect.Therefore, on the basis of forth generation delicate flavour flavouring, need employ new technology improves its quality, develops a kind of new
The delicate flavour substitute of type solves problem above.
Summary of the invention
The present invention is obtained using enzymatic hydrolysis material protein then using the microorganism direct fermentation enzymolysis product for producing delicate flavour material
Obtain natural fermented seasoning.Simple process, environmental protection, at low cost, taste is mellow, and nutritive value is high, and safety is good, for fermentation seasoning
The exploitation of product provides a new approach.
Corynebacterium glutamicum, bacillus subtilis mushroom are considered to have metabolite accumulation glutamic acid, nucleotide ability
Specified strain adds biotin, NH in the fermentation medium+、Mn2+、Mg2+The necessary factor needed for equal bacterial metabolisms, and control
Fermentation condition, such as temperature, pH, fermentation time factor can make bacterial metabolism generate the products such as glutamic acid, nucleotide.With industrialization
The delicate flavour flavoring agent of production is compared, and has natural food sense of taste by the delicate flavour ingredient that strain fermentation generates, as tasty agents energy
The natural taste of food is kept to the maximum extent, and safety is good, also has certain nutritive value.
Therefore, the solution of the present invention is to use animal protein source (including its hydrolysate) for substrate, by being inoculated with paddy ammonia
Sour corynebacteria or fermentation of bacillus subtilis and obtain rich in the delicate flavours such as glutamic acid, nucleotide, flavor peptide ingredient and special
The fermentation liquid or freeze-dried powder of ferment local-flavor can be used as seasoning use.
The preparation method of delicate flavour fermented flavoring material of the present invention, includes the following steps:
S1: digesting animal protein source using compound protease (AF) and neutral proteinase (AM), obtains enzymatic hydrolysis and produces
Object (hydrolyzate or enzymolysis liquid)；
S2: fermenting to hydrolysate using corynebacterium glutamicum and/or bacillus subtilis, obtains tunning
(fermentation liquid).
The above-mentioned mode for first digesting post-fermentation of the invention, can effectively improve delicate flavour ingredient and flavor intensity.It passes through first
Crossing the hydrolysate that enzymatic hydrolysis obtains has nutritional ingredient abundant, comprising small molecules ingredients such as amino acid, polypeptides, on the one hand can
Certain flavor components are provided, on the other hand can provide nutriment for bacterial metabolism in posterior fermentation process.Glutamic acid
This bacteroid of corynebacteria lay eggs white enzyme ability not as good as fungies and the producing enzyme bacterium such as aspergillus oryzae, yeast, direct fermentation animal
Albumen is difficult to make full use of protein, and digesting macro-molecular protein preferentially to add complex enzyme can be improved protein utilization rate, effectively
The degree of hydrolysis of albumen is improved, carries out adequate preparation for fermentation.Although also containing certain ammonia in the hydrolysate obtained through enzymatic hydrolysis
The ingredients such as base acid, polypeptide, carbohydrate, albumen can be used as a part of taste composition in final product, but its delicate flavour component content
With flavor deficiency.Enzymolysis protein is secondary objective in fermentation process, and main purpose is to make bacterial metabolism accumulation nucleotide and paddy ammonia
Acid, and flavor peptide is further accumulated, delicate flavour ingredient and flavor intensity can be greatly improved, is the master of delicate flavour ingredient in final product
Source is wanted, plays good reconciliation effect, natural sense of taste with ingredients such as unemployed remaining amino acid, polypeptide, carbohydrate, albumen
With delicate flavour flavor it is further preferred that.
Wherein, amino-acid nitrogen content is not less than 0.3g/100mL in the enzymolysis product；Using corynebacterium glutamicum into
In the tunning of row fermentation, the concentration of glutamic acid is greater than its threshold value (0.3g/L) in clear water；Using bacillus subtilis
In the tunning to ferment, the concentration of guanylic acid is greater than its threshold value (0.125g/L) in clear water；Preferably, using fermentation
Bacterial strain (corynebacterium glutamicum or bacillus subtilis) fermentation after target product (glutamic acid or guanylic acid) concentration compared to
10 times or more are improved before fermentation.The hydrolysis degree of control enzymolysis product is capable of in the optimization of amino-acid nitrogen content in enzymolysis product, hair
The accumulation degree of control delicate flavour flavor components is capable of in the optimization of target product concentration in ferment product, and final product is reached
The organoleptic requirements of target delicate flavour.
Wherein, the animal protein source is preferably the good shrimp of flavor, but not limited to this, also can be selected such as chicken,
The animal protein sources such as beef, the flesh of fish.Animal protein source is cleaned, drains, mashing, spare in the form of the meat of meat exquisiteness slurry.
Wherein, enzymatic hydrolysis condition in the enzymolysis process are as follows: 10~25LAPU compound protease (AF) 50LAPU/g, 0.05~
1~4h is digested in 6.0~7.0,50 DEG C of waters bath with thermostatic control of 0.075AU neutral proteinase (AM) 0.15AU/g, pH.
Wherein, the compound protease (AF) and neutral proteinase (AM) are to believe that (China) biotechnology is limited from Novi
The corynebacterium glutamicum 1276 and bacillus subtilis 1354 of company.
Wherein, the enzymolysis product before being fermented, is also added with composite nutrient, the composite nutrient composition
Are as follows: glucose 80；Corn pulp 3-12；K2HPO42-14；KH2PO40-4；MgSO4·7H2O 0-1.5；MgCl2·6H2O0-
1.5；MnSO4·7H2O 0.04-0.06；FeSO4·7H2O 0.0010-0.0020；CaCl2·2H2O 0-0.01；CuSO4·
5H2O 0-0.015；Half Guang hydrochloride 0-0.0020 of L-；(NH4)2SO45-20；Yeast extract (food-grade) 0-10；Unit:
g/L。
Wherein, fermentation condition in the fermentation process are as follows: 32 DEG C~38 DEG C of constant temperature, fluid nutrient medium shake culture
24~40h, midfeather adds appropriate ammonium hydroxide and biotin, for maintaining fermentation system pH while providing NH4+And provide bacterium
The necessary factor of body growth.
Wherein, before the fermentation process carries out, further include the steps that activating fermenting microbe: 30~32 DEG C of constant temperature shakes
Cultivation and fermentation agent, 130~150rpm/min, 8~12h are swung, fluid nutrient medium is continuously cultivated 2-3 times, by actication of culture, in 4 DEG C
Under the conditions of save；The bacterium fermented after activation number living is 107~108CFU/mL range.
The present invention uses technological parameter, reagent and the condition of above-mentioned optimization, enables thallus to the maximum extent in product
The delicate flavours ingredients such as middle accumulation glutamic acid, nucleotide, flavor peptide, keep the natural taste of food and mellow flavor, nutritive value equal
Weighing apparatus.The product of acquisition, since itself includes sugar, salt, organic acid, glutamic acid, nucleotide, flavor peptide, can be used as without extracting
Seasoning is directly applied, and can also be further processed into other products such as freeze-dried powder and be applied.
Compared with prior art, the method have the advantages that: fermentation substrate is native protein source, and source is wide
It is general, it can control production cost, its added value can be promoted after fermented.Fermentation time of the present invention is short, easy to operate, and production technology is held
It is easy to control, fermentation substrate is fully utilized, by-product and waste pollution are reduced, it is free from environmental pollution, it is applicable in industrialized production.Hair
Ferment product has both the characteristics of including delicate flavour taste composition and specific ferment local-flavor, is the natural fermented delicate flavour flavoring agent of production and base
The new method of material.
Detailed description of the invention
The attached drawing for constituting a part of the invention is used to provide further understanding of the present invention, schematic reality of the invention
It applies example and its explanation is used to explain the present invention, do not constitute improper limitations of the present invention.In the accompanying drawings:
Fig. 1 is that corynebacterium glutamicum 1276 of the present invention and the fermentation shrimp enzymolysis liquid of bacillus subtilis 1354 generate delicate flavour
Amino acid and nucleotide prepare the flow diagram of the process example of delicate flavour fermented flavoring material.
Fig. 2, Fig. 3 are corynebacterium glutamicum 1276 of the present invention and the fermentation shrimp enzymolysis liquid fermentation of bacillus subtilis 1354
Flavor mass spectrogram afterwards.
Specific embodiment
In order to better understand the present invention, below with reference to specific attached drawing, the present invention will be described in detail.
The preparation process of delicate flavour fermented flavoring material of the present invention is as shown in Figure 1, include following the description:
(1) pretreatment of raw material: using the good animal protein source shrimp of flavor as fermentation substrate, but not limited to this, it can be selected
The animal protein sources such as chicken, beef, the flesh of fish, the present invention is by taking shrimp as an example.
It takes appropriate shrimp cleaning, drain, be beaten, obtain the meat slurry of meat exquisiteness.
(2) it digesting: taking the shrimp slurry obtained in step (1), 2:5 adds water to mix in mass ratio, pH 6.0~7.0 is adjusted,
10~25LAPU compound protease (AF) 50LAPU/g, 0.05~0.075AU neutral proteinase (AM) 0.15AU/g are added respectively,
1~4h is digested in 50 DEG C of waters bath with thermostatic control.
(3) high pressure sterilization: take the enzymolysis liquid in step (2) in 115 DEG C of 15~30min of sterilizing.
(4) source of enzyme: protease used in step (2) believes (China) Bioisystech Co., Ltd from Novi；Its
Feature are as follows: 1LAPU, leucine aminopeptidase unit, Novi's letter internal rules, thorough hydrolysis substrate suggestion additive amount 10~
50LAPU/g；AF, AM, containing two kinds of activity of endo protease and exoproteinase, can be used to take off made from being fermented as aspergillus oryzae
Except rudimentary degree of hydrolysis product --- bitter taste protein hydrolysis degree bitter taste improves hydrolyzate flavor.
(5) nutrients: the battalion such as glucose, corn pulp, potassium dihydrogen phosphate, ammonium sulfate, yeast extract (food-grade) is added
Form point and the microorganisms such as half Guang hydrochloride of L-, magnesium ion, iron ion grow necessary to inorganic ions, prepare fermented and cultured
Base, and the 15~30min that sterilizes under the conditions of 115 DEG C.
(6) it prepares fermentation medium: taking animal protein enzymatic hydrolysis liquid after above-mentioned sterilizing, being rich in after addition step (5) sterilizing
The culture medium of nutritional ingredient is uniformly mixed, prepares fermentation medium, the mixing of two kinds of culture mediums need to be carried out aseptically.
(7) Spawn incubation: 30~32 DEG C of isothermal vibration cultivation and fermentation agent, fluid nutrient medium are continuously cultivated 2 times, and strain is living
Change, is saved under the conditions of 4 DEG C, for fermenting.
(8) ferment: the good fermenting microbe bacterium of activation culture lives number 10 for adjustment7~108CFU/mL range is inoculated into step
Suddenly in the culture medium of (6), 32 DEG C~38 DEG C of constant temperature is maintained the temperature at, fluid nutrient medium 24~40h of shake culture is intermediate
Appropriate ammonium hydroxide is added every a few hours and maintains fermentation system pH, while NH being provided4+, and add biotin provide thalli growth must
It must the factor.
(9) fermentation liquid detects: take appropriate fermentation liquid in centrifuge tube, 8000~10000rpm/min is centrifuged 1~10min,
Take supernatant, detection glutamic acid, nucleotide, inorganic ions and organic acid content.Fermentation liquid is taken to be uniformly mixed, 50~55 DEG C of conditions
Lower heat preservation 30~40min, 30~40min of headspace solid-phase microextraction detect volatile flavor component.
(10) sensory evaluation: taking appropriate fermentation liquid in triangular flask, 80~100 DEG C of 10~20min of heating water bath, sterilizing,
It is cooled to after room temperature and fermentation liquid is lyophilized into powder, carry out sensory evaluation.
(11) wherein, in fermentation liquid most of delicate flavour ingredient origin in bacterial metabolism.
(12) wherein, corynebacterium glutamicum belongs to facultative anaerobic bacteria, can be fermented, be passed through using carbon sources such as glucose
It controls fermentation temperature and improves isocitric dehydrogenase and Pidolidone dehydrogenase activity, make α-ketoglutaric acid in glutamte dehydrogenase
Catalysis and have NH4+ it is existing under the conditions of, generate glutamic acid.
(13) wherein, bacillus subtilis 5'-NT is energetic, metabolizable purine biosynthesis nucleotide.Utilize nitrogen source
With the carbon sources such as glucose, using 5 '-ribose phosphates as starting point by fully synthetic approach or by purine bases by glycosylation and phosphorylation
Nucleotide is generated by remedial pathway.
(14) production method of the good fermenting microbe of activation culture: ampoule bottle Kaifeng adds appropriate physiology salt water-soluble
Bacterium powder is solved, Agar Plating is inoculated in, is activated 2~3 times on agar medium, be inoculated into shaking flask, isothermal vibration later
(130~150rpm/min) 8~12h is cultivated, corynebacterium glutamicum 1276,1354 seed culture of bacillus subtilis is made
Liquid.
(15) wherein, since the culture medium of the nutritional ingredients such as shrimp protein enzymatic hydrolyzate and addition glucose contains a large amount of water
Point, supplemental oxygen in shake culture, conducive to the breeding of thallus and the accumulation of metabolite.
The present invention is further elaborated combined with specific embodiments below.
It is prepared by fermentation liquid of the embodiment 1 rich in glutamic acid:
Step (1): it obtains animal protein and digests liquid
35g shrimp is weighed to clean, drain, be beaten, obtain meat exquisiteness meat slurry, weigh 8 shrimp slurry, add 20mL go from
Sub- water, stirs evenly, adjust mixture ph be 7.0, respectively add 25LAPU compound protease (AF) 50LAPU/g,
0.075AU neutral proteinase (AM) 0.15AU/g digests 3h in 50 DEG C of waters bath with thermostatic control.
Step (2): high pressure sterilization
Take the enzymolysis liquid in step (1) in triangular flask, 115 DEG C of sterilizing 15min.
Step (3): addition nutrients (g/L):
Glucose 80；Corn pulp 4；K2HPO412；MgSO4·7H2O 1.0；MnSO4·7H2O 0.05；FeSO4·
7H2O0.0018；(NH4)2SO418, preparation is rich in the culture medium of nutritional ingredient, and the 20min that sterilizes under the conditions of 115 DEG C.
Step (4): fermentation medium is prepared
Animal protein after above-mentioned sterilizing is taken to digest liquid 20mL, the nutrient medium after addition 30mL step (3) sterilizing, mixing
Uniformly, fermentation medium is prepared, the mixing of two kinds of culture mediums need to be carried out aseptically.
Step (5): Spawn incubation
Two ring thallus of picking, is inoculated in 20mL fluid nutrient medium from Agar Plating, 32 DEG C of isothermal vibration trainings
12h is supported, twice, actication of culture saves under the conditions of 4 DEG C, for fermenting for continuous culture.
Step (6): fermentation
Adjustment the good fermenting microbe of activation culture with 8% inoculum concentration (initial bacterium number living: 2.12 × 107CFU/mL it) connects
Fermentation flask is placed on patrolling shaking table (165r/min), into the culture medium of step (4) originally in 32 DEG C of constant incubators by kind
Middle culture.It heats up 1 DEG C to 35 DEG C in fermentation process every 6h, and maintains this temperature fermentation 36h；200 μ L are added every 6h
HN3·H2O (25%) adjusts pH and maintains 7.0~7.5；The biotin for being 10 μ g/L every 6h addition 1mL concentration, provides thallus
Growth factor necessary to metabolism.
Step (7): fermentation liquor treatment
The triangular flask for filling fermentation liquid is heated into 15min in boiling water bath, thallus is inactivated, is cooled to room temperature, and in -20
DEG C save.
It is prepared by fermentation liquid of the embodiment 2 rich in nucleotide:
Step (1): it obtains animal protein and digests liquid
35g shrimp is weighed to clean, drain, be beaten, obtain meat exquisiteness meat slurry, weigh 8 shrimp slurry, add 20mL go from
Sub- water, stirs evenly, adjust mixture ph be 7.0, respectively add 25LAPU compound protease (AF) 50LAPU/g,
0.075AU neutral proteinase (AM) 0.15AU/g digests 3h in 50 DEG C of waters bath with thermostatic control.
Step (2): high pressure sterilization
Take the enzymolysis liquid in step (1) in triangular flask, 115 DEG C of sterilizing 15min.
Step (3): addition nutrients (g/L):
Glucose 80；Corn pulp 10；K2HPO43；KH2PO43；Yeast extract (food-grade) 8；MgCl2·6H2O1.0；
(NH4)2SO47；MnSO4·7H2O 0.05；FeSO4·7H2O 0.0018；CaCl2·2H2O 0.09；CuSO4·5H2O 0.01；
Half Guang hydrochloride 0.0018 of L-.
Step (4): fermentation medium is prepared
Animal protein after above-mentioned sterilizing is taken to digest liquid 20mL, the nutrient medium after addition 30mL step (3) sterilizing, mixing
Uniformly, fermentation medium is prepared, the mixing of two kinds of culture mediums need to be carried out aseptically.
Step (5): Spawn incubation
Two ring thallus of picking, is inoculated in 20mL fluid nutrient medium from Agar Plating, 32 DEG C of isothermal vibration trainings
10h is supported, twice, actication of culture saves under the conditions of 4 DEG C, for fermenting for continuous culture.
Step (6): fermentation
The good fermenting microbe of activation culture is connect with 10% inoculum concentration (initial bacterium number living: 1.0 × 108CFU/mL) for adjustment
Fermentation flask is placed on patrolling shaking table (165r/min), into the culture medium of step (4) originally in 32 DEG C of constant incubators by kind
Middle culture.It heats up 1 DEG C to 38 DEG C in fermentation process every 10h, and maintains this temperature fermentation 60h；Appropriate 2mol/ is added every 6h
The NaOH solution of L adjusts pH and maintains .6.8~7.0；The biotin for being 10 μ g/L every 6h addition 1mL concentration, provides thallus
Growth factor necessary to metabolism.
Step (7): fermentation liquor treatment
The triangular flask for filling fermentation liquid is heated into 15min in boiling water bath, thallus is inactivated, is cooled to room temperature, and in -20
DEG C save.
The non-volatile flavor analysis of 3 fermentation liquid of embodiment
Step (1): analysis content
Centrifuging and taking fermented liquid supernatant liquid, dilution, to it includes bacterial metabolism generate glutamic acid, nucleotide, organic acid
Content is detected, and the organic acid of measurement includes: tartaric acid, malic acid, lactic acid, succinic acid, citric acid, fumaric acid.And to hair
The variation of inorganic ion content added by before and after ferment is detected.
Step (2): it obtains fermentation liquid and analyzes result
Fermentation front and back taste compound changes of contents as can be seen from Table 1.The glutamic acid after corynebacterium glutamicum is fermented
Content is promoted, and provides delicate flavour ingredient, reduces tartaric acid, fumaric acid content before fermenting, and generates malic acid, lactic acid, citric acid.Withered grass
Guanylic acid, Inosine Content increase after fermentation of bacillus, provide delicate flavour ingredient, generate malic acid, lactic acid, improve lemon
Acid, fumaric acid content.There are Na in fermentation liquid+、K+Equal inorganic ions, can be auxiliary in a salt form in conjunction with amino acid or nucleotide
Help the formation of delicate flavour.
The fermentation of table 1 front and back taste composition changes of contents
Note: G indicates corynebacterium glutamicum fermentation, and B indicates fermentation of bacillus subtilis
The analysis of 4 fermentation liquid volatile flavor component of embodiment
According to the tunning that embodiment 1 and embodiment 2 obtain, headspace solid-phase microextraction and gas-chromatography and mass spectrum are utilized
Technology used in conjunction (SPME-GC-MS) analyzes the fermentation liquid of shrimp enzymolysis liquid, determines flavored active ingredient, returns by peak area
One change method finds out the relative amount of volatile chemical component, is indicated in the form of percentage.
(1) there are following aroma compounds: 2,3,5- trimethylpyrazine after corynebacterium glutamicum 1276 is fermented
(64.29%), 2,3,5,6- Tetramethylpyrazine (22.53%), long alkane ketone (1.21%).
(2) there are following aroma compounds after fermentation of bacillus subtilis: 2 Ethylbutanoic acid (12.74%), 2,3,5,6-
Tetramethylpyrazine (46.32%), 3- hydroxy-2-butanone (23.14%).
Secondly Pyrazine content highest after (2) two kinds of bacterium fermentations, the main flavor after contributing to fermentation are ketone chemical combination
Object.The Pyrazine flavor substance of high-content also reflects the apparent burnt odor taste of fermentation liquid and strong nutty flavor after fermentation,
Reflect fermentation process bacterial metabolism and Maillard reaction joint contribution flavor components.
(3) flavor mass spectrogram is as shown in attached drawing 2, attached drawing 3.
5 sensory evaluation of embodiment
10 trained valuation officer (6 female/4 male, ages 26~48 are chosen in Tianjin Chunfa Food Additives Co., Ltd
Year), sensory evaluation is carried out to the fresh shrimp fragrance of fermentation liquid, delicate flavour, concordance according to listed content in table 2, and with commercial product original
Delicious (the happy Co., Ltd of Shanghai Mrs) is as control.
Step (1): the preparation of fermentation liquid freeze-dried powder
Fermentation liquid is prepared according to the step of embodiment 1, embodiment 2, after fermentation liquid is cooled to room temperature, 15~20mL is taken to incline
In plate, pre-freeze for 24 hours, is lyophilized using freeze dryer in -80 DEG C of refrigerators, and Cheng Fen is saved in drier.
Step (2): evaluation method
After obtaining fermentation liquid freeze-dried powder, it is configured to 0.5% (w/v) concentration with 60 DEG C of warm water dissolved freeze-dried powders, is contained in size
In identical sensory evaluation cup, with 10~15mL entrance, 10~20s is stopped in mouth, is gargled after discharge with clear water, is given a mark, it is right
Sample is evaluated, and every a sample evaluates interval 2min, while fresh as control with commercially available original flavor.
Step (3): evaluation criterion
The fermentation shrimp enzymolysis liquid freeze-dried powder sensory evaluation criteria of table 2
Step (4): evaluation result
3 yeast powder evaluation result of table
Note: G indicates corynebacterium glutamicum fermentation, and B indicates fermentation of bacillus subtilis
Using the carry out sensory evaluation fresh to corynebacterium glutamicum fermentation liquid freeze-dried powder and original flavor of clear water evaluation assessment, by table 2
Fresh low compared with the happy original flavor of Mrs, the astringent taste scoring it is found that untreated yeast powder scores in terms of delicate flavour, concordance with table 3
It is fresh higher than original flavor.The fermentation liquid freeze-dried powder that corynebacterium glutamicum fermentation shrimp enzymolysis liquid obtains remains original shrimp flavor,
But its mouthfeel is sour and astringent；After two kinds of fermentation liquid freeze-dried powders are by 1:1 compounding, the more independent yeast powder of concordance is improved, and astringent taste is reduced.But from
On the whole, tunning contributes to delicate flavour ingredient, mentions fresh without other flavoring substances auxiliary, can be used to delicate flavour base-material or tune
Taste substance preparation, substantially close to the flavor effect of commercially available flavoring products, and at the same time in product natural sex, trophism and technique letter
The property changed aspect is better than commercial product.
Two plants of bacterium ferment respectively after product can by it is concentration, liquid or powdered etc. it is any in the form of use, can be with
It individually adds, addition or flavouring related to other are used in mixed way in the form of any proportion.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Within mind and principle, any modification, equivalent replacement, improvement and so on be should all be included in the protection scope of the present invention.
Claims (8)
1. a kind of preparation method of delicate flavour fermented flavoring material, includes the following steps:
S1: digesting animal protein source using compound protease and neutral proteinase, obtains enzymolysis product；
S2: fermenting to hydrolysate using corynebacterium glutamicum and/or bacillus subtilis, obtains tunning.
2. the preparation method of delicate flavour fermented flavoring material according to claim 1, which is characterized in that ammonia in the enzymolysis product
Ground state nitrogen content is not less than 0.3g/100mL；In the tunning to be fermented using corynebacterium glutamicum, glutamic acid it is dense
Degree is greater than its threshold value 0.3g/L in clear water；In the tunning to be fermented using bacillus subtilis, guanylic acid it is dense
Degree is greater than its threshold value 0.125g/L in clear water；Preferably, the target product concentration after being fermented using fermentation strain is compared to hair
10 times or more are improved before ferment.
3. the preparation method of delicate flavour fermented flavoring material according to claim 1, which is characterized in that the animal protein source is
Shrimp, chicken, beef or the flesh of fish；Animal protein source is cleaned, drains, mashing, spare in the form of the meat of meat exquisiteness slurry.
4. the preparation method of delicate flavour fermented flavoring material according to claim 1, which is characterized in that enzyme in the enzymolysis process
Solution condition are as follows: 10~25LAPU compound protease 50LAPU/g, 0.05~0.075AU neutral proteinase 0.15AU/g, pH 6.0
1~4h is digested in~7.0,50 DEG C of waters bath with thermostatic control.
5. the preparation method of delicate flavour fermented flavoring material according to claim 1, which is characterized in that the compound protease and
Neutral proteinase is corynebacterium glutamicum 1276 and the bacillus subtilis that (China) Bioisystech Co., Ltd is believed from Novi
Bacterium 1354.
6. the preparation method of delicate flavour fermented flavoring material according to claim 1, which is characterized in that the enzymolysis product into
Before row fermentation, it is also added with composite nutrient, the composite nutrient composition are as follows: glucose 80；Corn pulp 3-12；K2HPO4 2-
14；KH2PO40-4；MgSO4·7H2O 0-1.5；MgCl2·6H2O 0-1.5；MnSO4·7H2O 0.04-0.06；FeSO4·
7H2O 0.0010-0.0020；CaCl2·2H2O 0-0.01；CuSO4·5H2O 0-0.015；Half Guang hydrochloride 0-0.0020 of L-；
(NH4)2SO45-20；Yeast extract (food-grade) 0-10；Unit: g/L.
7. the preparation method of delicate flavour fermented flavoring material according to claim 1, which is characterized in that sent out in the fermentation process
Ferment condition are as follows: 32 DEG C~38 DEG C of constant temperature, fluid nutrient medium 24~40h of shake culture, midfeather add appropriate ammonium hydroxide
And biotin.
8. the preparation method of delicate flavour fermented flavoring material according to claim 1, which is characterized in that the fermentation process carries out
Before, further include the steps that activating fermenting microbe: 30~32 DEG C of isothermal vibration cultivation and fermentation agent, 130~150rpm/min,
8~12h, fluid nutrient medium are continuously cultivated 2-3 times, and actication of culture saves under the conditions of 4 DEG C；The bacterium fermented after activation
Number living is 107~108CFU/mL range.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110742260A (en) * | 2019-11-25 | 2020-02-04 | 青岛科素生物科技有限公司 | Corn-derived flavoring liquid and preparation method thereof |
| CN111642728A (en) * | 2020-06-30 | 2020-09-11 | 广东百味佳味业科技股份有限公司 | Preparation method and application of high-thickness chicken powder |
| CN113142542A (en) * | 2021-02-25 | 2021-07-23 | 仇俊鹏 | Fermented seasoning sauce using grains as raw materials, preparation method and application |
| WO2022016141A1 (en) * | 2020-07-17 | 2022-01-20 | International Flavors & Fragrances Inc. | Taste-enhancing composition |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103504368A (en) * | 2013-09-18 | 2014-01-15 | 中国科学院天津工业生物技术研究所 | Kelp-flavored food ingredient and preparation method thereof |
| CN104366427A (en) * | 2014-11-25 | 2015-02-25 | 烟台新海水产食品有限公司 | Production process of functional sea-intestine seasoning particles |
| CN104855918A (en) * | 2015-05-06 | 2015-08-26 | 河南科技大学 | Beef-flavored essence preparation method |
| CN104878062A (en) * | 2015-05-18 | 2015-09-02 | 华南理工大学 | Microbiological method for preparing sea-fish protein antihypertensive peptides |
| US20150272186A1 (en) * | 2013-07-23 | 2015-10-01 | Cj Cheiljedang Corporation | Method For Preparing Natural Neutral Flavor |
| CN106119173A (en) * | 2016-08-22 | 2016-11-16 | 广东轻工职业技术学院 | A kind of shrimps low molecular peptide without bitterness and preparation method and application |
| CN107173776A (en) * | 2017-05-02 | 2017-09-19 | 兰溪富晟食品科技有限公司 | A kind of prawn flavoring and process technology |
| CN109497492A (en) * | 2018-12-05 | 2019-03-22 | 阳江喜之郎果冻制造有限公司 | Method for preparing flavor-developing peptide through secondary fermentation |
| CN110229852A (en) * | 2019-06-20 | 2019-09-13 | 东莞理工学院 | A method of utilizing soybean protein hydrolysate fermenting and producing gamma-polyglutamic acid |
-
2019
- 2019-09-17 CN CN201910876200.6A patent/CN110447880A/en active Pending
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20150272186A1 (en) * | 2013-07-23 | 2015-10-01 | Cj Cheiljedang Corporation | Method For Preparing Natural Neutral Flavor |
| CN103504368A (en) * | 2013-09-18 | 2014-01-15 | 中国科学院天津工业生物技术研究所 | Kelp-flavored food ingredient and preparation method thereof |
| CN104366427A (en) * | 2014-11-25 | 2015-02-25 | 烟台新海水产食品有限公司 | Production process of functional sea-intestine seasoning particles |
| CN104855918A (en) * | 2015-05-06 | 2015-08-26 | 河南科技大学 | Beef-flavored essence preparation method |
| CN104878062A (en) * | 2015-05-18 | 2015-09-02 | 华南理工大学 | Microbiological method for preparing sea-fish protein antihypertensive peptides |
| CN106119173A (en) * | 2016-08-22 | 2016-11-16 | 广东轻工职业技术学院 | A kind of shrimps low molecular peptide without bitterness and preparation method and application |
| CN107173776A (en) * | 2017-05-02 | 2017-09-19 | 兰溪富晟食品科技有限公司 | A kind of prawn flavoring and process technology |
| CN109497492A (en) * | 2018-12-05 | 2019-03-22 | 阳江喜之郎果冻制造有限公司 | Method for preparing flavor-developing peptide through secondary fermentation |
| CN110229852A (en) * | 2019-06-20 | 2019-09-13 | 东莞理工学院 | A method of utilizing soybean protein hydrolysate fermenting and producing gamma-polyglutamic acid |
Non-Patent Citations (2)
| Title |
|---|
| 王艳玲等: "蛋白酶制剂对发酵牛肉酱风味物质的影响", 《中国酿造》 * |
| 赵小惠等: "菌酶协同处理金枪鱼暗色肉制备饲用肽的研究", 《中国饲料》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110742260A (en) * | 2019-11-25 | 2020-02-04 | 青岛科素生物科技有限公司 | Corn-derived flavoring liquid and preparation method thereof |
| CN111642728A (en) * | 2020-06-30 | 2020-09-11 | 广东百味佳味业科技股份有限公司 | Preparation method and application of high-thickness chicken powder |
| CN111642728B (en) * | 2020-06-30 | 2022-08-23 | 广东百味佳味业科技股份有限公司 | Preparation method and application of high-thick chicken powder |
| WO2022016141A1 (en) * | 2020-07-17 | 2022-01-20 | International Flavors & Fragrances Inc. | Taste-enhancing composition |
| CN113142542A (en) * | 2021-02-25 | 2021-07-23 | 仇俊鹏 | Fermented seasoning sauce using grains as raw materials, preparation method and application |
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