CN102220243B - Production of cheese-flavor substance and application of cheese-flavor substance in food - Google Patents
Production of cheese-flavor substance and application of cheese-flavor substance in food Download PDFInfo
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- CN102220243B CN102220243B CN2010101456988A CN201010145698A CN102220243B CN 102220243 B CN102220243 B CN 102220243B CN 2010101456988 A CN2010101456988 A CN 2010101456988A CN 201010145698 A CN201010145698 A CN 201010145698A CN 102220243 B CN102220243 B CN 102220243B
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Abstract
The invention relates to production of a cheese-flavor substance and application of the cheese-flavor substance in food. According to the invention, a novel microorganism capable of generating cheese flavor is screened by the inventor, and the microorganism is fermented by taking a substance containing milk, oil and lipid as a substrate to obtain the cheese-flavor substance; and the cheese-flavor substance can be widely applied to the field of food.
Description
Technical field
The invention belongs to microbiology and bromatology field; More specifically, the present invention relates to production and the Applications in food thereof of cheese flavor material.
Background technology
Cheese flavor (cheese flavor also claims cheese or cheesy flavour) is a kind of polycomponent mixing local flavor system.The nearly different volatile and non-volatile compounds of kind more than 3000 that surpasses in cheese, local flavor is produced by volatile matter, also claims the local flavor volatile matter.Typical local flavor volatile matter in the dissimilar cheese can be divided into 6 groups: fatty acid (such as acetic acid, propionic acid, butyric acid), ketone (such as 2-heptanone, methyl n-heptyl ketone, 2-butanone), ester class (such as ethyl butyrate, ethyl decylate), aldehydes (such as 3-methyl-butyraldehyde-n, 2-methyl-butyraldehyde-n, phenyl aldehyde), alcohols (such as n-butyl alcohol, 3-methyl-1-butanol, phenylethyl alcohol) and sulfide (such as disulphide, thiomethyl alcohol).
The way of production of cheese flavor material mainly is that fat, the protein in the butterfat raw material produces the zestful product of tool (usually also comprising the flavour compositions that carries in the raw dairy in the product) under the effect of enzyme.General complicated cheese flavor material then is to produce under the effect of microorganism (being actually the enzyme that microbial metabolism produces).
The formation of cheese flavor material and raw material substrate and the starter (microorganism) that uses when producing are relevant.
Common several cheese flavor materials, such as most widely used blue cheese (Blue Cheese, also claim blue line cheese, bleu cheese etc.) flavour substances, also have than right cheese (Brie Cheese) in addition, horse Soviet Union lira cheese (Mozzarella Cheese), car reaches cheese (Cheddar Cheese), Palma cheese (Parmesan Cheese) flavour substances etc.
So far, the research that utilizes mould to carry out fermentative production cheese flavor material only limits to the blue cheese flavour substances.Disclosed a kind of method of utilizing penicillium roqueforti and mutant strain penicillium camemberti liquid submerged fermentation thereof to produce Blue Cheese flavour substances among the US3100153 (1963/08/06).
Show after deliberation, the key component that produces typical blue cheese local flavor is methyl ketone, referring to document JOHN E.KINSELLA and DAN HWANG.Biosynthesis of Flavors by Penicilliumroqueforti.BIOTECHNOLOGY AND BIOENGINEERING, VOL.XVIII, PAGES927-938 (1976), specifically, mould spores, mycelium is secreted complicated enzyme in the growth metabolism process, these enzyme systems are with protein in the milk-product, the macromolecular substance such as fat are degraded to flavor precursors and local flavor base-material material, as: lipase can be lipid acid and glycerine with lipolysis, and lipid acid is methyl ketone by the metabolism of classical β-oxidation approach under the tryptophan side-chain alpha effect.
In these years, the technician is to producing only two kinds of mould bacterial classification system (penicillum requeforti Penicillium roqueforti of blue cheese flavour substances; Penicillum glaucum Penicillium glaucum) carried out replenishing and perfect.Yet along with people require to improve constantly to the each side of food, especially taste becomes more diverse, and obtains novel cheese flavour substances, and perhaps screening obtains to produce the bacterial strain of original or new cheese flavor material/kind always be the heat subject of the art.
Yet along with people require to improve constantly to the each side of food, especially taste becomes more diverse, and obtains novel cheese flavor, and perhaps screen the bacterial strain that obtains the original or new cheese flavor material of production is the heat subject of the art always.
Summary of the invention
The object of the present invention is to provide the bacterial strain of new production cheese flavor material.
The present invention also aims to provide production and the Applications in food thereof of cheese flavor material.
In a first aspect of the present invention, a kind of screening method of the bacterial strain for the production of the cheese flavor material is provided, this screening method comprises that directed screening goes out the step of mould from yak milk and fabricated product (for example natural butter) thereof, and described mould can come fermentative production methyl letones take the material that contains breast, oil, lipid and/or its enzymolysis product as substrate.
Natural butter is to extract the rare cream (lipid content 30-80%) that obtains, concentrated butter (lipid content 80-98%), anhydrous butter (lipid content 98-100%) etc. from yak milk, such as the yak milk from Nepal, Mongolia, India, China (Qinghai, Gansu, Xinjiang, Sichuan, Tibet, Yunnan, the Inner Mongol) and other places.
The contriver finds first can be from yak milk and the new microorganism that can produce the cheese flavor material of fabricated product (for example natural butter) screening thereof.
In another aspect of this invention, provide arbitrary bacterial strain or the application of its variant aspect production cheese flavor material in Java penicillium (Eupenicillium javanicum), light spore mould (Penicillium glabrum), Switzerland's wood mould (Trichoderma reesei), deep mould (Penicillium paxilli), scarlet mould (Penicillium chermesinum), aspergillus (Aspergillus sp), the branch spore mould (Cladosporiumsp).
In a preference, described bacterial strain or the screening of its variant are from the material that contains breast, oil, lipid.
In another preference, the described material that contains breast, oil, lipid is natural butter.
In another aspect of this invention, provide separated bacterial strain, it is
Java penicillium CFR0802, its preserving number are CGMCC NO.3551; Or its variant;
Light spore mould CFR0801, its preserving number is CGMCC NO.3552; Or its variant;
The mould CFR0803 of Switzerland's wood, its preserving number is CGMCC NO.3553; Or its variant.
The contriver has filtered out the new bacterial strain that above-mentioned three strains can be produced typical cheese flavor from natural butter.
In another aspect of this invention, provide the purposes of aforementioned arbitrary separated bacterial strain, be used for fermentative production cheese flavor material.
In another aspect of this invention, provide a kind of method of producing the cheese flavor material, the method comprises:
Adopt spore or the mycelium of aforementioned arbitrary separated bacterial strain or arbitrarily several combination, take the material that contains breast, oil, lipid and/or its enzymolysis product as fermentation substrate, through liquid fermenting or solid fermentation, obtain to have the tunning of cheese flavor.
In a preference, described mycelium is through the mycelium of 2-3 level enlarged culturing.
In a preference, described liquid fermenting comprises:
The spore suspension or the hypha fluid that prepare first described arbitrary bacterial strain or arbitrarily several combination, in the fermentation substrate that again spore suspension or the hypha fluid of significant quantity is inoculated into liquid form, obtain the tunning of liquid form, leavening temperature is 25-35 ℃, and fermentation time is 24-48h.
In another preference, described method also comprises:
Collect tunning or its concentrated solution of described liquid form, as the flavour substances product; Perhaps,
Described liquid form tunning is carried out degerming process, resulting liquid with cheese flavor after the collection and treatment is as the flavour substances product; Perhaps,
Extract volatile matter in the described liquid form tunning as the flavour substances product; Perhaps,
Described liquid form tunning is carried out drying treatment, and resulting solid with cheese flavor after the collection and treatment is as the flavour substances product.
In another preference, described degerming is processed and is adopted centrifugal, as to filter any one or two kinds combination.
In another preference, extract volatile matter in the described liquid form tunning and adopt any one or a few combination in distillation extraction, adsorbing and extracting, the extraction.
In another preference, described drying treatment adopts freeze-drying or spraying drying.
In another preference, described solid fermentation comprises:
The spore suspension or the hypha fluid that prepare first described arbitrary bacterial strain or its combination, spore suspension or hypha fluid with significant quantity is inoculated in the process that solid-state form fermentation substrate surface obtains the solid-state form tunning again, leavening temperature is 15-25 ℃, and fermentation time is 10-15 days.
In another preference, described method also comprises:
Collect described solid-state form fermentation substrate, as the flavour substances product; Perhaps,
Extract volatile matter in the described solid-state form tunning as the flavour substances product; Perhaps,
Described solid-state form fermentation substrate is carried out drying treatment, and resulting solid with cheese flavor after the collection and treatment is as the flavour substances product.
In another preference, extract volatile matter in the described liquid form tunning and adopt any one or a few combination in distillation extraction, adsorbing and extracting, the extraction.
In another preference, described drying treatment adopts freeze-drying or spraying drying.
In another preference, comprise at least 2-heptanone, 2 pentanone and methyl n-heptyl ketone in the described tunning.
In another preference, the described material that contains breast, oil, lipid is selected from: milk-product, animal-plant oil or microbial oil.
In another aspect of this invention, provide the cheese flavor material that adopts aforementioned either method to obtain.
In another aspect of this invention, provide the application of described cheese flavor material at field of food.
In another aspect of this invention, provide a kind of method for making food, the method comprises: before the cheese flavor material that the aforementioned either method of appropriate amount is obtained adds in the foodstuff base material, simultaneously or afterwards, described cheese flavor material is sterilized.
In another aspect of this invention, the application of cheese flavor material aspect fatty foods that provides fungi fermentation to produce.
In another preference, described fatty foods is oleomargarine, butter, crisp skin oil.
In another aspect of this invention, provide the manufacture method of fatty foods, the method comprises the step that the cheese flavor material that fungi fermentation is produced and the base-material of making fatty foods mix and sterilize.
In a preference, described manufacture method is: the cheese flavor material that first fungi fermentation is produced carries out mixing with the base-material of making fatty foods behind the homogeneous again.
Other side of the present invention is because the disclosure of this paper is apparent to those skilled in the art.
Description of drawings
The GC-MS detected result collection of illustrative plates of the local flavor volatile matter in the tunning that Fig. 1, embodiment 1 obtain;
The GC-MS detected result collection of illustrative plates of the local flavor volatile matter in the tunning that Fig. 2, embodiment 2 obtain;
The GC-MS detected result collection of illustrative plates of the local flavor volatile matter in the tunning that Fig. 3, embodiment 3 obtain;
In above-mentioned each collection of illustrative plates, transverse axis represents time/min, and the longitudinal axis represents abundance.
Embodiment
The inventor has found a kind of effective screening method through studying widely and testing, and screens the novel microorganism that can produce cheese flavor, and this microorganism obtains to have the material of cheese flavor take the material that contains breast, oil, lipid etc. as fermenting substrate production; This cheese flavor material can be widely used in food, essence and flavoring agent production field.
Bacterial strain screening method
The invention provides a kind of screening method of the bacterial strain for the production of the cheese flavor material, this screening method comprises that directed screening goes out the step of mould from yak milk and fabricated product (for example natural butter) thereof, and described mould can come fermentative production methyl letones take the material that contains breast, oil, lipid and/or its enzymolysis product as substrate.
In a specific embodiment of the present invention, from commercially available natural butter (sample center) sampling, prepare the bacteria suspension (for example 10 of different concns with the sterilized water mixing that has added tensio-active agent (being generally tween-80)
-3, 10
-4, 10
-5, 10
-6Bacteria suspension); Again the different concns bacteria suspension is evenly coated pre-prepd mould selective medium (namely added PDA or the rose-bengal solid medium of Streptomycin sulphate, only mould can survive); Cultivate through for some time, survive single bacterium colony of the different shape that gets off of picking lines on PDA or the rose-bengal slant medium and cultivates; Again respectively with each inoculation in suitable fermentation substrate (material and/or its enzymolysis product that contain breast, oil, lipid, whole milk powder solution for example) fermentation for some time is (better in, 24h-48h), the tunning that obtains is carried out local flavor sensory evaluation test and GC-MS detection, choose the bacterial strain that has obtained cheese flavor, preservation is for subsequent use.
Preferably, choose the bacterial strain that has obtained typical cheese flavor (particularly ketone local flavor), in other words screening can produce the bacterial strain of ketone local flavor volatile matter; Preferred, choose the bacterial strain that has obtained methyl ketone local flavor (a kind of very typical cheese flavor), namely can produce the bacterial strain of methyl ketone local flavor volatile matter.
The invention provides Java penicillium (Eupenicillium javanicum), light spore mould (Penicilliumglabrum), Switzerland's wood mould (Trichoderma reesei), deep mould (Penicillium paxilli), scarlet mould (Penicillium chermesinum), aspergillus (Aspergillus sp), arbitrary bacterial strain or its variant application aspect production cheese flavor material in the branch spore mould (Cladosporium sp).
Preferably, described microorganism at the preserving number of China Committee for Culture Collection of Microorganisms common micro-organisms center (CGMCC) is: CGMCC NO.3551 (its called after Java penicillium CFR0802; Belong to Eupenicillium sp, the Eupenicillium javanicum kind), CGMCC NO.3552 (its called after light spore mould CFR0801; Belong to Penicillium, light spore mould bacterial classification) or CGMCC NO.3553 (its called after Switzerland wood mould (Trichoderma reesei) CFR0803; Belong to Trichoderma, Switzerland's Trichoderma kind).This three strains bacterial strain obtains through above-mentioned bacterial strain screening method.
Identification of strains
Three strain bacterial strains of above-mentioned preservation are delivered to Guangdong Province microbiological analysis inspection center and (are entrusted registration number: the little inspection in Guangdong (2009) ZD0191 number) carry out the evaluation of form and molecular biology aspect.Appraisal basis: " fungi identification handbook " (super posthumous work of Wei Jing, Shanghai science tech publishing house), " form of fungi and classification " (Dai Fang posthumous work of calumniating, Science Press), and " molecular cloning experiment guide " (third edition, counterfeit, Li Mengfeng of gold winter etc. translates, 2002 Science Presses), microscope picture to single bacterium colony of above-mentioned bacterial strains carries out morphological analysis, and the total DNA that extracts is carried out ITS analyze, and qualification result is as follows:
1、CGMCC?NO.3551
Thalli morphology:
Cleistothecium presents spherical or subsphaeroidal, diameter 70～200 μ m, tawny; Ascus Dan Sheng, sphere, subsphaeroidal or near ellipse, thecaspore is oval; Conidiophore betides aerial hyphae, falx stem 50～120 * 2.0～2.5 μ m, and wall is smooth; The penicillus single-wheel is given birth to; Bottle stalk 7.0～11 * 2.0～2.5 μ m, ampuliform; Conidium is spherical or subsphaeroidal, 2.5～3.0 * 2.1～2.5 μ m, and wall is smooth.
Colonial morphology:
1., czapek's solution is cultivated the form description of bacterium colony:
Bacterium colony was cultivated after 7 days, and diameter is about 30～50mm, and the radioactivity wrinkle is arranged, the velvet-like cotton-shaped and particulate state of holding concurrently of quality; Conidium face pewter is bordering on smoky gray, and light ash is olive colour, and mycelium is faint yellow, red-purple; The reverse side tawny, the soluble pigment yellowish brown.
2., the form of malt extract medium bacterium colony is described:
Bacterium colony was cultivated 7 days, diameter 40～60mm, and radial wrinkle is more, and quality is velvet-like, particulate state; Conidium face greyish-green is bordering on pea green; The reverse side tawny is without soluble pigment.
3., the form of PDA substratum bacterium colony is described:
Bacterium colony was cultivated after 7 days, diameter 20mm, and smooth, quality is velvet-like to particulate state; Conidium face greyish-green is without transudate; Reverse side is khaki, without soluble pigment.
4., thalline is described in the microscopically form:
Cleistothecium presents spherical or subsphaeroidal, diameter 70～200 μ m, tawny; Ascus Dan Sheng, sphere, subsphaeroidal or near ellipse, thecaspore is oval; Conidiophore betides aerial hyphae, falx stem 50～120 * 2.0～2.5 μ m, and wall is smooth; The penicillus single-wheel is given birth to; Bottle stalk 7.0～11 * 2.0～2.5 μ m, ampuliform; Conidium is spherical or subsphaeroidal, 2.5～3.0 * 2.1～2.5 μ m, and wall is smooth.
To sum up, thalline, colonial morphology are the most similar to Java penicillium (Eupenicilliumjavanicum).
Molecular biology identification result:
The 18S rDNA sequence homology of the 18S rDNA sequence of CGMCC No.3551 and Java penicillium (Eupenicillium javanicum) AFT0L-ID 429 reaches 98.9%.
2、CGMCC?NO.3552
Thalli morphology:
Falx stem wall is level and smooth, ultimate swelling; The penicillus single-wheel is given birth to, and idol has metulae shape branch; The bottle stalk is every takes turns 8～16; 8.0～11 * 2.5～3.0 μ m, ampuliform; Conidium is spherical or subsphaeroidal, 2.5～3.5 μ m, and wall is smooth or be bordering on level and smooth; Conidia chain presents cylindric closely usually.
Colonial morphology:
1., czapek's solution is cultivated the form description of bacterium colony:
Bacterium colony was cultivated after 7 days, and diameter is about 25mm, and is smooth, and quality is velvet-like double cotton-shaped; Conidium face ash is olive colour, and white mycelium is without transudate; Reverse side is khaki, without soluble pigment.
2., the form of malt extract medium bacterium colony is described:
Bacterium colony was cultivated after 7 days, diameter 35～55mm, and there is radial wrinkle at the middle part, and quality is velvet-like double cotton-shaped, and the conidium structure is more, and the conidium face is olive colour, white mycelium, without transudate, the reverse side light tan is without soluble pigment.
3., the form of PDA substratum bacterium colony is described:
Bacterium colony was cultivated after 7 days, diameter 20mm, and smooth, quality is velvet-like to particulate state; Conidium face greyish-green is without transudate; Reverse side is khaki, without soluble pigment.
4., thalline is described in the microscopically form:
Falx stem wall is level and smooth, ultimate swelling; The penicillus single-wheel is given birth to, and idol has metulae shape branch; The bottle stalk is every takes turns 8～16; 8.0～11 * 2.5～3.0 μ m, ampuliform; Conidium is spherical or subsphaeroidal, 2.5～3.5 μ m, and wall is smooth or be bordering on level and smooth; Conidia chain presents cylindric closely usually.
To sum up, thalline, colonial morphology are the most similar to light spore mould (Penicillium glabrum).
Molecular biology identification result:
The 18S rDNA sequence homology of the 18S rDNA sequence of CGMCC No.3552 and light spore mould (Penicillium glabrum) ALI218 reaches 99.7%.
3、CGMCC?NO.3553
Thalli morphology:
Conidiophore is the short lateral branch of mycelia, on it to branch estranged, the crotch angle almost right angle, to giving birth to phialide, the taper of bottle stalk, 5.0～8 * 2.5～3.7 μ m, base portion is slightly narrow, the middle part is wider.The conidium great majority are spherical, and are inferior spherical, obovate, diameter 2.0～3.0 μ m, light green; Give birth between chlamydospore or push up and be born in the mycelia, wall is smooth, and diameter can reach 12 μ m.
Colonial morphology:
1., czapek's solution is cultivated the form description of bacterium colony:
It is poor to grow, and mycelia is sparse, thin, cultivates the gonimoblast bundle district that occurs being scattered in media surface after 10 days.
2., the form of malt extract medium bacterium colony is described:
Colony growth is rapid, cultivates 7 days, and bacterium colony is paved with whole 9cm culture dish, and the bacterium colony surface is the fine and close smooth mycelia of white, flocculence, and it is slightly slow to produce spore, cultivates after 9 days to occur green gonimoblast bundle district in media surface by edge, lines up center of circle wheel line; The bacterium colony reverse side is colourless.
3., the form of PDA substratum bacterium colony is described:
Fast growth was cultivated 7 days, and bacterium colony is paved with whole 9cm culture dish, and the bacterium colony surface is the fine and close smooth mycelia of white, flocculence; It is slightly slow to produce spore, cultivates after 9 days green gonimoblast bundle district, bacterium colony reverse side pistac to occur in media surface.
4., thalline is described in the microscopically form:
Conidiophore is the short lateral branch of mycelia, on it to branch estranged, the crotch angle almost right angle, to giving birth to phialide, the taper of bottle stalk, 5.0～8 * 2.5～3.7 μ m, base portion is slightly narrow, the middle part is wider.The conidium great majority are spherical, and are inferior spherical, obovate, diameter 2.0～3.0 μ m, light green; Give birth between chlamydospore or push up and be born in the mycelia, wall is smooth, and diameter can reach 12 μ m.
To sum up, thalline, colonial morphology and Switzerland's wood mould (Trichoderma reesei) is the most approaching.
Molecular biology identification result:
The 18S rDNA of CGMCC No.3553 and Hypocrea jecorina Hypocrea jecorina homology reach 99.9%, and (imperfect stage of Hypocrea jecorina Hypocrea jecorina is Switzerland's wood mould (Trichodermareesei).
Produce the method for cheese flavor material
Therefore, the method of production cheese flavor material of the present invention comprises: will be selected from Java penicillium (Eupenicillium javanicum), light spore mould (Penicillium glabrum), Switzerland's wood mould (Trichoderma reesei), deep mould (Penicillium paxilli), scarlet mould (Penicilliumchermesinum), aspergillus (Aspergillus sp), arbitrary bacterial strain or its variant are (preferably in the branch spore mould (Cladosporium sp), be selected from CGMCC NO.3551, CGMCC NO.3552 or CGMCC NO.3553) join and carry out fermentative production in the fermentation substrate, obtain tunning, described tunning has local flavor, and described local flavor is cheese flavor.
Described fermentation substrate generally provides the material that contains breast, oil, lipid; Or the enzymolysis product that behind lipase enzymolysis above-mentioned substance, forms; Or both mixtures.
The material that contains breast, oil, lipid includes but not limited to milk-product, animal-plant oil or microbial oil.
The material that contains breast, oil, lipid is preferred, marine organisms grease, for example algae oil.
In another preference, described milk-product are whole milk powder solution (preferably being 5-40% whole milk powder solution), cream (cream), butter (Butter), butterfat (milk fat), Mammals (such as ox, sheep) milk etc.
In another preference, described Vegetable oil lipoprotein can be Oleum Cocois, soybean oil, Semen Maydis oil, Oleum Cocois, rape oil, soya-bean oil, Thistle oil, high oleic safflower oil, sunflower seed oil, high oleic sunflower oil, sweet oil, Oenothera oil, Oleum Gossypii semen, Rice pollard oil, raisin seed oil, linseed oil, peanut oil, Prunus amygdalus oil, Walnut oil., Wheat germ oils, sesame wet goods.Described animal grease can be butter etc.
Above-mentioned animal-plant oil, microbial oil etc. also can use this quasi-grease through or without mixture or the monomer of physics or chemical modification (minute carry, hydrogenation, transesterify); It also can be the transgenosis grease.
Described microorganism can join in the fermentation substrate with spore shape, also can join in the fermentation substrate with mycelium morphology; Fermentation substrate can be liquid substrate (carrying out liquid fermenting) or solid-state substrate (carrying out solid fermentation).
Preferably, described microorganism through seed culture, thereby forms mycelium before joining substrate.Cultivate in the described seed bacterium culture medium, form first order seed (mycelium); Again first order seed is inoculated in the seed culture medium, forms secondary seed (mycelium); Preferably, also can further form three grades of seeds (mycelium).
It is significant quantity to the consumption in the fermentation substrate that spore or mycelium add (inoculation).Described significant quantity is to be enough to produce fermentation reaction, is enough to produce the consumption of the tunning of cheese flavor.Those skilled in the art can determine this significant quantity according to the characteristic of what and bacterial strain of fermentation substrate.In a preferred embodiment of the present invention, the concentration when the pityrosporion ovale suspension is 10
8-10
9Or mycelial dry weight is when 15-25g/L, and the volume ratio of pityrosporion ovale suspension or hypha fluid and liquid-type fermentation substrate is 5-20%; In another preferred implementation of the present invention, when the concentration of pityrosporion ovale suspension is 10
8-10
9Or mycelial dry weight is smeared 0.05-1ml pityrosporion ovale suspension or hypha fluid on the surface of every square centimeter of solid-state version fermentation substrate when 15-25g/L.
Preferably, when solid fermentation, make a plurality of shrinkage pools on the surface of solid fermentation substrate, best, shrinkage pool outer rim and consecutive position are smoothly connected.
Cultivating the temperature of (fermentation) and lasting time decides with the dependency of cheese flavor based on free fatty acids (FFA).Free fatty acids FFA content and flavor intensity are closely-related.Those skilled in the art can determine to cultivate the time that continues according to needed flavor intensity.
Preferably, the liquid fermenting temperature is 25-35 ℃, and fermentation time is 24-48h.
Preferably, the solid fermentation temperature is 15-25 ℃, and fermentation time is 10-15 days.
Unless otherwise indicated, other condition of cultivating (fermentation) has no particular limits, and can adopt those skilled in the art to cultivate the condition of the routine of mould (better, Trichoderma, Eupenicillium sp or Penicillium) microorganism.
The present invention also comprises the tunning that adopts aforesaid method to obtain, and described tunning has local flavor, and described local flavor is cheese flavor.Described tunning can be used for preparing the zestful food of tool, and described local flavor is cheese flavor.
Preferably, collect tunning or its concentrated solution of described liquid form, as the flavour substances product; Perhaps, described liquid form tunning is carried out degerming process, resulting liquid with cheese flavor after the collection and treatment is as the flavour substances product; Perhaps, extract volatile matter in the described liquid form tunning as the flavour substances product; Perhaps, described liquid form tunning is carried out drying treatment, resulting solid with cheese flavor after the collection and treatment is as the flavour substances product.
Described degerming is processed can adopt technology well known in the art.For example, carry out centrifugal or filtration to tunning, remove thalline wherein, get clear liquid as flavour substances.The local flavor volatile matter that extracts in the tunning also can adopt mode well-known to those skilled in the art, such as distillation extraction, adsorbing and extracting, extraction etc.Preferably, supercritical extraction is adopted in extraction.The solvent that distillation, adsorbing and extracting or extraction are adopted adopts the solvent that can extract volatile flavor substance well-known to those skilled in the art.Described drying treatment can adopt technology well known in the art.Preferably, adopt freeze-drying or spraying drying.
Preferably, can collect described solid-state form fermentation substrate, as the flavour substances product; Perhaps, extract volatile matter in the described solid-state form tunning as the flavour substances product; Perhaps, described solid-state form fermentation substrate is carried out drying treatment, resulting solid with cheese flavor after the collection and treatment is as the flavour substances product.
The local flavor volatile matter that extracts in the tunning also can adopt mode well-known to those skilled in the art, such as distillation extraction, adsorbing and extracting, extraction etc.Preferably, supercritical extraction is adopted in extraction.The solvent that distillation, adsorbing and extracting or extraction are adopted adopts the solvent that can extract volatile flavor substance well-known to those skilled in the art.Described drying treatment can adopt technology well known in the art.Preferably, adopt freeze-drying or spraying drying.
In a preference of the present invention, comprise at least 2-heptanone, 2 pentanone and methyl n-heptyl ketone in the described tunning.Namely can adopt bacterial strain production of the present invention to have methyl ketone local flavor (a kind of very typical cheese flavor) material.
The cheese flavor material is in the application of field of food
In a preference, described cheese flavor material is applied to the production of food grease.Preferably, the mixed oil that making family or foodstuffs industry are used, oleomargarine, anhydrous butter, crisp skin oil, liquid butter, butter, table butter etc., these product forms can be liquid state, bulk, powdery, sheet, particulate state etc.
In another preference, described cheese flavor material is applied to the bakery grease, and curing of food such as bread, cake, biscuit, toffee, puffed rice, vegetable fat powder, candy, puffed food used grease.
Preferably, described cheese flavor material is applied to produce Tibetan butter, buttered tea, yak milk powder, hides cake, fermented milk, sour milk, cheese.
In another preference, described cheese flavor material can be used as essence and flavoring agent to be added in the food.
Another aspect of the present invention also provides a kind of method for preparing the zestful food of tool, and described method comprises: before adding the cheese flavor material of appropriate amount in the foodstuff base material, simultaneously or afterwards, described cheese flavor material is sterilized.Described " appropriate amount " refers to behind the food prepared therefrom, be enough to form cheese flavor in joining foodstuff base material.For example, described cultured products accounts for the 1-60% (w/w) of amount of the mixture with after foodstuff base material mixes; 2-50% more preferably; 5-30% more preferably.Described foodstuff base material also namely prepares the raw material of food, and it can be but be not limited to: milk, plant wet goods.Described foodstuff base material can be solid-state, liquid state or emulsus.
In another aspect of this invention, the application of cheese flavor material aspect the manufacturing fatty foods that provides fungi fermentation to produce.
Fungi comprises Molds and yeasts, disclose multiple can the fermentation in the prior art and produced the Molds and yeasts of cheese flavor material, for example produce the mould commonly used of white mould cheese (White Mould Cheese) flavour substances, such as white mould Penicillium candidum, geotrichum candidum Geotrichum candidum, penicillium cammenberti Penicillium camemberti); Mould commonly used in the production blue cheese flavour substances is such as penicillum requeforti Penicillium roqueforti, Penicillum glaucum Penicillium glaucum; For example produce Danisco cheese flavour substances yeast commonly used, such as the inferior Dbaly yeast Debaryomyceshansenii of the Chinese, Kluyveromyces lactis Kluyveromyces lactis, Candida utilis Candida utilis etc.
In the prior art, for preventing local flavor, fragrance taint of odour or the mutually punching between each food raw material, the cheese flavor material that usually Molds and yeasts is produced only adds in cheese or the similar raw material with cheese flavor to be produced.The contriver finds existing cheese flavor material and cheese flavor material of the present invention to be applied in the fatty foods (add in the fatty foods as flavour agent and produce).
In a preference, described fatty foods comprises oleomargarine, butter, crisp skin oil.
Preferably, the oleomargarine that making family or foodstuffs industry are used, anhydrous butter, crisp skin oil, liquid butter, table butter etc., these product forms can be bulk, powdery, sheet, particulate state etc.
In another preference, described cheese flavor material is applied to bakery with grease and the food that utilizes these greases to process, such as bread, cake, biscuit, candy, and puffed rice, puffed food etc.
Preferably, described cheese flavor material is applied to produce Plateau Characteristic food such as yak milk powder, liquid or solid beverage, Tibetan butter, milk slag, hides cake, buttered tea, fermented milk, sour milk etc.
In another aspect of this invention, provide the manufacture method of flavor oil fat foods, the method comprises the step that the cheese flavor material that fungi fermentation is produced and the base-material of making fatty foods mix and sterilize.
In a preference, described manufacture method is: the cheese flavor material that first fungi fermentation is produced carries out mixing with the base-material of making fatty foods behind the homogeneous again.
Inventive point of the present invention mainly is:
(1) find first and can from natural butter, screen the new microorganism that can produce the cheese flavor material, different from existing thinking of only from cheese, screening.
(2) filtered out three new bacterial strains from natural butter, it acts on the material that can produce the cheese flavor feature behind the substrate, and this material can be applied to field of food industry.
(3) contriver finds and the cheese flavor material that fungi fermentation produces can be applied in the fatty foods, and is different from the existing thinking that only is applied in the cheese.
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used for explanation the present invention and be not used in and limit the scope of the invention.The experimental technique of unreceipted actual conditions in the following example, or the condition of advising according to manufacturer.Unless otherwise indicated, otherwise per-cent and umber calculate by weight.
Unless otherwise defined, the same meaning that employed all specialties and scientific words and one skilled in the art are familiar with in the literary composition.In addition, any method similar or impartial to described content and material all can be applicable among the present invention.The usefulness that better implementation method described in the literary composition and material only present a demonstration.
Embodiment 1, Java penicillium (Eupenicillium javanicium) CFR0802 (CGMCCNo.3551) liquid fermenting are produced the cheese flavor material
First step: obtain its enzymolysis product by the lipase hydrolysis Oleum Cocois;
(1) the 200g cochin oil is positioned in the triangular flask of 500ml, places shaking water bath pot (40 ℃) concussion water bath with thermostatic control 30 minutes;
(2) according to the addition of 0.1g/100ml, add lipase (PALATASE 20000L) and be hydrolyzed; Per 2 hours sampling and measuring acid values (AV value) and free fatty acid content (FFA) in the hydrolytic process;
(3) finish hydrolysis during without considerable change when the AV value; Hydrolysate is warming up to 85 ℃ again, is incubated and carries out deactivation after 15 minutes; Hydrolysate after the deactivation can place 4 ℃ of refrigerator storages for subsequent use.
Second step: microbial inoculant in fermentation substrate, is produced the cheese flavor material through liquid fermenting;
From cultivating on 3-5 days Java penicillium (Eupenicillium javanicium) the CFR0802 fresh PDA inclined-plane gently scraping spore one ring, be inoculated in eggplant bottle inclined-plane, 28 ℃ of static cultivations are after 3-5 days, drawing an amount of sterilized water drips in eggplant bottle inclined-plane, with transfering loop scraping slant pore gently, pour in the aseptic inoculation bottle, the preparation spore concentration is 10
8-10
9The suspension of individual/mL, be that 15% volume ratio (spore suspension/fermentation substrate) is inoculated in and is mounted with the 3.5L fermentation substrate and (or claims fermention medium by inoculum size, it consists of: the Oleum Cocois zymolyte of the above-mentioned first step gained of 175g, 5L fermentor tank 3325g fresh yak milk) (available from newbrunswick scientific, model: BIOFLO 110FERMENTOR) carry out fermentative production; The processing condition of fermentative production are as follows: 28 ℃ of culture temperature, stir revolution 200rpm/min, and ventilate 1: 1v/v, pH is automatic, and dissolved oxygen (DO%) is automatic, cultivates 40-48 hour;
Tunning is carried out local flavor sensory evaluation test, and the local flavor Analyses Methods for Sensory Evaluation Results: the tunning of acquisition has typical cheese flavor, and flavor quality is good, and peculiar smell intensity is low.Specifically, this local flavor mainly is that letones (typical cheese flavor volatile matter) produces, it is stronger to smell overall local flavor, the heptanone flavor is apparent in view, cheese flavor (cheesy) is strong, substantially can't smell milk flavor (milky), fermented bean curd flavor, slightly fatty tart flavour (fatty savory) is arranged, without other peculiar smell.
Adopt the method for solid-phase microextraction that the local flavor volatile matter in the tunning is extracted, and the sample that extraction obtains is carried out GC-MS (gas chromatography and mass spectromentry) detect; Gas chromatograph-mass spectrometer is available from Agilent, and model is 6890-5973.GC-MS detects and all adopts this instrument among the embodiment 2,3.
The GC-MS test result collection of illustrative plates of the local flavor volatile matter in the tunning that obtains is referring to Fig. 1, and as can be seen from Figure 1 main volatile matter is the typical local flavor volatile matter of cheese: methyl ketone (2-heptanone, methyl n-heptyl ketone, 2-undecyl ketone, 2 pentanone).
Embodiment 2, Switzerland's wood mould (Trichoderma reesei) CFR0803 (CGMCC No.3553) liquid fermenting are produced the cheese flavor material
From cultivating on 3-5 days the PDA inclined-plane of Switzerland's wood mould (Trichoderma reesei) CFR0803 gently scraping spore one ring, be inoculated in the 250mL triangular flask of the fresh yak milk that 50mL is housed, back and forth concussion is (28 ℃, 120rpm/min) cultivated 40 hours, obtain primary seed solution (being the one-level hypha fluid).
Draw the 2.5mL primary seed solution, be inoculated in the 500mL triangular flask that 100mL fresh yak milk is housed, (28 ℃ 120rpm/min) were cultivated 24-26 hour in back and forth concussion, obtain secondary seed solution (being the secondary hypha fluid), mycelium dry weight is about 15-25g/L.
With the secondary seed solution that obtains, be that 10% volume ratio (secondary seed solution/fermentation substrate) is inoculated in the 50L fermentor tank that is mounted with 35L fermentation substrate (it consists of: fresh yak milk) by inoculum size, the processing condition of fermentative production are as follows: 28 ℃ of culture temperature, stir revolution 200rpm/min, ventilate 1: 1v/v, tank pressure 0.02MPa, pH is automatic, DO% is automatic, cultivates 23-30 hour.
The tunning that fermentative production is obtained carries out local flavor sensory evaluation test, and the local flavor Analyses Methods for Sensory Evaluation Results: the tunning of acquisition has typical cheese flavor, and is good at flavor quality, and peculiar smell intensity is low.Specifically, this local flavor mainly is that letones (typical cheese flavor volatile matter) produces, smell overall flavor coordination, balanced, cheese flavor (cheesy) is pure and fresh, soft, and substantially can't smell milk flavor (milky), fatty tart flavour (fatty savory), fermented bean curd flavor, rancid flavor, also without other peculiar smell.
Adopt the method for solid-phase microextraction that the local flavor volatile matter in the tunning is extracted, and the sample that extraction obtains is carried out GC-MS (gas chromatography and mass spectromentry) detect;
The GC-MS test result collection of illustrative plates of the local flavor volatile matter in the tunning that obtains is referring to Fig. 2, and as can be seen from Figure 2 main volatile matter is the typical local flavor volatile matter of cheese: methyl ketone (2-heptanone, methyl n-heptyl ketone, 2-undecyl ketone, 2 pentanone).
Embodiment 3, light spore mould (Penicillium glabrum) CFR0801 (CGMCC No.3552) liquid fermenting are produced the cheese flavor material
Get commercially available New Zealand and pacify good cream 500g, at aseptic indoor aseptic cutter cream is cut into the high 1cm that is, length and width are 4 of flaps about 5cm, put in the aseptic dish, prick several apertures (the about 0.5-1mm of each hole diameter) with instrument at the upper surface of flap;
With transfering loop gently scraping cultivate ripe spore on 3-5 days the PDA inclined-plane of light spore mould (Penicillium glabrum) CFR0801, pour aseptic salting bottle into, then vibration evenly evenly trickles down on the good cream of peace surface.
(the upper grand experimental installation of Nereid company limited SHP-250) cultivated 1-2 days, and the upper surface of butter flap is covered with the white hypha body to be positioned over 25 ℃ of biochemical cultivation cases; Cultivated 10-15 days, the upper surface of butter flap and inside all are covered with the white hypha body, are embedded with the blue-greenish colour spore in the mycelium.
The tunning that fermentative production is obtained carries out local flavor sensory evaluation test, and the local flavor Analyses Methods for Sensory Evaluation Results: the tunning of acquisition has typical cheese flavor, and score is high aspect flavor quality, and peculiar smell intensity aspect score is low.Specifically, this local flavor mainly is that letones (typical cheese flavor volatile matter) produces, it is strong to smell overall local flavor, cheese flavor (cheesy) is strong, slightly acescency (acid taste) and milk flavor (milky) are arranged, fat-free tart flavour (fatty savory), fermented bean curd flavor, rancid flavor, and other peculiar smell.
Adopt the method for solid-phase microextraction that the local flavor volatile matter in the tunning is extracted, and the sample that extraction obtains is carried out GC-MS (gas chromatography and mass spectromentry) detect.
The GC-MS test result collection of illustrative plates of the local flavor volatile matter in the tunning that obtains is referring to Fig. 3, and as can be seen from Figure 3 main volatile matter is the typical local flavor volatile matter of cheese: methyl ketone (2-heptanone, methyl n-heptyl ketone, 2-undecyl ketone, 2 pentanone).
The application of embodiment 4, cheese flavor material
1, makes butter and the buttered tea of cheese flavor
1., the tunning 0.5kg with gained among the embodiment 1 carries out homogeneous first, together joins in the water tank with the 1kg yak milk, adds simultaneously the 2g citric acid, the 9g potassium sorbate is heated to 80 ℃, kept 15 minutes, while stirring the sterilization.
2., 8.5kg New Zealand anhydrous milkfat is joined in the oil phase tank, heat while stirring, add simultaneously the 68g mono-glycerides, 10g Yelkin TTS, the 0.004g β-carotene heats 60 ℃, stirs and keeps 15 minutes.
3., feed liquid in the water tank is filtered by deep bed filter, join the oil phase tank, emulsification 15 minutes forms emulsion.
4., emulsion is again through after the deep bed filter filtration, through the plate-type heat exchanger sterilization.
5., chilling is mediated: milk sap pumps into Chiller (A unit) by ram pump or toothed gear pump under certain pressure intensity, rapidly crystallization in the crystallization cylinder, enter kneader (B unit), the B unit is mediated the material vigorous stirring, and slowly forms 10kg cheese flavor butter again.
6., place 15 ℃, slaking.
Also can make oil and fat product such as oleomargarine by above-mentioned method or based on the mode of texturing of aforesaid method.
Although the basic making method of margarine oil's wet goods oil and fat product is well-known to those skilled in the art, yet mycetogenetic cheese flavor material can't be directly used in when producing oil fat prod, (flavour substances that fermentation produces generally contains fungi pellet and mycelium certain problem on the technique, in the technique of the oil and fat products such as manufacturing oleomargarine of routine, for guaranteeing that impurity does not enter system of processing, often before entering system of processing, dispose the filter bag of 10-100um, so oleomargarine moulding process routinely, mycelium in the fermented flavour material can be filtered bag and hold back, and causes the finished product flavour intensity greatly to reduce).
The present invention has adopted the method for carrying out homogeneous before entering system of processing dexterously, can effectively pulverize the mycelium in the fermented flavour, thereby guarantee that the local flavor material can smoothly by the forming production line of margarine oil's wet goods oil and fat product, guarantee that the local flavor of the finished product reaches design requirements.
In the present embodiment, before joining the cheese flavor material that obtains in the foodstuff base material it is carried out the homogeneous operation.
In the present embodiment, the cheese flavor material that utilizes new strain fermentation production of the present invention to obtain is made the zestful butter of tool; In addition, also can use existing cheese flavor material (such as blue cheese flavour substances) to make the zestful margarine oil's wet goods of tool oil and fat product (need to before joining the cheese flavor material in the foodstuff base material, carry out the homogeneous operation to it).
7., buttered tea preparation
In Stainless Steel Kettle, add 1000g water, be heated at electromagnetic oven and boil, add brick tea 3g, salt 2.5g stirs and boiled 10 minutes, gets tea with filtered through gauze, get tea 500g, pour in the high-shear homogenizer, add the cheese flavor butter 20g of above-mentioned slaking, high-speed stirring 1.5 minutes buttered tea.
2, as baking and banking up with earth food grease (making biscuit)
Cheese flavor butter 150g and the egg 45g of aforementioned preparation are stirred, add 150g flour and 120g Icing Sugar again, said mixture is kneaded, the baking oven baking is put in die sinking, obtains the local flavor biscuit.
3, make cheese
Get the 4L fresh milk, through 63 ℃ of sterilizations 30 minutes, 10% tunning (through sterilization, need not homogeneous) that adds embodiment 2 gained by volume after the cooling, rennin adds in the milk with 10 times of sterilized water dilutions, stirred 3-5 minute, the curdled milk time is about 45-60 minute, begins to stir after granular curd reaches suitable size, whole churning time was controlled in 20 minutes, the whey of draining, 4-8 ℃ ripe preservation 10-30 days, obtain to have the cheese of typical cheese flavor.
4, as food flavor(ing) spices
The tunning 30kg (through sterilization, need not homogeneous) that gets embodiment 3 carries out spraying drying, control water evaporation quantity 5kg/h, inlet temperature: 150 ℃, air outlet temperature 80-100 ℃, obtain 2.4kg powder cheese spices, it can add in the varieties of food items to obtain the cheese flavor feature.
Culture presevation
The Java penicillium of production cheese flavor material of the present invention (Eupenicillium javanicium) CFR0802 has been preserved in the common micro-organisms center (CGMCC of China Committee for Culture Collection of Microorganisms on December 28th, 2009, the BeiJing, China), preservation registration number is CGMCC No.3551.
Light spore mould (Penicillium glabrum) CFR0801 of production cheese flavor material of the present invention has been preserved in the common micro-organisms center (CGMCC of China Committee for Culture Collection of Microorganisms on December 28th, 2009, the BeiJing, China), preservation registration number is CGMCC No.3552.
Switzerland's wood of production cheese flavor material of the present invention mould (Trichoderma reesei) CFR0803 has been preserved in the common micro-organisms center (CGMCC of China Committee for Culture Collection of Microorganisms on December 28th, 2009, the BeiJing, China), preservation registration number is CGMCC No.3553.
All quote in this application as a reference at all documents that the present invention mentions, just as each piece document is quoted separately as a reference.Should be understood that in addition those skilled in the art can make various changes or modifications the present invention after having read above-mentioned teachings of the present invention, these equivalent form of values fall within the application's appended claims limited range equally.
Claims (16)
1. preserving number is the application of Java penicillium (Eupenicillium javanicum) aspect production cheese flavor material of CGMCC NO.3551.
2. separated bacterial strain, it is Java penicillium CFR0802, its preserving number is CGMCC NO.3551.
3. method of producing the cheese flavor material is characterized in that the method comprises:
Adopt spore or the mycelium of the described bacterial strain of claim 2, take the material that contains breast, oil, lipid and/or its enzymolysis product as fermentation substrate, through liquid fermenting or solid fermentation, obtain to have the tunning of cheese flavor.
4. method as claimed in claim 3 is characterized in that, described liquid fermenting comprises:
The spore suspension or the hypha fluid that prepare first bacterial strain described in the claim 2, in the fermentation substrate that again spore suspension or the hypha fluid of significant quantity is inoculated into liquid form, obtain the tunning of liquid form, leavening temperature is 25-35 ℃, and fermentation time is 24-48h.
5. method as claimed in claim 4 is characterized in that, also comprises:
Collect tunning or its concentrated solution of described liquid form, as the flavour substances product; Perhaps,
Described liquid form tunning is carried out degerming process, resulting liquid with cheese flavor after the collection and treatment is as the flavour substances product; Perhaps,
Extract volatile matter in the described liquid form tunning as the flavour substances product; Perhaps,
Described liquid form tunning is carried out drying treatment, and resulting solid with cheese flavor after the collection and treatment is as the flavour substances product.
6. method as claimed in claim 3 is characterized in that, described solid fermentation comprises:
The spore suspension or the hypha fluid that prepare first the bacterial strain described in the claim 2, spore suspension or hypha fluid with significant quantity is inoculated in the process that solid-state form fermentation substrate surface obtains the solid-state form tunning again, leavening temperature is 15-25 ℃, and fermentation time is 10-15 days.
7. method as claimed in claim 6 is characterized in that, also comprises:
Collect described solid-state form tunning, as the flavour substances product; Perhaps,
Extract volatile matter in the described solid-state form tunning as the flavour substances product; Perhaps,
Described solid-state form tunning is carried out drying treatment, and resulting solid with cheese flavor after the collection and treatment is as the flavour substances product.
8. method as described in arbitrary such as claim 3-7 is characterized in that: comprise at least 2-heptanone, 2 pentanone and methyl n-heptyl ketone in the described tunning.
9. method as described in arbitrary such as claim 3-7, it is characterized in that: the described material that contains breast, oil, lipid is selected from: milk-product, animal-plant oil or microbial oil.
10. adopt arbitrary such as claim 4-7 as described in the cheese flavor material that obtains of method.
11. the described cheese flavor material of claim 10 is in the application of field of food.
12. a method for making food is characterized in that the method comprises: before the cheese flavor material that the arbitrary described method of the claim 3-9 of appropriate amount is obtained adds in the foodstuff base material, simultaneously or afterwards, described cheese flavor material is sterilized.
13. the application of cheese flavor material in fatty foods that fungi fermentation produces; Described fungi is that preserving number is the Java penicillium of CGMCC NO.3551.
14. application as claimed in claim 13 is characterized in that: described fatty foods is oleomargarine, butter, crisp skin oil.
15. the manufacture method of flavor oil fat foods is characterized in that: the method comprises the step that cheese flavor material that fungi fermentation is produced and the base-material of fatty foods mix and sterilize; Described fungi is that preserving number is the Java penicillium of CGMCC NO.3551.
16. manufacture method as claimed in claim 15 is characterized in that: the cheese flavor material that first fungi fermentation is produced carries out mixing with the base-material of fatty foods behind the homogeneous again.
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| CN102703527A (en) * | 2012-05-16 | 2012-10-03 | 深圳波顿香料有限公司 | Preparation method for C5-C13 aliphatic series methyl ketone |
| CN104543924A (en) * | 2013-10-15 | 2015-04-29 | 丰益(上海)生物技术研发中心有限公司 | Method for producing flavor substance through fermenting fruit and vegetable residue and flavor substance obtained therefrom |
| CN103719794B (en) * | 2013-12-05 | 2016-08-17 | 杭州艾菲曼普香精香料有限公司 | A kind of milk flavor formula |
| CN107860872B (en) * | 2017-11-29 | 2018-08-31 | 北京工商大学 | Measure the method that volatile flavor substance in cheese perceives threshold value |
| CN111235061B (en) * | 2020-02-28 | 2022-04-15 | 东莞波顿香料有限公司 | Bacillus coagulans, milk-flavored essence base and preparation method and application thereof |
| CN111387295A (en) * | 2020-03-23 | 2020-07-10 | 新疆天牛乳业有限公司 | Instant cheese snack food produced rapidly by biological enzyme conversion method |
| CN112980694B (en) * | 2020-12-30 | 2022-04-15 | 中国中医科学院中药研究所 | Penicillium chrysogenum SF-85 strain and application thereof |
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