CN104472904B - The application in improving Lac Bovis seu Bubali flavor substance, vitamin or essential amino acids content of the Cordyceps strain fermentation thing - Google Patents

Abstract

The invention discloses the application in improving Lac Bovis seu Bubali flavor substance, vitamin or essential amino acids content of the Cordyceps strain fermentation thing.When scale of feeding is 30g/ head/sky, feeds 1 month, in test group Lac Bovis seu Bubali, the kind of flavor substance is more than the twice of matched group, and flavor substance total amount is up to more than 20 times of matched group；Vitamin A content improves more than 25%, and content of vitamin E improves more than 73.3%, it is necessary to amino acid content improves 12.96%.Show that Cordyceps strain fermentation thing can be effectively improved the kind of flavor substance in Lac Bovis seu Bubali and content and the content of nutrient substance, significantly improve Lac Bovis seu Bubali local flavor and quality.

Description

The application in improving Lac Bovis seu Bubali flavor substance, vitamin or essential amino acids content of the Cordyceps strain fermentation thing

Technical field

The present invention relates to the method for breeding of a kind of milch cow, be specifically related to Cordyceps strain fermentation thing and improving Lac Bovis seu Bubali flavor substances Application in matter, vitamin or essential amino acids content.

Background technology

Along with improving further and the enhancing of health care consciousness of people's living standard, consumers in general not only focus on Lac Bovis seu Bubali Nutritive value, and the most more pay attention to the quality of Lac Bovis seu Bubali, local flavor and health care.People are to the consumption of Lac Bovis seu Bubali Through from physiologically the increase in demand of various nutritional labelings and hygienic quality being enjoyed to psychological.Excellent or peculiar flavour It is joyful that Lac Bovis seu Bubali and milk product thereof can make people obtain on sense organ, and directly affects digestion and the absorption of nutrient substance, people Wish to obtain the agreeable to the taste good Lac Bovis seu Bubali of local flavor delicate fragrance, be more desirable to obtain the Gao Pin of homovitamin, high amino acid content simultaneously Matter Lac Bovis seu Bubali.

At present Lac Bovis seu Bubali local flavor oneself become the important indicator evaluating Lac Bovis seu Bubali grade, the most gradually change people and the tradition of Lac Bovis seu Bubali disappeared Take idea.But dairy obtain milk yield significantly promote with the high price of deed while, also along with Milk Quality The negative effect declined, adds a large amount of uses of antibiotic and chemical synthetic drug in production, more exacerbate Milk Quality, The decline of local flavor.Thus, consumer usually complains that some Lac Bovis seu Bubali palatability is poor, and it is to allowing people be difficult to accept sometimes Milk fishy smell, milk stink.

Characterization of Odor-active Compounds in Lac Bovis seu Bubali is of a great variety, varies, distinct, affects the factor of Lac Bovis seu Bubali local flavor very Many, but feed ingredient plays Main Function to the impact of milk local flavor.

Flavor substance in Lac Bovis seu Bubali by have flavour and fragrance activity become to be grouped into.Its flavor substance forming process is main It is to react to each other between protein in milk, fat, this three major types mass degradation of lactose or the derivant of each class material Generate.

Research shows, in butterfat flavor substance source has 4 kinds: (1) flavor substance without any change directly from feedstuff In entered in Lac Bovis seu Bubali by blood；(2) aroma compound through enzyme, aoxidize or heat make oils and fats decompose and formed；(3) Aroma compound be in the cud of ruminant, there is biochemical reaction by feedstuff after produce through digesting and assimilating；(4) Aroma compound be cattle protein of milk and carbohydrate reaction and generate.The aroma compound in Ruzhong mainly has Fatty acid, carbonyl compound, esters, terpenoid and sulfide (Liu Fang, Huo Guicheng, dairy industry science and technology, 2001(4):14-17)。

In Lac Bovis seu Bubali, volatile flavor substance generally can be summarized as two big classes: a class be hydrocarbon, alcohol, aldehyde, ketone, acid, The simple compounds such as ester, lactone；Another kind of is containing aerobic, nitrogen, the heterocyclic compound of sulfur.Such as Furan and its derivatives And thiophene and derivatives.Its concrete composition is: acetaldehyde, ethanol, isoamyl alcohol, Ethyl formate, ethyl acetate, second Acid isobutyl ester, isoamyl acetate and n-butyl acetate etc..Research finds, strong volatile material has: octane, epoxy Ethane, caproic acid, benzene hexanone, aldehyde C-9, octanoic acid, dodecane, tridecane, 2 one capraldehyde, 2.4 (E) laurylene aldehyde, 2 One methylene one cyclopentanol, capric acid etc..Being all the important component of flavor substance in Lac Bovis seu Bubali, these compounds are except there being uniqueness Local flavor outside, the also health to human body cuts much ice (Nursten H E, International Journal o f Diary Teach,1997(50):48-55)。

Raising dairy cattle is a complicated problem, and factors affects Milk Quality, local flavor.Therefore, research and development were both Can guarantee that milk yield and feed efficiency, Milk Quality, the feed additive of local flavor or feed formula can be improved again, Production safety high-quality Lac Bovis seu Bubali, oneself becomes the focus of research and development both at home and abroad.

Fungi-medicine that Dong Chongshi China is famous and precious and senior tonic, before the Ming Dynasty of China, it is known that road application Cordyceps, From the beginning of mid-Ming Dynasty, just liked by the Southeast Asia people, enjoyed a very good reputation in the international market.

Sensu lato Cordyceps refers to that entomogenous fungi infects insecticide or arthropod, the fungus sporophore (son eventually formed Seat) and insecticide or arthropodan complex.Cordyceps sinensis fungus there are about more than 400 kinds, and the famousst and precious is the worm summer in winter Grass (Cordyceps sinensis), is that the linear Cordyceps sinensis fungus of Clavicipitaceae (Ophiocordyceps) parasitizes Genus Hepialus The complex formed in the larva body of (Hepialus spp.), is the endemic species of high altitude localities, Qinghai-Tibet Platean, is called for short " Cordyceps ", is the rare Chinese medicine of Chinese tradition.Have been found that the Cordyceps existing 305 belonging to " Cordyceps " together at present in the world Ten kinds more than, wherein had been found that more than 60 planted in China, comprised Cordyceps (C.sinensis), Cordyceps militaris (L.) Link. (C.militaris), agglomerate Cordyceps (C.ophioglossoides), Cordyceps hawkesii Gary (C.hawkesii), Cordyceps gunnii (Berk.) Berk. (C.gunnii), Cordyceps martialis Speg. (C.martialis), sickle shaped Cordyceps (C.falcata), Taishan Cordyceps sp (C.taishanensis), Cordyceps shanxiensis (C.shanxiensis), Cordyceps liangshanensis Zang Hu et Liu (C.liangshanensis), Xinjiang Cordyceps (C.gracilis), incense stick cordycep (C.barnesii), Periostracum cicadae (C.sobolifera) etc..Although have been found that is various in style, but Cordyceps has the strongest mostly The effect of raising immunocompetence.

The worm grass product successively successfully developed has: (1) paecilomyces hepiall chen, listing in 1987, is to use from green grass or young crops In the Cordyceps of sea, separating obtained peacilomyce hepiahi (Paecilomyces hepiali) Cs-4 bacterial strain is prepared from； (2) Caps Bailing, be with Shen Nanying etc. at nineteen eighty-three isolated a kind of fungus China pilose spore (also known as China's synnema Spore (Synnematum sinense Y in et Shen sp.now), aweto-cephalosporin, Hirsutella hapial) it is that strain is raw Produce；(3) zhiling capsules, is to ferment with from the isolated and purified mortierella sp (Mortierella sp) out of Cordyceps To mycelium make；(4) Ningxinbao Capsules, for the Clavicipitaceae fungus of isolated from fresh Cordyceps Cordyceps cephalo (Cephalosporium sinensis Chen.sp.nov), through liquid submerged fermentation gained；(5) conscience is precious Capsule, be 1986 with the cephalosporium sinensis separated in Cordyceps fresh specimens, be a kind of fungi autoeciousness mycopowder Red glue mould (Gliocladium roseum (Link) Bain).

Worm grass product application on animal also obtain some achievements.Particularly mortierella Diding culture has multiple life Thing function, such as, (Wei Jianzhong etc., mortierella Diding culture is to child care weaned piglets and immunity for China's document The impact of level, Animal Nutrition and Feed Science, 2009,36 (2): 33-35.) disclose mortierella Diding culture energy Enough improve production performance and the immune level of child care piglet.

Notification number is the producer that the Chinese patent literature of CN101797014B discloses a kind of nutritive eggs with low cholesterol Method and products thereof, this patent finds that Cordyceps can improve quality of egg intension, produces the high-quality fowl egg of low cholesterol.

Notification number is that the Chinese patent of CN101002592A reports a kind of paecilomyces gunniliang novel nourishing type Chinese caterpillar fungus feed The production method of additive, but this patent application is to disclose the preparation of this Chinese caterpillar fungus feed additives and in animal growth-promoting Length is applied.

The Chinese patent of Publication No. CN 101066090B reports a kind of aweto activated reinforced immunological feed additive, Using Cordyceps militaris strain and Cordyceps strain mixed culture, gained compound criteria thing is this feed additive after drying, This patent is not the most pointed out or implies Chinese caterpillar fungus feed additives application in milch cow.

The patent of relevant Cordyceps application at present is not involved with to change Lac Bovis seu Bubali local flavor and improving the content of quality, does not also have Pertinent literature is had to report.

Summary of the invention

The invention provides Cordyceps strain fermentation thing and improve Lac Bovis seu Bubali flavor substance, vitamin or essential amino acids content In application, this Cordyceps strain fermentation thing is fed lactating cow, it is possible to significantly improve the kind of flavor substance in Lac Bovis seu Bubali Class and content and vitamin and the content of essential amino acids.

The application in improving Lac Bovis seu Bubali flavor substance, vitamin or essential amino acids content of the Cordyceps strain fermentation thing.

Described application includes: described Cordyceps strain fermentation thing feeds lactating cow, obtains Lac Bovis seu Bubali.

As preferably, in terms of dry weight, the scale of feeding of described Cordyceps strain fermentation thing is at least 5g/ head/sky.

Test finds, when the scale of feeding of Cordyceps strain fermentation thing is more than 5g/ head/sky, can improve local flavor in Lac Bovis seu Bubali The content of material, vitamin and essential amino acids, and, it is gradually increased with the scale of feeding of Cordyceps strain fermentation thing, The effect above is the most obvious.

As further preferably, in terms of dry weight, the scale of feeding of described Cordyceps strain fermentation thing is 20～40g/ heads/sky. In the range of this scale of feeding, in Lac Bovis seu Bubali, the content of flavor substance, vitamin and essential amino acids dramatically increases, Lac Bovis seu Bubali Local flavor and quality can obtain to be improved largely.Continue to increase scale of feeding, flavor substance in Lac Bovis seu Bubali, vitamin with And the content of essential amino acids likely continues to improve, it is also possible to no longer improve after arriving a certain value.Due to the present invention's Cordyceps strain fermentation thing, will not to the production of milch cow itself so increasing scale of feeding to milch cow without any side effects There is any impact.For obtaining corresponding feeding effect, those skilled in the art can join according to rearing conditions and feedstuff Side selects concrete scale of feeding.

Respectively, the invention provides the application in improving Lac Bovis seu Bubali flavor substance content of the Cordyceps strain fermentation thing.

Described flavor substance includes at least one in following (1), (2) or (3):

(1) in GC-MS collection of illustrative plates, retention time is the rudimentary alkanoic acid of 8～15min, such as: lactic acid, caproic acid etc.；

(2) in GC-MS collection of illustrative plates, retention time is alkyl acid, alkyl aldehydes or the alkyl ketone thing of 15～20min Matter, such as: octanoic acid, capraldehyde, maltol series material (including maltol, 5-hydroxyl maltol), furanone series matter Matter (including 4-hydroxyl dihydrofuran-2 (3H)-one, 5-(methylol) dihydrofuran-2 (3H)-one) etc.；

(3) in GC-MS collection of illustrative plates, retention time be 20～40min higher alkane, higher alkene, senior alkane Base aldehyde, senior alkyl amide or higher alkanoic acid and Ester thereof, such as: Squalene, certain herbaceous plants with big flowers octadecenoic acid sour, cis-, Palmic acid series derivates (including Palmic acid, palmital, palmitamide, tripalmitin) etc..

The testing conditions of described GC-MS collection of illustrative plates is:

INSTRUMENT MODEL: Thermo ISQ 1301；

GC conditions: specification is the HP-5 capillary column of 30 × 0.32mm × 0.5 μm；

Temperature programming: initial temperature is 40 DEG C, is incubated 4min, then rises to 100 DEG C with 5 DEG C/min, retains 2min, Rise to 220 DEG C with 10 DEG C/min again, retain 5min injector temperature 250 DEG C, detector temperature 280 DEG C, carrier gas For high pure nitrogen.

Compared with the milch cow not feeding described Cordyceps strain fermentation thing, after feeding Cordyceps strain fermentation thing, milch cow institute Producing in Lac Bovis seu Bubali and occur in that new flavor substance, these flavor substances are mainly maltol series material, furanone series matter Matter and Palmic acid series derivates.

As do not made specified otherwise, in the present invention, described rudimentary alkanoic acid refer to carbon atom number 1～6 alkanoic acid, described Higher alkane, higher alkene or higher alkanoic acid and Ester thereof refer to carbon atom number more than 6 alkane, alkene or Alkanoic acid and Ester thereof.Test finds, when scale of feeding is 30g/ head/sky, feed 1 month, from test group Lac Bovis seu Bubali sample Product examine measures 22 flavor substance characteristic peaks, and 10 flavor substance characteristic peaks only detected from matched group milk sample, Flavor substance total amount in test group milk sample is significantly larger than matched group；During feed 2 months, from test group Lac Bovis seu Bubali sample Product examine measures 28 flavor substance characteristic peaks, and 13 flavor substance characteristic peaks only detected from matched group milk sample. Flavor substance total amount in test group milk sample is significantly larger than matched group.Show that Cordyceps strain fermentation thing can be effective Increase kind and the content of flavor substance in Lac Bovis seu Bubali, significantly improve Lac Bovis seu Bubali local flavor.In addition, in Lac Bovis seu Bubali vitamin and The content of essential amino acids also dramatically increases.Therefore, present invention also offers Cordyceps strain fermentation thing and improve Lac Bovis seu Bubali Application in vitamin or essential amino acids content.

As preferably, described vitamin is at least one in VitAVitE or vitamin D.Test is sent out Existing, when scale of feeding is 30g/ head/sky, feed 1 month, the vitamin A content in test group milk sample compares matched group Improve 25%, content of vitamin E improves 73.3% than matched group；During feed 2 months, test group milk sample In vitamin A content improve 36.8% than matched group, content of vitamin E improves 46.7% than matched group；Raise Feed period, measure less than vitamin D from matched group milk sample, and dimension can be measured to from test group milk sample Raw element D.Show that Cordyceps strain fermentation thing can be effectively increased the content of vitamin in Lac Bovis seu Bubali, significantly improve Milk Quality.

As preferably, described essential amino acids be aspartic acid, glycine, alanine, cysteine, valine, At least one in leucine, phenylalanine, lysine or histidine.Test find, scale of feeding be 30g/ head/sky, During feed 1 month, in test group milk sample, essential amino acids content improves 12.96% than matched group, wherein, and sky Winter propylhomoserin, glycine, alanine, cysteine, valine, leucine, phenylalanine, lysine and group ammonia The content of acid improves 43.6% than matched group；Local flavor aminoacid (aspartic acid, glutamic acid, glycine, alanine) Content improve 41.87% than matched group.Show that Cordyceps strain fermentation thing can be effectively increased essential amino acids in Lac Bovis seu Bubali Content, significantly improve Lac Bovis seu Bubali local flavor and quality.

In the present invention, described Cordyceps strain fermentation thing is obtained through liquid fermentation or solid fermentation by Cordyceps sinensis fungus； Can be liquid fermentation bacterium solution, can be the mycopowder utilizing liquid fermentation solid content to be prepared as, it is also possible to be solid fermentation Thing.

Described Cordyceps microbial strain culture can be prepared by following fermentation process:

(1) liquid seeds fermentation: the Cordyceps sinensis fungus after activation is inoculated in one grade fermemtation tank, carries out seed culture, Obtain seed liquor；

Described inoculum concentration is preferably 8～12%, and the state modulator of described seed culture exists: temperature 25 DEG C, ventilation 1.00L/ (L min), incubation time 84h.

(2) liquid fermentation: seed liquor is inoculated in second order fermentation tank, ferments, it is thus achieved that Cordyceps sinensis fungus liquid Zymocyte liquid；

The inoculum concentration of described seed liquor is preferably 8～12%, described fermentation state modulator exist: temperature 25 DEG C, ventilation Amount 1.00L/ (L min), incubation time 84h.

(3) solid fermentation: described Cordyceps sinensis fungus liquid fermentation bacterium solution is inoculated in solid medium, tiles laggard Row solid fermentation, it is thus achieved that Cordyceps sinensis fungus solid culture.

The inoculum concentration of described Cordyceps sinensis fungus liquid fermentation bacterium solution is preferably 15%, and after tiling, material thickness is maintained at 3cm Left and right, the state modulator of described solid fermentation exists: temperature constant 25 DEG C, humidity 70%, fermentation time 165～170h； After mycelia covers with solid medium, cultivation terminates.

Described Cordyceps sinensis fungus liquid fermentation bacterium solution can be used for feeding milk directly as the Cordyceps strain fermentation thing of the present invention Cattle；Also it can be centrifuged, collect mycelia and solid content, this mycelia and solid content are placed in 45 DEG C of hot air dryings extremely Moisture is down to below 8wt%, pulverizes, it is thus achieved that Cordyceps sinensis fungus liquid fermentation mycopowder, this mycopowder also can be as the present invention's Cordyceps strain fermentation thing thing, for feeding cow.

Described Cordyceps sinensis fungus solid culture is down to below 8wt% through 60 DEG C of hot air drying to moisture, pulverizes, it is possible to make For the Cordyceps strain fermentation thing of the present invention, for feeding cow.

Following liquid culture based formulas can be used to carry out liquid seeds fermentation and liquid fermentation: sucrose 2.0%, peptone 0.4%, yeast extract powder 0.3%, Semen Maydis powder 0.5%, MgS0₄7H₂O 0.02%, K₂HP0₄0.04%.Natural pH。

Following solid culture based formulas can be used to carry out solid fermentation: potato starch 83%, Semen Maydis powder 10%, white sand Sugar 2%, peptone 2%, dried silkworm chrysalis meal 2.5%, magnesium sulfate 0.2%, ammonium citrate 0.2%, potassium dihydrogen phosphate 0.1%, VB11 00mg, water, pH value 5～7.

Described Cordyceps sinensis fungus is Cordyceps gunnii (Berk.) Berk. (Cordyceps.gunnii) strain, Cordyceps (C.sinensis) bacterium Kind, Cordyceps militaris (L.) Link. (C.militaris) strain, Cordyceps hawkesii Gary (C.hawkesii) strain, Xinjiang Cordyceps (C.gracilis) At least one in strain, agglomerate Cordyceps (C.ophioglossoides) strain or Periostracum cicadae (C.sobolifera) strain.

As do not made specified otherwise, indication Cordyceps sinensis fungus of the present invention refers to the parasitical fungi of isolated from Cordyceps, should Parasitical fungi forms the Stroma in corresponding Cordyceps.

This six classes Cordyceps is Cordyceps the most most commonly seen, the most representational, it is found by the applicant that this six classes Cordyceps Belong to fungus and all can be effectively improved the content of flavor substance in Lac Bovis seu Bubali.

Described Cordyceps gunnii (Berk.) Berk. strain can be selected for mortierella Diding (Acremonium terricola), and described Cordyceps strain can Selecting China pilose spore (Synnematium sinense sp.), described Coragyceps militaris bacterium can be selected for Cordyceps militaris (L.) Link. Paecilomyces varioti (Paecilomyces militaris), described Cordyceps hawkesii Gary strain can be selected for Paecilomyces hawkesii (Paecilomyces Hawkesii), described Xinjiang cordyceps species can be selected for Paraisaria mould (Paraisaria sp.), described agglomerate cordyceps species Can be selected for false truffle (Elaphomyces granulatus), described Cordyceps cicadae strain can be selected for Cicadae muscardine (Beauveria sobolifera)。

As preferably, described Cordyceps sinensis fungus is mortierella Diding (Acremonium terricola).True with other Cordyceps The fermented product of bacterium is compared, and mortierella Diding fermented product is obtained in that more preferable feeding effect.

Present invention also offers a kind of method improving Lac Bovis seu Bubali local flavor and quality, including: Cordyceps strain fermentation thing is raised Feed lactating cow, obtain flavor substance and nutrition content all improves, the Lac Bovis seu Bubali of flavor substance kind increase；

Described Cordyceps strain fermentation thing is obtained through liquid fermentation or solid fermentation by Cordyceps sinensis fungus；

Described Cordyceps sinensis fungus is Cordyceps gunnii (Berk.) Berk. (Cordyceps.gunnii) strain, Cordyceps (C.sinensis) bacterium Kind, Cordyceps militaris (L.) Link. (C.militaris) strain, Cordyceps hawkesii Gary (C.hawkesii) strain, Xinjiang Cordyceps (C.gracilis) At least one in strain, agglomerate Cordyceps (C.ophioglossoides) strain or Periostracum cicadae (C.sobolifera) strain.

As preferably, in terms of dry weight, the scale of feeding of described Cordyceps strain fermentation thing is no less than 5g/ head/sky.

Compared with prior art, the invention have the benefit that

The present invention utilizes and feeds lactating cow containing the feedstuff of described Chinese caterpillar fungus feed additives, scale of feeding be 30g/ head/sky, When feeding 1 month, in test group Lac Bovis seu Bubali, the kind of flavor substance is more than the twice of matched group, and flavor substance total amount is high Reach more than 20 times of matched group；In test group Lac Bovis seu Bubali, vitamin A content improves more than 25% than matched group, and dimension is raw Element E content improves more than 73.3% than matched group, and in test group Lac Bovis seu Bubali, essential amino acids content improves than matched group 12.96%, wherein, aspartic acid, glycine, alanine, cysteine, valine, leucine, phenylalanine, The content of lysine and histidine improves 43.6% than matched group；Local flavor aminoacid (aspartic acid, glutamic acid, Glycine, alanine) content improve 41.87% than matched group.Show that Cordyceps strain fermentation thing can effectively carry In high Lac Bovis seu Bubali, the kind of flavor substance and content and the content of nutrient substance, significantly improve Lac Bovis seu Bubali local flavor and quality.

Figure of description

Fig. 1 is the GC-MS figure of local flavor substance-measuring in test group milch cow produced Lac Bovis seu Bubali during the glad health of feed Cordyceps 1 month Spectrum；

Wherein, Relative Abundance represents relative abundance, time (min) express time (minute), lower with；

Fig. 2 is the GC-MS figure of local flavor substance-measuring in matched group milch cow produced Lac Bovis seu Bubali during the glad health of feed Cordyceps 1 month Spectrum；

Fig. 3 is produced essential amino acids in Lac Bovis seu Bubali by test group milch cow during the glad health of feed Cordyceps 1 month and is measured collection of illustrative plates；

Retention Time represents that retention time, minutes represent minute, and Name represents (at corresponding retention time) Amino acid name, wherein, Asp, Thr, Ser, Glu, Pro, Gly, Ala, Cys, Val, Met, Ile, Leu, Tyr, Phe, Lys, His, Arg represent aspartic acid, threonine, serine, glutamic acid, proline, sweet successively Propylhomoserin, alanine, methionine, isoleucine, leucine, tyrosine, phenylalanine, histidine, arginine, NH3 represents amino, lower same；

Fig. 4 is produced essential amino acids in Lac Bovis seu Bubali by matched group milch cow during the glad health of feed Cordyceps 1 month and is measured collection of illustrative plates.

Detailed description of the invention

Below in conjunction with detailed description of the invention, the present invention is described in further detail.

The preparation of embodiment 1 mortierella Diding culture (the glad health of Cordyceps)

1, the preparation of mortierella Diding culture

(1) prepared by mortierella Diding liquid fermentation bacterium solution

1. strain: mortierella Diding (Acremonium terricola)

2. liquid fermentation medium

First order seed liquid fermentation medium: 600g white sugar, 200g yeast powder, 10g MgSO₄、10g KH₂PO₄, It is dissolved in 20L water, natural pH；121 DEG C of high pressure steam sterilization 15min, standby.

Secondary seed liquid fermentation medium: 6000g white sugar, 2000g analysis for soybean powder, 2000g yeast powder, 100g MgSO₄,100g KH₂PO₄, it is dissolved in 200L water, natural pH；121 DEG C of high pressure steam sterilization 15min, standby.

(3) liquid fermentation

Liquid seeds ferments: it is temperature 25 DEG C that liquid fermentation controls parameter, ventilation 1.00L/ (L min), inoculum concentration 10%, 300L airlift fermentor liquid amount 67%, fermentation time 84h.

Liquid fermentation: it is temperature 25 DEG C that liquid fermentation controls parameter, ventilation 1.00L/ (L min), inoculum concentration 10%, 300L airlift fermentor liquid amount 67%, fermentation time 84h.

(4) prepared by mortierella Diding solid culture

1. solid fermentation culture medium: Semen Maydis powder 55%, wheat-middlings of Semen Tritici aestivi 5%, bean cake 25, Semen arachidis hypogaeae dregs 5%, Testa Tritici 9.8%, Magnesium sulfate 0.05%, zinc sulfate 0.05%, potassium dihydrogen phosphate 0.05%, sodium dihydrogen phosphate 0.05%；Natural pH, 100 DEG C of steam sterilization 6h, standby.

2. solid fermentation: fresh mortierella Diding bacterium solution and solid medium are stirred inoculation according to weight ratio 15%, tiling At tray top fermentation, material thickness 3cm, temperature constant 25 DEG C, humidity 70%, fermentation time 168h.

3. drying and crushing: collecting mortierella Diding solid culture, 60 DEG C of hot air drying to moisture are down to below 8wt%, powder Broken pack, seals, and lucifuge is stored.

4. quality analysis

Measurement result is shown in Table 1:

Table 1 mortierella Diding solid culture quality test results

Detection project	Detected value
		Moisture %	5.8
Protein %	27.8
		Cordyceps polysaccharide %	3.3
Adenosine mg/kg	652
		Ergosterol mg/kg	627

The embodiment 2 mortierella Diding culture (the glad health of the Cordyceps) impact on Lac Bovis seu Bubali local flavor

(1) experimental animal: test group and matched group 10 cow heads respectively, parity is consistent with milk yield.

(2) scale of feeding

Test group: every adult dairy cattle day raises 10kg concentrated feed, adds the mortierella Diding solid training of 0.3% in concentrated feed Support thing (the glad health of Cordyceps)；

Matched group: every adult dairy cattle day raises 10kg concentrated feed, without the glad health of Cordyceps in concentrated feed；

Day concentrated feed formula is: Semen Maydis 52%, Testa Tritici 5%, skin of Semen Maydis 5%, bean cake 8%, cottonseed meal 12%, dish The seed dregs of rice 8%, DDGS3%, stone powder 2%, magnesium oxide 0.3%, carbamide 1%, Sal 0.7%, sodium bicarbonate 1%, phosphorus Acid hydrogen calcium 1%, premix material 1%.

(3) test procedure

1., before starting to feed, observe lactating cow 3 days, record its food-intake, milk yield；

2. according to content feeding experiment group and the matched group of (2nd) part, in feeding process, according to plant's managing system Degree, record often organize milch cow milk yield, search for food, feces, health status.

3. after experiment starts, within the 30th, 60 days, take milk sample respectively, measure local flavor content of material in Lac Bovis seu Bubali.

(4) local flavor substance-measuring method in Lac Bovis seu Bubali

INSTRUMENT MODEL: Thermo ISQ 1301；

GC conditions: HP-5 (30 × 0.32mm × 0.5um) capillary column；

Temperature programming: initial temperature is 40 DEG C, is incubated 4min, then rises to 100 DEG C with 5 DEG C/min, retains 2min, Rise to 220 DEG C with 10 DEG C/min again, retain 5min injector temperature 250 DEG C, detector temperature 280 DEG C, carrier gas For high pure nitrogen.

Sample treatment: use the volatile ingredient in solid-phase microextraction (SPME) technology separation Lac Bovis seu Bubali, take Lac Bovis seu Bubali sample Product 10g, places in hermetic container, adds 3g NaCl, 40 DEG C of heating 40min, sampling, uses gas chromatography mass spectrometry chromatograph Measure volatile ingredient.

(5) measurement result

Flavor substance determination data during feed 1 month is shown in Table 2 respectively, the GC-MS figure that test group flavor substance measures Spectrum is shown in Fig. 1, and Fig. 2 is shown in by the GC-MS collection of illustrative plates that matched group flavor substance measures, and determination data during feed 2 months is shown in Table 3.

Flavor substance measurement result during table 2 feed 1 month

Flavor substance measurement result during table 3 feed 2 months

Result above is found out:

(1), during feed 1 month, 22 flavor substance characteristic peaks are detected from test group milk sample, and from matched group Milk sample only detects 10 flavor substance characteristic peaks.Flavor substance total amount in test group milk sample is significantly larger than It it is matched group.

(2), during feed 2 months, 28 flavor substance characteristic peaks are detected from test group milk sample, and from matched group Milk sample only detects 13 flavor substance characteristic peaks.Flavor substance total amount in test group milk sample is significantly larger than It it is matched group.

Twice measurement result shows, the flavor substance of control sample detects about 10-13, test group flavor substances Quality inspection measures 22-28, and the flavor substance of test group is more than matched group, is more than 2 times of matched group.Test group contains Amount is more than matched group several times.

Illustrating that the flavor substance in test group Lac Bovis seu Bubali is more than matched group, Lac Bovis seu Bubali local flavor is better than matched group.

Embodiment 3 mortierella Diding culture (the glad health of Cordyceps) is on vitamin in Lac Bovis seu Bubali and the impact of essential amino acids

(1) experimental animal: test group and matched group 10 cow heads respectively, parity is consistent with milk yield.

(2) scale of feeding

Test group: every adult dairy cattle day raises 10kg concentrated feed, adds the mortierella Diding solid training of 0.3% in concentrated feed Support thing (the glad health of Cordyceps)；

Matched group: every adult dairy cattle day raises 10kg concentrated feed, without the glad health of Cordyceps in concentrated feed；

Day concentrated feed formula is: Semen Maydis 52%, Testa Tritici 5%, skin of Semen Maydis 5%, bean cake 8%, cottonseed meal 12%, dish The seed dregs of rice 8%, DDGS3%, stone powder 2%, magnesium oxide 0.3%, carbamide 1%, Sal 0.7%, sodium bicarbonate 1%, phosphorus Acid hydrogen calcium 1%, premix material 1%.

(3) test procedure

1., before starting to feed, observe lactating cow 3 days, record its food-intake, milk yield；

2. according to content feeding experiment group and the matched group of (2nd) part, in feeding process, according to plant's managing system Degree, record often organize milch cow milk yield, search for food, feces, health status.

3. within 30,60 days, remove milk sample respectively in experiment beginning, measure vitamin and essential amino acids content in Lac Bovis seu Bubali.

(4) assay method of vitamin A, E

1. chromatograph reference conditions:

Chromatographic column: C18 post, 250mm × 4.6mm, 5 μm, or have the chromatographic column of equal performance.

Flowing phase: methanol.

Flow velocity: 1.0mL/min.

Detection wavelength: vitamin A: 325nm；Vitamin E: 294nm.

Column temperature: 35 DEG C ± 1 DEG C.

Sample size: 20 μ L.

2. vitamin A, the drafting of E standard curve

The most accurately draw vitamin A standard reserving solution 0.50mL, 1.00mL, 1.50mL, 2.00mL, 2.50mL In 50mL brown volumetric flask, it is settled to scale mixing with ethanol.This standard series working solution concentration is respectively 1.00 μg/mL、2.00μg/mL、3.00μg/mL、4.00μg/mL、5.00μg/mL。

The most accurately draw vitamin E standard reserving solution 1.00mL, 2.00mL, 3.00mL, 4.00mL, 5.00mL In 50mL brown volumetric flask, it is settled to scale mixing with ethanol.This standard series working solution concentration is respectively 10.0 μg/mL、20.0μg/mL、30.0μg/mL、40.0μg/mL、50.0μg/mL。

Respectively vitamin A, E standard working solution are injected in chromatograph of liquid, obtain peak height (or peak area).With peak height (or peak area) is vertical coordinate, with vitamin A, E standard working solution concentration as abscissa, draw respectively vitamin A, E standard curve.

3. vitamin A, the mensuration of E sample

Test solution (A pipe) is injected in chromatograph of liquid, obtains peak height (or peak area), obtain according to respective standard curve The concentration of vitamin A, E in solution to be measured.

(5) mensuration of vitamin D

1. the purification of vitamin D liquid to be measured

Chromatograph reference conditions:

Chromatographic column: silicagel column, 150mm × 4.6mm, or have the chromatographic column of equal performance.

Flowing phase: hexamethylene mixes with normal hexane 1:1 by volume, and by volume mark 0.8% adds isopropanol.

Flow velocity: 1mL/min.

Wavelength: 264nm.

Column temperature: 35 DEG C ± 1 DEG C.

Sampling volume: 500 μ L.

Purification method: take about 0.5mL vitamin D standard reserving solution and fill in test tube in 10mL tool, at 40 DEG C ± 2 DEG C Nitrogen evaporator on dry up.Residue 5mL normal hexane vibration is dissolved.Take this solution 50 μ L and inject survey in chromatograph of liquid Fixed, determine vitamin D retention time.

Then 500 μ L liquid to be measured (B pipe) are injected in chromatograph of liquid, when retaining according to vitamin D standard solution Between collect vitamin D fraction in test tube C.Test tube C is placed in the Nitrogen evaporator under the conditions of 40 DEG C ± 2 DEG C and dries up, Taking out and accurately add 1.0mL methanol, residue vibration is dissolved, and is vitamin D and measures liquid.

2. vitamin D measures the mensuration of liquid

Reference color spectral condition:

Chromatographic column: C18 post, 250mm × 4.6mm, 5 μm, or have the chromatographic column of equal performance.

Flowing phase: methanol (4.7).

Flow velocity: 1mL/min.

Detection wavelength: 264nm.

Column temperature: 35 DEG C ± 1 DEG C.

Sample size: 100 μ L.

3. the drafting of standard curve

The most accurately draw vitamin D₂(or D₃) standard reserving solution 0.20mL, 0.40mL, 0.60mL, 0.80mL, 1.00mL, in 100 brown volumetric flasks, is settled to scale mixing with ethanol.This standard series working solution concentration is respectively Be 0.200 μ g/mL, 0.400 μ g/mL, 0.600 μ g/mL, 0.800 μ g/mL, 1.000 μ g/mL.

Respectively by vitamin D₂(or D₃) standard working solution injects in chromatograph of liquid, obtains peak height (or peak area). With peak height (or peak area) as vertical coordinate, with vitamin D₂(or D₃) standard working solution concentration is that abscissa is drawn respectively Standard curve.

4. the mensuration of vitamin D sample

Draw vitamin D to measure in liquid (C pipe) 100 μ L injection chromatograph of liquid, obtain peak height (or peak area), Obtain vitamin D according to standard curve and measure vitamin D in liquid₂(or D₃) concentration.

Vitamin D determination of recovery rates result is designated as response rate correction factor f, substitutes into measurement result computing formula (2), Vitamin D content measurement result is corrected.

(6) statement of analysis result

1. the calculating of vitamin A content

Vitamin A content is calculated by (1):

$X_A=\frac{\mathrm{Cs}\×10/2\×5\×100}{m}...\left(1\right);$

In formula:

X_A: the content of vitamin A in sample, unit is the every hectogram of microgram (μ g/100g)；

Cs: from the concentration of the vitamin A liquid to be measured that standard curve obtains, unit is micrograms per millilitre (μ g/mL)；

The quality of m: sample, unit is gram (g).

Note: 1 μ g retinol=3.33IU vitamin A.

The arithmetic mean of instantaneous value of twice independent measurement result to obtain under the conditions of repeatability represents, result retains three effectively Numeral.

2. the calculating of vitamin D content

Vitamin D content is calculated by (2):

$X_D=\frac{\mathrm{Cs}\×10/7\×2\×2\×100}{m\×f}...\left(2\right);$

In formula:

X_D: vitamin D in sample₂(or D₃) content, unit is the every hectogram of microgram (μ g/100g)；

Cs: the vitamin D obtained from graticule₂(or D₃) concentration of liquid to be measured, unit is micrograms per millilitre (μ g/mL)；

The quality of m: sample, unit is gram (g)；

F: response rate correction factor.

Note: in sample, the content of vitamin D is with vitamin D₂And D₃Content summation meter.

The arithmetic mean of instantaneous value of twice independent measurement result to obtain under the conditions of repeatability represents, result retains three effectively Numeral.

3. the calculating of content of vitamin E

Content of vitamin E is calculated by (3):

$X_E=\frac{\mathrm{Cs}\×10/2\×5\×100}{m\×1000}...\left(3\right);$

In formula:

X_E: the content of vitamin E (alpha-tocopherol) in sample, unit is the every hectogram of milligram (mg/100g)；

Cs: from the concentration of the vitamin E liquid to be measured that standard curve obtains, unit is micrograms per millilitre (μ g/mL)；

The quality of m: sample, unit is gram (g).

The arithmetic mean of instantaneous value of twice independent measurement result to obtain under the conditions of repeatability represents, result retains three effectively Numeral.

(7) in Lac Bovis seu Bubali, essential amino acids measures

1. sample collecting

Gather dairy cow milk, with refrigerated centrifuger at 4 DEG C, 3000r min^-1Centrifugal 20min, discard upper strata fat and Precipitation at the bottom of centrifuge tube, takes middle milk surum and carries out aminoacid detection.

2. instrument

High performance liquid chromatograph (waters company), C18 chromatographic column: venusil-AA analytical column (4.6mm × 250mm ×5μm)。

3. 17 kinds of aminoacid of acid hydrolysis detection

Sample treatment: accurately measure 200 μ L milk samples in long-neck test tube, adds 6mol L^-1Concentrated hydrochloric acid 10mLmL, alcohol blast burner calcination is sealed.Sample sealing is put in baking oven (110 DEG C) 24h, test tube mouth is broken, fall Go out hydrolyzed solution in beaker, adjust between pH to 8～9 with sodium hydroxide.Then move in 50mLmL volumetric flask constant volume, The abundant rinse of beaker.Constant volume liquid is filtered to conical flask, with 5mLmL needle tubing sucking-off filtrate, uses microporous filter membrane mistake Filter to 2mLmL tubule.

Derivatization: take 200 μ L sample and put in 1mL sample cell, adds after 100 μ L triethylamines-second eyeball solution shakes up Static, add 100 μ L phenyl isothiocyanates-nitrile sealed membrane and seal standing and reacting 1h, add 400 μ L normal hexane, Static 10min after mixing, draws lower floor solution 200 μ L in sample bottle, adds 800 μ L diluents after layering, mixed Static 5min after even.After instrument balances, take 40 μ L sample introduction analyses, take standard substance 5 μ L sample introduction simultaneously.

The BTI protection reaction of glutamine: take 100 μ L-Alas-glutamine dipeptide standard substance or sample solution, Add the BTI-acetonitrile solution that 100 μ L newly prepare, and add 25 μ L give a tongue-lashing pyridine aqueous solution (50mmol L^-1), mixing Nitrogen charging after uniformly, 50 DEG C of reaction 4h, room temperature dries up standby.

Hydrolysis: add the 6mol L of 200 μ L in the sample that BTI processed^-1Concentrated hydrochloric acid is in pipe, after high temperature tube sealing, 110 DEG C of hydrolysis 24h, cut tube sealing mouth, and room temperature under nitrogen dries up.

Isothiocyanic acid benzene fat (PITC) derives: the sour water desampling acetonitrile of 100 μ L-pyridine-triethylamine-water (10:5:2:3) delays Rushing liquid to dissolve, nitrogen adds the same buffer of 250 μ L, and adds 20 μ L isothiocyanic acid benzene fat, 50 DEG C of reactions after drying up After 1h, add normal hexane 200 μ L, mixing, stratification, draw 200 μ L from lower floor and move into 3mL sample cell, Adding 2800 μ L diluents, be i.e. available on the machine after mixing analysis.

(8) measurement result

1. the measurement result on vitamin content impact, the results are shown in Table 4,5.

Vitamin content measurement result in milk sample during table 4 feed 1 month

Sample	Vitamin A (μ g/kg)	Vitamin E (μ g/kg)	Vitamin D (μ g/mL)
				Test group milk sample	25	260	Sample peak detected
Matched group milk sample	20	150	0

From table 4, compared with matched group, the vitamin A content in test group milch cow sample improves 25%, and dimension is raw Element E content improves 73.3%, and in matched group and test group milch cow sample, vitamin D content is the most relatively low, but from matched group Measuring less than vitamin D in milch cow sample, and can be measured to vitamin D from test group milch cow sample, vitamin D contains Measuring near detection limit, in prompting test group sample, the content existence of vitamin D significantly improves.

Vitamin content measurement result in milk sample during table 5 feed 2 months

Sample	Vitamin A (μ g/kg)	Vitamin E (μ g/kg)	Vitamin D (μ g/mL)
				Test group milk sample	26	245	Sample peak detected
Matched group milk sample	19	167	0

From table 5, compared with matched group, the vitamin A content in test group milch cow sample improves 36.8%, dimension Raw element E content improves 46.7%, and in matched group and test group milch cow sample, vitamin D content is the most relatively low, but from comparison Group milch cow sample measures less than vitamin D, and vitamin D can be measured to from test group milch cow sample, vitamin D Content is near detection limit, and in prompting test group sample, the content existence of vitamin D significantly improves.

In sum, the glad health of Cordyceps can effectively change the local flavor of Lac Bovis seu Bubali, improves vitamin A, the content of E, D.

2. the measurement result on essential amino acids content impact

Test group essential amino acids measures collection of illustrative plates and sees Fig. 3, and matched group essential amino acids measures collection of illustrative plates and sees Fig. 4, feed 1 month Time essential amino acids content measurement result be shown in Table 6, during feed 1 month, the amino acid whose assay of local flavor the results are shown in Table 7.

During table 6 feed 1 month, the glad health of Cordyceps is on the impact of essential amino acids level in Lac Bovis seu Bubali

From table 6, comparing with matched group, in test group milk sample, TEAA content is improved largely, Comparing with matched group, in test group milk sample, total amino acids content improves 12.96%.

During table 7 feed 1 month, the glad health of Cordyceps is on the impact of local flavor amino acid levels in Lac Bovis seu Bubali

From table 7, in test group milk sample, local flavor amino acid content is improved largely, and compares with matched group, In test group milk sample, local flavor amino acid content improves 41.87%.

Claims (6)

1. Cordyceps strain fermentation thing contains at preparation raising Lac Bovis seu Bubali flavor substance content, vitamin content or essential amino acids Application in amount feedstuff, it is characterised in that including: described Cordyceps strain fermentation thing is fed lactating cow, obtains Lac Bovis seu Bubali；

Described Cordyceps strain fermentation thing is obtained through liquid fermentation or solid fermentation by Cordyceps sinensis fungus；

Described Cordyceps sinensis fungus is Cordyceps gunnii (Berk.) Berk. (Cordyceps.gunnii) strain, Cordyceps (C.sinensis) bacterium Kind, Cordyceps militaris (L.) Link. (C.militaris) strain, Cordyceps hawkesii Gary (C.hawkesii) strain, Xinjiang Cordyceps (C.gracilis) At least one in strain, agglomerate Cordyceps (C.ophioglossoides) strain or Periostracum cicadae (C.sobolifera) strain；

The culture medium of described liquid fermentation is: sucrose, peptone, yeast extract powder, Semen Maydis powder, MgS0₄·7H₂O、 K₂HP0₄；

The culture medium of described solid fermentation is: potato starch, Semen Maydis powder, white sugar, peptone, dried silkworm chrysalis meal, sulphuric acid Magnesium, ammonium citrate, potassium dihydrogen phosphate, vitamin B₁。

Apply the most as claimed in claim 1, it is characterised in that described flavor substance include following (1), (2) or (3) at least one in:

(1) in GC-MS collection of illustrative plates, retention time is the rudimentary alkanoic acid of 8～15min；

(2) in GC-MS collection of illustrative plates, retention time is alkyl acid, alkyl aldehydes or the alkyl ketone thing of 15～20min Matter；

(3) in GC-MS collection of illustrative plates, retention time be 20～40min higher alkane, higher alkene, senior alkane Base aldehyde, senior alkyl amide or higher alkanoic acid and Ester thereof.

Apply the most as claimed in claim 1, it is characterised in that described vitamin is VitAVitE Or at least one in vitamin D.

Apply the most as claimed in claim 1, it is characterised in that described essential amino acids be aspartic acid, glycine, At least one in alanine, cysteine, valine, leucine, phenylalanine, lysine or histidine.

Apply the most as claimed in claim 1, it is characterised in that in terms of dry weight, described Cordyceps strain fermentation thing Scale of feeding is at least 5g/ head/sky.

6. prepare flavor substance, vitamin or a method for essential amino acids content feedstuff, its feature in raising Lac Bovis seu Bubali It is, including: Cordyceps strain fermentation thing is fed lactating cow, obtains Lac Bovis seu Bubali；

Described Cordyceps strain fermentation thing is obtained through liquid fermentation or solid fermentation by Cordyceps sinensis fungus；

Described Cordyceps sinensis fungus is Cordyceps gunnii (Berk.) Berk. (Cordyceps.gunnii) strain, Cordyceps (C.sinensis) bacterium Kind, Cordyceps militaris (L.) Link. (C.militaris) strain, Cordyceps hawkesii Gary (C.hawkesii) strain, Xinjiang Cordyceps (C.gracilis) At least one in strain, agglomerate Cordyceps (C.ophioglossoides) strain or Periostracum cicadae (C.sobolifera) strain；

The culture medium of described liquid fermentation is: sucrose, peptone, yeast extract powder, Semen Maydis powder, MgS0₄·7H₂O、 K₂HP0₄；

The culture medium of described solid fermentation is: potato starch, Semen Maydis powder, white sugar, peptone, dried silkworm chrysalis meal, sulphuric acid Magnesium, ammonium citrate, potassium dihydrogen phosphate, vitamin B₁。