CN102660520B - Fermentative nutrition aid, application of nutrition aid to preparation of xylanase and application method for nutrition aid - Google Patents
Fermentative nutrition aid, application of nutrition aid to preparation of xylanase and application method for nutrition aid Download PDFInfo
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Abstract
The invention discloses a fermentative nutrition aid, application of the nutrition aid to the preparation of xylanase and an application method for the nutrition aid. Each liter of fermentative nutrition aid comprises the following nutritional ingredients: 80 to 120 g of betaine, 0.1 to 0.4 g of vitamin A, 0.1 to 0.4 g of vitamin B1, 0.05 to 0.15 g of vitamin B6, 0.05 to 0.15 g of vitamin B12, 0.1 to 0.3 g of vitamin D3, 0.1 to 0.6 g of vitamin E, 0.1 to 0.3 g of vitamin K3 and 0.5 to 0.8 g of nicotinic acid. The xylanase is prepared by the steps of adding 300 to 500 g of fermentative nutrition aid into a fermentation culture medium based on each Kg of solid matrix, and performing fermentation culture for 4 to 5 days. The fermentative nutrition aid, which is used for the fermentation culture of the xylanase, can effectively promote the growth of thalli of an Aspergillus niger-spore suspension and the metabolism of the xylanase, remarkably improves a fermentation effect and the yield of the xylanase, and shortens fermentation time.
Description
Technical field
The present invention relates to fermentation technical field, relate in particular to a kind of Nutrious fermented auxiliary agent and fermentation process be used to improving solid-state zytase fermentation yield.
Background technology
Xylan is the important component of hemicellulose in plant cell wall, accounts for 1/3rd of plant carbohydrates total amount, is the abundant polysaccharide of content second after Mierocrystalline cellulose, and its main chain is to be connected with the wood sugar glycosidic bond by a plurality of β-D-xylopyranosyl to form.
The zytase of broad sense refers to the general name of one group of enzyme that can the degradation of hemicellulose xylan.Usually said zytase refers to inscribe β-Isosorbide-5-Nitrae-D zytase, is responsible for the degraded of xylan skeleton carbochain.Xylan is after xylanase treatment, and Partial digestion can obtain xylo-oligosaccharide, degradable obtaining take wood sugar as five main carbon monose.
There is a large amount of hemicelluloses in plant feed.Non-ruminant animal almost can not digest hemicellulose owing to lacking corresponding digestive enzymes.The indigested hemicellulose that exists in animal intestinal can make the viscosity of food increase, and hinders digesting and assimilating of the nutritive substances such as fat and protein, has reduced the utilization ratio of feed.If add zytase in feed, just can improve the utilization ratio of feed by the xylan in the degraded feed, improve the immunizing power of body, improve the production performance of animal.
Tradition papermaking bleaching process is to adopt the acid bleaching agent such as chlorine and derivative thereof to remove xylogen, but contains the strong carcinogenic poisonous organic chloride of teratogenesis such as a large amount of Dioxins in this paper waste that can cause discharging, and causes serious environmental pollution.If before chemical bleaching, paper pulp is carried out pre-treatment with zytase, can accelerate the release of xylogen in paper pulp, the SYNTHETIC OPTICAL WHITNER such as promotion chlorine more effectively act on, thereby reduce the usage quantity of harmful chemical SYNTHETIC OPTICAL WHITNER, and have shortened the time of pulp processing.
Skin at liquor-making raw material has Mierocrystalline cellulose and hemicellulose layer, and this can affect the utilization of raw material internal starch.By the effect of double cellulose layer of zytase, be conducive to amylase to the degraded of raw material internal starch, improve the utilization ratio of starch, increase the productive rate of alcohol.In addition, in the beer brewing process, due to the impact of the xylan in raw materials for production and dextran, can cause the wort filtration difficulty, beer is muddy, the problems such as beer filter membrane obstruction.If add zytase in the process of preparation wort, can make xylan degrading and destroy plant cell wall, thereby effectively discharge lytic enzyme and the entocytes such as proteolytic enzyme, amylase, saccharification time is shortened, the viscosity of wort is reduced, improve filtration velocity and the yield of wheat juice.
In sum, zytase has all demonstrated huge application potential at feedstuff industry, pulping and paper-making, liquor industry etc.China was studied zytase since the eighties in last century, but up to the present, because the output of zytase is lower, caused the production cost of zytase still higher, and deficiency in economic performance has seriously restricted its application in above industrial circle.
Summary of the invention
First technical problem to be solved by this invention is: for the deficiency that prior art exists, provide a kind of Nutrious fermented auxiliary agent that can significantly improve ferment effect in being applied to ferment.
Second technical problem to be solved by this invention is: for the deficiency that prior art exists, provide the application of a kind of Nutrious fermented auxiliary agent in the zytase preparation.
The 3rd technical problem to be solved by this invention is: for the deficiency that prior art exists, provide in a kind of fermentation that Nutrious fermented auxiliary agent is applied to zytase, can significantly improve the method for zytase output.
For solving above-mentioned first technical problem, technical scheme of the present invention is:
A kind of Nutrious fermented auxiliary agent comprises following nutritive ingredient (g/L): trimethyl-glycine 80～120, vitamin A 0.1～0.4, vitamins B
10.1～0.4, vitamins B
60.05～0.15, vitamins B
120.05～0.15, vitamins D
30.1～0.3, vitamin-E 0.1～0.6, vitamin K
30.1～0.3 and nicotinic acid 0.5～0.8.
As a kind of preferred, described trimethyl-glycine is one or more the mixture in BETAINE anhydrous, a water trimethyl-glycine, hydrochloride trimethyl-glycine, sulphonic acid betaine, phosphoric acid salt trimethyl-glycine.
For solving above-mentioned second technical problem, technical scheme of the present invention is:
The application of Nutrious fermented auxiliary agent in the zytase preparation joins the addition of described Nutrious fermented auxiliary agent with 300～500g/Kg solid substrate in fermention medium.
For solving above-mentioned the 3rd technical problem, technical scheme of the present invention is:
The application method of a kind of Nutrious fermented auxiliary agent in fermentation preparation zytase comprises the following steps:
(1) seed culture: with seed culture medium 121 ℃ of sterilizations 20 minutes, be cooled to 28～32 ℃ after access aspergillus niger spore suspension, cultivated under agitation condition 2～3 days.
(2) preparation fermention medium: be the ratio preparation solid substrate of 50～70wt% vinasse and 30～50wt% wheat bran according to butt, growth limiting factor is added in described solid substrate after with a small amount of water dissolution, 121 ℃ of sterilizations 20 minutes, relatively every 1Kg solid substrate adds the Nutrious fermented auxiliary agent of 300～500g again, and then replenishing sterilized water to fermention medium water content is 35～50wt%.
(3) fermentation culture: the seed liquor that step (1) is obtained, inoculate in the fermention medium that step (2) obtains according to the inoculum size of 100～200mL/Kg solid substrate, be 28～32 ℃ of beginning fermentation culture in temperature, 3.5～4.5 days total times of fermentation.
Wherein, described Nutrious fermented auxiliary agent comprises following nutritive ingredient (g/L): trimethyl-glycine 80～120, vitamin A 0.1～0.4, vitamins B
10.1～0.4, vitamins B
60.05～0.15, vitamins B
120.05～0.15, vitamins D
30.1～0.3, vitamin-E 0.1～0.6, vitamin K
30.1～0.3 and nicotinic acid 0.5～0.8.
Preferably, described trimethyl-glycine is one or more the mixture in BETAINE anhydrous, a water trimethyl-glycine, hydrochloride trimethyl-glycine, sulphonic acid betaine, phosphoric acid salt trimethyl-glycine.
Preferably, the formula of described seed culture medium is glucose 45～55g/L, peptone 8～12g/L, yeast extract paste 12～18g/L, KH
2PO
41.5～3g/L and MgSO
47H
2O 0.5～2g/L.
As further preferred, the formula of described seed culture medium is glucose 50g/L, peptone 10g/L, yeast extract paste 15g/L, KH
2PO
42g/L and MgSO
47H
2O1g/L.
Preferably, add in every 1Kg solid substrate and comprise Dried Corn Steep Liquor Powder 25～40g, MgSO
47H
2O0.5～1.0g, CuSO
45H
2O 1.0～2.0g, MnSO
42H
2O 0.5～1.5g and FeSO
47H
2The growth limiting factor of O 0.05～1.5g.
As a kind of improvement, during described fermentation culture, to 36～48h, again add Nutrious fermented auxiliary agent according to the addition of 300～500g/Kg solid substrate in fermentation, and the loose fermention medium that shakes up of button.
Owing to having adopted technique scheme, the invention has the beneficial effects as follows:
1, in Nutrious fermented auxiliary agent of the present invention, contain trimethyl-glycine and multivitamin, trimethyl-glycine is as a kind of alkaloid, the methyl of providing is provided, promote the effect of metabolism of fat and protein synthesis, in addition, trimethyl-glycine can be regulated born of the same parents' inner penetration and press, improve the permeability of cell, promote the outer of enzyme to secrete, and the indispensable factor of the VITAMIN normal function that is microorganism and Metabolic activity, participate in the regulation and control of a lot of microbial metabolisms as cofactor, it is a key link in microorganism metabolism controlling, the present invention is by adopting rational nutritive ingredient formula, change the size of certain enzyme vigor by the VITAMIN of adding different levels, thereby redistribute the carbon skeleton material in the intracellular flow direction, can effectively promote especially xylan Enzyme Production of enzyme.
2, the present invention adds Nutrious fermented auxiliary agent in fermention medium when the fermentation culture of zytase, can effectively promote the growth of aspergillus niger spore suspension thalline and the metabolism of zytase, significantly improved the output of ferment effect and zytase, shorten fermentation time, reduced production cost.
3, the present invention's main component take industrial waste vinasse, wheat bran as fermention medium when the fermention medium of preparation zytase has reduced the cost of fermentation, has reduced simultaneously the pollution of vinasse to environment.
4, the present invention's auxiliary agent that in good time supplements the nutrients during the fermentation, the moisture of wall losses and produce the required nutritive substance of enzyme before afterfermentation that can be suitable for the production by biological enzyme provides suitable environment and enough nutrition, has further guaranteed ferment effect.
Embodiment
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.Should be understood that in addition those skilled in the art can make various changes or modifications the present invention after the content of having read the present invention's instruction, these equivalent form of values fall within the application's appended claims limited range equally.
Embodiment 1
After each nutritive ingredient water was fully dissolved, the micro-filtration degerming was prepared into the Nutrious fermented auxiliary agent of following content: BETAINE anhydrous 80g/L, vitamin A 0.1g/L, vitamins B
10.4g/L, vitamins B
60.05g/L, vitamins B
120.15g/L, vitamins D
30.1g/L, vitamin-E 0.6g/L, vitamin K
30.1g/L and nicotinic acid 0.8g/L.
Embodiment 2
After each nutritive ingredient water was fully dissolved, the micro-filtration degerming was prepared into the Nutrious fermented auxiliary agent of following content: a water trimethyl-glycine 120g/L, vitamin A 0.4g/L, vitamins B
10.1g/L, vitamins B
60.15g/L, vitamins B
120.05g/L, vitamins D
30.3g/L, vitamin-E 0.1g/L, vitamin K
30.3g/L and nicotinic acid 0.5g/L.
Embodiment 3
After each nutritive ingredient water was fully dissolved, the micro-filtration degerming was prepared into the Nutrious fermented auxiliary agent of following content: hydrochloride trimethyl-glycine 100g/L, vitamin A 0.2g/L, vitamins B
10.3g/L, vitamins B
60.10g/L, vitamins B
120.10g/L, vitamins D
30.2g/L, vitamin-E 0.4g/L, vitamin K
30.2g/L and nicotinic acid 0.6g/L.
Embodiment 4
After each nutritive ingredient water was fully dissolved, the micro-filtration degerming was prepared into the Nutrious fermented auxiliary agent of following content: sulphonic acid betaine 100g/L, vitamin A 0.3g/L, vitamins B
10.25g/L, vitamins B
60.09g/L, vitamins B
120.09g/L, vitamins D
30.25g/L, vitamin-E 0.3g/L, vitamin K
30.25g/L and nicotinic acid 0.7g/L.
Embodiment 5
After each nutritive ingredient water was fully dissolved, the micro-filtration degerming was prepared into the Nutrious fermented auxiliary agent of following content: the mixture 100g/L of hydrochloride trimethyl-glycine and phosphoric acid salt trimethyl-glycine, vitamin A 0.25g/L, vitamins B
10.3g/L, vitamins B
60.10g/L, vitamins B
120.10g/L, vitamins D
30.25g/L, vitamin-E 0.35g/L, vitamin K
30.25g/L and nicotinic acid 0.65g/L.
Embodiment 6
(1) preparation seed culture medium: glucose 45g/L, peptone 12g/L, yeast extract paste 12g/L, KH
2PO
43g/L and MgSO
47H
2O 0.5g/L.Seed culture medium was sterilized 20 minutes at 121 ℃, be cooled to 28 ℃, add the 5ml sterilized water in aspergillus niger spore suspension slant strains, scrape gently media surface with inoculating needle, the bacteria suspension of making is moved into access aspergillus niger spore suspension in liquid seed culture medium, cultivated 3 days under agitation condition.
(2) preparation fermention medium: be the proportioning preparation solid substrate of 550g vinasse and 450g wheat bran according to butt, will comprise Dried Corn Steep Liquor Powder 30g, MgSO
47H
2O 0.8g, CuSO
45H
2O 1.5g, MnSO
42H
2O1.5g and FeSO
47H
2The growth limiting factor of O 1.5g adds in solid substrate after with a small amount of water dissolution, 121 ℃ of sterilizations 20 minutes, the more Nutrious fermented auxiliary agent 400g that adds embodiment 1 to obtain, then replenishing sterilized water to fermention medium water content is 45wt%.
(3) fermentation culture: get the seed liquor 150mL that step (1) obtains, inoculation enters in the fermention medium that step (2) obtains, be 28 ℃ of beginning fermentation culture in temperature, fermenting to 36h, the Nutrious fermented auxiliary agent 400g that again adds embodiment 1 to obtain, and the loose fermention medium that shakes up of button, 4.5 days total times of fermentation.
Embodiment 7
(1) preparation seed culture medium: glucose 55g/L, peptone 8g/L, yeast extract paste 18g/L, KH
2PO
41.5g/L and MgSO
47H
2O 2g/L.Seed culture medium was sterilized 20 minutes at 121 ℃, be cooled to 32 ℃, add the 10ml sterilized water in aspergillus niger spore suspension slant strains, scrape gently media surface with inoculating needle, the bacteria suspension of making is moved into access aspergillus niger spore suspension in liquid seed culture medium, cultivated 2 days under agitation condition.
(2) preparation fermention medium: be the proportioning preparation solid substrate of 500g vinasse and 500g wheat bran according to butt, will comprise Dried Corn Steep Liquor Powder 40g, MgSO
47H
2O 0.7g, CuSO
45H
2O 1.8g, MnSO
42H
2O1.3g and FeSO
47H
2The growth limiting factor of O 1.0g adds in solid substrate after with a small amount of water dissolution, 121 ℃ of sterilizations 20 minutes, the more Nutrious fermented auxiliary agent 450g that adds embodiment 2 to obtain, then replenishing sterilized water to fermention medium water content is 50wt%.
(3) fermentation culture: get seed liquor 180mL that step (1) obtains and inoculate in the fermention medium that step (2) obtains, be 32 ℃ of beginning fermentation culture in temperature, fermenting to 48h, the Nutrious fermented auxiliary agent 450g that again adds embodiment 2 to obtain, and the loose fermention medium that shakes up of button, 4 days total times of fermentation.
Embodiment 8
(1) preparation seed culture medium: glucose 50g/L, peptone 10g/L, yeast extract paste 15g/L, KH
2PO
42g/L and MgSO
47H
2O 1g/L.Seed culture medium was sterilized 20 minutes at 121 ℃, be cooled to 30 ℃, add the 8ml sterilized water in aspergillus niger spore suspension slant strains, scrape gently media surface with inoculating needle, the bacteria suspension of making is moved into access aspergillus niger spore suspension in liquid seed culture medium, cultivated 2.5 days under agitation condition.
(2) preparation fermention medium: be the proportioning preparation solid substrate of 600g vinasse and 400g wheat bran according to butt, will comprise Dried Corn Steep Liquor Powder 25g, MgSO
47H
2O 0.5g, CuSO
45H
2O 1.3g, MnSO
42H
2O0.5g and FeSO
47H
2The growth limiting factor of O 0.7g adds in solid substrate after with a small amount of water dissolution, 121 ℃ of sterilizations 20 minutes, the more Nutrious fermented auxiliary agent 300g that adds embodiment 3 to obtain, then replenishing sterilized water to fermention medium water content is 48wt%.
(3) fermentation culture: get the seed liquor 100mL that step (1) obtains, inoculation enters in the fermention medium that step (2) obtains, be 30 ℃ of beginning fermentation culture in temperature, fermenting to 40h, the Nutrious fermented auxiliary agent 300g that again adds embodiment 3 to obtain, and the loose fermention medium that shakes up of button, 4 days total times of fermentation.
Embodiment 9
(1) seed culture medium was sterilized 20 minutes at 121 ℃, be cooled to 30 ℃, add the 5ml sterilized water in aspergillus niger spore suspension slant strains, scrape gently media surface with inoculating needle, the bacteria suspension of making is moved into access aspergillus niger spore suspension in liquid seed culture medium, cultivated 3 days under agitation condition.
(2) preparation fermention medium: be the proportioning preparation solid substrate of 700g vinasse and 300g wheat bran according to butt, will comprise Dried Corn Steep Liquor Powder 28g, MgSO
47H
2O 0.9g, CuSO
45H
2O 1.2g, MnSO
42H
2O0.9g and FeSO
47H
2The growth limiting factor of O 0.9g adds in solid substrate after with a small amount of water dissolution, 121 ℃ of sterilizations 20 minutes, the more Nutrious fermented auxiliary agent 480g that adds embodiment 4 to obtain, then replenishing sterilized water to fermention medium water content is 44wt%.
(3) fermentation culture: get the seed liquor 140mL that step (1) obtains, inoculation enters in the fermention medium that step (2) obtains, be 30 ℃ of beginning fermentation culture in temperature, fermenting to 42h, the Nutrious fermented auxiliary agent 480g that again adds embodiment 4 to obtain, and the loose fermention medium that shakes up of button, 4 days total times of fermentation.
Comparative Examples 1
(1) preparation seed culture medium: glucose 50g/L, peptone 10g/L, yeast extract paste 15g/L, KH
2PO
42g/L and MgSO
47H
2O 1g/L.Seed culture medium was sterilized 20 minutes at 121 ℃, be cooled to 30 ℃, add the 8ml sterilized water in aspergillus niger spore suspension slant strains, scrape gently media surface with inoculating needle, the bacteria suspension of making is moved into access aspergillus niger spore suspension in liquid seed culture medium, cultivated 2.5 days under agitation condition.
(2) preparation fermention medium: vinasse (in butt) 650g, wheat bran 350g.Dried Corn Steep Liquor Powder 30g, MgSO
47H
2O 0.9g, CuSO
45H
2O1.2g, MnSO
42H
2O 0.9g, FeSO
47H
2O 0.9g, above growth limiting factor add in solid substrate after with a small amount of water dissolution.Make up water to the whole water content of substratum is 45wt%.In 121 ℃ of sterilization 20min.
(3) fermentation culture: get the seed liquor 170mL that step (1) obtains, inoculation enters in the fermention medium that step (2) obtains, and is 30 ℃ of beginning fermentation culture in temperature, 5 days total times of fermentation.
Comparative Examples 2
(1) preparation seed culture medium: glucose 45g/L, peptone 12g/L, yeast extract paste 12g/L, KH
2PO
43g/L and MgSO
47H
2O 0.5g/L.Seed culture medium was sterilized 20 minutes at 121 ℃, be cooled to 28 ℃, add the 5ml sterilized water in aspergillus niger spore suspension slant strains, scrape gently media surface with inoculating needle, the bacteria suspension of making is moved into access aspergillus niger spore suspension in liquid seed culture medium, cultivated 3 days under agitation condition.
(2) preparation fermention medium: vinasse (in butt) 600g, wheat bran 400g.Dried Corn Steep Liquor Powder 35g, MgSO
47H
2O 1.0g, CuSO
45H
2O2.0g, MnSO
42H
2O 1.5g, FeSO
47H
2O0.9g, above growth limiting factor add in solid substrate after with a small amount of water dissolution.Replenishing sterilized water to the whole water content of substratum is 50wt%.In 121 ℃ of sterilization 20min.
(3) fermentation culture: get the seed liquor 180mL that step (1) obtains, inoculation enters in the fermention medium that step (2) obtains, and is 28 ℃ of beginning fermentation culture in temperature, 5.5 days total times of fermentation.
Comparative Examples 3
(1) preparation seed culture medium: glucose 50g/L, peptone 10g/L, yeast extract paste 15g/L, KH
2PO
42g/L and MgSO
47H
2O 1g/L.Seed culture medium was sterilized 20 minutes at 121 ℃, be cooled to 30 ℃, add the 8ml sterilized water in aspergillus niger spore suspension slant strains, scrape gently media surface with inoculating needle, the bacteria suspension of making is moved into access aspergillus niger spore suspension in liquid seed culture medium, cultivated 2.5 days under agitation condition.
(2) preparation fermention medium: vinasse (in butt) 650g, wheat bran 350g.Dried Corn Steep Liquor Powder 30g, MgSO
47H
2O 0.9g, CuSO
45H
2O1.2g, MnSO
42H
2O0.9g, FeSO
47H
2O 0.9g, above growth limiting factor add in solid substrate after with a small amount of water dissolution.Cultivation is based on 121 ℃ of sterilization 20min.The Nutrious fermented auxiliary agent 480g that adds embodiment 3 to obtain.Replenishing sterilized water to the whole water content of substratum is 50wt%.
(3) fermentation culture: get the seed liquor 170mL that step (1) obtains, inoculation enters in the fermention medium that step (2) obtains, and is 30 ℃ of beginning fermentation culture in temperature, 4.8 days total times of fermentation.
Comparative Examples 4
(1) preparation seed culture medium: glucose 45g/L, peptone 12g/L, yeast extract paste 12g/L, KH
2PO
43g/L and MgSO
47H
2O 0.5g/L.Seed culture medium was sterilized 15 minutes at 121 ℃, be cooled to 28 ℃, add the 5ml sterilized water in aspergillus niger spore suspension slant strains, scrape gently media surface with inoculating needle, the bacteria suspension of making is moved into access aspergillus niger spore suspension in liquid seed culture medium, cultivated 3 days under agitation condition.
(2) preparation fermention medium: vinasse (in butt) 600g, wheat bran 400g.Dried Corn Steep Liquor Powder 35g, MgSO
47H
2O 1.0g, CuSO
45H
2O2.0g, MnSO
42H
2O 1.5g, FeSO
47H
2O 0.9g, above growth limiting factor add in solid substrate after with a small amount of water dissolution.Make up water to the whole water content of substratum is 50wt%.Cultivation is based on 121 ℃ of sterilization 20min.
(3) fermentation culture: get the seed liquor 150mL that step (1) obtains, inoculation enters in the fermention medium that step (2) obtains, be 28 ℃ of beginning fermentation culture in temperature, fermenting to 40h, the Nutrious fermented auxiliary agent 500g that adds embodiment 3 to obtain, and the loose fermention medium that shakes up of button, 5.2 days total times of fermentation.
The enzyme situation of living of the zytase of fermentation time to the Comparative Examples 4 of embodiment 6 to embodiment 9, Comparative Examples 1, preparation sees Table 1.
Table 1
Produce the 1 needed enzyme amounts of μ mol wood sugar at 40 ℃ of lower per minute catalysis, 1% oat xylans and be defined as the enzyme unit (U) that lives.
As can be seen from Table 1, Comparative Examples 1 and Comparative Examples 2 fermentation times that do not add Nutrious fermented auxiliary agent obviously extend, fermentation yield obviously reduces; And Comparative Examples 3 and the Comparative Examples 4 of respectively adding the one time fermentation nutritional auxiliary respectively in the middle of fermentation beginning and fermentation, fermentation time is not short when adding, fermentation yield is high.
Claims (8)
1. a Nutrious fermented auxiliary agent, is characterized in that being comprised of following nutritive ingredient: trimethyl-glycine 80～120g/L, vitamin A 0.1～0.4g/L, vitamins B
10.1～0.4g/L, vitamins B
60.05～0.15g/L, vitamins B
120.05～0.15g/L, vitamins D
30.1～0.3g/L, vitamin-E 0.1～0.6g/L, vitamin K
30.1～0.3g/L and nicotinic acid 0.5～0.8g/L.
2. Nutrious fermented auxiliary agent as claimed in claim 1 is characterized in that: described trimethyl-glycine is one or more the mixture in BETAINE anhydrous, a water trimethyl-glycine, hydrochloride trimethyl-glycine, sulphonic acid betaine, phosphoric acid salt trimethyl-glycine.
3. the application of Nutrious fermented auxiliary agent as claimed in claim 1 in the zytase preparation is characterized in that: the addition of described Nutrious fermented auxiliary agent with 300～500g/Kg solid substrate joined in fermention medium.
4. a Nutrious fermented auxiliary agent as claimed in claim 1 at the application method that ferments in preparing zytase, is characterized in that comprising the following steps:
(1) seed culture: with seed culture medium 121 ℃ of sterilizations 20 minutes, be cooled to 28～32 ℃ after access aspergillus niger spore suspension, cultivated under agitation condition 2～3 days;
(2) preparation fermention medium: be the ratio preparation solid substrate of 50～70wt% vinasse and 30～50wt% wheat bran according to butt, growth limiting factor is added in described solid substrate after with a small amount of water dissolution, add by Dried Corn Steep Liquor Powder 25～40g, MgSO in every 1Kg solid substrate
47H
2O0.5～1.0g, CuSO
45H
2O1.0～2.0g, MnSO
42H
2O0.5～1.5g and FeSO
47H
2The growth limiting factor that O0.05～1.5g forms, 121 ℃ of sterilizations 20 minutes, relatively every 1Kg solid substrate adds the Nutrious fermented auxiliary agent of 300～500g again, then replenishing sterilized water to fermention medium water content is 35～50wt%, and described Nutrious fermented auxiliary agent is comprised of following nutritive ingredient: trimethyl-glycine 80～120g/L, vitamin A 0.1～0.4g/L, vitamins B
10.1～0.4g/L, vitamins B
60.05～0.15g/L, vitamins B
120.05～0.15g/L, vitamins D
30.1～0.3g/L, vitamin-E 0.1～0.6g/L, vitamin K
30.1～0.3g/L and nicotinic acid 0.5～0.8g/L;
(3) fermentation culture: the seed liquor that step (1) is obtained, inoculate in the fermention medium that step (2) obtains according to the inoculum size of 100～200mL/Kg solid substrate, be 28～32 ℃ of beginning fermentation culture in temperature, 3.5～4.5 days total times of fermentation.
5. the application method of Nutrious fermented auxiliary agent as claimed in claim 4 in fermentation preparation zytase is characterized in that: described trimethyl-glycine is one or more the mixture in BETAINE anhydrous, a water trimethyl-glycine, hydrochloride trimethyl-glycine, sulphonic acid betaine, phosphoric acid salt trimethyl-glycine.
6. Nutrious fermented auxiliary agent as claimed in claim 4 is at the application method that ferments in preparing zytase, and it is characterized in that: the formula of described seed culture medium is glucose 45～55g/L, peptone 8～12g/L, yeast extract paste 12～18g/L, KH
2PO
41.5～3g/L and MgSO
47H
2O0.5～2g/L.
7. Nutrious fermented auxiliary agent as claimed in claim 6 is at the application method that ferments in preparing zytase, and it is characterized in that: the formula of described seed culture medium is glucose 50g/L, peptone 10g/L, yeast extract paste 15g/L, KH
2PO
42g/L and MgSO
47H
2O1g/L.
8. any Nutrious fermented auxiliary agent as described in claim 4 to 7 is at the application method that ferments in preparing zytase, it is characterized in that: during described fermentation culture, fermenting to 36～48h, addition according to 300～500g/Kg solid substrate adds Nutrious fermented auxiliary agent as claimed in claim 1 again, and the loose fermention medium that shakes up of button.
Priority Applications (1)
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| CN105039271A (en) * | 2015-06-25 | 2015-11-11 | 山东祥维斯生物科技有限公司 | Method for increasing yield of various enzyme preparations |
| CN108440034A (en) * | 2018-05-08 | 2018-08-24 | 山东祥维斯生物科技股份有限公司 | Application of the glycine betaine in fermentation, the application of compound betaine auxiliary agent, microbial-bacterial fertilizer and the microbial-bacterial fertilizer based on the application |
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| CN101182502A (en) * | 2007-11-23 | 2008-05-21 | 河南天冠企业集团有限公司 | Fermentation production process of solid body acidic xylanase |
| CN101705262A (en) * | 2009-12-03 | 2010-05-12 | 天津科技大学 | New process for improving fermentation and acid production rate of glutamic acid temperature sensitive mutant by utilizing glycine betaine |
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| CN101182502A (en) * | 2007-11-23 | 2008-05-21 | 河南天冠企业集团有限公司 | Fermentation production process of solid body acidic xylanase |
| CN101705262A (en) * | 2009-12-03 | 2010-05-12 | 天津科技大学 | New process for improving fermentation and acid production rate of glutamic acid temperature sensitive mutant by utilizing glycine betaine |
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Effective date of registration: 20160513 Address after: 261061, No. 36, No. two, 211 hi tech Zone, Weifang hi tech Zone, Shandong, China Patentee after: Shandong Xiang Weiss Medical Technology Co., Ltd. Address before: 261061, No. 36, No. two, 206 hi tech Zone, Weifang hi tech Zone, Shandong, China Patentee before: Sunwin Chemicals Co., Ltd. |