CN109136285A - A kind of catechin bioconversion nutrient solution and a kind of method for promoting catechin bioconversion - Google Patents
A kind of catechin bioconversion nutrient solution and a kind of method for promoting catechin bioconversion Download PDFInfo
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Abstract
The invention discloses a kind of catechin bioconversion nutrient solution and a kind of method for promoting catechin bioconversion, the catechin bioconversion nutrient solution includes raw material containing catechin, nitrogen source compound and water；The raw material containing catechin is tea polyphenols or catechin；The mass percentage of the nitrogen source compound is 0.01% ~ 5%.Catechin bioconversion nutrient composition provided by the invention is simple, without other complicated inorganic salts, will not interfere to catechin EGCG etc.；Simple and efficient, environmentally friendly, cost of material is low, can be widely applied to food, medicine field；The nitrogen source compound wherein added can not only improve the activity of tannase, rapid conversion catechin EGCG, additionally it is possible to promote aspergillus to the bioconversion of other catechins ECGs, GCG, EGC, EC, GA, GC, CG, C etc..
Description
Technical field
The present invention relates to technical field of bioengineering, more particularly, to a kind of catechin bioconversion nutrient solution and one
The method that kind promotes catechin bioconversion.
Background technique
Catechin is the more rich a kind of functional activity ingredient of content in tea polyphenols, accounts for tea polyphenols total amount 65%~80%,
It mainly include ester catechin and non-ester catechin, ester catechin has: Epigallo-catechin gallate (EGCG)
(Epigallocatechin gallate, EGCG), nutgall catechin gallic acid ester (Gallocatechin gallate,
GCG), L-Epicatechin gallate (Epicatechin gallate, ECG), catechin and gallate (Catechin
Gallate, CG)；Non-ester catechin: epicatechin (Epicatechin, EC), epigallocatechin
(Epigallocatechin, EGC), nutgall catechin (Gallocatechin, GC), gallic acid (Gallocate
Acid, GA).
Catechin, especially EGCG, have remove free radical, anti-aging, it is anti-radiation, anti-cancer and anticancer, anti-bacteria and anti-virus,
Reduce cholesterol and a series of pharmacological functions such as blood lipid, hypoglycemic, weight-reducing.Just because of this special oxidation resistance, youngster
Theine becomes at present the natural additive of most application prospect, is widely used in medicine, food service industry, agricultural production etc..
But EGCG is very low in the intracorporal bioavilability of people, and after EGCG enters mouse intestinal, only 0.1% ~ 1.6% is absorbed, big portion
After dividing EGCG to enter colon, is utilized by the microorganism decomposition in colon, finally excreted with excrement.
Studies have reported that its bioavilability and bioactivity can be improved in EGCG after bioconversion.At present to EGCG
Bioconversion most study is intestinal flora, and Roowi discovery EGCG is converted into EGC and GA in mouse intestinal environment, then
EGC is further converted to other small-molecule substances, these small-molecule substances are easier by small intestinal absorption.Takagaki has found EGCG
Many converted products are generated after mouse intestinal flora acts on EGC, the anti-tumor activity of these converted products significantly improves.So
And the research of intestinal flora In vitro metabolism is extremely complex.
In recent years, fungi microbe mediates the research of EGCG bioconversion to receive significant attention, and fungi microbe possesses rich
Rich enzyme system converts condition of providing convenience to probe into external biological.ZL201510085412.4 describes a kind of aspergillus niger Jie
The method for leading EGCG production epigallocatechin and gallic acid, however the bioconversion nutrient composition that this method uses is multiple
It is miscellaneous, higher cost in actual production, and isolating and purifying for later period catechin can be impacted；In addition, this method only needle
EGC and GA are converted to EGCG, EGC is not related to and GA is further converted, and the biology for being not directed to other catechins turns
Change, there is certain limitation.
Therefore, it is badly in need of developing the biology that ingredient is simple and aspergillus can be promoted to carry out a variety of catechins bioconversion
Convert nutrient solution；
In addition, it is also necessary to develop the method that can carry out bioconversion to a variety of catechins.
Summary of the invention
The present invention is to overcome bioconversion nutrient composition described in the above-mentioned prior art complicated and only convert EGCG
At the defect of EGC and GA, a kind of catechin bioconversion nutrient solution is provided, the bioconversion nutrient composition provided is simply, just
It is isolated and purified after culture, moreover it is possible to aspergillus be promoted to carry out bioconversion to a variety of catechins.
Another object of the present invention is to provide a kind of methods for promoting catechin bioconversion, and the method provided is convenient for training
It is isolated and purified after supporting, moreover it is possible to which bioconversion is carried out to a variety of catechins.
In order to solve the above technical problems, the technical solution adopted by the present invention is that:
A kind of catechin bioconversion nutrient solution, including raw material containing catechin, nitrogen source compound and water；The raw material containing catechin
For tea polyphenols or catechin；The mass percentage of the nitrogen source compound is 0.01% ~ 5%.
Above-mentioned catechin bioconversion nutrient solution generally passes through 121 DEG C of sterilizing 20min processing.
In catechin conversion process of the present invention, any time point, obtained product can be known as " converted product ", also
It is to say that " converted product " can be the mixture of various catechins, can also be the mixed of various catechins and other small-molecule substances
Object is closed, can also be the mixture of various small-molecule substances.
Catechin bioconversion nutrient composition provided by the invention is simple, will not be to catechu without other complicated inorganic salts
Plain EGCG etc. is interfered, convenient for culture after isolate and purify, overcome traditional chemical synthesis environmental pollution caused by pressure
Power has higher safety, is further widely applied and provides the foundation to catechin.
The catechin bioconversion nutrient solution is added with nitrogen source compound, can not only improve the activity of tannase, quickly
Catechin EGCG is converted, EGCG converts under the action of tannase, the antiproliferative of antioxidant activity after conversion, tumour cell
Effect significantly improves；Aspergillus can also be promoted to the bioconversion of other catechins ECGs, GCG, EGC, EC, GA, GC, CG, C etc..
Whole Cell Bioconversion mild condition of the aspergillus in the catechin bioconversion nutrient solution, low in cost, operating condition is simple,
It can be widely applied to food and medicine and other fields.
When increasing the content of nitrogen source compound, is conducive to the ability for improving Aspergillus transformation catechin, accelerates the life of catechin
Object conversion；But when the too high levels of nitrogen source compound, aspergillus thalli growth can be too fast, autolysis occurs, and increase production
Cost.
Preferably, the nitrogen source compound be yeast extract powder, tryptone, soy peptone, amino acid, sodium nitrate,
One or more of sodium nitrite, sodium glutamate, ammonium sulfate or ammonium nitrate.
The yeast extract powder can use commercially available yeast extract powder.
Preferably, the nitrogen source compound is one of yeast extract powder, tryptone, sodium nitrate or ammonium sulfate or several
Kind.
Inventor the study found that nitrogen source compound using yeast extract powder, tryptone, sodium nitrate or when ammonium sulfate, it is bent
The ability of mould conversion catechin is stronger, and the bioconversion speed of catechin is faster.
Preferably, the mass percentage of the nitrogen source compound is 0.01% ~ 5%.
Preferably, the mass percentage of the nitrogen source compound is 0.1% ~ 2%.
Preferably, the mass percentage of the nitrogen source compound is 0.1% ~ 1%.
Preferably, the catechin is one or more of EGCG, ECG, GCG, EGC, EC, GA, GC, CG or C.
Preferably, the mass percentage of the raw material containing catechin is 0.1% ~ 10%.
Preferably, the mass percentage of the raw material containing catechin is 1% ~ 5%.
The present invention protects a kind of method for promoting catechin bioconversion simultaneously, and described method includes following steps:
S1. aspergillus spore suspension is prepared；
S2. aspergillus spore suspension is added in above-mentioned catechin bioconversion nutrient solution, is cultivated；
S3. aspergillus hyphae is removed after cultivating, obtains converted product.
Method provided by the invention uses Aspergillus strain, and bioconversion nutrient composition is simple, without other complicated inorganic salts
Catechin EGCG etc. is interfered, pressure caused by traditional chemical synthesis environmental pollution is overcome, there is higher safety
Property, catechin is further widely applied and is provided the foundation；
Moreover, nitrogen source compound is added into bioconversion nutrient solution in the journey of aspergillus bioconversion catechin, it can not only
Improve tannase activity, rapid conversion catechin EGCG, additionally it is possible to promote aspergillus to other catechins ECGs, GCG, EGC, EC,
The bioconversion of GA, GC, CG, C etc.；Whole Cell Bioconversion mild condition, low in cost, operating condition is simple, can answer extensively
For food and medicine and other fields.
Preferably, the aspergillus be aspergillus niger (Aspergillus niger) RAF106 or Brazil's aspergillus
(Aspergillus tubingensis) ATCC16404.
Aspergillus niger (Aspergillus niger) RAF 106 the deposit date is on August 27th, 2014, deposit number is
CGMCC NO.9608, depositary institution are China Committee for Culture Collection of Microorganisms's common micro-organisms center；Depositary institution
Location is city of BeiJing, China Chaoyang District North Star West Road 1 institute 3.
Brazil's aspergillus (Aspergillus tubingensis) ATCC16404 be reference culture.
Preferably, the preparation method of the aspergillus spore suspension includes the following steps:
Step1. Aspergillus strain, plate stationary culture are inoculated in seed culture medium；
Step2. it uses the sterile water containing Tween-80 gently to scrape aspergillus spore, aspergillus spore suspension is made.
Preferably, the formula of the seed culture medium are as follows: potato 200g, glucose 20g, agar 20g, distilled water
1000mL, pH are natural.Commercially available potato sucrose agar (PDA) culture medium can also be used in seed culture medium.
Preferably, the seed culture medium is handled through 121 DEG C of sterilizing 20min.
Preferably, in step Step1. aspergillus seed culture condition are as follows: 20~35 DEG C of cultivation temperature, incubation time 3~
10d。
Preferably, in step Step2. aspergillus spore suspension acquisition methods are as follows: using sterile containing 0.02% Tween-80
Water washing covers with the plate of black spore, gently scrapes media surface with liquid transfer gun head, obtains aspergillus spore suspension.
Preferably, the concentration of aspergillus spore is 10 in catechin bioconversion nutrient solution in step S2.4~109A/mL.
Preferably, the condition cultivated in step S2. is 20~45 DEG C of temperature, 100~200rpm/min of shaking speed.
Preferably, incubation time is 6 ~ 120h in step S2..
Preferably, using centrifuge separation removal aspergillus hyphae in step S3..
Preferably, the rate being centrifuged in step S3. is 8000~12000rpm/min, and centrifugation time is 5~10min.
Compared with prior art, the beneficial effects of the present invention are:
Catechin bioconversion nutrient composition provided by the invention is simple, will not be to catechin without other complicated inorganic salts
EGCG etc. is interfered, and will not be interfered to catechin EGCG etc.；Simple and efficient, environmentally friendly, cost of material is low, can be widely applied
In food, medicine field；
The nitrogen source compound wherein added can not only improve the activity of tannase, rapid conversion catechin EGCG, additionally it is possible to promote
Into aspergillus to the bioconversion of other catechins ECGs, GCG, EGC, EC, GA, GC, CG, C etc..
Detailed description of the invention
Fig. 1 is yeast extract powder in embodiment 1 and sodium nitrate to each catechu in aspergillus niger RAF106 bioconversion tea polyphenols
The influence situation of cellulose content.
Fig. 2 is tryptone in embodiment 2 and ammonium sulfate to each youngster in Brazil's aspergillus ATCC16404 bioconversion tea polyphenols
The influence situation of catechin content.
Fig. 3 is sodium nitrate, tryptone and tryptophan in embodiment 3 to each youngster in aspergillus niger RAF106 bioconversion EGCG
The influence situation of catechin content.
Fig. 4 is ammonium sulfate in embodiment 4 and yeast extract powder to each in Brazil's aspergillus ATCC16404 bioconversion tea polyphenols
The influence situation of catechin content.
Fig. 5 is that tryptophan and sodium nitrate contain each catechin in aspergillus niger RAF106 bioconversion tea polyphenols in embodiment 5
The influence situation of amount.
Fig. 6 is yeast extract powder in embodiment 6 and tryptophan to each catechin in aspergillus niger RAF106 bioconversion EGCG
The influence situation of content.
Fig. 7 is tryptone in embodiment 7 and soy peptone to each in Brazil's aspergillus ATCC16404 bioconversion EGCG
The influence situation of catechin content.
Fig. 8 is sodium nitrate in embodiment 8 and sodium glutamate to each catechu in Brazil's aspergillus ATCC16404 bioconversion EGCG
The influence situation of cellulose content.
Fig. 9 is tryptone, yeast extract powder, sodium nitrate, soy peptone in embodiment 9 respectively to aspergillus niger RAF106
The active influence of tannase during bioconversion EGCG.
Figure 10 is tryptophan, ammonium sulfate, yeast extract powder, sodium nitrate in embodiment 10 respectively to Brazil's aspergillus
The active influence of tannase during ATCC16404 bioconversion tea polyphenols.
Specific embodiment
The present invention is further illustrated With reference to embodiment.
The equal cocoa of raw material in embodiment is by being commercially available；
Unless stated otherwise, the present invention uses reagent, method and apparatus is the art conventional reagents, method and apparatus.
Aspergillus niger (Aspergillus niger) RAF106 bacterial strain, the deposit date is on August 27th, 2014, deposit number
For CGMCC NO.9608, depositary institution is China Committee for Culture Collection of Microorganisms's common micro-organisms center；Depositary institution
Address is city of BeiJing, China Chaoyang District North Star West Road 1 institute 3.
Brazil's aspergillus (Aspergillus tubingensis) ATCC16404 be reference culture.
High performance liquid chromatography detection condition are as follows: sample introduction 10 μ L, wavelength 256nm, flow velocity 0.8mL/min, mobile phase A: 0.5%
Formic acid water.Mobile phase B: acetonitrile.Elution process: 0~5min A phase keeps 95%；5~20min A phase 95% drops to 50%；20~
30min A phase keeps 50%；30~35min A phase rises to 95% from 50%.
Tannin enzyme assay method are as follows:
Prepare 3 test tubes, number is blank tube, control tube and measurement pipe respectively；0.5mL crude enzyme liquid is added in every test tube,
It is subsequently placed into 40 DEG C of water-baths and preheats 15min；
1mL dehydrated alcohol (inactivating tannase) is added in control tube；The Citrate buffer of 0.5mL is added in blank tube
Liquid (pH 5.0)；0.5mL propylgallate solution, 40 DEG C of reaction 25min are all added in control tube and measurement pipe；
In addition to control tube, 1mL dehydrated alcohol is all added in remaining each pipe, then is settled to 25mL with citrate buffer；
It is compareed with blank tube, 520nm measures the absorbance of each pipe.
Embodiment 1
Bioconversion nutrient solution: 2% tea polyphenols is prepared by formula as below, 0.5% yeast extract powder, 0.5% sodium nitrate, surplus is water,
121 DEG C of sterilizing 20min.
The inoculated aspergillus niger RAF106 bacterial strain in PDA culture medium, 28 DEG C of stationary culture 5d；
The plate that black aspergillus niger spore is covered with the sterile water washing containing 0.05% Tween-80, training is gently scraped with liquid transfer gun head
Primary surface is supported, aspergillus niger spore suspension is obtained.
Spore suspension is adjusted to 1 × 10 with sterile water5A/mL then takes 1mL spore suspension that bioconversion nutrient solution is added
In, 28 DEG C of 160 r/min cultivates 50h.
Every 10h takes out appropriate converted product, and 8000rpm/min is centrifuged 5min, and 1mL supernatant ultrapure water is taken to dilute 100
Times, with 0.22 μm of membrane filtration.
Filtrate moderately dilution after with HPLC detection conversion after each catechin changes of contents, if Fig. 1 show, 20h EGCG with
ECG is all converted, and 30h GCG and GA all degrade, and 40h EGC, EC are totally converted, GC Partial digestion, until 50h whole catechu
Element realizes conversion.
Embodiment 2
Bioconversion nutrient solution liquid: 3% tea polyphenols is prepared by formula as below, 0.8% tryptone, 0.8% ammonium sulfate, surplus is water,
121 DEG C of sterilizing 20min.
Brazil's aspergillus ATCC16404,28 DEG C of stationary culture 3d are inoculated in PDA culture medium.
The plate of black Brazil's aspergillus spore is covered with the sterile water washing containing 0.05% Tween-80, gently with liquid transfer gun head
Media surface is scraped, Brazil's aspergillus spore suspension is obtained.
Spore suspension is adjusted to 1 × 10 with sterile water6A/mL then takes 1mL spore suspension that bioconversion nutrient solution is added
In, 32 DEG C of 160 r/min cultivates 80h.
Every 10h takes out appropriate converted product, and 10000rpm/min is centrifuged 3min, and 1mL supernatant ultrapure water is taken to dilute 100
Times, with 0.22 μm of membrane filtration.
The changes of contents of each catechin after filtrate is converted after moderately diluting with HPLC detection, if Fig. 2 is shown, 20h ECG is complete
Portion is converted, and 30h EGCG is all converted, and GCG has almost been degraded, 50h EGC and GA Partial digestion, until 80h whole catechu
Element is thoroughly converted.
Embodiment 3
Bioconversion nutrient solution: 3%EGCG is prepared by formula as below, and 1.0% sodium nitrate, 1.0% tryptone, 1.0% tryptophan is remaining
Amount is water, 121 DEG C of sterilizing 20min.
The inoculated aspergillus niger RAF106 bacterial strain in PDA culture medium, 35 DEG C of stationary culture 4d.
The plate that black aspergillus niger spore is covered with the sterile water washing containing 0.05% Tween-80, is gently scraped with liquid transfer gun head
Media surface is taken, aspergillus niger spore suspension is obtained.
Spore suspension is adjusted to 1 × 10 with sterile water7A/mL then takes 1mL spore suspension that bioconversion nutrient solution is added
In, 32 DEG C of 160 r/min cultivates 80h.
It chooses suitable time point and takes out appropriate converted product, 12000rpm/min is centrifuged 2min, takes 1mL supernatant to use super
Pure water dilutes 100 times, with 0.22 μm of membrane filtration.
With the changes of contents of each catechin after HPLC detection conversion, if Fig. 3 is shown, 40h EGCG, ECG are all converted,
50h EGCG thoroughly degrades, and 60h EGC, GA and EC start to degrade, until 80h whole catechin thoroughly converts.
Embodiment 4
Bioconversion nutrient solution: 4% tea polyphenols is prepared by formula as below, 1.5% ammonium sulfate, 1.5% yeast extract powder, surplus is water,
121 DEG C of sterilizing 20min.
Brazil's aspergillus ATCC16404 bacterial strain, 35 DEG C of stationary culture 5d are inoculated in PDA culture medium.
The plate of black Brazil's aspergillus spore is covered with the sterile water washing containing 0.05% Tween-80, gently with liquid transfer gun head
Media surface is scraped, aspergillus niger spore suspension is obtained.
Spore suspension is adjusted to 1 × 10 with sterile water8A/mL then takes 1mL spore suspension that bioconversion nutrient solution is added
In, 37 DEG C of 160 r/min cultivates 60h.
It chooses suitable time point and takes out appropriate converted product, 8000rpm/min is centrifuged 10min, takes 1mL supernatant to use super
Pure water dilutes 100 times, with 0.22 μm of membrane filtration.
With the changes of contents of each catechin after HPLC detection conversion, if Fig. 4 is shown, 20h ECG has been totally converted, 30h
EGCG and GCG have been totally converted, and all degradation is complete by 40h GA, EGC, EC, GC Partial digestion, until 60h whole catechin is thorough
Conversion.
Embodiment 5
Bioconversion nutrient solution: 2% tea polyphenols is prepared by formula as below, 1.0% tryptophan, 1.0% sodium nitrate, surplus is water, 121
DEG C sterilizing 20min.
The inoculated aspergillus niger RAF106 bacterial strain in PDA culture medium, 32 DEG C of stationary culture 3d.
The plate that black aspergillus niger spore is covered with the sterile water washing containing 0.05% Tween-80, is gently scraped with liquid transfer gun head
Media surface is taken, aspergillus niger spore suspension is obtained.
Spore suspension is adjusted to 1 × 10 with sterile water5A/mL then takes 1mL spore suspension that bioconversion nutrient solution is added
In, 30 DEG C of 160 r/min cultivates 90h.
Every 10h takes out appropriate converted product, and 10000rpm/min is centrifuged 5min, and 1mL supernatant ultrapure water is taken to dilute 100
Times, with 0.22 μm of membrane filtration.
With the changes of contents of each catechin after HPLC detection conversion, if Fig. 5 is shown, 20h ECG is totally converted, 30h
EGCG, GCG are totally converted, and 50h GA all degrades, EGC and EC Partial Conversion, until 90h whole catechin is thoroughly degraded.
Embodiment 6
Bioconversion nutrient solution: 3%EGCG is prepared by formula as below, 1.5% yeast extract powder, 1.5% tryptophan, surplus is water,
121 DEG C of sterilizing 20min.
The inoculated aspergillus niger RAF106 bacterial strain in PDA culture medium, 28 DEG C of stationary culture 5d.
The plate that black aspergillus niger spore is covered with the sterile water washing containing 0.05% Tween-80, is gently scraped with liquid transfer gun head
Media surface is taken, aspergillus niger spore suspension is obtained.
Spore suspension is adjusted to 1 × 10 with sterile water7A/mL then takes 1mL spore suspension that bioconversion nutrient solution is added
In, 40 DEG C of 160 r/min cultivates 60h.
It chooses suitable time point and takes out appropriate converted product, 12000rpm/min is centrifuged 4min, takes 1mL supernatant to use super
Pure water dilutes 100 times, with 0.22 μm of membrane filtration.
With the changes of contents of each catechin after HPLC detection conversion, if Fig. 6 is shown, 20h GCG has been totally converted, 30h
EGCG, ECG have been totally converted, EGC, GA Partial digestion, and 60h whole catechin thoroughly converts.
Embodiment 7
Bioconversion nutrient solution: 5%EGCG is prepared by formula as below, 1.5% tryptone, 1.5% soy peptone, surplus is water,
121 DEG C of sterilizing 20min.
Brazil's aspergillus ATCC16404 bacterial strain, 35 DEG C of stationary culture 5d are inoculated in PDA culture medium.
The plate of black Brazil's aspergillus spore is covered with the sterile water washing containing 0.05% Tween-80, gently with liquid transfer gun head
Media surface is scraped, aspergillus niger spore suspension is obtained.
Spore suspension is adjusted to 1 × 10 with sterile water7A/mL then takes 1mL spore suspension that bioconversion nutrient solution is added
In, 42 DEG C of 160 r/min cultivates 60h.
It chooses suitable time points and takes out appropriate converted product, 11000rpm/min is centrifuged 6min, takes 1mL supernatant with ultrapure
Water dilutes 100 times, with 0.22 μm of membrane filtration.
With the changes of contents of each catechin after HPLC detection conversion, if Fig. 7 is shown, 20h ECG has been totally converted, 40h
EGCG, GCG have been totally converted, EGC, GA and EC Partial Conversion, until 60h whole catechin is degradable.
Embodiment 8
Bioconversion nutrient solution: 1%EGCG is prepared by formula as below, 0.5% sodium nitrate, 0.5% sodium glutamate, surplus is water, 121
DEG C sterilizing 20min.
Brazil's aspergillus ATCC16404 bacterial strain, 28 DEG C of stationary culture 5d are inoculated in PDA culture medium.
The plate of black Brazil's aspergillus spore is covered with the sterile water washing containing 0.05% Tween-80, gently with liquid transfer gun head
Media surface is scraped, aspergillus niger spore suspension is obtained.
Spore suspension is adjusted to 1 × 10 with sterile water5A/mL then takes 1mL spore suspension that bioconversion nutrient solution is added
In, 45 DEG C of 160 r/min cultivates 100h.
It chooses suitable time point and takes out appropriate converted product, 10000rpm/min is centrifuged 8min, takes 1mL supernatant to use super
Pure water dilutes 100 times, with 0.22 μm of membrane filtration.
With the changes of contents of each catechin after HPLC detection conversion, if Fig. 8 is shown, 30h ECG has been totally converted, 60h
EGCG, GCG have been totally converted, and 80h GA, EGC, EC and GC are largely degraded, until 100h whole catechin thoroughly converts.
Embodiment 9
Using 5%EGCG as common component, it is separately added into 1.0% tryptone, yeast extract powder, sodium nitrate, soy peptone, surplus
For water, it is made into the bioconversion nutrient solution of different nitrogen sources compound, 121 DEG C of sterilizing 20min.
Aspergillus niger RAF106 spore suspension is adjusted to 1 × 10 with sterile water7A/mL then takes 1mL spore suspension that life is added
Object converts in nutrient solution, 28 DEG C of shake culture 40h, samples in 0h, 10h, 20h, 30h, 40h, and filtering removal mycelia is to get thick enzyme
Liquid；According to the activity of the tannase in above-mentioned tannin enzyme assay method measurement crude enzyme liquid, turned so that the biology of nitrogen source compound is not added
Changing nutrient solution is control.
If Fig. 9 is shown, in the bioconversion nutrient solution of addition nitrogen source compound, the activity of tannase is than control group
Height illustrates that nitrogen source compound can remarkably promote aspergillus niger RAF106 strain secretes tannase.
Embodiment 10
Using 3% tea polyphenols as common component, it is separately added into 0.5% tryptophan, yeast extract powder, sodium nitrate, ammonium sulfate, surplus is
Water is made into the bioconversion nutrient solution of different nitrogen sources compound, 121 DEG C of sterilizing 20min.
Brazil's aspergillus ATCC16404 spore suspension is adjusted to 1 × 10 with sterile water7A/mL then takes 1mL spore suspension
It is added in bioconversion nutrient solution, 32 DEG C of shake culture 60h, is sampled in 0h, 10h, 30h, 40h, 60h, filtering removal mycelia, i.e.,
Obtain crude enzyme liquid；According to the activity of the tannase in above-mentioned tannin enzyme assay method measurement crude enzyme liquid, nitrogen source compound is not added
Bioconversion nutrient solution is control.
If Figure 10 is shown, in the bioconversion nutrient solution of nitrogen source compound, the activity of tannase is higher than control group, says
Bright nitrogen source compound can remarkably promote Brazil's aspergillus ATCC16404 strain secretes tannase.
Obviously, the above embodiment of the present invention be only to clearly illustrate example of the present invention, and not be pair
The restriction of embodiments of the present invention.For those of ordinary skill in the art, may be used also on the basis of the above description
To make other variations or changes in different ways.There is no necessity and possibility to exhaust all the enbodiments.It is all this
Made any modifications, equivalent replacements, and improvements etc., should be included in the claims in the present invention within the spirit and principle of invention
Protection scope within.
Claims (9)
1. a kind of catechin bioconversion nutrient solution, which is characterized in that including raw material containing catechin, nitrogen source compound and water；Institute
Stating raw material containing catechin is tea polyphenols or catechin；The mass percentage of the nitrogen source compound is 0.01% ~ 5%.
2. catechin bioconversion nutrient solution according to claim 1, which is characterized in that the nitrogen source compound is yeast
In extract powder, tryptone, soy peptone, amino acid, sodium nitrate, sodium nitrite, sodium glutamate, ammonium sulfate or ammonium nitrate
It is one or more of.
3. catechin bioconversion nutrient solution according to claim 1, which is characterized in that the nitrogen source compound is yeast
One or more of extract powder, tryptone, sodium nitrate or ammonium sulfate.
4. catechin bioconversion nutrient solution according to claim 1, which is characterized in that the quality of the nitrogen source compound
Percentage composition is 0.01% ~ 5%.
5. catechin bioconversion nutrient solution according to claim 4, which is characterized in that the quality of the nitrogen source compound
Percentage composition is 0.1% ~ 1%.
6. catechin bioconversion nutrient solution according to claim 1, which is characterized in that the catechin be EGCG,
One or more of ECG, GCG, EGC, EC, GA, GC, CG or C.
7. a kind of method for promoting catechin bioconversion, which comprises the steps of:
S1. aspergillus spore suspension is prepared；
S2. aspergillus spore suspension is added in the described in any item bioconversion nutrient solutions of claim 1 ~ 6, is cultivated；
S3. aspergillus hyphae is removed after cultivating, obtains converted product.
8. the method according to the description of claim 7 is characterized in that the aspergillus be aspergillus niger (Aspergillus niger)
RAF106 or Brazil's aspergillus (Aspergillus tubingensis) ATCC16404.
9. the method according to the description of claim 7 is characterized in that the condition cultivated in step S2. is, 20~45 DEG C of temperature,
100~200rpm/min of shaking speed.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113186104A (en) * | 2021-03-16 | 2021-07-30 | 四川大学 | Aspergillus niger and application and method thereof |
| CN113303386A (en) * | 2021-05-08 | 2021-08-27 | 华南农业大学 | Fermented tea helpful for delaying organism aging and preparation method and application thereof |
| CN115777811A (en) * | 2022-12-29 | 2023-03-14 | 华南农业大学 | Novel fermented sour tea and preparation method thereof |
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| CN104673845A (en) * | 2015-02-15 | 2015-06-03 | 华南农业大学 | Method for producing epigallocatechin and gallic acid through transformation of aspergillus niger whole cell |
| CN104673844A (en) * | 2015-02-15 | 2015-06-03 | 华南农业大学 | Method for preventing transformation of epigallocatechin and gallic acid |
| CN106978350A (en) * | 2016-01-15 | 2017-07-25 | 北京大学 | One plant of aspergillus niger and its application in the preparation of Pu'er tea chlorins compound |
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| CN104673845A (en) * | 2015-02-15 | 2015-06-03 | 华南农业大学 | Method for producing epigallocatechin and gallic acid through transformation of aspergillus niger whole cell |
| CN104673844A (en) * | 2015-02-15 | 2015-06-03 | 华南农业大学 | Method for preventing transformation of epigallocatechin and gallic acid |
| CN106978350A (en) * | 2016-01-15 | 2017-07-25 | 北京大学 | One plant of aspergillus niger and its application in the preparation of Pu'er tea chlorins compound |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113186104A (en) * | 2021-03-16 | 2021-07-30 | 四川大学 | Aspergillus niger and application and method thereof |
| CN113303386A (en) * | 2021-05-08 | 2021-08-27 | 华南农业大学 | Fermented tea helpful for delaying organism aging and preparation method and application thereof |
| CN115777811A (en) * | 2022-12-29 | 2023-03-14 | 华南农业大学 | Novel fermented sour tea and preparation method thereof |
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Application publication date: 20190104 |