CN110093380A - A kind of resveratrol process for extracting, separating and purifying - Google Patents
A kind of resveratrol process for extracting, separating and purifying Download PDFInfo
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- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2434—Glucanases acting on beta-1,4-glucosidic bonds
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- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
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Abstract
This application involves separating and purifying technology fields, and in particular to a kind of resveratrol process for extracting, separating and purifying includes the following steps: S1: plant sample processing；S2: beta-glucosidase enzyme solution is prepared；S3: immobilised enzymes；S4: immobilized β-glucosidase enzyme solution is added into plant sample suspension；S5: conversion fluid is collected: S6: the extraction purification of resveratrol.Beta-glucosidase is fixed by investment, immobilised enzymes is reusable, improves the service efficiency of enzyme, use cost reduces；Immobilised enzymes is easily separated with reaction system, simplifies purifying technique, and product yield is high, high-quality；In most cases, enzyme is improved through immobilization rear stability；The catalytic reaction process of immobilised enzymes is easier to control.In addition, suspension, which is made, in plant sample sufficiently to react with immobilised enzymes, accelerates reaction speed, improve the utilization rate of plant sample.
Description
Technical field
The present invention relates to separating and purifying technology fields, more specifically, it relates to a kind of resveratrol extraction separation and purification
Technique.
Background technique
Resveratrol (chemistry entitled resvertrol) is the non-flavones polyphenol compound containing stilbene class formation, be plant by
To stress when itself synthesis a kind of antitoxin, be insoluble in water, being soluble in methanol, ethyl alcohol, acetone, ethyl acetate, chloroform etc. has
Solvent.Mainly there are antitumor, anti-inflammatory, antibacterial, anti-oxidant, free radical resisting, protection liver, protection angiocarpy and resists myocardial ischemia
Etc. functions, be widely used in health care of food product, cosmetics and field of medicaments.
The production of resveratrol is mainly plant extraction method at present.The plant for extracting resveratrol has grape, peanut, mulberry
Shen, polygonum cuspidate etc..However, the Resveratrol content in plant is very low, so that it isolates and purifies high expensive.And resveratrol class
Like 7~12 times that content of the object polydatin in plant is resveratrol, if converting white black false hellebore for polydatin
The yield of alcohol, resveratrol is greatly improved.Enzymatic hydrolysis polydatin is converted into resveratrol, because of mild condition, reaction
Product is more single, and yield is high, and is widely adopted.But in traditional enzymatic hydrolysis polydatin method, enzyme is dispersed in solution
Middle easy in inactivation, is not easily recycled, at high cost.
Summary of the invention
The purpose of the present invention is to provide a kind of resveratrol process for extracting, separating and purifying, have low cost, can be repeatedly
Extract resveratrol.
Above-mentioned technical purpose of the invention has the technical scheme that a kind of resveratrol extracts separation
Purifying process, characterized by the following steps:
S1: plant sample processing: plant is milled crushings, and the deionized water that polyploid product is added is made plant sample and hangs
Supernatant liquid；
S2: prepare beta-glucosidase enzyme solution: the strain for producing beta-glucosidase be inoculated in culture medium, 25 DEG C~
50 DEG C, under conditions of 120~220rpm of shaking speed, producing enzyme is fermented 36~72 hours, then by all fermentation materials 5000~
It is centrifuged 15~40min under the revolving speed of 10000r/min, collects supernatant and obtains beta-glucosidase crude enzyme liquid, by β-glucose
Glycosides enzyme crude enzyme liquid is further centrifuged 15min under the revolving speed of 4000~12000r/min, removes supernatant, the β-Portugal purified
Polyglycoside enzyme enzyme solution；
S3: immobilised enzymes: taking 5ml beta-glucosidase enzyme solution, and it is molten to be added to the sodium alginate that 3mL mass fraction is 3.5%
In liquid, it is uniformly mixed, adds appropriate glutaraldehyde solution, the constant temperature oscillation in shaking table draws mixed liquor with 5mL syringe, with
In 10cm or so height CaCl2 solution that implantation quality score is 2% dropwise, smooth gel bead is formed immediately, is coagulated out
Glue bead replaces CaCl2 solution, is placed in 4 DEG C of refrigerators and stands hardening 2h；Gelled pill is filtered out again, is with mass fraction
0.9% NaCl solution is washed 3 times and is used distilled water flushing 2 times again, and immobilized β-glucosidase is obtained；
S4: immobilized β-glucosidase enzyme solution is added into plant sample suspension, in 25 DEG C~45 after being sufficiently stirred
Conversion reaction 15~24 hours are stood under the conditions of DEG C；Or it is sufficiently stirred and is placed on 25 DEG C~45 DEG C, 80~180rpm of shaking speed
Under the conditions of carry out conversion reaction；
S5: conversion fluid is collected: by all materials after S4 conversion reaction with 4000~8000r/min pelleted by centrifugation, being collected
Supernatant is up to the solution containing resveratrol；
S6: its 1 times of volume the extraction purification of resveratrol: is added in the solution containing resveratrol obtained to step S5
50~95% ethyl alcohol, carry out heating and refluxing extraction at a temperature of 50 DEG C~100 DEG C, obtain pure beta-glucosidase conversion produce
Object-resveratrol liquid.
By using above-mentioned technical proposal, beta-glucosidase to be fixed by investment, immobilised enzymes is reusable,
Improve the service efficiency of enzyme, use cost reduces；Immobilised enzymes is easily separated with reaction system, simplifies purifying technique, and
And product yield is high, high-quality；In most cases, enzyme is improved through immobilization rear stability；The catalysis of immobilised enzymes is anti-
Answer process easier to control.In addition, suspension, which is made, in plant sample sufficiently to react with immobilised enzymes, accelerates reaction speed, mention
The high utilization rate of plant sample.
Further, grape, peanut, mulberries or polygonum cuspidate can be selected in the plant sample in S1.
Further, three times are added in S1 or plant sample suspension is made in the deionized water of four times of volumes.
Further, the bacterium for beta-glucosidase being produced described in S2 selects Penicillium citrinum (Penicillium citrinum)
CGMCC 3.2938, Rhizopus arrhizus (Rhizopus arrhizus) CGMCC No.1963, Trichoderma viride
(Trichodermaviride) CGMCC No.1962, Mucor (Mucor sp.) CGMCC NO.0151, Aspergillus terreus
One of (Aspergillus terreus) CGMCC No.0569.
Further, the immobilized β-glucosidase being prepared in S3 blots bead surface moisture with blotting paper, storage
It is stored in spare in 4 DEG C of refrigerators.
Further, centrifugal speed is 10~15min in S5.
Further, the temperature of heating and refluxing extraction is 60 DEG C~100 DEG C in S6.
Further, the time of conversion reaction is 5~24 hours in S4.
In conclusion the invention has the following advantages:
Beta-glucosidase is fixed by investment, immobilised enzymes is reusable, and the service efficiency of enzyme is made to improve, make
It is reduced with cost；Immobilised enzymes is easily separated with reaction system, simplifies purifying technique, and product yield is high, high-quality；?
In most cases, enzyme is improved through immobilization rear stability；The catalytic reaction process of immobilised enzymes is easier to control.In addition, will
Suspension, which is made, in plant sample sufficiently to react with immobilised enzymes, accelerate reaction speed, improve the utilization rate of plant sample.And
Using the polidatin in microbial enzyme method hydrolyzing plant, it is translated into resveratrol, there is mild condition, reaction product
Many advantages, such as more single, efficiency of pcr product is high, environmental-friendly needs so as to overcome traditional glycoside hydrolysis to react in high temperature
Under high pressure, the shortcomings that being completed, the high requirements on the equipment, and polluted the environment by acid or base catalysis.
Specific embodiment
Embodiment 1:
A kind of resveratrol process for extracting, separating and purifying, characterized by the following steps:
S1: plant sample processing: plant is milled crushings, and the deionized water that three times volume is added is made plant sample and hangs
Supernatant liquid, Plants selection grape；
S2: it prepares beta-glucosidase enzyme solution: the Penicillium citrinum (Penicillium of beta-glucosidase will be produced
Citrinum) CGMCC 3.2938 is inoculated in culture medium, under conditions of 25 DEG C, shaking speed 120rpm, producing enzyme fermentation 36
Hour, then all fermentation materials are centrifuged 15min under the revolving speed of 5000r/min, collect supernatant and obtain beta-glucosidase
Beta-glucosidase crude enzyme liquid is further centrifuged under the revolving speed of 4000r/min 15min, removes supernatant, obtain by enzyme crude enzyme liquid
To the beta-glucosidase enzyme solution of purification；
S3: immobilised enzymes: taking 5ml beta-glucosidase enzyme solution, and it is molten to be added to the sodium alginate that 3mL mass fraction is 3.5%
In liquid, it is uniformly mixed, adds appropriate glutaraldehyde solution, the constant temperature oscillation in shaking table draws mixed liquor with 5mL syringe, with
In 10cm or so height CaCl2 solution that implantation quality score is 2% dropwise, smooth gel bead is formed immediately, is coagulated out
Glue bead replaces CaCl2 solution, is placed in 4 DEG C of refrigerators and stands hardening 2h；Gelled pill is filtered out again, is with mass fraction
0.9% NaCl solution is washed 3 times and is used distilled water flushing 2 times again, obtains immobilized β-glucosidase, blotting paper blots bead table
Face moisture is stored in spare in 4 DEG C of refrigerators；
S4: immobilized β-glucosidase enzyme solution is added into plant sample suspension, in 25 DEG C of conditions after being sufficiently stirred
Lower standing conversion reaction 15 hours；Or be sufficiently stirred and be placed on 25 DEG C, that conversion reaction 12 is carried out under the conditions of shaking speed 80rpm is small
When；
S5: conversion fluid is collected: by all materials after S4 conversion reaction with 4000r/min pelleted by centrifugation 10min, in collection
Clear liquid is up to the solution containing resveratrol；
S6: its 1 times of volume the extraction purification of resveratrol: is added in the solution containing resveratrol obtained to step S5
50% ethyl alcohol, heating and refluxing extraction is carried out at a temperature of 60 DEG C DEG C, obtains the white black false hellebore of converted product-of pure beta-glucosidase
Alcohol liquid.
Embodiment 2:
A kind of resveratrol process for extracting, separating and purifying, characterized by the following steps:
S1: plant sample processing: plant is milled crushings, and the deionized water that three times volume is added is made plant sample and hangs
Supernatant liquid, Plants selection peanut；
S2: it prepares beta-glucosidase enzyme solution: the Rhizopus arrhizus (Rhizopus arrhizus) of beta-glucosidase will be produced
CGMCC No.1963 is inoculated in culture medium, and under conditions of 30 DEG C, shaking speed 140rpm, producing enzyme is fermented 40 hours, then
All fermentation materials are centrifuged 20min under the revolving speed of 6000r/min, supernatant is collected and obtains beta-glucosidase crude enzyme liquid,
Beta-glucosidase crude enzyme liquid is further centrifuged 15min under the revolving speed of 6000r/min, supernatant is removed, is purified
Beta-glucosidase enzyme solution；
S3: immobilised enzymes: taking 5ml beta-glucosidase enzyme solution, and it is molten to be added to the sodium alginate that 3mL mass fraction is 3.5%
In liquid, it is uniformly mixed, adds appropriate glutaraldehyde solution, the constant temperature oscillation in shaking table draws mixed liquor with 5mL syringe, with
In 10cm or so height CaCl2 solution that implantation quality score is 2% dropwise, smooth gel bead is formed immediately, is coagulated out
Glue bead replaces CaCl2 solution, is placed in 4 DEG C of refrigerators and stands hardening 2h；Gelled pill is filtered out again, is with mass fraction
0.9% NaCl solution is washed 3 times and is used distilled water flushing 2 times again, obtains immobilized β-glucosidase, blotting paper blots bead table
Face moisture is stored in spare in 4 DEG C of refrigerators；
S4: immobilized β-glucosidase enzyme solution is added into plant sample suspension, in 30 DEG C of conditions after being sufficiently stirred
Lower standing conversion reaction 17 hours；Or it is sufficiently stirred and is placed on 30 DEG C, carry out conversion reaction 16 under the conditions of shaking speed 100rpm
Hour；
S5: conversion fluid is collected: by all materials after S4 conversion reaction with 5000r/min pelleted by centrifugation 11min, in collection
Clear liquid is up to the solution containing resveratrol；
S6: its 1 times of volume the extraction purification of resveratrol: is added in the solution containing resveratrol obtained to step S5
60% ethyl alcohol, carry out heating and refluxing extraction at a temperature of 70 DEG C, obtain converted product-resveratrol of pure beta-glucosidase
Liquid.
Embodiment 3:
A kind of resveratrol process for extracting, separating and purifying, characterized by the following steps:
S1: plant sample processing: plant is milled crushings, and the deionized water that three times volume is added is made plant sample and hangs
Supernatant liquid, Plants selection mulberries；
S2: it prepares beta-glucosidase enzyme solution: Mucor (Mucor sp.) CGMCC of beta-glucosidase will be produced
NO.0151 is inoculated in culture medium, and under conditions of 37.5 DEG C, shaking speed 170rpm, producing enzyme is fermented 54 hours, then by institute
There is fermentation material to be centrifuged 27.5min under the revolving speed of 7500r/min, collects supernatant and obtain beta-glucosidase crude enzyme liquid, it will
Beta-glucosidase crude enzyme liquid is further centrifuged 15min under the revolving speed of 7000r/min, removes supernatant, the β-purified
Glucuroide enzyme solution；
S3: immobilised enzymes: taking 5ml beta-glucosidase enzyme solution, and it is molten to be added to the sodium alginate that 3mL mass fraction is 3.5%
In liquid, it is uniformly mixed, adds appropriate glutaraldehyde solution, the constant temperature oscillation in shaking table draws mixed liquor with 5mL syringe, with
In 10cm or so height CaCl2 solution that implantation quality score is 2% dropwise, smooth gel bead is formed immediately, is coagulated out
Glue bead replaces CaCl2 solution, is placed in 4 DEG C of refrigerators and stands hardening 2h；Gelled pill is filtered out again, is with mass fraction
0.9% NaCl solution is washed 3 times and is used distilled water flushing 2 times again, obtains immobilized β-glucosidase, blotting paper blots bead table
Face moisture is stored in spare in 4 DEG C of refrigerators；
S4: immobilized β-glucosidase enzyme solution is added into plant sample suspension, in 35 DEG C of conditions after being sufficiently stirred
Lower standing conversion reaction 19.5 hours；Or it is sufficiently stirred and is placed on 35 DEG C, carry out conversion reaction under the conditions of shaking speed 130rpm
18 hours；
S5: conversion fluid is collected: by all materials after S4 conversion reaction with 6000r/min pelleted by centrifugation 12.5min, being collected
Supernatant is up to the solution containing resveratrol；
S6: its 1 times of volume the extraction purification of resveratrol: is added in the solution containing resveratrol obtained to step S5
72.5% ethyl alcohol, carry out heating and refluxing extraction at a temperature of 80 DEG C, obtain the white black false hellebore of converted product-of pure beta-glucosidase
Alcohol liquid.
Embodiment 4:
A kind of resveratrol process for extracting, separating and purifying, characterized by the following steps:
S1: plant sample processing: plant is milled crushings, and the deionized water that three times volume is added is made plant sample and hangs
Supernatant liquid, Plants selection polygonum cuspidate；
S2: it prepares beta-glucosidase enzyme solution: the Aspergillus terreus (Aspergillus terreus) of beta-glucosidase will be produced
CGMCC No.0569 is inoculated in culture medium, and under conditions of 37.5 DEG C, shaking speed 170rpm, producing enzyme is fermented 54 hours, so
Afterwards all fermentation materials are centrifuged 27.5min under the revolving speed of 7500r/min, collect supernatant and obtain the thick enzyme of beta-glucosidase
Beta-glucosidase crude enzyme liquid is further centrifuged under the revolving speed of 7000r/min 15min, removes supernatant, purified by liquid
Beta-glucosidase enzyme solution；
S3: immobilised enzymes: taking 5ml beta-glucosidase enzyme solution, and it is molten to be added to the sodium alginate that 3mL mass fraction is 3.5%
In liquid, it is uniformly mixed, adds appropriate glutaraldehyde solution, the constant temperature oscillation in shaking table draws mixed liquor with 5mL syringe, with
In 10cm or so height CaCl2 solution that implantation quality score is 2% dropwise, smooth gel bead is formed immediately, is coagulated out
Glue bead replaces CaCl2 solution, is placed in 4 DEG C of refrigerators and stands hardening 2h；Gelled pill is filtered out again, is with mass fraction
0.9% NaCl solution is washed 3 times and is used distilled water flushing 2 times again, obtains immobilized β-glucosidase, blotting paper blots bead table
Face moisture is stored in spare in 4 DEG C of refrigerators；
S4: immobilized β-glucosidase enzyme solution is added into plant sample suspension, in 35 DEG C of conditions after being sufficiently stirred
Lower standing conversion reaction 19.5 hours；Or it is sufficiently stirred and is placed on 35 DEG C, carry out conversion reaction under the conditions of shaking speed 130rpm
18 hours；
S5: conversion fluid is collected: by all materials after S4 conversion reaction with 6000r/min pelleted by centrifugation 12.5min, being collected
Supernatant is up to the solution containing resveratrol；
S6: its 1 times of volume the extraction purification of resveratrol: is added in the solution containing resveratrol obtained to step S5
72.5% ethyl alcohol, carry out heating and refluxing extraction at a temperature of 80 DEG C, obtain the white black false hellebore of converted product-of pure beta-glucosidase
Alcohol liquid.
Embodiment 5:
A kind of resveratrol process for extracting, separating and purifying, characterized by the following steps:
S1: plant sample processing: plant is milled crushings, and the deionized water that three times volume is added is made plant sample and hangs
Supernatant liquid, Plants selection polygonum cuspidate；
S2: it prepares beta-glucosidase enzyme solution: the Aspergillus terreus (Aspergillus terreus) of beta-glucosidase will be produced
CGMCC No.0569 is inoculated in culture medium, and under conditions of 50 DEG C, shaking speed 220rpm, producing enzyme is fermented 72 hours, then
All fermentation materials are centrifuged 40min under the revolving speed of 10000r/min, supernatant is collected and obtains beta-glucosidase crude enzyme liquid,
Beta-glucosidase crude enzyme liquid is further centrifuged 15min under the revolving speed of 12000r/min, supernatant is removed, is purified
Beta-glucosidase enzyme solution；
S3: immobilised enzymes: taking 5ml beta-glucosidase enzyme solution, and it is molten to be added to the sodium alginate that 3mL mass fraction is 3.5%
In liquid, it is uniformly mixed, adds appropriate glutaraldehyde solution, the constant temperature oscillation in shaking table draws mixed liquor with 5mL syringe, with
In 10cm or so height CaCl2 solution that implantation quality score is 2% dropwise, smooth gel bead is formed immediately, is coagulated out
Glue bead replaces CaCl2 solution, is placed in 4 DEG C of refrigerators and stands hardening 2h；Gelled pill is filtered out again, is with mass fraction
0.9% NaCl solution is washed 3 times and is used distilled water flushing 2 times again, obtains immobilized β-glucosidase, blotting paper blots bead table
Face moisture is stored in spare in 4 DEG C of refrigerators；
S4: immobilized β-glucosidase enzyme solution is added into plant sample suspension, in 45 DEG C of conditions after being sufficiently stirred
Lower standing conversion reaction 24 hours；Or it is sufficiently stirred and is placed on 45 DEG C, carry out conversion reaction 24 under the conditions of shaking speed 180rpm
Hour；
S5: conversion fluid is collected: by all materials after S4 conversion reaction with 8000r/min pelleted by centrifugation 15min, in collection
Clear liquid is up to the solution containing resveratrol；
S6: its 1 times of volume the extraction purification of resveratrol: is added in the solution containing resveratrol obtained to step S5
95% ethyl alcohol, carry out heating and refluxing extraction at a temperature of 100 DEG C, obtain the white black false hellebore of converted product-of pure beta-glucosidase
Alcohol liquid.
The above is only a preferred embodiment of the present invention, protection scope of the present invention is not limited merely to above-mentioned implementation
Example, all technical solutions belonged under thinking of the present invention all belong to the scope of protection of the present invention.It should be pointed out that for the art
Those of ordinary skill for, several improvements and modifications without departing from the principles of the present invention, these improvements and modifications
It should be regarded as protection scope of the present invention.
Claims (8)
1. a kind of resveratrol process for extracting, separating and purifying, characterized by the following steps:
S1: plant sample processing: plant is milled crushings, and the deionized water that polyploid product is added is made plant sample and suspends
Liquid；
S2: it prepares beta-glucosidase enzyme solution: the strain for producing beta-glucosidase is inoculated in culture medium, 25 DEG C~50
DEG C, under conditions of 120~220rpm of shaking speed, producing enzyme is fermented 36~72 hours, then by all fermentation materials 5000~
It is centrifuged 15~40min under the revolving speed of 10000r/min, collects supernatant and obtains beta-glucosidase crude enzyme liquid, by β-glucose
Glycosides enzyme crude enzyme liquid is further centrifuged 15min under the revolving speed of 4000~12000r/min, removes supernatant, the β-Portugal purified
Polyglycoside enzyme enzyme solution；
S3: immobilised enzymes: taking 5ml beta-glucosidase enzyme solution, is added in the sodium alginate soln that 3mL mass fraction is 3.5%,
It is uniformly mixed, adds appropriate glutaraldehyde solution, the constant temperature oscillation in shaking table draws mixed liquor with 5mL syringe, with the left side 10cm
In the right height CaCl2 solution that implantation quality score is 2% dropwise, smooth gel bead is formed immediately, out gelled pill,
CaCl2 solution is replaced, is placed in 4 DEG C of refrigerators and stands hardening 2h；Gelled pill is filtered out again, is 0.9% with mass fraction
NaCl solution is washed 3 times and is used distilled water flushing 2 times again, and immobilized β-glucosidase is obtained；
S4: immobilized β-glucosidase enzyme solution is added into plant sample suspension, in 25 DEG C~45 DEG C items after being sufficiently stirred
Conversion reaction 15~24 hours are stood under part；Or it is sufficiently stirred and is placed on 25 DEG C~45 DEG C, shaking speed 80~180rpm condition
Lower carry out conversion reaction；
S5: conversion fluid is collected: by all materials after S4 conversion reaction with 4000~8000r/min pelleted by centrifugation, collecting supernatant
Liquid is up to the solution containing resveratrol；
S6: the 50 of its 1 times of volume the extraction purification of resveratrol: is added in the solution containing resveratrol obtained to step S5
~95% ethyl alcohol carries out heating and refluxing extraction at a temperature of 50 DEG C~100 DEG C, obtains the converted product-of pure beta-glucosidase
Resveratrol liquid.
2. resveratrol process for extracting, separating and purifying according to claim 1, it is characterised in that: the plant sample in S1
Grape, peanut, mulberries or polygonum cuspidate can be selected in product.
3. resveratrol process for extracting, separating and purifying according to claim 2, it is characterised in that: three times or four are added in S1
Plant sample suspension is made in the deionized water of times volume.
4. resveratrol process for extracting, separating and purifying according to claim 2, it is characterised in that: produce β-grape described in S2
The bacterium of glycosidase selects Penicillium citrinum (Penicillium citrinum) CGMCC 3.2938, Rhizopus arrhizus (Rhizopus
Arrhizus) CGMCC No.1963, Mucor (Mucor sp.) CGMCC NO.0151, Aspergillus terreus (Aspergillus
Terreus) one of CGMCC No.0569.
5. resveratrol process for extracting, separating and purifying according to claim 3, it is characterised in that: what is be prepared in S3 consolidates
Surely change beta-glucosidase and blot bead surface moisture with blotting paper, be stored in spare in 4 DEG C of refrigerators.
6. resveratrol process for extracting, separating and purifying according to claim 3, it is characterised in that: centrifugal speed is 10 in S5
~15min.
7. resveratrol process for extracting, separating and purifying according to claim 3, it is characterised in that: heating and refluxing extraction in S6
Temperature be 60 DEG C~100 DEG C.
8. resveratrol process for extracting, separating and purifying according to claim 3, it is characterised in that: in S4 conversion reaction when
Between be 12~24 hours.
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| CN201910341202.5A Pending CN110093380A (en) | 2019-04-25 | 2019-04-25 | A kind of resveratrol process for extracting, separating and purifying |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114657073A (en) * | 2022-03-30 | 2022-06-24 | 中溶科技股份有限公司 | Penicillium citrinum strain capable of producing cellobiase at high yield and application thereof |
| CN116764157A (en) * | 2023-01-12 | 2023-09-19 | 苏州盛仓生物环保科技有限公司 | Method for degrading toluene by enzyme oxidation |
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2019
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114657073A (en) * | 2022-03-30 | 2022-06-24 | 中溶科技股份有限公司 | Penicillium citrinum strain capable of producing cellobiase at high yield and application thereof |
| CN116764157A (en) * | 2023-01-12 | 2023-09-19 | 苏州盛仓生物环保科技有限公司 | Method for degrading toluene by enzyme oxidation |
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Application publication date: 20190806 |