EP0402416A1 - Process for the preparation of tannase intended for the production of gallic acid, with the aid of an aspergillus culture - Google Patents

Process for the preparation of tannase intended for the production of gallic acid, with the aid of an aspergillus culture

Info

Publication number
EP0402416A1
EP0402416A1 EP19890904502 EP89904502A EP0402416A1 EP 0402416 A1 EP0402416 A1 EP 0402416A1 EP 19890904502 EP19890904502 EP 19890904502 EP 89904502 A EP89904502 A EP 89904502A EP 0402416 A1 EP0402416 A1 EP 0402416A1
Authority
EP
European Patent Office
Prior art keywords
tannase
process according
fermentation
tannins
production
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
EP19890904502
Other languages
German (de)
French (fr)
Inventor
Erick Jérome Maurice Cornelius VANDAMME
Maria Magdaléna VERMEIRE Annie
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Universiteit Gent
Original Assignee
Universiteit Gent
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Universiteit Gent filed Critical Universiteit Gent
Publication of EP0402416A1 publication Critical patent/EP0402416A1/en
Pending legal-status Critical Current

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • C12N9/18—Carboxylic ester hydrolases (3.1.1)

Definitions

  • the present invention relates to a process for the preparation of a tannase from microorganisms, mainly by means of a particular strain of Aspergillus.
  • the invention extends to the tannase prepared by means of this process and to the use of this tannase for the preparation of gallic acid.
  • Tannase an enzyme discovered by Scheele in 1786, exhibits the property of hydrolysing the ester bond as well as the depside bond of tannic acid and glucose.
  • the Applicant Company has found that, surpris ⁇ ingly, the genus Asper ⁇ illus tamarii. in particular the CBS 10413 strain, is characterized by a very highly efficient production of tannase.
  • the said strain is characterized by a high specificity together with a high volumetric activity in respect of tannase during the fermentation process.
  • tannase is obtained by fermentation of Asper ⁇ illus tamarii in a medium con ⁇ taining tannins intended to induce the synthesis of tannase.
  • the fungal mycelium can be isolated from the fermentation medium and employed repeatedly in the form of a tannase enzyme system immobilized on a support, in situ, for the bioconversion of tannins.
  • the biomass can be recycled and reused without addition of any special nutrient agent.
  • the gallic acid obtained has the con ⁇ ventional properties of commercial gallic acids and is in the form of a practically colourless crystalline powder which is soluble in water, in ethanol and in acetone. This compound is employed, inter alia, for the preparation of gallic esters for use in foodstuffs and in pharmaceutical compounds.
  • the starting tannins form a complex mixture of polyphenolic compounds which are present in many plants. They are categorized into condensed tannins and into hydrolysable tannins. Gallotannins are hydrolysable either by an enzyme route or by a chemical route, producing gallic acid and releasing the central molecule of the nucleus (consisting of glucose or guinic acid) .
  • a TU unit is the quantity of tan- nase which hydrolyses one micromole of ester bonds of gallic acid in one minute after incubation under optimum reaction conditions.
  • this technique has been slightly modified as follows; 1 gram of fungal mycelium is incubated in a 1-2% strength solution of tara-tannin at pH 5.5 (0.1 M cit ⁇ rate buffer) for 30 minutes at 30°C. Before and after incubation, 50 ⁇ l of the tannin solution are trans ⁇ ferred with a pipette into 10 ml of 90% ethanol. The drop in the absorption is measured at 310 nm.
  • the enzyme activity can be calculated from a purified tannase standard used as reference.
  • the refer ⁇ ence tannase activity is determined according to Iibuchi et al. Process for the fermentation of Asper ⁇ illus tamarii CBS 10413 with a view to the production of tannase.
  • tannins Since the biosynthesis of tannase constitutes an operation which must be induced, it is appropriate to add tannins to the fermentation media. An addition of tannins to the preculture as well as to the germination medium is recommended in order to obtain fast subsequent growth and production stages.
  • the concentration of tannins in the fermentation liquid can vary between 1 and 30% (weight/volume) , although the germination and preculture media must contain only low levels of tannins (1 to 5% - weight/volume) for the inductio .
  • the compounds resulting from the hydrolysis of the tannin can be metabolized by the mycelium.
  • the fermentation operation can be accelerated by the addition of sugars to the fermentation media.
  • Sucrose, glucose, fructose and its industrial sources, namely molasses and glucose and fructose syrups can be added in concentrations of the order to 2 to 4% (weight/volume) depending on the level of the nitrogen which is fed.
  • a high C/N ratio (of the order of 10 to 15) promotes the production of a highly active biomass.
  • the nitrogen can be fed in the form of inorganic ammonium and nitrate salts such as: (NH 4 ) 2 S0 4 , (NH 4 ) 2 C0 3 , (NH 4 ) 2 HP0 4 , NH 4 C1, NH 4 N0 3 , NO3 and NaN0 3 .
  • inorganic ammonium and nitrate salts such as: (NH 4 ) 2 S0 4 , (NH 4 ) 2 C0 3 , (NH 4 ) 2 HP0 4 , NH 4 C1, NH 4 N0 3 , NO3 and NaN0 3 .
  • K, Mg, Zn, P and S are added to the culture medium in the form of inorganic sulphate or phosphate salts. Folic acid and pantothenic acid may be added as additional growth agents.
  • Tannase-producing fermentation starting with Asper ⁇ illus tamarii CBS 10413 has been investigated in a conventional stirred reactor (Applikon, Biolaffite) .
  • tannase The production of tannase is carried out at 30°C. During the fermentation (in particular during the growth and exponential production stage) , the pH changes rapidly. NaOH (5 N) and H 2 S0 4 (5 N) are added during the fermentation process in order to maintain the pH between 3.5 and 4.5.
  • the fermentation liquid is aerated by blowing in 0.5 to 1 wm of oxygen-saturated air into the cul- ture medium.
  • the rate of agitation is adjusted to the rate of oxygen transfer.
  • the reactor (10 1) is inocu ⁇ lated with 10% (v/v) of preculture or of germination medium, which, in its turn, is prepared from a fresh spore suspension (5% volume/volume) collected from a malted agar medium.
  • the composition of the liquid medium is identical for the various fermentation steps.
  • the biomass is separated from the fer ⁇ mentation broth by vacuum filtration. The biomass is washed and kept at -18°C.
  • K 2 HP0 1.0 MgS0 4 .7H 2 0 0.5 ZnS0 4 .7H 2 0 0.5 NaCl 1.0 is inoculated with a 10% (volume/volume) suspension of the said spores.
  • the germination medium is adjusted to pH 5 before sterilisation. After 16 to 24 hours' incu ⁇ bation of the culture in flasks on a mechanical shaker at 30° , the germination medium is transferred into the agitated reactor containing 5 1 of preculture (at a concentration of 10% volume/volume), the total volume of the reactor being 10 1.
  • the preculture medium (g/1) is made up as follows: tannins 50 glucose 30
  • the fermentation reactor (containing the medium) is sterilized at 121 oC for 30 minutes. Tannin is added to the medium after sterilization to avoid chemical hydrolysis of the induction molecules.
  • the preculture is incubated for 48 h at 30 o .
  • An aeration of 1 wm and an agitation of 400 rev/min are maintained.
  • Variations in the pH values of the fermentation medium of between 3.0 and 4.5 are permitted. Adjustments of pH are performed us ⁇ ing sulphuric acid and sodium hydroxide.
  • the preculture is transferred into a 100-litre fermenter (70 1 working capacity) .
  • the composition of the medium, as well as the preparation and the sterilization, are identical with those shown for the preculture.
  • the fermentation is conducted at 30°C, 1 w , 300 rev/min and a pH of 3.0 to 4.5. Frothing is controlled by adding a silicone- based foam suppressor. After 38 h of incubation the fermentation is stopped; all the available carbon sources have been metabolized. The fermentation medium is filtered and washed with demineralized water. The biomass (1.125 kg of dried cell mass) is stored at - 18 o . The total intercellular activity was 1.125 x IO 6 units. The extracellular activity was approximately 1.5 x 10 ⁇ or 12% of the total volumetric activity.
  • Germination medium (spore suspension 5%)

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

La production de tannase se caractérise en ce que l'on prépare cette dernière par fermentation d'Aspergillus tamarii, notamment la souche CBS 10413. On obtient la tannase avec un rendement de production élevé et on l'emploie à la production d'acide galliqueThe production of tannase is characterized in that the latter is prepared by fermentation of Aspergillus tamarii, in particular the strain CBS 10413. The tannase is obtained with a high production yield and it is used for the production of gallic acid

Description

Process for the preparation of tannase intended for the production of σallic acid, with the aid of an Asperσillus culture.
SUBJECT OF THE INVENTION
The present invention relates to a process for the preparation of a tannase from microorganisms, mainly by means of a particular strain of Aspergillus.
The invention extends to the tannase prepared by means of this process and to the use of this tannase for the preparation of gallic acid.
SUMMARY OF THE STATE OF THE ART
Tannase, an enzyme discovered by Scheele in 1786, exhibits the property of hydrolysing the ester bond as well as the depside bond of tannic acid and glucose.
The preparation of tannase from various types of Asperσillus or Penicillium has already been described.
Freudenberg et al. (Z.Physiol.Chem. 164, 262, 1927); have prepared tannase from Asperσillus niσer. and
K. Yamada et al. (J. Ferment. Tech., 45, 233, 1967); (Agr. Biol. Chem. 31, 513, 1967) have pre¬ pared tannase from Asperσillus orvzae.
The use of Asperσillus oryzae is also described in Chemical Abstracts, vol.83, No. 9, 1 September 1975, page 508, abstract no. 76990g, Columbus, Ohio, US; and JP-A-75/25,786 (Kikkoman Shoyu Co., Ltd) 18-03-1975.
Document GB-A-1,249,932 (Tenco Brooke Bond Ltd) describes the use of a tannase derived from Asperσillus, niσer, oryzae or flavus for the treatment of "tea cream".
CHARACTERISTIC FEATURES OF THE INVENTION
The Applicant Company has found that, surpris¬ ingly, the genus Asperσillus tamarii. in particular the CBS 10413 strain, is characterized by a very highly efficient production of tannase. The said strain is characterized by a high specificity together with a high volumetric activity in respect of tannase during the fermentation process. According to the invention, tannase is obtained by fermentation of Asperσillus tamarii in a medium con¬ taining tannins intended to induce the synthesis of tannase. The fungal mycelium can be isolated from the fermentation medium and employed repeatedly in the form of a tannase enzyme system immobilized on a support, in situ, for the bioconversion of tannins. The biomass can be recycled and reused without addition of any special nutrient agent. The gallic acid obtained has the con¬ ventional properties of commercial gallic acids and is in the form of a practically colourless crystalline powder which is soluble in water, in ethanol and in acetone. This compound is employed, inter alia, for the preparation of gallic esters for use in foodstuffs and in pharmaceutical compounds. The starting tannins form a complex mixture of polyphenolic compounds which are present in many plants. They are categorized into condensed tannins and into hydrolysable tannins. Gallotannins are hydrolysable either by an enzyme route or by a chemical route, producing gallic acid and releasing the central molecule of the nucleus (consisting of glucose or guinic acid) .
It is appropriate to note that it is already known and proven that fermentation processes can succeed or fail, depending on the suitable or unsuitable choice of an appropriate strain. The Applicant Company has noticed, in particular, that among the many cultures selected and isolated on agar culture plates and having received a preparation of tannins, very few compounds exhibit a highly efficient production of tannase in a liquid culture medium.
The particular choice of the Asperσillus tamarii species for the preparation of tannase makes it possible, surprisingly, to obtain tannase production efficiency characteristics which are superior to the result obtained according to the state of the art.
PREFERRED EMBODIMENTS OF THE INVENTION The invention will be described in greater detail with the aid of specific examples of the uses of individual strains according to the invention of the Asperσillus genus.
Measurement of the activity in respect of tannase The activity in respect of tannase can be measured by various techniques which have already been described in literature. A particular technique has been developed by Iibuchi et al. (Agr.Biol.Chem. 31, 513, 1967). To do this, the hydrolysis of a standardized solution of tannins is followed by spectrophotometry and is directly related to the activity in respect of tannase. The activity in respect of tannase is expressed in terms of a tannase unit (tannase units = TU) . A TU unit is the quantity of tan- nase which hydrolyses one micromole of ester bonds of gallic acid in one minute after incubation under optimum reaction conditions. As described initially in the above reference, this technique has been slightly modified as follows; 1 gram of fungal mycelium is incubated in a 1-2% strength solution of tara-tannin at pH 5.5 (0.1 M cit¬ rate buffer) for 30 minutes at 30°C. Before and after incubation, 50 μl of the tannin solution are trans¬ ferred with a pipette into 10 ml of 90% ethanol. The drop in the absorption is measured at 310 nm.
The enzyme activity can be calculated from a purified tannase standard used as reference. The refer¬ ence tannase activity is determined according to Iibuchi et al. Process for the fermentation of Asperσillus tamarii CBS 10413 with a view to the production of tannase.
Since the biosynthesis of tannase constitutes an operation which must be induced, it is appropriate to add tannins to the fermentation media. An addition of tannins to the preculture as well as to the germination medium is recommended in order to obtain fast subsequent growth and production stages. The concentration of tannins in the fermentation liquid can vary between 1 and 30% (weight/volume) , although the germination and preculture media must contain only low levels of tannins (1 to 5% - weight/volume) for the inductio .
In the absence of additional carbon sources, the compounds resulting from the hydrolysis of the tannin (gallic acid, glucose and quininic acid) can be metabolized by the mycelium. The fermentation operation can be accelerated by the addition of sugars to the fermentation media. Sucrose, glucose, fructose and its industrial sources, namely molasses and glucose and fructose syrups can be added in concentrations of the order to 2 to 4% (weight/volume) depending on the level of the nitrogen which is fed. A high C/N ratio (of the order of 10 to 15) promotes the production of a highly active biomass. The nitrogen can be fed in the form of inorganic ammonium and nitrate salts such as: (NH4)2S04, (NH4)2C03, (NH4)2HP04, NH4C1, NH4N03, NO3 and NaN03.
Organic nitrogenous sources (beef extract, soya peptone, peptone, slop, dried soluble distillation residues, molasses extract liquor, blood flour, soya flour) cannot be employed because, in the case of all these compounds, a precipitation reaction is observed with tannins. Asperσillus tamarii, in particular the CBS
10413 strain, is relatively undemanding in respect of minerals. K, Mg, Zn, P and S are added to the culture medium in the form of inorganic sulphate or phosphate salts. Folic acid and pantothenic acid may be added as additional growth agents.
EXAMPLES
The invention will be described in greater de¬ tail with reference to two particular examples of implementation of the invention:
Example 1. Tannase fermentation starting with
Asperσillus tamarii CBS 10413 Composition of the medium (g/1)
Tannins 10 to 300
Sucrose 30 N03 8.5
KC1 1.0
(NH4)2HP04 1.0
MgS04.7H20 0.5
ZnS0 .7H20 0.3
Folic acid 1 mg/1
Pantothenic acid lmg/1
Tannase-producing fermentation starting with Asperσillus tamarii CBS 10413 has been investigated in a conventional stirred reactor (Applikon, Biolaffite) .
The production of tannase is carried out at 30°C. During the fermentation (in particular during the growth and exponential production stage) , the pH changes rapidly. NaOH (5 N) and H2S04 (5 N) are added during the fermentation process in order to maintain the pH between 3.5 and 4.5.
The fermentation liquid is aerated by blowing in 0.5 to 1 wm of oxygen-saturated air into the cul- ture medium. The rate of agitation is adjusted to the rate of oxygen transfer. The reactor (10 1) is inocu¬ lated with 10% (v/v) of preculture or of germination medium, which, in its turn, is prepared from a fresh spore suspension (5% volume/volume) collected from a malted agar medium. The composition of the liquid medium is identical for the various fermentation steps. At the end of the fermentation (exhaustion of the car¬ bon substrate) the biomass is separated from the fer¬ mentation broth by vacuum filtration. The biomass is washed and kept at -18°C. An intercellular tannase activity of 15,000 units/l can be obtained. Approxi¬ mately 3,000 units/1 are secreted into the fermentation broth. Example 2. Sequence for producing tannase from Asperσillus tamarii CBS 10413 The preparative sequence is illustrated in table I, appended. The inoculum is prepared as follows: spores originating from an initial spore culture, which have been grown on agar/malt extract (Roux flasks) are transferred into a physiological saline solution (150ml). The germination medium (g/1) made up as follows: tannins 50 glucose 30 KN03 8.5
K2HP0 1.0 MgS04.7H20 0.5 ZnS04.7H20 0.5 NaCl 1.0 is inoculated with a 10% (volume/volume) suspension of the said spores. The germination medium is adjusted to pH 5 before sterilisation. After 16 to 24 hours' incu¬ bation of the culture in flasks on a mechanical shaker at 30° , the germination medium is transferred into the agitated reactor containing 5 1 of preculture (at a concentration of 10% volume/volume), the total volume of the reactor being 10 1. The preculture medium (g/1) is made up as follows: tannins 50 glucose 30
NH4C1 3.75 KC1 1.0
(NH4)2HP04 1.0
MgSO4.7H20 0.5
ZnS04.7H20 0.5
NaCl 1.0 Folic acid 1 mg/1 pantothenic acid 1 mg/1
The fermentation reactor (containing the medium) is sterilized at 121oC for 30 minutes. Tannin is added to the medium after sterilization to avoid chemical hydrolysis of the induction molecules.
After inoculation, the preculture is incubated for 48 h at 30o . An aeration of 1 wm and an agitation of 400 rev/min are maintained. Variations in the pH values of the fermentation medium of between 3.0 and 4.5 are permitted. Adjustments of pH are performed us¬ ing sulphuric acid and sodium hydroxide.
After 36 hours' incubation, the preculture is transferred into a 100-litre fermenter (70 1 working capacity) . The composition of the medium, as well as the preparation and the sterilization, are identical with those shown for the preculture.
The fermentation is conducted at 30°C, 1 w , 300 rev/min and a pH of 3.0 to 4.5. Frothing is controlled by adding a silicone- based foam suppressor. After 38 h of incubation the fermentation is stopped; all the available carbon sources have been metabolized. The fermentation medium is filtered and washed with demineralized water. The biomass (1.125 kg of dried cell mass) is stored at - 18o . The total intercellular activity was 1.125 x IO6 units. The extracellular activity was approximately 1.5 x 10^ or 12% of the total volumetric activity.
Table 1 (Example 2)
Agar + malt extract
30°C, 3 days
Asperσillus tamarii CBS 10413 spores
physiological saline solution (0.75% NaCl) spore suspension
Germination medium (spore suspension 5%)
Preculture - 10 1
Fermenter - 100 1
Filtration
Filtrate
Biomass - tannase

Claims

Claims
1. Process for the preparation of tannase, charac¬ terized in that the genus Asperσillus tamarii, in par¬ ticular the CBS 10413 strain, is employed for the production.
2. Process according to Claim 1, characterized in that the tannase is obtained by fermentation of Asperσillus tamarii in a medium containing tannins intended to induce the synthesis of tannase, it being possible for the fungal mycelium to be isolated from the fermentation medium and employed repeatedly in the form of a tannase enzyme system immobilized on a sup¬ port, in situ, for the bioconversion of tannins and it being possible for the biomass to be recycled and re¬ used without addition of any special nutrient agent.
3. Process according to Claim 2, characterized in that tannins are added to the preculture and to the germination medium to obtain fast subsequent growth and production stages.
4. Process according to Claim 3, characterized in that the concentration of tannins in the fermentation liquid is between 1 and 30% (weight/volume), while the germination and preculture media contain only low levels of tannins (1 to 5% weight/volume) for the induction.
5. Process according to any one of Claims 1 to 4, characterized in that the fermentation operation is accelerated by the addition of sugars to the fermentation media.
6. Process according to Claim 5, characterized in that sucrose, glucose, fructose or their industrial sources, namely molasses and glucose and fructose syrups, are added in concentrations of the order of 2 to 4% (weight/volume) depending on the level of nitrogen which is fed.
7. Process according to Claim 4 or 5, character¬ ized in that a high C/N ratio (of the order of 10 to 15) is produced to promote the production of a highly active biomass.
8. Process according to Claim 7, characterized in that the nitrogen is fed in the form of inorganic ammo¬ nium and nitrate salts such as : (NH4) S04, (NH4)2C03, (NH )2HP0 , NH4C1, NH N03, KNO3 and NaN03-
9. Process according to any one of Claims 2 to 8, characterized in that K, Mg, Zn, P and S minerals are added to the culture medium in the form of sulphate or inorganic phosphate salts.
10. Process according to any one of Claims 2 to 9, characterized in that folic acid and pantothenic acid are added as additional growth agents.
11. Tannase obtained by the process of any one of Claims 1 to 10.
12. Use of tannase according to Claim 11 for the production of gallic acid.
EP19890904502 1988-04-19 1989-04-17 Process for the preparation of tannase intended for the production of gallic acid, with the aid of an aspergillus culture Pending EP0402416A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
LU87203 1988-04-19
LU87203A LU87203A1 (en) 1988-04-19 1988-04-19 PROCESS FOR THE PREPARATION OF TANNASE FOR THE PRODUCTION OF GALLIC ACID USING A CULTURE OF ASPERGILLUS, TANNASE THUS OBTAINED AND USE THEREOF FOR THE PRODUCTION OF GALLIC ACID

Publications (1)

Publication Number Publication Date
EP0402416A1 true EP0402416A1 (en) 1990-12-19

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EP89870053A Withdrawn EP0339011A1 (en) 1988-04-19 1989-04-17 Process for the preparation of tannase intended for the production of gallic acid, with the aid of an aspergillus culture
EP19890904502 Pending EP0402416A1 (en) 1988-04-19 1989-04-17 Process for the preparation of tannase intended for the production of gallic acid, with the aid of an aspergillus culture

Family Applications Before (1)

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LU (1) LU87203A1 (en)
WO (1) WO1989010400A1 (en)

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DK42092D0 (en) * 1992-03-27 1992-03-27 Novo Nordisk As
WO2003000912A1 (en) * 2001-06-22 2003-01-03 Indian Institute Of Technology A process for the preparation of gallic acid by co-culture
CN102719413A (en) * 2012-04-28 2012-10-10 中山大学 Novel tannase and application thereof
CN103589758B (en) * 2013-11-12 2015-07-22 遵义市倍缘化工有限责任公司 Method for preparing gallic acid by using gallnut tannin as raw material and utilizing fermentation separation coupling technology
CN112852780B (en) * 2021-02-05 2023-03-24 泸州品创科技有限公司 Monascus flavus and application thereof in preparing tannase and degrading tannin
CN116515647B (en) * 2023-06-25 2023-09-12 华南农业大学 Aspergillus flavus and application thereof in preparing tannase and/or degrading tannin

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Publication number Priority date Publication date Assignee Title
GB1249932A (en) * 1969-07-10 1971-10-13 Tenco Brooke Bond Ltd Enzymatic solubilisation of tea cream

Non-Patent Citations (1)

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Title
See references of WO8910400A1 *

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EP0339011A1 (en) 1989-10-25
LU87203A1 (en) 1989-11-14

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