CN113969295B - Regulation and control method for reducing impurity content in beta-phenethyl alcohol fermentation liquor - Google Patents
Regulation and control method for reducing impurity content in beta-phenethyl alcohol fermentation liquor Download PDFInfo
- Publication number
- CN113969295B CN113969295B CN202010728363.2A CN202010728363A CN113969295B CN 113969295 B CN113969295 B CN 113969295B CN 202010728363 A CN202010728363 A CN 202010728363A CN 113969295 B CN113969295 B CN 113969295B
- Authority
- CN
- China
- Prior art keywords
- beta
- phenethyl alcohol
- fermentation
- dissolved oxygen
- carbon source
- 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.)
- Active
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
- C12P7/22—Preparation of oxygen-containing organic compounds containing a hydroxy group aromatic
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C29/00—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom not belonging to a six-membered aromatic ring
- C07C29/74—Separation; Purification; Use of additives, e.g. for stabilisation
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C29/00—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom not belonging to a six-membered aromatic ring
- C07C29/74—Separation; Purification; Use of additives, e.g. for stabilisation
- C07C29/76—Separation; Purification; Use of additives, e.g. for stabilisation by physical treatment
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C29/00—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom not belonging to a six-membered aromatic ring
- C07C29/74—Separation; Purification; Use of additives, e.g. for stabilisation
- C07C29/76—Separation; Purification; Use of additives, e.g. for stabilisation by physical treatment
- C07C29/80—Separation; Purification; Use of additives, e.g. for stabilisation by physical treatment by distillation
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
Landscapes
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Distillation Of Fermentation Liquor, Processing Of Alcohols, Vinegar And Beer (AREA)
Abstract
The invention discloses a regulating and controlling method for reducing impurity content in beta-phenethyl alcohol fermentation liquor, which comprises controlling dissolved oxygen and carbon source regulation according to thallus concentration, and feeding substrate and nitrogen source according to alkali addition in a fermentation tank.
Description
Technical Field
The invention belongs to the technical field of biological fermentation, and particularly relates to a regulating and controlling method for reducing impurity content in beta-phenethyl alcohol fermentation liquid.
Description of the background
Beta-phenethyl alcohol is a natural spice widely existing in plants such as roses/jasmine, is a second largest essence spice with the use amount inferior to vanillin, and is widely used in the fields of daily chemicals, foods, medicines and the like.
At present, the synthesis of beta-phenethyl alcohol is mainly divided into two modes of chemical synthesis and biological synthesis, and the chemical synthesis of beta-phenethyl alcohol has certain difference with the fragrance of natural beta-phenethyl alcohol due to the existence of some impurities which are difficult to remove, so that the application of the beta-phenethyl alcohol in the high-end field is limited. In recent years, with the improvement of living standard, people are more and more in favor of green and healthy living modes, and the pursuit of products is more and more in favor of natural products. The production method of the natural beta-phenethyl alcohol mainly comprises two methods of extraction and microbial fermentation. The extraction method has low efficiency and high cost, and is difficult to realize industrial production. The microbial method adopts the substrate from fermentation source for production, retains the natural characteristics, has low production cost and has the potential of large-scale production. However, in the process of synthesizing beta-phenethyl alcohol by a microbiological method, thallus is metabolized to generate impurities such as peculiar smell and heteroacid (butyric acid, valeric acid and isovaleric acid), and the substances generally have special pungent smell, and if the substances are high in content, the substances remain in a separated product and seriously influence the fragrance quality of the beta-phenethyl alcohol. In addition, when the acid impurities are more, the acid impurities can further react with beta-phenethyl alcohol to generate ester impurities such as phenethyl acetate, phenethyl butyrate and the like, the boiling point of the ester impurities is close to that of the beta-phenethyl alcohol, and the ester impurities are difficult to remove in the later period, so that the purity of the product is affected. While there are few current studies focused on this problem, most of them focus on how to increase the yield of beta-phenylethanol. Only chinese patent application CN104498539a discloses a method for preparing natural perfume 2-phenethyl alcohol by batch fermentation with L-phenylalanine as substrate, glucose as carbon source, and antioxidant with a certain concentration under the alternating oxidation degradation system of glucose and ethanol. The method adopts a method of adding the antioxidant at one time, can reduce the content of impurities such as butanol, isobutyric acid, butyric acid, isovaleric acid and the like in the biological conversion process, and further improves the fragrance quality of the 2-phenethyl alcohol. Chinese patent application CN102392055a mentions that ethanol (ethanol is used as a carbon source) is degraded by oxidation to reduce the content of organic impurities in the fermentation process, but at present, the beta-phenylethanol polysaccharide produced by fermentation uses saccharides as a carbon source, and fewer strains capable of utilizing ethanol are available.
One key indicator of a fragrance product is the aroma quality of the final product, and the impurity content and the difficulty of later removal play a key role in the quality of the product. Thus, solving the accumulation of impurity content during fermentation is a key to improving the aroma quality of the final product, and there are few studies focused on this problem.
Disclosure of Invention
The invention provides a regulating and controlling method for reducing the impurity content in beta-phenethyl alcohol fermentation liquor, reducing the generation of by-product hetero acid and hetero alcohol, reducing the separation and purification difficulty and optimizing the product aroma so as to be more similar to a natural product, aiming at solving the problems that impurities (hetero acid, hetero alcohol and corresponding esters) generated in the fermentation process and the quality of the product are difficult to remove in the later separation and purification process.
In order to achieve the above purposes and the above technical effects, the technical scheme adopted by the invention is as follows:
a regulation method for reducing impurity content in beta-phenethyl alcohol fermentation liquor uses yeast as production strain, adds adsorbent in fermentation tank and phenylalanine as substrate to make fermentation so as to prepare beta-phenethyl alcohol.
In a specific embodiment, the yeast is one of kluyveromyces marxianus, saccharomyces cerevisiae, yarrowia lipolytica, and the adsorbent is a macroporous resin; preferably, the macroporous resin adsorbent is any one of HZ818, SP825 and H103, and the adding amount of the macroporous resin adsorbent is 10-35% of the initial fermentation volume; the temperature in the fermentation tank is 25-37 ℃, the pH is 3.5-6, and the ventilation is kept at 0.5-3vvm.
In a preferred embodiment, sufficient carbon source and dissolved oxygen are provided to stimulate the growth of the cells during the cell growth phase such that the yeast concentration reaches the target biomass; preferably, the carbon source supplementing strategy is to maintain the concentration of the carbon source in the tank to be more than 2g/L and supplement the carbon source when the concentration of the carbon source in the tank is less than 2 g/L; the dissolved oxygen regulation strategy is to correlate the rotating speed with dissolved oxygen, so that the volume fraction of the dissolved oxygen in the fermentation tank is kept between 10% and 50%; more preferably, the dissolved oxygen volume fraction in the fermentation tank is kept between 20% and 40%, the single additional amount of the additional carbon source is 50g/L (based on the total volume of fermentation), and the additional carbon source is a carbon source solution with the mass percentage of 60%. The carbon source is any one of glucose, sucrose and maltose.
Wherein the yeast is required to accumulate to a biomass of 5-40g/L (calculated by dry weight of thallus); preferably, the yeast should accumulate to a biomass (based on dry weight of the cells) of 15-35 g/L.
In a preferred embodiment, in the product synthesis stage, after the yeast accumulates to a certain concentration, substrate phenylalanine is added, dissolved oxygen is adjusted according to the change of the concentration of yeast thallus, and a constant-rate feed is adopted for carbon source regulation; preferably, when the fluctuation range of the biomass of the thalli is larger than 5g/L (based on the dry weight of the thalli), the dissolved oxygen regulation strategy is to correlate the rotating speed with the dissolved oxygen, so that the volume fraction of the dissolved oxygen in the fermentation tank is kept between 10% and 50%; more preferably, the dissolved oxygen volume fraction in the fermenter is maintained at 20% to 40%; when the fluctuation range of the biomass of the thalli is smaller than 5g/L (calculated by the dry weight of the thalli), the fixed rotating speed is 400-700rpm; the carbon source supplementing rate is 10-20 g/L.h (the volume is calculated by the total volume of initial fermentation), and the supplementing carbon source is a carbon source solution with the mass percent of 60%. The carbon source is any one of glucose, sucrose and maltose.
Wherein, in the product synthesis stage, the change of pH and alkali addition amount is monitored in real time, and the supplementation of phenylalanine and nitrogen sources is adjusted according to the alkali addition rate; preferably, when the rate of addition of base is less than 5g/h (based on 100% base), only phenylalanine is added; and when the alkali adding rate is more than 5g/h, adding the mixed solution of phenylalanine and the nitrogen source.
Wherein the alkali is any one of sodium hydroxide, potassium hydroxide and ammonia water, and the nitrogen source is an inorganic nitrogen source; preferably, the inorganic nitrogen source is one or more of urea, ammonium sulfate, ammonium chloride and sodium nitrate, and the concentration of the alkali is 5-20% by mass; more preferably, the concentration of the phenylalanine only is 10-20g/L (based on the volume of the feed); the mixed solution of phenylalanine and nitrogen source is supplemented, the concentration of phenylalanine is 5-10g/L, the concentration of nitrogen source is 0.5-5g/L (calculated by the volume of the supplementing material), and the supplementing flow rate of phenylalanine and the mixed solution is 10-20mL/h.
Wherein, in the product synthesis stage, the biological quality of the yeast is controlled to be 20-50g/L, preferably 20-35g/L (calculated by dry weight of thallus).
In a specific embodiment, the separation and purification process comprises the steps of filtering fermentation liquor obtained after fermentation is finished, separating adsorbent from the fermentation liquor, analyzing the adsorbent, and carrying out reduced pressure distillation and rectification on the analyzed liquor to finally obtain the pure beta-phenethyl alcohol with the purity of more than 99.5%.
Wherein the filtering condition is 0.1mPa, the pore diameter of the filter cloth is 1000-1500 meshes, the resolving solvent is ethanol, methanol or ethyl acetate, the reduced pressure distillation condition is 50-80 ℃, the pressure is-0.1 to-0.5 MPa, and the rectification condition is-0.01 to-0.05 MPa.
Compared with the prior art, the invention has the following beneficial effects:
(1) In the growth stage of the thalli, the invention provides sufficient dissolved oxygen and carbon source, ensures that the thalli can be rapidly accumulated to reach the target concentration, and provides a large amount of thalli for the generation of beta-phenethyl alcohol; in the product synthesis stage, the growth speed of the thalli is slowed down by controlling dissolved oxygen and a carbon source, and the final biomass is kept stable, so that the substrate conversion is mainly carried out in the stage, and the efficient synthesis of beta-phenethyl alcohol is facilitated.
(2) In the product synthesis stage, when the pH value in the fermentation tank is changed drastically, the acid production is obvious, and the alkali supplementing rate is accelerated. When more acidic substances are present, the difficulty of separation and purification in the later stage is increased, and the impurities possibly react with beta-phenethyl alcohol further to generate ester impurities, and the boiling point of the impurities is close to that of the beta-phenethyl alcohol, so that the impurities are difficult to remove in the later stage. The invention well solves the problem by adjusting the feed supplement through the change of the alkali adding rate, inhibits the generation of the mixed acid, reduces the impurity in the fermentation liquor, reduces the separation difficulty and saves the production cost.
(3) The method is adopted for fermentation, the thallus metabolism is mainly used for product synthesis, the impurity content is low, and the production efficiency is high. After separation and purification in the later stage, the purity of the product can reach more than 99.5%, and the fragrance of the product is optimized so that the product is more similar to a natural product.
Drawings
FIG. 1 is a gas chromatogram of a purified beta-phenethyl alcohol product after separation and purification.
Detailed Description
The following examples will further illustrate the method provided by the present invention for a better understanding of the technical solution of the present invention, but the present invention is not limited to the examples listed but should also include any other known modifications within the scope of the claims of the present invention.
A regulation and control method for reducing impurity content in beta-phenethyl alcohol fermentation liquor comprises the steps of using yeast as a production strain, adding an adsorbent into a fermentation tank, fermenting in a certain range by regulating and controlling temperature, pH and ventilation, and adopting the following dissolved oxygen and feeding strategies: the adding amount of dissolved oxygen and carbon source is regulated and controlled according to the concentration of thallus in the fermentation tank, and the adding amount of substrate and nitrogen source is feedback controlled according to the adding amount of alkali in the fermentation tank, so that the generation of byproduct impurities is reduced, and the pure beta-phenethyl alcohol is obtained after separation and purification.
Specifically, in different stages of fermentation, dissolved oxygen and carbon source supplementing amounts are regulated and controlled according to the concentration of thalli in the fermentation tank respectively, and substrate and nitrogen source supplementing amounts are feedback controlled according to the alkali adding amount in the fermentation tank, so that the technical effect of reducing the generation of byproduct impurities is achieved.
(1) And a thallus growth stage: providing sufficient carbon source and dissolved oxygen to stimulate the growth of thallus so that the concentration of yeast reaches the target biological quality.
(2) Product synthesis stage: when the yeast is accumulated to a certain concentration, a substrate phenylalanine is added, dissolved oxygen is adjusted according to the change of the thallus concentration, and the carbon source is regulated and controlled by adopting constant-rate feed. The pH and the alkali addition amount are monitored in real time, and the addition of phenylalanine and nitrogen sources is regulated according to the alkali addition rate. This stage controls the biomass of the yeast within a suitable range.
(3) And (3) separating and purifying: and filtering the fermentation liquor obtained after the fermentation is finished, separating the adsorbent from the fermentation liquor, analyzing the adsorbent, and carrying out reduced pressure distillation and rectification on the analysis liquor to finally obtain the beta-phenethyl alcohol pure product.
The method provides sufficient dissolved oxygen and carbon source in the growth stage of the thalli, and ensures that the thalli grow rapidly to reach the target biological quality. In the product accumulation stage, due to the addition of phenylalanine, the phenylalanine can serve as a substrate and supplement a part of nitrogen source, and can stimulate the growth of thalli, so that when the biomass of the thalli is greatly changed, sufficient dissolved oxygen is provided to maintain good growth of the thalli; when the biomass of the thalli is stable, the rotating speed is fixed, and the constant oxygen demand is maintained, so that the thalli is not accumulated any more. Meanwhile, the carbon source is regulated and controlled by adopting constant-rate feed supplement, so that the bacterial volume is gradually reduced and kept stable, and the synthesis of the product is mainly ensured at the stage. The alkali adding rate in the fermentation process reflects the rate of producing the hybrid acid, under the aerobic condition, the concentration of the carbon source is constant, and the hybrid acid can be produced when the amino acid is the only nitrogen source, so that when the alkali adding rate is accelerated, a certain inorganic nitrogen source is supplied, and meanwhile, the supply concentration of phenylalanine is reduced, and the production of the hybrid acid can be inhibited. The impurities in the fermentation liquor are reduced, the separation difficulty is reduced, the production cost is saved, and the high-purity product with the purity of more than 99.5% can be obtained after further treatment in the later stage.
The yeast is any one of Kluyveromyces marxianus, saccharomyces cerevisiae and yarrowia lipolytica, and is preferably Saccharomyces cerevisiae. The fermentation temperature is controlled at 25-37 ℃, the pH value is 3.5-6, and ventilation is kept at 0.5-3vvm.
In the bacterial growth stage, the dissolved oxygen regulation strategy is to correlate the rotating speed with the dissolved oxygen, so that the dissolved oxygen is kept between 10% and 50%, and preferably kept between 20% and 40%. The related operation method is a common operation method in the fermentation field, and the related rotation speed can be selected by recording the dissolved oxygen in the matched software of the fermentation tank, and only the corresponding parameters are required to be set. In particular, the rotational speed is associated with dissolved oxygen, for example, the rotational speed is controlled between 200-1000rpm, such that the dissolved oxygen content in the fermenter is maintained between 10% -50%. When the dissolved oxygen content is larger, the rotating speed is correspondingly reduced; conversely, the rotational speed will increase accordingly.
In the thallus growth stage, the carbon source supplementing strategy is to maintain the carbon source concentration in the tank to be greater than 2g/L and to supplement carbon source once less than 2 g/L. The carbon source content in the fermentation tank is achieved through DNS chromogenic method period detection, when the carbon source concentration in the tank is detected to be less than 2g/L, the carbon source is supplemented, the single supplementing amount of the carbon source is 50g/L (calculated by total fermentation volume), and the carbon source supplementing amount is a carbon source solution with the mass percentage of 60%. The carbon source is any one of glucose, sucrose and maltose, but is not limited thereto, and other commonly used carbon sources should be considered as equivalent substitutes of the present invention and should be within the scope of the present invention. Wherein the total volume of the fermentation is equal to the sum of the total volume of the culture medium, the inoculation volume and the resin added in the fermentation tank, and in the actual fermentation process, the total volume of the fermentation is usually calculated by referring to the preset value of the fermentation scheme, wherein various materials are added in proportion based on the total volume of the fermentation.
In the thallus growth stage, the thallus is ensured to grow fast to reach the target biological quality by providing sufficient dissolved oxygen and carbon source. Specifically, the yeast is allowed to accumulate to a biomass (based on dry weight of the cells) of at least 5-40 g/L; preferably, the biomass of the yeast to be accumulated at this stage is preferably 15-35g/L (based on dry weight of the cells).
In the product synthesis stage, the dissolved oxygen control method comprises the following steps: when the fluctuation range of the biomass of the thalli is larger than 5g/L (calculated by the dry weight of the thalli), the dissolved oxygen regulation strategy is the same as that of the thalli in the growth stage, namely the rotating speed is related to the dissolved oxygen, so that the dissolved oxygen is kept between 10% and 50%, and preferably kept between 20% and 40%. When the fluctuation range of the biomass of the thalli is smaller than 5g/L (calculated by the dry weight of the thalli), the fixed rotating speed is 400-700rpm; the carbon source supplementing rate is 10-20 g/L.h (the volume is calculated by the total volume of initial fermentation), and the supplementing carbon source is a carbon source solution with the mass percent of 60%. The carbon source is any one of glucose, sucrose and maltose.
In the product synthesis stage of the invention, the strategy for supplementing the substrate phenylalanine and the nitrogen source is as follows: when the alkali addition rate is less than 5g/h (based on 100% alkali), only phenylalanine is added. Wherein the base mass calculated as 100% base means that the base mass calculated as 100% base without water, for example 5g/h is 5g sodium hydroxide without water added per hour; if an alkaline solution is added, this is converted to the corresponding non-aqueous 100% alkali, for example, at an alkaline rate of 5g/h, if 5% sodium hydroxide solution is added, 100g per hour of this solution is required, and if 20% sodium hydroxide solution is added, 25g per hour of this solution is required. When the alkali adding rate is more than 5g/h, adding the mixed solution of phenylalanine and nitrogen source. Wherein the nitrogen source is an inorganic nitrogen source; specifically, the inorganic nitrogen source is one or more of urea, ammonium sulfate, ammonium chloride and sodium nitrate; the alkali is one of sodium hydroxide, potassium hydroxide and ammonia water; preferably, the concentration of the alkali is 5-20% by mass. When only phenylalanine is added, the feeding concentration of the phenylalanine is 10-20g/L (based on the feeding volume); when the mixed solution of phenylalanine and nitrogen source is added, the concentration of phenylalanine is 5-10g/L, and the concentration of nitrogen source is 0.5-5g/L (based on the volume of the feed). Preferably, the additional flow rate of the phenylalanine and the mixed solution is 10-20mL/h.
In the product synthesis stage, after yeast accumulates to a certain concentration, substrate phenylalanine is added, dissolved oxygen is adjusted according to the change of the concentration of yeast thallus, and constant-rate feed is adopted for carbon source regulation; and simultaneously, the adding amount of the substrate and the nitrogen source is feedback controlled according to the adding amount of the alkali in the fermentation tank. At this stage, the biomass of the yeasts according to the invention is controlled to be 20-50g/L, preferably 20-35g/L (based on the dry weight of the cells).
The separation and purification process of the invention is to filter the fermentation liquor obtained after fermentation, separate the adsorbent from the fermentation liquor, analyze the adsorbent, decompress and distill and rectify the analysis liquor to finally obtain the pure beta-phenethyl alcohol with purity more than 99.5%, optimize the product aroma and make the product aroma more similar to the natural product. The invention adopts an in-situ separation and purification technology, and an adsorbent is added into a fermentation tank, wherein the adsorbent is used for adsorbing beta-phenethyl alcohol, so that the inhibition effect of the beta-phenethyl alcohol is solved. Specifically, the adsorbent is macroporous resin; preferably, the adsorbent macroporous resin is any one of HZ818, SP825 and H103; more preferably, the adsorbent macroporous resin is added in an amount of 10% -35% of the initial fermentation volume.
Specifically, the filtering condition is 0.1mPa, the pore diameter of the filter cloth is 1000-1500 meshes, the resolving solvent is ethanol, methanol or ethyl acetate, the reduced pressure distillation condition is 50-80 ℃, the pressure is-0.1 to-0.5 MPa, and the rectification condition is-0.01 to-0.05 MPa.
The invention is further illustrated by the following more specific examples, which do not limit the scope of the invention in any way.
Raw materials:
phenylalanine is purchased from the net of alaa Ding Shiji, 99% or more;
macroporous resin SP825 was purchased from Mitsubishi chemical corporation (ORgano), specific surface area 1000;
macroporous resin SP850 was purchased from mitsubishi chemical (ORgano), specific surface area 1000;
macroporous resin H103 is purchased from Nankai university chemical plant, and the specific surface area is 1000-1100;
the fermenter was purchased from infors and had a volume of 10L.
The strain is yeast, wherein Kluyveromyces marxianus is purchased from Guangdong microbiological institute, the number is GIM2.119, saccharomyces cerevisiae is autonomously screened, and the Kluyveromyces marxianus is preserved to China Center for Type Culture Collection (CCTCC) in 12 months and 20 days in 2019, the number is M20191077, and yarrowia lipolytica is purchased from China Center for General Microbiological Culture Collection (CGMCC), and the number is CGMCC2.1305.
Fermentation culture:
the fermentation culture comprises three steps of plate culture, seed culture and fermentation tank culture.
Plate culture: inoculating yeast into a planar culture medium to obtain planar culture strain, wherein the planar culture medium is a solid YPD culture medium, namely, 10g/L of yeast powder, 20g/L of peptone, 20g/L of glucose and 20g/L of agar powder, and culturing in a biological incubator (Shanghai Zhizheng) at 25-37 ℃ for 15-24h.
Seed culture: a ring of strain is selected from the flat plate and inoculated into a seed culture medium, wherein the culture medium is a YPD culture medium based on 10g/L of yeast powder, 20g/L of peptone and 20g/L of glucose, and the culture medium is placed in a shaking table (Infors shaking table) for culturing for 15-24 hours at 25-37 ℃.
Fermentation culture: inoculating the seed liquid into a fermentation tank for culturing according to the inoculation amount of 10% (based on the total fermentation volume), wherein the fermentation medium comprises 15-30g/L of carbon source, 1-10g/L of nitrogen source and 0.2-8g/L of potassium salt, and meanwhile, conventional microelements are properly added. The fermenter used for the fermentation culture was an info 10L fermenter.
The method for detecting the purity of the beta-phenethyl alcohol comprises the following steps:
chromatographic column: the capillary column is kept at an initial temperature of 50 ℃ for 2min by using nitrogen as carrier gas, then heated to 80 ℃ at 5 ℃/min, heated to 240 ℃ at 15 ℃/min and kept for 10min.
Monitoring a carbon source: DNS color development.
And (3) detecting dry weight of the bacterial cells: taking 1mL of fermentation liquor, centrifuging at 8000rpm for 5min, taking the lower layer of thalli, and drying in a baking oven at 60 ℃ until the mass of the thalli is no longer changed, and weighing the thalli.
Example 1:
and a thallus growth stage:
the saccharomyces cerevisiae is used as a strain, the strain is inoculated into a fermentation tank containing 2.7L of fermentation medium (3L of total fermentation volume) according to an inoculation amount of 10 percent (based on the total fermentation volume) after activation, 30 percent of resin SP825 of the initial total fermentation volume is added into the fermentation tank, the temperature is controlled to be 25 ℃ in the fermentation process, the pH is controlled to be 5.5, and the ventilation is controlled to be 3vvm. The rotating speed is related to the dissolved oxygen, and the dissolved oxygen is controlled to be about 50%; the concentration of the carbon source in the detection tank is sampled every 3 hours, and when the concentration is detected to be more than 2g/L, the carbon source is not added, otherwise, 50g/L (based on the total fermentation volume) of glucose solution with the mass percentage of 60% is added at one time.
Product synthesis stage:
sampling every 3 hours to measure dry weight of cells of the thalli, adding phenylalanine when the detected thalli amount is accumulated to 40g/L, monitoring the thalli amount, when the fluctuation range of the thalli amount is more than 5g/L, correlating dissolved oxygen with the rotating speed, controlling the dissolved oxygen to be more than 30%, otherwise, fixing the rotating speed to be 450rpm; the carbon source is supplemented at a constant rate of 15 g/L.h (60% glucose solution by mass); regulating the pH value by using 10% potassium hydroxide solution, wherein the early alkali adding rate is less than 5g/h (calculated by 100% alkali), and 12g/L phenylalanine solution is fed in; when the alkali adding rate is more than 5g/h, a mixed solution of sodium nitrate and phenylalanine is fed, wherein the concentration of sodium nitrate is 5g/L (based on the fed-batch volume), and the concentration of phenylalanine is 8g/L (based on the fed-batch volume). The flow rate was 10mL/h. The final cell mass was kept at 50g/L.
And (3) separating and purifying:
filtering the fermentation broth with 1000-1500 mesh filter cloth under 0.1MPa, collecting resin SP825, and resolving the resin with methanol to obtain resolved solution. Distilling the resolving solution under reduced pressure at 50 deg.C and-0.2 MPa to remove resolving solvent, and rectifying the concentrated solution at-0.02 MPa to obtain high purity beta-phenethyl alcohol product with purity of 99.5%.
Example 2
And a thallus growth stage:
yarrowia lipolytica is used as a strain, activated and inoculated into a fermentation tank containing 2.7L of fermentation medium (3L of total fermentation volume) according to an inoculum size of 10% (based on the total fermentation volume), 50% of resin H103 of the initial total fermentation volume is added into the fermentation tank, the temperature is controlled to be 30 ℃ in the fermentation process, the pH is controlled to be 3.5, and the ventilation is carried out for 1vvm. The rotating speed is related to the dissolved oxygen, and the dissolved oxygen is controlled to be about 20 percent; the concentration of the carbon source in the detection tank is sampled every 3 hours, and when the concentration is detected to be more than 2g/L, the carbon source is not added, otherwise, 50g/L (based on the total fermentation volume) of the maltose solution with the mass percentage of 60% is added at one time.
Product synthesis stage:
sampling every 3h to measure dry weight of the cells of the bacteria, adding phenylalanine when the detected bacterial amount is accumulated to 15g/L, monitoring the bacterial amount, when the fluctuation range of the bacterial amount is more than 5g/L, correlating dissolved oxygen with the rotating speed, controlling the dissolved oxygen to be more than 20%, otherwise, fixing the rotating speed to be 600rpm; the carbon source is supplemented at a constant rate of 18 g/L.h (60% by mass of maltose solution); regulating the pH value by using a 20% sodium hydroxide solution, wherein the early alkali adding rate is less than 5g/h (calculated by 100% alkali), and 10g/L phenylalanine solution is fed in; when the alkali adding rate is more than 5g/h, a mixed solution of sodium nitrate and phenylalanine is fed, wherein the concentration of sodium nitrate is 0.5g/L (based on the fed-batch volume), and the concentration of phenylalanine is 5g/L (based on the fed-batch volume). The flow rate was 20mL/h. The final cell mass was kept at 30g/L.
And (3) separating and purifying:
filtering the fermentation liquor by using 1000-1500 mesh filter cloth under the condition of 0.1MPa, collecting resin H103, and analyzing the resin by using ethanol to obtain analysis liquid. Distilling the resolving solution under reduced pressure at 55 deg.C and-0.1 MPa to remove resolving solvent, and rectifying the concentrated solution under-0.035 MPa to obtain high purity beta-phenethyl alcohol product with purity of 99.8%.
Example 3
And a thallus growth stage:
kluyveromyces marxianus is used as a strain, after activation, the strain is inoculated into a fermentation tank containing 2.7L of fermentation medium (3L of total fermentation volume) according to an inoculum size of 10 percent (calculated by total fermentation volume), macroporous resin SP825 accounting for 20 percent of the initial total fermentation volume is added into the fermentation tank, the temperature is controlled to be 35 ℃ in the fermentation process, the pH is 6, and the ventilation is 3vvm. The rotating speed is related to the dissolved oxygen, and the dissolved oxygen is controlled to be about 40 percent; and sampling and detecting the concentration of the carbon source in the tank every 3 hours, and if the concentration is detected to be more than 2g/L, adding no carbon source, otherwise adding 50g/L (based on the total fermentation volume) of sucrose solution with the mass percentage of 60 percent at one time.
Product synthesis stage:
sampling every 3h to measure dry weight of the cells of the bacteria, adding phenylalanine when the detected bacterial amount is accumulated to 35g/L, monitoring the bacterial amount, when the fluctuation range of the bacterial amount is more than 5g/L, correlating dissolved oxygen with the rotating speed, controlling the dissolved oxygen to be more than 40%, otherwise, fixing the rotating speed to be 400rpm; the carbon source is supplemented at a constant rate of 10 g/L.h (60% sucrose solution by mass); regulating the pH value by using 5% ammonia water solution, wherein the early alkali adding rate is less than 5g/h (calculated by 100% alkali), and 15g/L phenylalanine solution is fed in; when the alkali adding rate is more than 5g/h, a mixed solution of ammonium sulfate and phenylalanine is fed, wherein the concentration of the ammonium sulfate is 5g/L (based on the fed-batch volume), and the concentration of the phenylalanine is 5g/L (based on the fed-batch volume). The flow rate was 12mL/h. The final cell mass was kept at 35g/L.
And (3) separating and purifying:
filtering the fermentation broth with 1000-1500 mesh filter cloth under 0.1MPa, collecting resin SP825, and resolving the resin with ethyl acetate to obtain resolved solution. Distilling the resolving solution under reduced pressure at 80 deg.C and-0.5 MPa to remove resolving solvent, and rectifying the concentrated solution at-0.05 MPa to obtain high purity beta-phenethyl alcohol product with purity of 99.5%.
Example 4
And a thallus growth stage:
the saccharomyces cerevisiae is used as a strain, the strain is inoculated into a fermentation tank containing 2.7L of fermentation medium (3L of total fermentation volume) according to 10 percent of inoculation amount (calculated by total fermentation volume) after activation, macroporous resin HZ818 accounting for 35 percent of the initial total fermentation volume is added into the fermentation tank, the temperature is controlled to be 32 ℃ in the fermentation process, the pH is controlled to be 5, and the ventilation is controlled to be 2vvm. The rotating speed is related to dissolved oxygen, and the dissolved oxygen is controlled to be about 30 percent; the concentration of the carbon source in the detection tank is sampled every 3 hours, and when the concentration is detected to be more than 2g/L, the carbon source is not added, otherwise, 50g/L (based on the total fermentation volume) of glucose solution with the mass percentage of 60% is added at one time.
Product synthesis stage:
sampling every 3h to measure dry weight of the cells of the bacteria, adding phenylalanine when the detected bacterial amount is accumulated to 25g/L, monitoring the bacterial amount, when the fluctuation range of the bacterial amount is more than 5g/L, correlating dissolved oxygen with the rotating speed, controlling the dissolved oxygen to be more than 30%, otherwise, fixing the rotating speed to be 500rpm; the carbon source is supplemented at a constant rate of 20 g/L.h (60% glucose solution by mass); regulating the pH value by using 10% sodium hydroxide solution, wherein the early alkali adding rate is less than 5g/h (calculated by 100% alkali), and 20g/L phenylalanine solution is fed in; when the alkali adding rate is more than 5g/h, a mixed solution of urea and phenylalanine is fed, wherein the concentration of the urea is 2g/L (based on the feeding volume), and the concentration of the phenylalanine is 10g/L (based on the feeding volume). The flow rate was 16mL/h. The final cell mass was kept at 25g/L.
And (3) separating and purifying:
filtering the fermentation broth with 1000-1500 mesh filter cloth under 0.1MPa, collecting resin HZ818, and resolving the resin with ethanol to obtain resolved liquid. Distilling the resolving solution under reduced pressure at 60 deg.C and-0.3 MPa to remove resolving solvent, rectifying the concentrated solution at-0.01 MPa to obtain high purity beta-phenethyl alcohol product, and detecting that the purity of the product is 99.9% as shown in figure 1.
Example 5
And a thallus growth stage:
kluyveromyces marxianus is used as a strain, after activation, the strain is inoculated into a fermentation tank containing 2.7L of fermentation medium (3L of total fermentation volume) according to an inoculum size of 10 percent (calculated by total fermentation volume), macroporous resin H103 accounting for 10 percent of the initial total fermentation volume is added into the fermentation tank, the temperature is controlled to be 37 ℃ in the fermentation process, the pH is controlled to be 4.5, and ventilation is carried out for 0.5vvm. The rotating speed is related to the dissolved oxygen, and the dissolved oxygen is controlled to be about 10 percent; and sampling and detecting the concentration of the carbon source in the tank every 3 hours, and if the concentration is detected to be more than 2g/L, adding no carbon source, otherwise adding 50g/L (based on the total fermentation volume) of sucrose solution with the mass percentage of 60 percent at one time.
Product synthesis stage:
sampling every 3 hours to measure dry weight of cells of the thalli, adding phenylalanine when the quantity of the thalli is detected to be accumulated to 5g/L, monitoring the quantity of the thalli, associating dissolved oxygen with a rotating speed when the fluctuation range of the quantity of the thalli is more than 5g/L, controlling the dissolved oxygen to be more than 10%, otherwise, fixing the rotating speed to be 700rpm; the carbon source is supplemented at a constant rate of 16 g/L.h (60% sucrose solution by mass); regulating the pH value by using 15% ammonia water solution, wherein the early alkali adding rate is less than 5g/h (calculated by 100% alkali), and 18g/L phenylalanine solution is fed in; when the alkali adding rate is more than 5g/h, a mixed solution of ammonium chloride and phenylalanine is fed, wherein the concentration of the ammonium chloride is 4g/L (based on the fed-batch volume), and the concentration of the phenylalanine is 6g/L (based on the fed-batch volume). The flow rate was 14mL/h. The final cell mass was maintained at 20g/L.
And (3) separating and purifying:
filtering the fermentation liquor by using 1000-1500 mesh filter cloth under the condition of 0.1MPa, collecting resin H103, and analyzing the resin by using ethyl acetate to obtain analysis liquid. Distilling the resolving solution under reduced pressure at 70 deg.C and-0.4 MPa to remove resolving solvent, and rectifying the concentrated solution at-0.02 MPa to obtain high purity beta-phenethyl alcohol product with purity of 99.6%.
Comparative example 1:
the comparative example uses a conventional continuous feed, with the substrate being added at the beginning of the fermentation in its entirety, and the other process being the same as in example 1.
The saccharomyces cerevisiae is used as a strain, the strain is inoculated into a fermentation tank containing 2.7L of fermentation medium (the total fermentation volume is 3L) according to the inoculation amount of 10 percent (based on the total fermentation volume), 30 percent of resin SP825 of the initial total fermentation volume is added into the fermentation tank, 5-20g/L phenylalanine is added, the temperature is controlled to be 25 ℃ in the fermentation process, the pH is controlled to be 5.5, and 4vvm is ventilated. The rotating speed is related to dissolved oxygen, the dissolved oxygen is controlled to be about 50%, and glucose solution with the mass percent of 60% is continuously added.
After the fermentation, the fermentation broth was filtered using 1000-1500 mesh filter cloth under 0.1MPa, and resin SP825 was collected and analyzed using methanol to obtain an analysis solution. Distilling the resolving solution under reduced pressure at 50 ℃ and minus 0.2MPa to remove resolving solvent, and rectifying the concentrated solution under minus 0.02MPa to obtain high purity beta-phenethyl alcohol product, wherein the purity is 98% through detection.
Compared with the conventional fermentation process, the method for regulating and controlling the impurity content in the beta-phenethyl alcohol fermentation liquid controls dissolved oxygen and carbon source by controlling the concentration of thalli, and feeds back and controls substrate and nitrogen source according to the alkali addition amount in the fermentation tank.
While the present invention has been described in detail through the foregoing description of the preferred embodiment, it should be understood that the foregoing description is not to be considered as limiting the invention. Those skilled in the art will appreciate that certain modifications and adaptations of the invention are possible and can be made under the teaching of the present specification. Such modifications and adaptations are intended to be within the scope of the present invention as defined in the appended claims.
Claims (15)
1. A regulation method for reducing impurity content in beta-phenethyl alcohol fermentation liquor uses yeast as production strain, adds adsorbent in fermentation tank and phenylalanine as substrate to make fermentation so as to prepare beta-phenethyl alcohol, and is characterized by that according to the thallus concentration in fermentation tank the dissolved oxygen and carbon source supplementing quantity can be regulated and controlled, and according to the alkali quantity in fermentation tank the supplementing quantity of substrate and nitrogen source can be fed back so as to reduce by-product impurity production, and after separation and purification the pure beta-phenethyl alcohol can be obtained;
providing sufficient carbon source and dissolved oxygen to stimulate the growth of the thalli in the thalli growth stage so that the concentration of the yeast reaches the target biological quality of 5-40g/L, based on the dry weight of the thalli;
in the product synthesis stage, after the yeast accumulates to target biological quality, adding substrate phenylalanine, regulating dissolved oxygen according to the change of yeast thallus concentration, and adopting constant-rate feed for regulating and controlling carbon source; when the fluctuation range of the biomass of the thalli is larger than 5g/L, the speed is adopted to correlate dissolved oxygen regulation based on the dry weight of the thalli, and when the fluctuation range of the biomass of the thalli is smaller than 5g/L, the speed is fixed based on the dry weight of the thalli;
in the product synthesis stage, monitoring the pH value in the fermentation tank and the change of the alkali adding amount in real time, and adjusting the supplementation of phenylalanine and a nitrogen source according to the alkali adding rate; when the alkali adding rate is less than 5g/h, only phenylalanine is added by 100% alkali, and when the alkali adding rate is more than 5g/h, the mixed solution of phenylalanine and nitrogen source is added.
2. The method for controlling the impurity content in beta-phenethyl alcohol fermentation broth according to claim 1, wherein the yeast is one of kluyveromyces marxianus, saccharomyces cerevisiae and yarrowia lipolytica, and the adsorbent is macroporous resin; the temperature in the fermentation tank is 25-37 ℃, the pH is 3.5-6, and the ventilation is kept at 0.5-3vvm.
3. The method for controlling the impurity content in beta-phenethyl alcohol fermentation broth according to claim 2, wherein the macroporous resin adsorbent is any one of HZ818, SP825 and H103, and the addition amount of the macroporous resin adsorbent is 10% -35% of the initial fermentation volume.
4. The method for controlling the impurity content in beta-phenethyl alcohol fermentation broth according to claim 1, wherein the carbon source supplementing strategy is to maintain the carbon source concentration in the tank to be more than 2g/L and supplement the carbon source to be less than 2g/L in the thallus growth stage; the dissolved oxygen regulation strategy is to correlate the rotating speed with dissolved oxygen, so that the volume fraction of the dissolved oxygen in the fermentation tank is kept between 10% and 50%.
5. The method for controlling the impurity content in beta-phenethyl alcohol fermentation broth according to claim 4, wherein the dissolved oxygen volume fraction in the fermentation tank is kept between 20% and 40% in the thallus growth stage, the single additional amount of the additional carbon source is 50g/L, and the additional carbon source is a carbon source solution with the mass percentage of 60% based on the total volume of fermentation.
6. The method for controlling the impurity content in beta-phenethyl alcohol fermentation broth according to claim 1, wherein the yeast is required to accumulate 15-35g/L of biomass in terms of dry weight of thalli.
7. The method for reducing impurity content in beta-phenethyl alcohol fermentation broth according to claim 1, wherein in the product synthesis stage, when the fluctuation range of biomass of thalli is greater than 5g/L, the dissolved oxygen regulation strategy is to correlate the rotation speed with dissolved oxygen based on the dry weight of thalli, so that the volume fraction of dissolved oxygen in the fermentation tank is kept between 10% and 50%; when the fluctuation range of the biomass of the thalli is smaller than 5g/L, the fixed rotating speed is 400-700rpm according to the dry weight of the thalli; the carbon source supplementing rate is 10-20 g/L.h, the volume is calculated by the total volume of initial fermentation, and the supplementing carbon source is a carbon source solution with the mass percentage of 60%.
8. The method for reducing impurity levels in a beta-phenylethanol fermentation broth of claim 7 wherein the dissolved oxygen volume fraction in the fermentor is maintained at 20% to 40%.
9. The method for controlling the impurity content in beta-phenethyl alcohol fermentation broth according to claim 1, wherein the alkali is any one of sodium hydroxide, potassium hydroxide and ammonia water, and the nitrogen source is an inorganic nitrogen source.
10. The method for reducing the impurity content in beta-phenethyl alcohol fermentation broth according to claim 9, wherein the inorganic nitrogen source is one or more of urea, ammonium sulfate, ammonium chloride and sodium nitrate, and the concentration of the alkali is 5-20% by mass.
11. The method for controlling the impurity content in beta-phenethyl alcohol fermentation broth according to claim 1, wherein the concentration is 10-20g/L based on the volume of the feed when only phenylalanine is fed; when the mixed solution of phenylalanine and nitrogen source is supplemented, the concentration of phenylalanine is 5-10g/L, the concentration of nitrogen source is 0.5-5g/L, and the supplementing flow rate of phenylalanine and mixed solution is 10-20mL/h based on the volume of the supplementing material.
12. The method for controlling the impurity content in beta-phenethyl alcohol fermentation broth according to claim 1, wherein the biomass of yeast is controlled to be 20-50g/L in terms of dry weight of thallus in the product synthesis stage.
13. The method for controlling the impurity content in beta-phenethyl alcohol fermentation broth according to claim 12, wherein the biomass of yeast is controlled to be 20-35g/L in terms of dry weight of thallus in the product synthesis stage.
14. The method for regulating and controlling the impurity content in the beta-phenethyl alcohol fermentation broth according to claim 1 or 2, wherein the separation and purification process is characterized in that the fermentation broth obtained after the fermentation is finished is filtered, an adsorbent and the fermentation broth are separated, the adsorbent is resolved, and the resolved fluid is distilled and rectified under reduced pressure, so that the purity of the beta-phenethyl alcohol pure product is finally obtained to be more than 99.5%.
15. The method for regulating and controlling the impurity content in the beta-phenethyl alcohol fermentation broth according to claim 14, wherein the filtering condition is 0.1mPa, the pore diameter of the filter cloth is 1000-1500 meshes, the resolving solvent is ethanol, methanol or ethyl acetate, the reduced pressure distillation condition is 50-80 ℃, the pressure is-0.1-0.5 mPa, and the rectifying condition is-0.01-0.05 mPa.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010728363.2A CN113969295B (en) | 2020-07-24 | 2020-07-24 | Regulation and control method for reducing impurity content in beta-phenethyl alcohol fermentation liquor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010728363.2A CN113969295B (en) | 2020-07-24 | 2020-07-24 | Regulation and control method for reducing impurity content in beta-phenethyl alcohol fermentation liquor |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113969295A CN113969295A (en) | 2022-01-25 |
| CN113969295B true CN113969295B (en) | 2023-05-30 |
Family
ID=79585961
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010728363.2A Active CN113969295B (en) | 2020-07-24 | 2020-07-24 | Regulation and control method for reducing impurity content in beta-phenethyl alcohol fermentation liquor |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113969295B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117740976B (en) * | 2023-12-07 | 2024-08-13 | 山东京卫制药有限公司 | Analysis method for detecting related substances of phenethyl alcohol by high performance liquid phase method |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101608190A (en) * | 2009-07-17 | 2009-12-23 | 上海应用技术学院 | The preparation method of natural spice 2-phenethyl alcohol |
| CN102168115A (en) * | 2010-12-23 | 2011-08-31 | 内蒙古金达威药业有限公司 | Industrialized production method of coenzyme Q10 |
| CN107022591A (en) * | 2017-05-11 | 2017-08-08 | 宜昌东阳光长江药业股份有限公司 | A kind of Pichia pastoris fermentation process for improving insulin and the like precursor expression |
| CN108624626A (en) * | 2018-05-15 | 2018-10-09 | 上海应用技术大学 | A kind of bacterial strain and method producing bata-phenethyl alcohol |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11613768B2 (en) * | 2017-07-25 | 2023-03-28 | Arizona Board Of Regents On Behalf Of Arizona State University | Microbial production of 2-phenylethanol from renewable substrates |
-
2020
- 2020-07-24 CN CN202010728363.2A patent/CN113969295B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101608190A (en) * | 2009-07-17 | 2009-12-23 | 上海应用技术学院 | The preparation method of natural spice 2-phenethyl alcohol |
| CN102168115A (en) * | 2010-12-23 | 2011-08-31 | 内蒙古金达威药业有限公司 | Industrialized production method of coenzyme Q10 |
| CN107022591A (en) * | 2017-05-11 | 2017-08-08 | 宜昌东阳光长江药业股份有限公司 | A kind of Pichia pastoris fermentation process for improving insulin and the like precursor expression |
| CN108624626A (en) * | 2018-05-15 | 2018-10-09 | 上海应用技术大学 | A kind of bacterial strain and method producing bata-phenethyl alcohol |
Non-Patent Citations (3)
| Title |
|---|
| Synergistic inhibition effect of 2-phenylethanol and ethanol on bioproduction of natural 2-phenylethanol by Saccharomyces cerevisiae and process enhancement;Hang Wang et al.;《Journal of Bioscience and Bioengineering》;第112卷(第1期);第26-31页 * |
| 溶氧量对酿酒酵母及其工程菌的β-苯乙醇合成代谢的影响及调控效应;王成涛 等;《中国食品学报》;第16卷(第8期);第78-86页 * |
| 酿酒酵母CWY132发酵罐小试生产2-苯乙醇;汪琨;沈情佳;魏秀燕;朱廷恒;崔志峰;;食品与发酵工业;37(4);第102-106页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113969295A (en) | 2022-01-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CA2786903C (en) | Novel bacteria and methods of use thereof for producing ethanol and acetate | |
| CN105112476B (en) | Method for producing lipopeptide biosurfactant by fermentation | |
| CN111304106B (en) | Bacillus clausii and method for producing tetrahydropyrimidine by using same | |
| CN107746861B (en) | Biological preparation method of (R) -1- (2-trifluoromethylphenyl) ethanol | |
| WO2009064200A2 (en) | Novel bacteria and methods of use thereof | |
| Amin et al. | Determination of by-products formed during the ethanolic fermentation, using batch and immobilized cell systems of Zymomonas mobilis and Saccharomyces bayanus | |
| US10150973B2 (en) | Apparatus and method for separating and refining product manufactured by microbial fermentation by using adsorbent | |
| US11060118B2 (en) | Production of vanillin by fermentation | |
| CN110616158A (en) | Method for producing dodecanedioic acid by fermentation of candida virustata | |
| CN113969295B (en) | Regulation and control method for reducing impurity content in beta-phenethyl alcohol fermentation liquor | |
| CN103695325A (en) | Candida tropicalis and method for preparing L-valine through microbiological method | |
| CN102382778A (en) | Strain for producing 3-hydroxyl propionic acid at high yield and application thereof | |
| CN112358986A (en) | Clostridium butyricum and application thereof in immobilized fermentation production of 1, 3-propylene glycol | |
| CN114350731B (en) | Preparation method of glutathione | |
| TW201525139A (en) | Method for producing 2,3-butanediol | |
| CN110904160B (en) | Fermentation method for increasing beta-phenethyl alcohol unit yield | |
| CN110628546A (en) | Method for increasing content of tetramethylpyrazine in base liquor of Maotai-flavor liquor | |
| CN113699081B (en) | Bacillus subtilis for producing (2R, 3R) -butanediol and acetoin through fermentation and application thereof | |
| CN113614221A (en) | Lactobacillus bioconversion process | |
| CN115975826B (en) | Saccharomyces cerevisiae mStr003 and application thereof in production of beta-arbutin | |
| Domenech et al. | Ethanol utilization for metabolite production by Candida utilis strains in liquid medium | |
| EP4177349A1 (en) | Saccharomyces cerevisiae and fermentation product and use thereof | |
| CN114277065B (en) | Method for co-production of lactic acid and succinic acid by mixed fermentation | |
| CN117801972A (en) | Preparation method of sefeverish ham saccharomycetes and tetraacetyl phytosphingosine | |
| CN115612654A (en) | Fermentation method for improving chemical purity of lactic acid |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |