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WO2002038791A1 - Procede de fabrication de biomasse contenant du lycopene et procede de fabrication de lycopene cristallin a partir de celle-ci - Google Patents

Procede de fabrication de biomasse contenant du lycopene et procede de fabrication de lycopene cristallin a partir de celle-ci Download PDF

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Publication number
WO2002038791A1
WO2002038791A1 PCT/RU2000/000464 RU0000464W WO0238791A1 WO 2002038791 A1 WO2002038791 A1 WO 2002038791A1 RU 0000464 W RU0000464 W RU 0000464W WO 0238791 A1 WO0238791 A1 WO 0238791A1
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WIPO (PCT)
Prior art keywords
biomass
lycopene
oil
liκοπina
fact
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.)
Ceased
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PCT/RU2000/000464
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English (en)
Russian (ru)
Inventor
Gulsina Akhmatnurovna Mullakhmetova
Malik Favzavievich Gaisin
Odis Favzavievich Gaisin
Semen Gennadievich Mlodik
Nina Alexandrovna Panova
Vera Ivanovna Medvedeva
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URALBIOFARM OTKRYTOE AKTSIONERNOE OBSCHESTVO
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URALBIOFARM OTKRYTOE AKTSIONERNOE OBSCHESTVO
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Application filed by URALBIOFARM OTKRYTOE AKTSIONERNOE OBSCHESTVO filed Critical URALBIOFARM OTKRYTOE AKTSIONERNOE OBSCHESTVO
Priority to AU2001234264A priority Critical patent/AU2001234264A1/en
Priority to PCT/RU2000/000464 priority patent/WO2002038791A1/fr
Publication of WO2002038791A1 publication Critical patent/WO2002038791A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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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
    • C12P23/00—Preparation of compounds containing a cyclohexene ring having an unsaturated side chain containing at least ten carbon atoms bound by conjugated double bonds, e.g. carotenes
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
    • C09B61/00—Dyes of natural origin prepared from natural sources, e.g. vegetable sources
    • 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
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor

Definitions

  • the method of producing organic biomass and the method of obtaining crystalline lycine from it The area of technology.
  • the invention is available for the field of biotechnology, and specifically for the methods of receiving a valuable food preservative and anti-oxidant - lycine; May be used in the food, medical and pharmaceutical industries.
  • Lykopypin is a natural compound (carotenoid) and is synthesized by plants, algae and pests.
  • the main property of licin-pin, which causes an increased interest, is its ability to run in the form of a very common antioxidant.
  • This property of lycopene used in recent years has been widely used in medicine.
  • this pigment is used for all kinds of non-hazardous food.
  • lipin to consumables, as well as vegetable oils, ointments and creams does not only treat the detergent, but it does not affect the health.
  • SIGNIFICANT FOX (DR. 26) 2 is also non-existent ⁇ and practical, which significantly complicates the purification of the target product.
  • the indicated disadvantages are deprived of the microbiological method for the generation of lycopene, which includes the method of obtaining the biocompositive biomass and the methods of isolation from non-degenerative organisms.
  • the process is aimed at solving the general problem - increasing the yield of liquid in the target product and reducing its cost.
  • the target product is an organic biomass
  • the second is an industrial lycine.
  • SIGNIFICANT FOX (DR. 26) 3 ethanol at a ratio of 1: 5 at 50 ° C and they produce crystals containing
  • the main disadvantage of this method is the low output of cypine, which negatively affects the cost-effectiveness of the device.
  • Is ⁇ lzuyu ⁇ s ⁇ edu, s ⁇ de ⁇ zhaschuyu is ⁇ chni ⁇ i ugle ⁇ da and az ⁇ a in ⁇ aches ⁇ ve ⁇ y ⁇ is ⁇ lzuyu ⁇ 4%> s ⁇ ev ⁇ y mu ⁇ i and 1.75% ⁇ u ⁇ u ⁇ uzn ⁇ y mu ⁇ i, ⁇ as ⁇ i ⁇ eln ⁇ e ( ⁇ ds ⁇ lnechn ⁇ e) masl ⁇ ⁇ liches ⁇ ve to 4%, in v ⁇ du ⁇ liches ⁇ ve 90.5%> is ⁇ chni ⁇ ⁇ s ⁇ a - ⁇ 2 ⁇ 4 in the amount of 0.05%; from stimulation of the form of alcohol - a derivative of pyridine and an acidic fraction in the amount of 0.01-0.005) and a tobacco margin of 1%.
  • the yield of lycopene is 0.4-0.7 g per 1 liter of medium.
  • ⁇ ed ⁇ s ⁇ a ⁇ m leave ⁇ g ⁇ me ⁇ da yavlyae ⁇ sya niz ⁇ y vy ⁇ d li ⁇ ina in bi ⁇ - weight b ⁇ lshaya dli ⁇ eln ⁇ s ⁇ ⁇ tsessa and is ⁇ lz ⁇ vanie in ⁇ aches ⁇ ve s ⁇ imu- lya ⁇ a li ⁇ in ⁇ b ⁇ az ⁇ vaniya znachi ⁇ eln ⁇ g ⁇ ⁇ ⁇ liches ⁇ va d ⁇ g ⁇ s ⁇ yascheg ⁇ s ⁇ edineniya ⁇ lassa amin ⁇ me ⁇ il ⁇ i ⁇ idin ⁇ v, ch ⁇ in sv ⁇ yu ⁇ che ⁇ ed s ⁇ azyvae ⁇ sya
  • SIGNIFICANT FOX (DR. 26 4 at an increase in the cost of lipin.
  • ⁇ -carotene is observed at the same time with lycopene, which reduces the content of lycopene in the culture fluid.
  • ⁇ The disadvantages of the system are also lacking in stability of biomass when stored, due to its high humidity.
  • the task for the solution of the shortcut sent by this invention, is the creation of a more effective means of receiving a private message
  • strains which are the cultures of the mycelial fungus ⁇ kakzezea
  • the regent belonging to the graves of the class of the United Kingdom, the family of the family of the United Kingdom.
  • the indicated mushroom is a heterogeneous mushroom, i.e. it has both (+), and (-) phores.
  • beta-carotene (lycopinum) is synthesized by (-) body fungus, (+) the form is added to stimulate carotogenesis (-) by the body.
  • a cultured ka ⁇ k ⁇ ea ⁇ sustaining is necessary
  • the strains have the following characteristics.
  • (+) 8 ⁇ The airy mycelium of gray-white color with a plentiful control, in the main expressed expressions. Departure for the first days has a dense center with ample diversions
  • (-) 8 ⁇ The airy mycelium is yellow, weakly develops, secrete a yellow-orange color pigment in the substrate. Compilation is average, mainly provided by endoscopes. Primary care has a dense center with ample diversification.
  • (+) 185 ⁇ , B Aerial mycelium of a dark-gray color with primary abundant sensation, in the main expressed expressions. Dwellings in the early days have a dense center with less abundant facilities compared to strain 8 ⁇ .
  • (-) 185 ⁇ , B The yellow mycelium of the yellow color, is poorly developed, does not secrete a yellow-orange color in the substrate in comparison with strain 8 ⁇ . Compilation is average, mainly provided by endoscopes.
  • the method of obtaining free biomass is as follows.
  • the process of processing is carried out at a temperature of 28 ⁇ 2 ° C during continuous stirring and aeration.
  • the culture fluid heats up at a temperature of 85 ° C ⁇ 5 ° C for 15 minutes.
  • the moist biomass in the filter process is blown out by compressed air and transferred to the drying stage. Drying of biomass is carried out by vacuum and with an operating mixer in steams of no more than 85 ° C with a sufficient humidity of> 10%>.
  • the proposed method for the production of free biomass increases the yield of lycopene in biomass by 2 times, i.e. s (0.4 - 0.7) to (0.9 - 1.5) g per 1 liter of ⁇ red; due to the optimization of the process of biosynthesis of lipin, it helps to reduce the duration of the activity from 72 hours to 58 hours, which significantly reduces the energy consumption and reduces the energy consumption
  • the obtained crystals are filtered and washed with an organic solvent, and the filter obtained with this filter is an oil separator - it is an oil-free device From ⁇ il ⁇ a ⁇ a, ⁇ luchenn ⁇ g ⁇ ⁇ i ⁇ bezzhi ⁇ ivanii bi ⁇ massy, ⁇ - g ⁇ nyayu ⁇ ⁇ ganiches ⁇ y ⁇ as ⁇ v ⁇ i ⁇ el and ⁇ s ⁇ avshiysya li ⁇ idny ⁇ m ⁇ le ⁇ s s ⁇ avya ⁇ ⁇ is ⁇ allizatsiyu ⁇ i ⁇ em ⁇ e ⁇ a ⁇ u ⁇ e at 20 - 25 ° C for 2-4 su ⁇ , ⁇ lu- chennye ⁇ is ⁇ ally ⁇ il ⁇ vyvayu ⁇ , ⁇ myvayu ⁇ ⁇ ganiches ⁇ im ⁇ as ⁇ v ⁇ - ⁇ i ⁇ elem, na ⁇ ime ⁇ e ⁇ il ⁇ vym s ⁇ i ⁇ m, d
  • ⁇ Kasezhea ⁇ zpoga obtained by cultivating strains or (+) 8 ⁇ (-) 8 ⁇ , or ⁇ (+) ⁇ (-), or 185 ⁇ (+) 185 ⁇ (-), or 185B (+) 185B (-) with humidity not> 10 %
  • SIGNIFICANT FOX 10 The proposed method for the disposal of primary biomass eliminates the need for lipid compounds that are known to be free from hazardous substances.
  • This method contributes to an additional 4-6% increase of the original biomass of crystalline lycopene from the lipid complex with 90% of the substance.
  • the total yield for the target product will be 80-85%> due to the result.
  • a consumer filter equipped with a dry and dry mixer does not dry> 10% of the organic biomass and adds a coolant of 11% : (3 - 4), or it is at a ratio of 1: 3 and it is safe to stir at a temperature of 16-25 ° C, 50-55 ° C, 18-25 ° C is Liko- pin, using in the quality of filtering material completely from BELTING.
  • the resulting filter is loaded into the process, equipped with a mixer and a device for stirring, and discourages the process coupler from the temperature of 20.
  • the resulting lipid complex put on installation at a temperature of 20-25 ° C for 2-4 days.
  • Za ⁇ em ⁇ luchennye ⁇ is ⁇ ally ⁇ il ⁇ vyvayu ⁇ on d ⁇ u ⁇ - ⁇ il ⁇ e, ⁇ my- vayu ⁇ e ⁇ il ⁇ vym s ⁇ i ⁇ m ⁇ i ⁇ em ⁇ e ⁇ a ⁇ u ⁇ e 50-78 ° C s ⁇ n ⁇ shenii 1: (3 - 35), and vyg ⁇ uzhayu ⁇ ⁇ luchayu ⁇ ⁇ is ⁇ alliches ⁇ y li ⁇ in with s ⁇ de ⁇ zhaniem ⁇ s- n ⁇ vn ⁇ g ⁇ vesches ⁇ va 95-99.9%>.
  • the yield of lycopene from the original biomass is 35–50%.
  • the filter is a waste product and can be used at the stage of separation for the partial replacement of vegetable oil for a healthy environment.
  • P ⁇ ime ⁇ 1 (. ⁇ abl 1) P ⁇ sevn ⁇ y ma ⁇ e ⁇ ial mitselialn ⁇ g ⁇ g ⁇ iba ⁇ akez ⁇ ea y ⁇ z ⁇ ga vy ⁇ aschivayu ⁇ ⁇ azdeln ⁇ on s ⁇ ede, s ⁇ de ⁇ zhaschey s ⁇ ev ⁇ y mu ⁇ i 4%> 5- ⁇ u ⁇ u ⁇ uzn ⁇ y mu ⁇ i 2.3> ⁇ ds ⁇ lnechn ⁇ e masl ⁇ 2%>, ⁇ ⁇ 0 4 2 - 0.05 %>.
  • the cultivation of the sowing material is carried out at a temperature of 28 ⁇ 2 ° C under conditions of stirring and aeration for 48 hours.
  • the process is eliminated by heating the culture fluid with a temperature of 85 ° C for 15 minutes.
  • the separation of the mycelium of the producer from the filter is transmitted to the filter.
  • the resulting biomass is dried to a moisture content of 3-7%>.
  • the yield of lycopene is 0.96 g per 1 liter.
  • Example 2 ( ⁇ Tab. 1). But, as in Example 1, but the pyridine is produced in quantities - 0.004%>. The yield of lycopene is 1.0 g / liter.
  • ⁇ 3 ( ⁇ Table 1). However, as in Example 1, but the pyridine is produced in quantities - 0.003% (preferable). The yield of lycopene is 1.04 g / liter.
  • Example 3 we changed the delivery of compressed air during the first hours of business.
  • the supply of the supplied air to the volume of the medium is (30-70): 1.
  • the yield of lycopene is 0.86 g per 1 liter. Duration of the operation is 70 hours.
  • Example 3 the supply of the supplied air to the volume of the medium is (70 - 100): 1.
  • the yield of the liquid is 1.04 g / liter.
  • the duration of the fragmentation is from 70 to 58 hours.
  • EXAMPLE 8 (Table H). However, as in Example 7, in the amount of oil in the composition of the medium, an oil is used. The yield of pin is 1.09 g / liter.
  • Example 8 75% of the vegetable oil was replaced with a liquid lipin in oil and the preparation of food medium for the preparation.
  • the yield of lycopene is 0.95 g / liter.
  • Example 12 ( ⁇ . 3). However, in the case of Example 8, 20% of the vegetable oil was replaced with a solution of lycopene in the oil when the food medium was processed. The yield of lycopene is 1.0 g / liter.
  • Example 13 ( ⁇ . 3). But in fact, in Example 8, 70% of the vegetable oil was replaced with a liquid lipin in oil and the preparation of food medium for processing. The yield of lycopene is 0.96 g / liter. EXAMPLES 14a, b, c, d ( ⁇ . 4). Well, that is, as in Example 9, but in the quality of producers use the strains 8 ⁇ (+) 8 ⁇ (-), or AP (+) AP (-), or
  • Example 15 ( ⁇ bl. 5). ⁇ Reactor equipped with a mixer and stirrer, load 300 kg of mushroom biomass (+) (-) 8 ⁇ or (+) (-) 185 ⁇ or (+) (-) 185B or (+) (-) 6.7 kg of lycopene, dried to a moisture content of 7>, and 1200 kg of vegetable oil.
  • composition of biomass the extractant is 1: 4.
  • the extracted mass is heated to a temperature of 9 ° ⁇ and transferred to the filter
  • Example 17 ( ⁇ bl. 5). But, also in Example 16, with the exception of that, in the quality of the agent used, ethyl acetate (ethyl ester) was eliminated in case of a 2-room discharge, and there was a decrease in the rate of dilution of 1: Extracts combined. Manufacturer 16 We have removed the vacuum. The crystals that were removed from the cube wastewater were washed out, and then washed with ethyl alcohol. Out of stock, crystalline lycopene makes up 39% of the original biomass. Contents of the Main Substance 90). ⁇ 18 18 (best ⁇ Mémul ⁇ ⁇ ).
  • the dried degassed biomass from two extracts is consumed in 1200 kg of vegetable oil at a temperature of 95 ° C (the biomass is added: the concentration is 4).
  • a saturated extract takes place at the filter and sets to crystallization at a temperature of 18-22 ° C for 3 days.
  • the obtained crystals do not filter, wash off at a temperature of 1: 35 at a temperature of 50-55 ° C, dry and receive 2.86 kg, which is only 99%.
  • the yield of crystalline glycopine from the original biomass is 41.9%>.
  • the filter with a content of lycopene of 1.1 g / kg, an acid number of 2.89 mg / g and a conversion number of 7.8 mmole is an active substance that is free from any kind of food.
  • the filter obtained by cleaning the biomass is also removed by removing the temperature from the temperature range of 25-50 ° C and by increasing the temperature
  • Received crystals are processed, washed, dried, and obtained from crystalline lycopene in the amount of 0.32 kg, which results in 90% of the waste being obtained.
  • Example 19 (Tab. 5). But, also, in the case of Example 18, with the exception of the fact that the degradation of the organic biomass was eliminated with ethyl alcohol at a temperature of 1: 4 at a temperature of 50 ° C. In this case, a crystalline lycopene with a content of 97% was obtained, the yield exceeded 35.3% of the original biomass.
  • Examples 21-27 (Tab. 6). But, as in Example 18, we changed the duration of the process of crystallization of lycopene from a saturated oil extract from 1 to 8 days. The best output results for lypopin from the original biomass are shown in an interval of 3–7 days (3 days are preferable).
  • the invention may be used in the food, medical, pharmaceutical, and commercial industries.

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Abstract

L'invention concerne un procédé de fabrication de biomasse contenant du lycopène; le procédé consiste à cultiver séparément pendant 48 heures le champignon mycélien Blakeslea trispora souche 8A (+) et 8A (-). On utilise comme agent producteur complémentaire des souches de Blakeslea trispora 3C (+) 3 C (-) ou 185A (+) 185A (-) ou 185B (+) 185MB (-); en complément, on introduit dans le milieu de culture du bromure ou de la thiamine de chlorure, des sels de pyridine et une solution de lycopène dans l'huile, et le procédé est conduit avec le mélangeage et l'aération, avec séparation ultérieure de la masse biologique et son séchage. Le procédé de fabrication de lycopène cristallin consiste à l'extraire au moyen de l'huile végétale à partir de la masse biologique du champignon Blakeslea trispora sur la base des souches indiquées ci-dessus au moyen d'un solvant organique non polaire pour obtenir un extrait saturé, cristalliser le lycopène et laver les cristaux; avant l'extraction du lycopène, la masse biologique dégraissée est séchée et, après son extraction avec un solvant organique non polaire, l'extrait du lycopène est cristallisé. Lors du lavage des cristaux de lycopène il se forme des déchets de fabrication qui peuvent s'utiliser dans la fabrication du plastique contenant du lycopène.
PCT/RU2000/000464 2000-11-13 2000-11-13 Procede de fabrication de biomasse contenant du lycopene et procede de fabrication de lycopene cristallin a partir de celle-ci Ceased WO2002038791A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
AU2001234264A AU2001234264A1 (en) 2000-11-13 2000-11-13 Method for producing lycopene-containing biomass and method for the production of crystalline lycopene from said biomass
PCT/RU2000/000464 WO2002038791A1 (fr) 2000-11-13 2000-11-13 Procede de fabrication de biomasse contenant du lycopene et procede de fabrication de lycopene cristallin a partir de celle-ci

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PCT/RU2000/000464 WO2002038791A1 (fr) 2000-11-13 2000-11-13 Procede de fabrication de biomasse contenant du lycopene et procede de fabrication de lycopene cristallin a partir de celle-ci

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100335642C (zh) * 2005-03-16 2007-09-05 佛山市英迈瑞生物技术服务有限公司 基因重组毕赤酵母生产番茄红素的方法

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU94017353A (ru) * 1994-05-11 1996-08-10 Товарищество с ограниченной ответственностью "Инвест" Способ получения ликопина и лекарственный препарат ликопина
RU2102416C1 (ru) * 1995-03-22 1998-01-20 Акционерное общество открытого типа "Уралбиофарм" Способ получения ликопина
US5858700A (en) * 1997-04-03 1999-01-12 Kemin Foods, Lc Process for the isolation and purification of lycopene crystals
RU2126806C1 (ru) * 1996-05-28 1999-02-27 Акционерное общество открытого типа "Уралбиофарм" Способ получения кристаллического ликопина
US5935808A (en) * 1997-07-29 1999-08-10 Yissum Research And Development Company Of The Hebrew University Of Jerusalem Carotenoid-producing bacterial species and process for production of carotenoids using same

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU94017353A (ru) * 1994-05-11 1996-08-10 Товарищество с ограниченной ответственностью "Инвест" Способ получения ликопина и лекарственный препарат ликопина
RU2102416C1 (ru) * 1995-03-22 1998-01-20 Акционерное общество открытого типа "Уралбиофарм" Способ получения ликопина
RU2126806C1 (ru) * 1996-05-28 1999-02-27 Акционерное общество открытого типа "Уралбиофарм" Способ получения кристаллического ликопина
US5858700A (en) * 1997-04-03 1999-01-12 Kemin Foods, Lc Process for the isolation and purification of lycopene crystals
US5935808A (en) * 1997-07-29 1999-08-10 Yissum Research And Development Company Of The Hebrew University Of Jerusalem Carotenoid-producing bacterial species and process for production of carotenoids using same

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100335642C (zh) * 2005-03-16 2007-09-05 佛山市英迈瑞生物技术服务有限公司 基因重组毕赤酵母生产番茄红素的方法

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