• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
  • 同族专利
  • 引用文献
  • 法律状态
  • 优先权
    • 专利摘要:
    Continuous fermentation of Alcaligenes micro-organisms capable of accumulating PHB under limitation of a nutrient required for growth, but not PHB accumulation, so that the PHB content is above 25% by weight. By means of this the carbon in the carbon and energy source, i.e. the substrate, is utilised more efficiently.
    公开号:SU1303035A3
    申请号:SU813326314
    申请日:1981-08-18
    公开日:1987-04-07
    发明作者:Хьюз Лорензо；Рэймонд Ричардсон Кеннет
    申请人:Империал Кемикал Индастриз Лимитед (Фирма)；
    IPC主号:
    专利说明:

    W
    15
    , 1303035
    The invention relates to biotechnology, in particular, to the method of fermentation of Alcaligenes entrophus microorganisms for the production of cells containing poly-α-hydroxybutyric acid (POM).
    The purpose of the invention is to increase the degree of carbon conversion.
    The method is carried out as follows.
    To increase the degree of conversion, the fermentation of Alcaligenes entrophus microorganisms is carried out under conditions where the simultaneous growth of microorganisms and POM accumulation occurs in them, namely, the fermentation is carried out continuously with a dilution rate from 0.05 to 0.30 h when the process is limited to nitrogen. The nitrogen content in the starting medium is from 0.4 to 1.5 g / l. At the same time, the content of POM in cells derived from the enzyme is not less than 25%. It is precisely with the observance of the indicated intervals of the fermentation regimes that the maximum degree of carbon conversion is achieved, both in POM and in other cellular compounds.
    Other fermentation conditions, namely pH, temperature, degree of aeration, concentration of mineral salts, are commonly used to cultivate Alcaligenes entrophus bacteria.
    The bacterial suspension removed from the fermenter can be processed to isolate the POM from the cells. However, it is advisable to add a carbon source to the suspension and re-ferment it.
     30 n
    20
    25
    40
    j to jg n
    For re-fermentation, the same carbon source can be used as for the first fermentation. In some cases, it is possible to increase the efficiency of the process if substrates are used in the first stage, for example, carbohydrates, which can be equally efficiently converted into PEM and other cell material, and in the second stage, substrates for example organic acids and their salts are used. POM output, and growth 1: the entrance to them slows down.
    Example 1. The nutrient medium for growing microorganisms Alca ligenes entrophus H 16 has the following composition (per liter of deionized water)
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    15
    3035


     (N11), H / 0 (1.1 M) Mf .SO 7 H about 11.80,
    Feso
    7
    2
    2 g 12 ml 0.8 g 0.45 g 7.5 mg
    trace element solution 24 ml, i.e. N content of 0.4 g / l.
    The trace element solution has the following composition:
    Calcium 720 mg / l
    Copper5 mg / l
    Manganese 24 mg / l
    Zinc22 mg / l
    3.5 liters of nutrient medium and 35 g of fructose are loaded into a fermenter with a stirrer with a capacity of 5 liters and the mixture is kept at 30 ° C. Fermentation is carried out under aerobic conditions at a partial pressure of dissolved oxygen of 60% from air saturation. During the fermentation process, the pI value of the medium is automatically maintained at 6.8 by adding 4MNaOH. An additional amount of fructose is periodically added to maintain the excess substrate.
    After the end of the periodic fermentation, the process is continued continuously, with feeding of the nutrient medium containing 16 g / l of fructose. After the establishment of a steady state, fermentation is continued for 2 weeks. The dilution rate of the medium is about 1 g.
    20
    25
    The yield of cells is 0.575 g per g of substrate, the content of POM in cells is 30%, the degree of carbon conversion is 71%. According to a known method, under similar conditions, a cell yield of 0.42 g per g of substrate, the content of POM in cells is 70%, the degree of carbon conversion is 54%,
    EXAMPLE 2. The method is carried out according to Example 1, but glucose is used as the substrate, and glucose-consuming mutant Alcaligenes entrophus S 301/05 is used as the microorganism. The fermentation temperature is 34 ° C, the nutrient medium, which is continuously fed to the fermenter, contains 15 g / l of glucose. The yield of cells is 0.542 g per g of substrate, the POM content is 47%, the degree of carbon conversion is 69%.
    Example 3: The method is carried out according to example 2.
    A suspension of bacteria continuously withdrawn from the fermenter is added with 15 g / l glucose and fed to a second 10 l fermenter, where fermentation is carried out at 34 ° C, pH 6.8 and the partial pressure of the dissolved
    oxygen 60% of air saturation. When the volume of medium supplied to the second fermenter reaches 7 liters, it is started to be withdrawn at a speed corresponding to the rate of its withdrawal from the first fermenter, so that the dilution rate is 0.05.
    After the steady state is established, the process is carried out for two weeks. The average content of cells in the medium removed from the second fermenter is 10.9 g / l with a POM concentration of 70%. The degree of carbon conversion in the second stage is 45%. The carbon conversion rate of the entire process is 57.9%.
    EXAMPLE 4. The method is carried out according to Example 2, but the nitrogen content in the initial medium is
    1.5 g / l, and a dilution rate of 0.07 g. 20 containing sources of nitrogen and phosphorus. The concentration of cells on a dry weight of 24.5 g / l, the concentration of POM in the cells is 49%, the degree of carbon conversion is 59%. Example 5: The method is carried out according to Example 2, the nitrogen content in the initial medium is 0.69 g / l, and the dilution rate is 0.05 g. The concentration of cells on a dry weight is 18.6 g / l, the concentration of POM in cells 65%, the degree of carbon conversion 63%.
    when the process is limited to nitrogen to obtain a cell suspension, characterized in that, in order to increase the degree of carbon conversion, 25 fermentation is carried out continuously at a dilution rate of 0.05-0.30 h so that the content of poly-oxoxylic acid in the cells it was not less than 25%, while the nitrogen content in the feed medium was 0.4–1.5 g / l.
    2. Method POP.1, characterized in that a carbon source is added to the cell suspension and
    thirty
    EXAMPLE 6. The method is carried out according to Example 2, but the content of azo. In the initial medium is 0.78 g / l. 35 re-fermentation is carried out.
    2. Method POP.1, characterized in that a carbon source is added to the cell suspension;
    Editor A.Vorovich
    Compiled by T. Melentyeva
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    this case of inventions and discoveries 113035, Moscow, Zh-35, Raushsk nab., 4/5
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    and a dilution rate of 0.3 g. The concentration of cells on a dry weight of 8.2 g / l, the concentration of POM in the cells is 25%, the degree of carbon conversion is 66%.
    Thus, the proposed method allows to increase the degree of carbon conversion and thereby increase the biomass yield.
    15
    权利要求:
    Claims (2)
    [1]
    1. The method of fermentation of Alcaligenes entrophus microorganisms for obtaining cells containing poly- (3-hydroxy acid) in the conditions of azlation on an aqueous nutrient medium containing a carbon source and energy, which includes carbon and hydrogen, as well as mineral salts.
    containing sources of nitrogen and phosphorus.
    re-fermentation is carried out.
    when the process is limited to nitrogen to obtain a cell suspension, characterized in that, in order to increase the degree of carbon conversion, the fermentation is carried out continuously at a dilution rate of 0.05-0.30 h so that the content of poly-oxoxylic acid in The cells were at least 25%, while the nitrogen content in the feed medium was 0.4–1.5 g / l.
    [2]
    2. Method POP.1, characterized in that a carbon source is added to the cell suspension and
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    同族专利:
    公开号 | 公开日
    DE3171017D1|1985-07-25|
    EP0046344A2|1982-02-24|
    JPH0220238B2|1990-05-08|
    US4433053A|1984-02-21|
    AU542807B2|1985-03-14|
    JPS5774084A|1982-05-10|
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    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    US3044942A|1960-09-27|1962-07-17|Grace W R & Co|Process for preparing poly-beta-hydroxybutyric acid|
    US4140741A|1976-01-14|1979-02-20|Agroferm A.G.|Use of cyclic carbonic acid esters as solvents for poly-|
    DE2733202A1|1976-08-04|1978-02-09|Agroferm Ag|PROCESS FOR THE PREPARATION OF D - 3-HYDROXYBUTTERIC ACID|
    US4336334A|1979-02-21|1982-06-22|Imperial Chemical Industries Limited|Microbiological process for the production of poly|EP0104731B1|1982-08-27|1987-11-25|Imperial Chemical Industries Plc|3-hydroxybutyrate polymers|
    GB8301344D0|1983-01-18|1983-02-16|Ici Plc|Poly|
    EP0145233B2|1983-11-23|1991-11-06|Imperial Chemical Industries Plc|Separation processfor a 3-hydroxybutyrate polymer|
    DE3343576A1|1983-12-01|1985-06-13|Lentia GmbH Chem. u. pharm. Erzeugnisse - Industriebedarf, 8000 München|METHOD FOR THE BIOTECHNOLOGICAL PRODUCTION OF POLY-D- 3-HYDROXYBUTTERIC ACID|
    DE3343551A1|1983-12-01|1985-06-13|Lentia GmbH Chem. u. pharm. Erzeugnisse - Industriebedarf, 8000 München|METHOD FOR THE BIOTECHNOLOGICAL PRODUCTION OF POLY-D- 3-HYDROXYBUTTERIC ACID|
    US4981794A|1984-01-27|1991-01-01|E. R. Squibb & Sons, Inc.|Method of preparing D-β-hydroxyisobutyric acid by fermentation|
    US5266470A|1985-05-28|1993-11-30|Imperial Chemical Industries Plc|Copolymer production|
    US4876331A|1987-08-18|1989-10-24|Mitsubishi Kasei Corporation|Copolyester and process for producing the same|
    US5569595A|1991-09-27|1996-10-29|Center For Innovative Technology|Production of poly-β-hydroxybutyrate in prokaryotic host cells|
    US5371002A|1989-06-07|1994-12-06|James Madison University|Method of production of poly-beta-hydroxyalkanoate copolymers|
    US5518907A|1989-06-07|1996-05-21|Center For Innovative Technology|Cloning and expression in Escherichia coli of the Alcaligenes eutrophus H16 poly-beta-hydroxybutyrate biosynthetic pathway|
    GB8927794D0|1989-12-08|1990-02-14|Ici Plc|Copolymer production|
    AT393134B|1989-12-27|1991-08-26|Danubia Petrochemie|METHOD FOR RELEASING POLY-3-HYDROXYCARBONIC ACIDS|
    US5334520A|1990-05-25|1994-08-02|Center For Innovative Technology|Production of poly-beta-hydroxybutyrate in transformed escherichia coli|
    US5268422A|1990-07-19|1993-12-07|Manssur Yalpani|Functionalized poly and methods of manufacturing same|
    DE4037325C2|1990-11-23|1993-08-19|Karl Mueller U. Co Kg, 7000 Stuttgart, De|
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    US7579176B2|2003-10-15|2009-08-25|Newlight Technologies, Llc|Method for the production of polyhydroxyalkanoic acid|
    US8735113B2|2003-10-15|2014-05-27|Newlight Technologies, Llc|Methods and systems for production of polyhydroxyalkanoate|
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    法律状态:
    优先权:
    申请号 | 申请日 | 专利标题
    GB8027004|1980-08-19|
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