• 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:
    METHOD OF PREPARATION OF 13 MOLKNAYA. Acids by fermenting glucose or lactose with bacteria of the genus LactobacHlOs in the presence of a nitrogen source, a pH regulator and other nutrient mineral salts under anaerobic conditions at a pH of 4.5-7.0 and a temperature of 3050 ° C with a post-release release of the target product, characterized in that , in order to increase the acid output, from bacteria of the genus Lactobaci Mus, a strain of .Lactobacillus bulgaricus DSM 2129 is used.
    公开号:SU1139375A3
    申请号:SU823476930
    申请日:1982-08-10
    公开日:1985-02-07
    发明作者:Фельсков Хартмут；Сукач Дитер
    申请人:Хехст Аг (Фирма)；
    IPC主号:
    专利说明:

    The invention relates to the microbiological industry, and specifically to methods for producing D-lactic acid by fermentation of sugars. A known method of producing; 0-lactic acid by fermenting glucose or beet sugar molasses in the presence of nutrient mineral salts by Lactobaciflus leichman nil AMCC 4797 strain. The yield in 60 hours is g D-lactic acid 1. The closest to the invention is the method for producing D- lactic acid, which consists in fermenting glucose or lactose of milk sugar with bacteria of the genus LactobacM lus on a nutrient medium in the presence of a nitrogen source, calcium carbonate, a pH regulating medium, and other nutrient mineral salts in anaerobic conditions preferably at a pH of 5.1 and temperature to 50 ° C followed by isolation of the 0-lactic acid. The yield of acid is up to / O g 2. The aim of the invention is to increase the yield of 0-lactic acid. This goal is achieved by the fact that according to the method of obtaining O-moxylic acid, which involves the fermentation of glucose or lactose by bacteria of the genus Lactobaci1lus in the presence of a nitrogen source, pH regulator and other nutrient mineral salts under anaerobic conditions at pH 4.5-7 , 0 and 30–50 ° C, followed by isolation of the target product from bacteria of the genus LactobaciI1 us using Lactobaci I lus bulgaricus DSM 2129 rlthamm.. The Lactobaci1lus bulgaricus DSM 2129 used to obtain D-molar acid was isolated from an acidified milk test and improved by systematic selection so that its productivity in 48 hours reaches 115 g / l of lactic acid. The strain is deposited in a non-megus collection of microorganisms under the number DSM 2129. Identification of the strain shows that it consists of long gram-positive rods that are negative for catalase, immobile, microaerophilic, Viohigi. Metabolism. Homofermentative lactic acid fermentation: 5, gas from the gas from glucose gluconate growth at 15 with a growth at 45 ° C + Acid formation from: ribozyarabinozy xylose, mannitol, sorbitol galaktozy- glucose + lactose + maltose sucrose, trehalose, melibiose cellobiose raffinose salicin amygdalin Cleavage of arginine: - configuration .: lactic acid: D / - /. Diaminopimelic acid in the cell wall: no. Stata-1m LactobacMlus bulgaricus DSM 2129 is not specialized in controlling glucose, but can ferment (ferment) milk sugar almost as quickly as glucose to lactic acid. Thus, available in large quantities, c. as a dairy waste, fresh whey or as a suspension of milk powder can be used as a source of raw material for the production of D-lactic acid. The nutrient media used to produce D-lactic acid are formulated in a known manner. In addition to glucose and / or milk aahar, they also contain a source of nitrogen, such as M51c extract, corn flour swelling in water (.Cqrnstupj or soybean flour. In addition, mineral salts, vitamins and surfactants are added. As a surfactant active agents use commercially available non-ionic surfactants, primarily liquid products. These surfactants contain polyoxide alkylate as the biologically active substance, for example, the products of the interaction of alcohols and acids with tilenoksidom and / or propylene oxide.
    Monohydric alcohols are used as alcohols. Such as fatty alcohols, resin alcohols, glycerin, pentaerythritol; or sugar alcohols, such as sorbitol, and mannitol; fatty and resin acids are primarily used as acids. Partial esters from such polyhydric alcohols and these acids, such as hydroxyethyl anhydrosorbitol monooleate, are also beneficial.
    Since LactobaciHus bulgaricus DSM 2129 is sensitive to acids, the resulting lactic acid must be bonded with hydroxides, or carbonates of alkali or alkaline earth metals, especially calcium carbonate. The pH value is thus maintained in the range of 4.5-7.0, preferably 6.56, 8.
    Any nutrient medium before sowing Lact.obaci 1 lus bulgaricus DSM 2129 is sterilized to eliminate possible contaminants due to foreign organisms. Dp it enough to heat the culture medium for 15 min at 121 ° C.
    As with all lactic acid fermentation (fermentation), anaerobic conditions are required for the proposed method. This is sufficiently formed by the neutralization of lactic acid with calcium carbonate, carbon dioxide or the enrichment of the fermentation medium with nitrogen.
    Lactic fermentation is carried out in the temperature range of 30-50 ° C. Especially favorable temperature range of 40-45 ° C.
    The 0-lactic acid salt precipitating in accordance with the inventive method can be used to obtain free 0 lactic acid using an ion exchanger or, in the case of calcium lactate, by acidifying with sulfuric acid.
    Example 1. The growth of the strain Lactobacillus bulqarlcus DSM 2129.
    Sample of fermented milk, LactobaciHus bulgaricus schismm is isolated and cultured in the following medium 1, g / l:
    Casein peptone
    (triptych | ski learned) 10
    M sleepy extract
    (Merck) JO
    Yeast Extract 5
    Glucose20
    2 5
    CGNRO
    Sodium Acetate MgS04.7H20
    0.2 MnSO .N30 0.05
    Non-ionic surfactant 1 ml / l. Then, samples of the strain are diluted and sown on agar plates prepared from the same medium supplemented with 1.8% agar. The inoculated agar plates are then kept under anaerobic conditions during the day under a thermostat. I 5 In general, 30 separate colonies are then removed and, under circumstances, introduced into a tube that contains the following medium 4, g / l:
    Glucose50
    0 СаСО70
    Yeast extract 7 corn flour swelling (dry) 15 sodium acetate 5
    5 Non-ionic surfactant .1 ppm / l Cultivation time is 24 hours at 45 ° C. After 24 hours, the difference between different, well-fermented cultures and the amount of O-moltoic acid was clearly visible. The tube with the largest amount of D-lactic acid is used for a new smear on glass. This one. the process is repeated regularly for 8 weeks, and with this, the sugar content in the medium is first increased to P, 87, and after the next 5 weeks. to 10%. Then the strain is selected following 0 12 yed. until he is able to metabolize 100 g of glucose / l of medium for 24 hours at 65%, with 55 g of D-lactic acid / l being formed. Complete destruction of glucose is observed after 36-40 hours.
    PRI mme R 2. The LaetobacF1lus bulgaricus strain obtained in accordance with Example 1 is placed in test tubes which contain I5 ml of medium. Tubes 0 for culture are placed vertically at 45 ° C for at least 8 hours and the most. more for 20 hours and then used as inoculum for cultures in Erlenmeyer flasks, which are roderzkat 5 in 250 ml of medium 1. They are also held at 45 ° C for 8-20 hours. The contents of 6 of these Erlenmeyer flasks then serve as inoculum for fermeite with a capacity of 30 l, in which the following medium 2 is preliminarily placed, g / l: Glucose30 Yeast extract Casein peptone Maize swelling flour (dry) 20 Sodium acetate 5 Nsionous surfactant 1 ml / l Fermenters are mixed at iOO rpm, p after 8-2 hours at 45 ° C in a fermenter zag Soot contains in the amount of 270 l, which contains either the said medium or 1 Wednesday, or the following medium 3, g / l: Glucose30 CaCOz 70 Yeast extract 7 Soybean flour15 Sodium acetate 5 Non-ionic surfactant 1 ml / l After growing the culture, t. e. after 7 hours, more glucose is added. When glucose is added, I am guided by the consumption of glucose and the formation of acid. In general, such an amount of glucose, which constitutes about 10% by weight of the nutrient medium, is added over about 45 hours. And 5 g of 0-lactic acid nutrient medium (equal to 166 g of calcium lactate) is formed from glucose. Isolation of free lactic acid is carried out by known methods, namely precipitation of calcium, sulfuric acid, filtration, evaporation and purification, by distillation in the form of ethyl or methyl ether. Use of medium 3 has the advantage that the target product does not contain a detectable amount of L -lactic acid. Wednesday 4 has the premise that after the farm {tation is better filtered than medium 3, since the soybean flour in media 3 is destroyed only. The product obtained with medium 4 contains 2% L-lactic acid, which is derived from corn flour swelling in the medium. After filtration, a fraction of 1 lactic acid can be split by passing the filtrate through a fermentation reactor, which contains an enzyme that specifically cleaves L-lactic acid. Systems with L-lactate oxidase, L-lactate dehydrogenate or cytochrome bj are suitable. Lactate dehydrogenase needs as a cofactor in NAD, which can also be bound to the carrier and then can be regenerated and reused. For cytochrome b, hexacyanoferrate is suitable as a cofactor, which can be re-oxidized again using an electric current on a noble metal electrode. Proper 3. According to the method indicated in npro-tepe 2, culture culture and inoculum are prepared, however, the following medium 5 is used as nutrient medium, which contains milk sugar as a fermentable substrate, g / l: Milk whey powder 60 Yeast Soya flour extract5 Sodium acetate. 5 Non-ionic surfactant 1 ml / l As in Example 2, after growing the culture, an additional sugar substrate is added, i.e. before. as long as the added total amount of milk sugar is 130 g / l. The formation of lactic acid lasts somewhat longer than in Example Z, but after 50 hours. is running out. The rest comes in the manner described in example 2. The yield of D-lactic acid is 10 g / l. Example 4. Strain culture and inoculum are prepared by saturating with nitrogen, as in Example 1. Media 3,4 or 5 are used, but they do not contain CaCOj at first. then, so that the pH does not fall below 5.5. After 48 hours, an additional dosage of the nutrient solution is started. The dilution rate / dilution is 0.01 L / h per liter of nutrient solution, with equal amounts of liquid being taken out of the fermenter. The dilution rate is slowly increased to a value of 0.04 l / h per liter of nutrient solution, and the fermenter drain has a D-Molic acid content of 5-7%. The cell mass of the non-mercury separator 5pcs is from the drain from the in-line separator and returned to the fermenter by 9%. In the final part, the pH is adjusted to 6.5 with calcium hydroxide and evaporated to 1/10 of its initial volume. In a low-temperature trap, the concentrate is cooled to, while calcium lactate crystallizes. The concentrated solution is fed uninterruptedly into the low-temperature trap, while precipitated calcium lactate is withdrawn at regular intervals using a gate. The exact solution is returned to the fermenter. The isolation of lactic acid from the calcium lactate obtained is carried out with sulfuric acid and by subsequent filtration. The yield is 110 g of D-lactic acid per liter of nutrient medium. P h meper 5. Repeat the continuous production of lactic acid
    according to the example, the difference is that sodium hydroxide is used instead of sodium chloride to adjust the pH. The advantage of this method is that it maintains the pH at 6.5-6.8, whereby the fermentation rate of LactobacI1lus Dulaaricus DbM is elevated. The resulting sodium lactate solution is passed through ion exchange columns that absorb lactic acid. As soon as the column is loaded with lactic acid, the latter is eluted with hydrochloric acid. After regeneration with a dilute solution of hydroxide and sodium, the column can again be used to adsorb lactic acid. The output of D-lactic acid 115 g per liter of nutrient medium.
    Using the proposed method provides a higher yield of the target product and its optically pure.
    权利要求:
    Claims (1)
    [1]
    METHOD FOR PRODUCING ID-MILK. ACIDS by fermenting glucose or lactose with bacteria of the genus Lactobacillus in the presence of a nitrogen source, pH regulator and other nutritious mineral salts under anaerobic conditions at pH 4.5 - 7.0 and a temperature of 3050 ° C, followed by isolation of the target product, distinguishing that, in order to increase the acid yield, bacteria of the genus' Lactobacil lus use the strain. Lacto bacillus buldaricus DSM 2129.
    smsii t ns
    J139375 i
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    同族专利:
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    EP0072010A3|1984-08-15|
    NO822723L|1983-02-14|
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    JPH04252177A|1992-09-08|
    BR8204684A|1983-08-02|
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    US4758345A|1986-02-28|1988-07-19|The United States Of America As Represented By The United States Department Of Energy|Anaerobic microbial dissolution of lead and production of organic acids|
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    法律状态:
    优先权:
    申请号 | 申请日 | 专利标题
    DE19813131717|DE3131717A1|1981-08-11|1981-08-11|LACTOBACILLUS BULGARICUS DSM 2129 AND ITS USE FOR THE PRODUCTION OF D-LACTIC ACID|
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