JPH07308195A - Production of l-aspartic acid - Google Patents
Production of l-aspartic acidInfo
- Publication number
- JPH07308195A JPH07308195A JP6102789A JP10278994A JPH07308195A JP H07308195 A JPH07308195 A JP H07308195A JP 6102789 A JP6102789 A JP 6102789A JP 10278994 A JP10278994 A JP 10278994A JP H07308195 A JPH07308195 A JP H07308195A
- Authority
- JP
- Japan
- Prior art keywords
- acid
- aspartic acid
- alkali metal
- enzyme
- ammonia
- 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.)
- Granted
Links
Classifications
-
- 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
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/52—Improvements relating to the production of bulk chemicals using catalysts, e.g. selective catalysts
Landscapes
- Enzymes And Modification Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Description
【０００１】[0001]
【産業上の利用分野】本発明はフマル酸とアンモニアか
らＬ−アスパラギン酸を生産する際、多量のアンモニウ
ムイオンを含んだ廃水を排出しないようにする方法に関
するものである。BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for preventing the discharge of waste water containing a large amount of ammonium ions when producing L-aspartic acid from fumaric acid and ammonia.
【０００２】[0002]
【従来の技術】従来、アスパルターゼ活性を有する微生
物を用いてフマル酸アンモニウムからＬ−アスパラギン
酸を製造する方法としては、α−アミノ酪酸に耐性を有
する微生物を好気的に培養後反応に供する方法（特公昭
６１ー２９７１８号公報）、フマル酸添加培地で培養し
た微生物菌体を用いる方法（特開昭６０ー１２０９８３
号公報）、天然物多糖由来のポリマー等に大腸菌(Esche
richia coli)を固定化した固定化微生物充填カラムを用
いる方法（特開昭５３ー６４８３号公報）など種々の方
法が知られている。2. Description of the Related Art Conventionally, as a method for producing L-aspartic acid from ammonium fumarate using a microorganism having aspartase activity, a microorganism resistant to .alpha.-aminobutyric acid is aerobically subjected to post-culture reaction. Method (Japanese Patent Publication No. 61-29718), method using microbial cells cultured in a fumaric acid-containing medium (Japanese Patent Laid-Open No. 60-120983).
Escherichia coli (Escherichia coli
Various methods are known, such as a method using an immobilized microbial packed column in which richia coli is immobilized (Japanese Patent Laid-Open No. 53-6483).
【０００３】通常、フマル酸とアンモニアの反応液から
Ｌ−アスパラギン酸を回収するためには、硫酸などの鉱
酸を用いて、反応液のｐＨをＬ−アスパラギン酸の等電
点であるｐＨ２．７程度に調節後、冷却することによっ
てＬ−アスパラギン酸の結晶を析出させ、これを濾別す
る方法がとられている。Usually, in order to recover L-aspartic acid from a reaction solution of fumaric acid and ammonia, the pH of the reaction solution is adjusted to pH 2. which is the isoelectric point of L-aspartic acid by using a mineral acid such as sulfuric acid. After adjusting to about 7, crystals of L-aspartic acid are precipitated by cooling, and the crystals are separated by filtration.
【０００４】この方法は安価な鉱酸を用いること、生産
物であるＬ−アスパラギン酸の結晶としての回収率が高
いこと、得られるＬ−アスパラギン酸の純度が高いこと
から非常に経済的に有利な方法である。This method is very economically advantageous because it uses an inexpensive mineral acid, has a high recovery rate of the product L-aspartic acid as crystals, and has a high purity of the obtained L-aspartic acid. That's the method.
【０００５】しかしながら、産業廃棄物という観点から
は、高濃度の硫酸アンモニウム等のアンモニウム塩を含
有した廃水が大量に排出されるという問題点を有してい
る。水溶液中のアンモニウムイオンの除去は廃水処理の
面でも非常に困難であり、湖沼や瀬戸内海などの内湾で
はアンモニウムイオンを含む窒素濃度が上昇することに
よる水質汚染などの問題が生じてきている。また最近、
工場廃水中の窒素濃度の規制についても各省庁で検討が
行われているようである。従って、Ｌ−アスパラギン酸
の製造においても硫安などの副生成物が多量に発生しな
い系の開発が望まれている。However, from the viewpoint of industrial waste, there is a problem that a large amount of waste water containing a high concentration of ammonium salt such as ammonium sulfate is discharged. The removal of ammonium ions from the aqueous solution is very difficult in terms of wastewater treatment, and problems such as water pollution have arisen in the inner bays such as lakes and the Seto Inland Sea due to an increase in the concentration of nitrogen containing ammonium ions. Also recently
It seems that ministries and agencies are also studying the regulation of nitrogen concentration in factory wastewater. Therefore, in the production of L-aspartic acid, development of a system in which a large amount of by-products such as ammonium sulfate does not occur is desired.
【０００６】米国特許４５６０６５３ではＬ−アスパラ
ギン酸の生産の際にアスパルターゼもしくはアスパルタ
ーゼ生産菌をフマル酸とアンモニアに作用させ、生成し
たアスパラギン酸アンモニウム水溶液にマレイン酸を添
加して酸性にすることによってＬ−アスパラギン酸を析
出させ、濾液を異性化することによって反応液のリサイ
クルを行う方法が提案されている。この方法は、硫安な
どの副生成物が発生しない方法である。しかしこの方法
ではＬ−アスパラギン酸の析出に用いたマレイン酸を、
臭素イオンを含んだ触媒を用いて、アスパルターゼが作
用できるフマル酸に異性化し、異性化後、触媒を除去す
る工程が含まれており、Ｌ−アスパラギン酸の製造工程
が煩雑になる欠点を有している。In US Pat. No. 4,560,653, aspartase or an aspartase-producing bacterium is allowed to act on fumaric acid and ammonia during the production of L-aspartic acid, and maleic acid is added to the resulting ammonium aspartate aqueous solution to make it acidic. A method has been proposed in which L-aspartic acid is deposited and the reaction solution is recycled by isomerizing the filtrate. This method does not generate by-products such as ammonium sulfate. However, in this method, the maleic acid used for the precipitation of L-aspartic acid is
It includes a step of isomerizing fumaric acid that can act aspartase using a catalyst containing bromide ion, and removing the catalyst after the isomerization, which has a drawback that the production process of L-aspartic acid becomes complicated. is doing.
【０００７】[0007]
【発明が解決しようとする課題】本発明の目的はフマル
酸とアンモニアからＬ−アスパラギン酸を生産する際、
多量のアンモニウム塩を排出しない、Ｌ−アスパラギン
酸の製造方法を提供しようとするものである。The object of the present invention is to produce L-aspartic acid from fumaric acid and ammonia.
An object of the present invention is to provide a method for producing L-aspartic acid which does not discharge a large amount of ammonium salt.
【０００８】本発明者らはこのような高濃度のアンモニ
ウムイオンを含有した廃水が大量に排出されない、簡易
なＬ−アスパラギン酸の製造方法について鋭意検討を行
った結果、この反応の基質であるフマル酸を中和するの
に従来用いられていたアンモニア単独の条件にかえてア
ルカリ金属イオンをあわせて用いると反応の転化率、選
択率共に通常工業的に行われている方法と遜色ない結果
が得られることを見いだし本発明を完成させるに至っ
た。The present inventors have conducted extensive studies on a simple method for producing L-aspartic acid in which a large amount of waste water containing such a high concentration of ammonium ions is not discharged, and as a result, fumarase which is a substrate for this reaction. When the alkali metal ion is used in combination with the ammonia alone condition conventionally used to neutralize the acid, the conversion rate and selectivity of the reaction are comparable to the methods usually used in the industry. The present invention has been completed and the present invention has been completed.
【０００９】[0009]
【課題を解決するための手段】本発明はフマル酸とアン
モニアおよびアルカリ金属イオンを含む基質媒体に、ア
スパルターゼ活性を有する酵素含有物を作用せしめるこ
とによりＬ−アスパラギン酸を生成せしめ、次にＬ−ア
スパラギン酸を含有する反応済媒体に鉱酸を加え、析出
したＬ−アスパラギン酸の結晶を濾別・回収すると共
に、鉱酸のアルカリ金属塩を主成分とする廃液を排出す
ることを特徴とするＬ−アスパラギン酸の製造方法に関
するものである。According to the present invention, L-aspartic acid is produced by allowing an enzyme-containing substance having aspartase activity to act on a substrate medium containing fumaric acid, ammonia and alkali metal ions, and then L-aspartic acid. -A mineral acid is added to a reacted medium containing aspartic acid, the precipitated L-aspartic acid crystals are filtered and recovered, and a waste liquid containing an alkali metal salt of mineral acid as a main component is discharged. The present invention relates to a method for producing L-aspartic acid.
【００１０】本発明に用いるフマル酸はフマル酸あるい
はフマル酸塩から選ばれるものであって、これらの混合
物でもよい。反応の際のフマル酸濃度は通常５〜３０重
量％が好ましいがフマル酸塩の溶解度と生産性の面から
特に１０〜２５重量％が好ましい。The fumaric acid used in the present invention is selected from fumaric acid or fumaric acid salt, and may be a mixture thereof. The concentration of fumaric acid in the reaction is usually preferably 5 to 30% by weight, but 10 to 25% by weight is particularly preferable from the viewpoint of solubility of fumaric acid salt and productivity.
【００１１】本発明に用いられるアンモニアは液体アン
モニア、アンモニア水溶液等が使用可能であるが、取扱
上、アンモニア水溶液が有利である。As the ammonia used in the present invention, liquid ammonia, aqueous ammonia solution or the like can be used, but aqueous ammonia solution is advantageous in handling.
【００１２】アンモニア水の濃度としては特に限定され
るものではないが、工業的には１０〜３５重量％が利用
するのに好ましい。The concentration of the ammonia water is not particularly limited, but industrially 10 to 35% by weight is preferable for use.
【００１３】本発明に用いられるフマル酸を中和するに
あたって使用するアルカリ金属イオンの量はフマル酸に
対して０．５〜１．５倍モル、好ましくは０．９〜１．
３倍モル、より好ましくは１．１０〜１．２５倍モル用
いるのがよい。The amount of alkali metal ion used for neutralizing the fumaric acid used in the present invention is 0.5 to 1.5 times mol, preferably 0.9 to 1.
It is good to use 3 times mol, more preferably 1.10 to 1.25 times mol.
【００１４】本発明に用いるアルカリ金属イオンとして
はナトリウムイオン、カリウムイオンのほか各種のアル
カリ金属イオンが使用できるが、経済的には水酸化ナト
リウムか水酸化カリウムをアルカリ金属イオンとして用
いるのが好ましい。またこれらのアルカリ金属水酸化物
は２種以上のものを混合して用いても差し支えない。さ
らにフマル酸をアルカリ金属イオンで中和するかわりに
フマル酸のアルカリ金属塩をそのまま用いても差し支え
ない。As the alkali metal ion used in the present invention, various alkali metal ions can be used in addition to sodium ion and potassium ion, but it is preferable to use sodium hydroxide or potassium hydroxide as the alkali metal ion economically. Further, these alkali metal hydroxides may be used as a mixture of two or more kinds. Further, instead of neutralizing fumaric acid with an alkali metal ion, an alkali metal salt of fumaric acid may be used as it is.
【００１５】反応液のｐＨは５から１０の範囲、好まし
くは７．５から９．０の範囲、さらに好ましくはアスパ
ルターゼの至適ｐＨである８．０〜８．５程度にアルカ
リ金属イオンおよびアンモニアを添加して調整すればよ
い。The pH of the reaction solution is in the range of 5 to 10, preferably in the range of 7.5 to 9.0, and more preferably in the range of 8.0 to 8.5 which is the optimum pH of aspartase. It may be adjusted by adding ammonia.
【００１６】本発明に用いるアスパルターゼ活性を有す
る酵素含有物は、アスパルターゼが高活性な微生物菌体
そのもの、あるいは超音波、摩砕、凍結融解、酵素処
理、界面活性剤処理などにより物理的または生化学的に
処理して破砕した菌体破砕物、さらに硫酸アンモニウム
塩析、アセトン沈殿等常法により得られる酵素のいずれ
でも使用できる。アスパルターゼ活性を有する微生物と
しては、例えばエッシェリシア（Escherichia ）属に属
する微生物（エッシェリシア・コリ（Escherichia col
i）ＡＴＣＣ１１３０３、ＡＴＣＣ９６３７、ＡＴＣＣ
２７３２５）、ブレビバクテリウム（Brevibacterium）
属に属する微生物などフマル酸よりＬ−アスパラギン酸
を収率よく生成する特徴を有する微生物であれば特に限
定されない。これらのアスパルターゼ活性を有する酵素
含有物を担体に固定化して用いることもできる。固定化
の担体としては、セルロース、アルギン酸、カラギー
ナ、ナンマンゲルなどの適当な天然系高分子、あるいは
イオン交換樹脂やポリビニルアルコール、ポリアクリル
アミドなどの適当な合成高分子などを常法により用いる
ことができる。The enzyme-containing substance having aspartase activity used in the present invention is a microbial cell itself having high aspartase activity, or is physically or ultrasonically treated by ultrasonication, grinding, freeze-thawing, enzyme treatment, surfactant treatment or the like. Any of the crushed cells obtained by biochemical treatment and crushing, and the enzyme obtained by a conventional method such as salting out with ammonium sulfate or acetone precipitation can be used. Examples of the microorganism having aspartase activity include microorganisms belonging to the genus Escherichia (Escherichia col).
i) ATCC11303, ATCC9637, ATCC
27325), Brevibacterium
There is no particular limitation as long as it is a microorganism belonging to the genus such as a microorganism having a characteristic of producing L-aspartic acid from fumaric acid in a high yield. These enzyme-containing substances having aspartase activity can also be used after being immobilized on a carrier. As a carrier for immobilization, a suitable natural polymer such as cellulose, alginic acid, carrageen, and Nanmann gel, or a suitable synthetic polymer such as an ion exchange resin, polyvinyl alcohol, and polyacrylamide can be used by a conventional method.
【００１７】また、反応に用いるアスパルターゼ活性を
有する酵素含有物中に含まれるフマラーゼ活性など該反
応の妨げになりうるアスパルターゼ活性以外の酵素を予
め失活させたものを反応に用いることも可能である。た
とえば酵素含有物を、予め、Ｌ−アスパラギン酸および
アンモニウムイオン存在下、アルカリ域で４０〜６０℃
に加熱処理を行い、フマラーゼ活性を予め失活させてお
くこともできる。It is also possible to use in the reaction a product in which an enzyme other than the aspartase activity such as the fumarase activity contained in the enzyme-containing substance having the aspartase activity used in the reaction, which may interfere with the reaction, is deactivated in advance. Is. For example, the enzyme-containing material is preliminarily subjected to the presence of L-aspartic acid and ammonium ions in the alkaline region at 40 to 60 ° C.
The fumarase activity can be deactivated in advance by heat treatment.
【００１８】フマル酸とアンモニアとの反応はそれらを
溶解した水性媒体、たとえば水または緩衝液中で行う。
反応の際の原料のフマル酸の濃度は５〜３０重量％好ま
しいが、フマル酸塩の溶解性と生体触媒の反応性を考え
ると特に１０〜２５重量％の範囲の水溶液で反応させる
のが効果的である。The reaction between fumaric acid and ammonia is carried out in an aqueous medium in which they are dissolved, such as water or a buffer solution.
The concentration of the raw material fumaric acid during the reaction is preferably 5 to 30% by weight, but considering the solubility of the fumarate and the reactivity of the biocatalyst, it is particularly effective to react with an aqueous solution in the range of 10 to 25% by weight. Target.
【００１９】また反応液にはさらに塩化マンガン、硫酸
マンガンなどのマンガン塩、または塩化マグネシウム、
硫酸マグネシウムなどのマグネシウム塩を０．１〜５０
ｍＭ、特に１〜１０ｍＭの濃度で添加することがことが
好ましい。The reaction solution further contains manganese salts such as manganese chloride and manganese sulfate, or magnesium chloride,
0.1-50 magnesium salts such as magnesium sulfate
It is preferable to add it at a concentration of mM, particularly 1 to 10 mM.
【００２０】本発明における反応槽の態様は特に限定さ
れないが、例えば、バッチ型反応装置、カラム型反応装
置など従来から知られている反応槽で反応を行うことが
できる。反応槽は１つであってもよいし、複数あっても
差し支えない。またカラム型の反応装置の場合には、通
液速度をカラムに充填されている酵素の量によって変え
て反応することも可能である。The mode of the reaction vessel in the present invention is not particularly limited, but the reaction can be carried out in a conventionally known reaction vessel such as a batch type reaction apparatus or a column type reaction apparatus. There may be one reaction tank or a plurality of reaction tanks. In the case of a column-type reaction device, it is also possible to change the liquid flow rate depending on the amount of enzyme packed in the column to carry out the reaction.
【００２１】反応の際の温度は低温では反応速度が低下
するため通常２０℃程度を下限とし、高温下ではアスパ
ルターゼの失活を招くため５０℃程度を上限とするのが
好ましく、より好ましくは２５〜４０℃の範囲で行うの
がよい。It is preferable that the temperature during the reaction has a lower limit of about 20 ° C. because it lowers the reaction rate at low temperature, and an upper limit of about 50 ° C. at high temperature because it causes inactivation of aspartase, and more preferably. It is preferable to carry out in the range of 25 to 40 ° C.
【００２２】反応後の液中のＬ−アスパラギン酸は常法
通り等電点沈殿法等により容易に回収できる。例えば反
応液に硫酸等の鉱酸を添加しｐＨをＬ−アスパラギン酸
の等電点である２．７７程度に低下させ、冷却すること
によって結晶を析出させれば良い。The L-aspartic acid in the liquid after the reaction can be easily recovered by an isoelectric focusing method or the like as usual. For example, crystals may be precipitated by adding a mineral acid such as sulfuric acid to the reaction solution to lower the pH to about 2.77 which is the isoelectric point of L-aspartic acid, and cooling.
【００２３】本発明に用いる鉱酸としては硫酸、塩酸、
リン酸などが使用できる。Mineral acids used in the present invention include sulfuric acid, hydrochloric acid,
Phosphoric acid or the like can be used.
【００２４】析出したＬ−アスパラギン酸の結晶は通常
の方法、例えば濾過、遠心分離、デカンテーションなど
の方法で液から分離し、通常の方法にしたがって乾燥さ
れる。斯くして結晶を分離した液中のアンモニウムイオ
ンの濃度は通常の工業的に実施されている方法に比べて
数分の一から数十分の一となっており、主成分は鉱酸の
アルカリ金属塩である。The precipitated L-aspartic acid crystals are separated from the liquid by a conventional method such as filtration, centrifugation or decantation, and dried according to a conventional method. Thus, the concentration of ammonium ions in the liquid obtained by separating the crystals is a fraction to a few tens of ten as compared with the method which is usually carried out industrially, and the main component is alkali of mineral acid. It is a metal salt.
【００２５】[0025]
【作用】本発明によれば、フマル酸とアンモニアからＬ
−アスパラギン酸の製造に際して、廃水の主成分を従来
の硫酸アンモニウムから硫酸のアルカリ金属塩にかえる
ことができ、近年の工業廃水に対する窒素規制に対応す
ることができる。According to the present invention, fumaric acid and ammonia are used to form L
In the production of aspartic acid, the main component of wastewater can be changed from conventional ammonium sulfate to alkali metal salt of sulfuric acid, and it is possible to comply with the recent nitrogen regulation for industrial wastewater.
【００２６】[0026]
【実施例】次に本発明の方法を実施例をあげて説明する
が、本発明はかかる実施例のみに限定されるものではな
い。EXAMPLES The method of the present invention will now be described with reference to examples, but the present invention is not limited to these examples.
【００２７】実施例１ ２Ｌジャーファーメンターにフマル酸２０ｇ、リン酸１
カリウム１ｇ、硫酸マグネシウム７水塩０．５ｇ、酵母
エキス２０ｇ、コーンスティープリカー２０ｇを水に溶
解し、ｐＨをアンモニアで６．８に調節した培地１Ｌを
仕込み滅菌した後、別に５００ｍｌ振盪フラスコに同上
の培地５０ｍｌをいれて培養しておいたエッシェリヒア
コリ（Escherichia coli ATCC 11303 ）を接種し、３７
℃で通気撹拌培養した。培地中のフマル酸が消失した時
点で、菌体培養液に酢酸を加え、ｐＨを約５に調整し、
４５℃で１時間放置した後、培養液を遠心分離にかけ、
菌体を分離した。この菌体を３等分し、−８０℃で凍結
して冷蔵した。Example 1 Fumaric acid 20 g and phosphoric acid 1 in a 2 L jar fermenter
Dissolve 1 g of potassium, 0.5 g of magnesium sulfate heptahydrate, 20 g of yeast extract, 20 g of corn steep liquor in water, charge 1 L of a medium whose pH was adjusted to 6.8 with ammonia, and sterilize it. Escherichia coli ATCC 11303 cultivated in 50 ml of the culture medium of
The culture was performed with aeration and stirring at ℃. When fumaric acid in the medium disappeared, acetic acid was added to the cell culture solution to adjust the pH to about 5,
After leaving it at 45 ° C for 1 hour, the culture solution is centrifuged,
The cells were separated. The cells were divided into 3 equal parts, frozen at -80 ° C and refrigerated.
【００２８】フマル酸２００ｇおよび硫酸マグネシウム
７水塩０．２５ｇを水６００ｍｌにいれ、水酸化ナトリ
ウムを８２．８ｇ（対フマル酸１．２倍モル）添加後、
２５％アンモニア水を用いてｐＨを８．３に調節し、水
を追加して１Ｌとし、これを反応基質溶液とした。200 g of fumaric acid and 0.25 g of magnesium sulfate heptahydrate were added to 600 ml of water, and after adding 82.8 g of sodium hydroxide (1.2 times mol of fumaric acid),
The pH was adjusted to 8.3 using 25% aqueous ammonia, and water was added to make 1 L, which was used as a reaction substrate solution.
【００２９】この基質液に先に３等分した凍結菌体の一
つを入れ、３７℃で緩やかに振盪しながら５時間反応さ
せた。この反応液中のＬ−アスパラギン酸は２２１．
３．ｇであった。この反応液を遠心分離して菌体を除い
た後、硫酸を添加し、ｐＨを２．７７に調節した。これ
を６０℃に加熱、その後冷却した。冷却後、吸引濾過器
で吸引濾過し、濾過器内の結晶を約１５０ｍｌの水で吸
引しながら洗浄し、この結晶を乾燥し重量、純度を調べ
たところ、重量２１６．４ｇ、純度９９．６％のＬ−ア
スパラギン酸を得た。濾液１Ｌ中のアンモニア濃度を測
定したところ、ＮＨ3 として約１．０ｇ／Ｌであった。One of the frozen bacterial cells that had been divided into three equal parts was put into this substrate solution, and the mixture was reacted at 37 ° C. for 5 hours with gentle shaking. L-aspartic acid in this reaction solution was 221.
3. It was g. The reaction solution was centrifuged to remove cells, and sulfuric acid was added to adjust the pH to 2.77. It was heated to 60 ° C. and then cooled. After cooling, suction filtration was carried out with a suction filter, and the crystals in the filter were washed with suction of about 150 ml of water, and the crystals were dried and the weight and purity were examined. As a result, the weight was 216.4 g and the purity was 99.6. % L-aspartic acid was obtained. When the ammonia concentration in 1 L of the filtrate was measured, it was about 1.0 g / L as NH 3 .
【００３０】実施例２ 実施例１において、反応基質に加える水酸化ナトリウム
の量を１００ｇ（対フマル酸１．４５倍モル）とした以
外は実施例１と同様な操作を行った。反応後の反応液中
のＬ−アスパラギン酸は１７８．１ｇであった。実施例
１と同様な方法でＬ−アスパラギン酸の晶析を行い、重
量２１９ｇ、純度８０％のＬ−アスパラギン酸を得た。
濾液中のアンモニア濃度を測定したところ、ＮＨ3 とし
て約０．８５ｇ／Ｌであった。Example 2 The same operation as in Example 1 was carried out except that the amount of sodium hydroxide added to the reaction substrate was changed to 100 g (1.45 times mol of fumaric acid). The amount of L-aspartic acid in the reaction solution after the reaction was 178.1 g. L-Aspartic acid was crystallized in the same manner as in Example 1 to obtain L-aspartic acid having a weight of 219 g and a purity of 80%.
When the ammonia concentration in the filtrate was measured, it was about 0.85 g / L as NH 3 .
【００３１】実施例３ 実施例１において、反応基質に水酸化ナトリウムを水酸
化カリウムにし、これを９６．７ｇ（対フマル酸１．０
倍モル）添加する以外は実施例１と同様な反応を行っ
た。反応後の反応液中のＬ−アスパラギン酸は２２５．
９ｇであった。実施例１と同様な方法でＬ−アスパラギ
ン酸の晶析を行い、重量２２２．０ｇ、純度９９．４％
のＬ−アスパラギン酸を得た。濾液中のアンモニア濃度
を測定したところ、ＮＨ3 として約５．７ｇ／Ｌであっ
た。Example 3 In Example 1, sodium hydroxide was changed to potassium hydroxide as a reaction substrate, and 96.7 g (1.0% fumaric acid) was added to the reaction substrate.
The same reaction as in Example 1 was carried out except that the addition amount was added. The L-aspartic acid in the reaction solution after the reaction was 225.
It was 9 g. Crystallization of L-aspartic acid was performed in the same manner as in Example 1 to obtain a weight of 222.0 g and a purity of 99.4%.
L-aspartic acid was obtained. When the ammonia concentration in the filtrate was measured, it was about 5.7 g / L as NH 3 .
【００３２】比較例１ 実施例１において、反応基質に水酸化ナトリウムを添加
せずに２５％アンモニア水を加え、ｐＨを８．３に調節
した以外は実施例１と同様の操作を行なった。反応後の
反応液中のＬ−アスパラギン酸は２２７．３ｇであっ
た。この反応液を実施例１と同様な方法で処理し結晶と
炉液を得た。結晶を乾燥し重量、純度を調べたところ、
重量２２３．３ｇ、純度９９．６％のＬ−アスパラギン
酸を得た。濾液中のアンモニア濃度を測定したところ、
ＮＨ3 として約３５．２ｇ／Ｌであった。Comparative Example 1 The same operation as in Example 1 was carried out except that 25% ammonia water was added to the reaction substrate without adding sodium hydroxide to adjust the pH to 8.3. The amount of L-aspartic acid in the reaction solution after the reaction was 227.3 g. This reaction liquid was treated in the same manner as in Example 1 to obtain crystals and a furnace liquid. When the crystals were dried and examined for weight and purity,
L-Aspartic acid having a weight of 223.3 g and a purity of 99.6% was obtained. When the concentration of ammonia in the filtrate was measured,
The amount of NH 3 was about 35.2 g / L.
【００３３】[0033]
【発明の効果】本発明によれば、高濃度の硫酸アンモニ
ウム水溶液などの環境上好ましくない副生成物を伴わず
にＬ−アスパラギン酸を効率よくフマル酸を原料に製造
することができる。Industrial Applicability According to the present invention, L-aspartic acid can be efficiently produced from fumaric acid as a raw material without environmentally unfavorable by-products such as a high-concentration ammonium sulfate aqueous solution.
───────────────────────────────────────────────────── フロントページの続き (51)Int.Cl.6 識別記号 庁内整理番号 ＦＩ 技術表示箇所 Ｃ１２Ｒ 1:13） ─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. 6 Identification code Internal reference number FI technical display C12R 1:13)
Claims (4)
金属イオンを含む基質媒体に、アスパルターゼ活性を有
する酵素含有物を作用せしめることによりＬ−アスパラ
ギン酸を生成せしめ、次にＬ−アスパラギン酸を含有す
る反応済媒体に鉱酸を加え、Ｌ−アスパラギン酸の結晶
を濾別・回収すると共に、鉱酸のアルカリ金属塩を主成
分とする廃液を排出することを特徴とするＬ−アスパラ
ギン酸の製造方法。1. L-aspartic acid is produced by reacting an enzyme-containing substance having aspartase activity with a substrate medium containing fumaric acid, ammonia and an alkali metal ion, and then a reaction containing L-aspartic acid. A method for producing L-aspartic acid, which comprises adding a mineral acid to a used medium, filtering and collecting L-aspartic acid crystals, and discharging a waste liquid containing an alkali metal salt of the mineral acid as a main component.
物が、酵素活性を有する微生物菌体、菌体破砕物、部分
精製酵素もしくは精製酵素、またはこれらを含んでなる
固定化物である請求項１記載の方法。2. The enzyme-containing material having aspartase activity is a microbial cell having enzymatic activity, a disrupted cell, a partially purified enzyme or a purified enzyme, or an immobilized product containing these. Method.
ルのアルカリ金属イオンを含む請求項１〜２のいずれか
に記載の方法。3. The method according to claim 1, which contains 0.5 to 1.5 times the molar amount of alkali metal ions with respect to fumaric acid.
ンおよび／またはカリウムイオンである請求項１〜３の
いずれかに記載の方法。4. The method according to claim 1, wherein the alkali metal ion is sodium ion and / or potassium ion.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP6102789A JP2798886B2 (en) | 1994-05-17 | 1994-05-17 | Method for producing L-aspartic acid |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP6102789A JP2798886B2 (en) | 1994-05-17 | 1994-05-17 | Method for producing L-aspartic acid |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH07308195A true JPH07308195A (en) | 1995-11-28 |
| JP2798886B2 JP2798886B2 (en) | 1998-09-17 |
Family
ID=14336894
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP6102789A Expired - Fee Related JP2798886B2 (en) | 1994-05-17 | 1994-05-17 | Method for producing L-aspartic acid |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2798886B2 (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1998010008A1 (en) * | 1996-09-06 | 1998-03-12 | Bayer Aktiengesellschaft | Process for preparing polymers with recurring succinyl units |
| EP0832982A2 (en) * | 1996-09-20 | 1998-04-01 | DSM Chemie Linz GmbH | Process for the preparation of the disodium salt of z-l-aspartic acid from fumaric acid |
| EP0945517A2 (en) * | 1998-02-02 | 1999-09-29 | DSM Fine Chemicals Austria GmbH | Process for the preparation of the disodium salt of z-l-aspartic acid from fumaric acid |
| EP0952225A2 (en) * | 1998-02-13 | 1999-10-27 | Nippon Shokubai Co., Ltd. | Process for production of l-aspartic acid from fumaric acid with aspartase |
| EP0994189A1 (en) * | 1998-09-30 | 2000-04-19 | Nippon Shokubai Co., Ltd. | Methods for producing L-aspartic acid crystals |
-
1994
- 1994-05-17 JP JP6102789A patent/JP2798886B2/en not_active Expired - Fee Related
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1998010008A1 (en) * | 1996-09-06 | 1998-03-12 | Bayer Aktiengesellschaft | Process for preparing polymers with recurring succinyl units |
| EP0832982A2 (en) * | 1996-09-20 | 1998-04-01 | DSM Chemie Linz GmbH | Process for the preparation of the disodium salt of z-l-aspartic acid from fumaric acid |
| KR19980024723A (en) * | 1996-09-20 | 1998-07-06 | 하르트만 미카엘 | Method for preparing disodium B-L-aspartate from fumaric acid |
| EP0832982A3 (en) * | 1996-09-20 | 1998-09-02 | DSM Chemie Linz GmbH | Process for the preparation of the disodium salt of z-l-aspartic acid from fumaric acid |
| EP0945517A2 (en) * | 1998-02-02 | 1999-09-29 | DSM Fine Chemicals Austria GmbH | Process for the preparation of the disodium salt of z-l-aspartic acid from fumaric acid |
| EP0945517A3 (en) * | 1998-02-02 | 2000-08-02 | DSM Fine Chemicals Austria GmbH | Process for the preparation of the disodium salt of z-l-aspartic acid from fumaric acid |
| EP0952225A2 (en) * | 1998-02-13 | 1999-10-27 | Nippon Shokubai Co., Ltd. | Process for production of l-aspartic acid from fumaric acid with aspartase |
| EP0952225A3 (en) * | 1998-02-13 | 2000-08-02 | Nippon Shokubai Co., Ltd. | Process for production of l-aspartic acid from fumaric acid with aspartase |
| EP0994189A1 (en) * | 1998-09-30 | 2000-04-19 | Nippon Shokubai Co., Ltd. | Methods for producing L-aspartic acid crystals |
| US6821760B1 (en) | 1998-09-30 | 2004-11-23 | Nippon Shokubai Co., Ltd. | Methods for producing L-aspartic acid |
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| Publication number | Publication date |
|---|---|
| JP2798886B2 (en) | 1998-09-17 |
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