KR960007617B1 - Recovery process of 5'-inosinic acid by using ion exchange resin - Google Patents

Recovery process of 5'-inosinic acid by using ion exchange resin Download PDF

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KR960007617B1
KR960007617B1 KR1019920024171A KR920024171A KR960007617B1 KR 960007617 B1 KR960007617 B1 KR 960007617B1 KR 1019920024171 A KR1019920024171 A KR 1019920024171A KR 920024171 A KR920024171 A KR 920024171A KR 960007617 B1 KR960007617 B1 KR 960007617B1
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eluent
imp
concentration
circulating
elution
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KR940014798A (en
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박종호
최재효
이종균
황이남
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주식회사 미원
유영학
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Abstract

The circulating extraction agent 1 with 0.9-1.1% of chlorine ion is formed by mixing the first-stage extraction soln. 2 with inosinic acid(IMP) of 0.15% obtained after the first extraction and the last-stage extraction soln. 2 with IMP of 0.05%. The circulating extraction agent 2 with 3.0-3.5% conc. of chlorine ion is formed by mixing the middle-stage extraction soln. 1 with IMP of 2.0-2.5% obtained after the first extraction and the middle-stage extraction soln. 3 with IMP of 2.0-2.5%. The last-stage extraction soln. 1 with IMP of 0.2-0.4% is used for the first- and middle-stage extractions, and then 2.0% HCL is used for the last-stage extraction.

Description

이온 교환수지에 의한 5'-이노신산의 회수방법Recovery of 5'-inosinic acid by ion exchange resin

제1도는 종래 방법에 따른 이온 교환수지탑의 운전 공정도이고 ;1 is an operation flowchart of an ion exchange resin tower according to a conventional method;

제2도는 본 발명의 방법에 따른 이온 교환수지탑의 운전 공정도이다.2 is a flowchart of the operation of the ion exchange resin tower according to the method of the present invention.

본 발명은 통상의 발효법에 의하여 생산한 이노신산 모노포스페이트(이하 IMP라 칭한다)-함유 발효액으로부터 이온교환수지를 이용하여 IMP를 회수하는 방법에 관한 것이다. 보다 상세히 설명하면, 본 발명은 IMP 발효액을 원심분리하여 얻은 상징액을 이온교환수지 컬럼, 특히 Cl형 음이온 교환수지에 통액시킨 후 용출을 행함에 있어서, 1차 용출 후에 얻어지는 분획들 중 대부분의 IMP가 포함되어 있는 중기 분획은 다음 정제단계를 거쳐 정제하고, 소량의 IMP를 함유하는 초기 및 말기 용출분획을 재차 이온교환수지의 용리액으로 재사용함으로써 이차로 수지에 통액되는 IMP 발효액중의 IMP가 중기 용출분획중에 집중적으로 용출되어 고농도로 회수되도록 하고, 용리시간을 단축하며, 용리제의 재순환에 따른 폐수 발생량을 감소시킬 수 있도록 하는 IMP 회수 방법에 관한 것이다.The present invention relates to a method for recovering IMP from an inosinic acid monophosphate (hereinafter referred to as IMP) -containing fermentation broth produced by a conventional fermentation method using an ion exchange resin. In more detail, in the present invention, the supernatant obtained by centrifuging the IMP fermentation broth is passed through an ion exchange resin column, particularly a Cl type anion exchange resin, and then eluted. The intermediate fraction contained is purified through the following purification steps, and the IMP from the IMP fermentation liquor, which is passed through the resin secondly, is reused as an eluate of the ion exchange resin again and again. The present invention relates to a method for recovering IMP, which can be eluted intensively to recover a high concentration, shorten elution time, and reduce the amount of wastewater generated by recycling of the eluent.

종래 이온교환수지를 이용한 IMP의 회수방법은 다단계수지탑을 이용하는 것으로 원심분리 상징액을 직접 음이온 교환수지(Cl형)탑, 또는 양이온 교환수지탑에 통액하여 각종 유기물, 무기질을 제거한 후 다음 단계의 수지탑을 이용하여 정제한 후 농축 및 결정 과정을 거쳐 IMP 결정을 얻고 있다(일본특허공개 昭40-39715, 昭 35-34088).The conventional method of recovering IMP using ion exchange resin is to use a multi-stage resin tower. The centrifugal supernatant is directly passed through an anion exchange resin (Cl type) column or a cation exchange resin tower to remove various organics and minerals, followed by the resin of the next step. After purification using a column, IMP crystals are obtained through concentration and crystallization (JP-A-40-39715, A-35-34088).

상기 서술된 방법에서는 이온교환수지에 흡착된 IMP를 용출시키기 위하여 일정한 능도의 용리제, 예를들어 음이온 교환수지(Cl형)탑의 경우 약 2% 농도의 Cl-를 이용하고 있다. 한편, 당업계에서는 용리제의 사용량을 줄이고 IMP의 회수율을 높이고자 한번 사용된 용리제를 재차 재순환하여 사용하고자 하는 시도가 있어왔으며, 이렇게 재순환되는 용리제를 통상 순환 용리제가 불러오고 있다. 순환 용리제를 사용함에 있어서, 초기 순환 용리제의 Cl-농도를 균일하게 하기 위하여 고농도 IMP 용리후의 마지막 용리단계에서 과량의 액량을 용리하여 다음번 초기 용리제로 사용하는데 이때 IMP가 용리되기전 공극수를 포함하여 초기용리 1(Pre Cut 1)의 액량과 폐수 발생량이 많고, 다량의 유틸리티(물)을 사용하여야 하고, 수지탑 운전시간이 길고, 수지의 보충 비용부담이 클 뿐만 아니라 IMP 회수율이 현저히 떨어지며, 다량의 용리제 사용에 의한 운전비용이 많은 것이 단점으로 지적되었다.In the above-described method, to elute the IMP adsorbed on the ion exchange resin, an eluent having a certain ability, for example, Cl- at a concentration of about 2% in the case of an anion exchange resin (Cl type) column, is used. On the other hand, there has been an attempt in the art to recycle the eluent once used to reduce the amount of the eluent and to increase the recovery rate of the IMP, and the recirculating eluent is generally called a circulating eluent. In using the circulating eluent, in order to make the Cl- concentration of the initial circulating eluent uniform, the excess elution is eluted in the last elution step after the high concentration of IMP elution, and the pore water before the IMP is eluted. Pre-Elution 1 (Pre Cut 1) has a large amount of liquid and waste water generation, a large amount of utility (water) must be used, the operation time of the resin tower is long, the cost of replenishing the resin is high, and the recovery rate of IMP is remarkably decreased. It was pointed out that a high operating cost by using a large amount of eluent was a disadvantage.

또한 고농도외 IMP를 얻는 중기 용출단계에서 회수되는 IMP 농도의 편차가 심하고, 부피가 너무 많기 때문에 전체적으로 볼때 IMP의 농도가 낮아지는 단점이 있다.In addition, there is a disadvantage in that the concentration of IMP in the medium-term dissolution step of obtaining a high concentration of IMP is severe, and the IMP concentration is lowered as a whole because of too much volume.

따라서, 순환 용리제를 사용하여 IMP 발효액의 원심분리 상징액으로부터 IMP를 회수하는 방법에 있어서, 상기한 단점을 갖지 않고, 효율적이고 경제적인 방법으로 고농도의 IMP를 회수할 수 있는 방법의 제공이 요망되어 왔다.Therefore, in the method of recovering IMP from the centrifugation supernatant of an IMP fermentation broth using a circulating eluent, it is desired to provide a method capable of recovering high concentrations of IMP in an efficient and economical manner without having the aforementioned disadvantages. come.

이러한 상황하에서 본 발명자들은 예의 연구한 결과 1차 이온교환수지 용출에 있어서, 용리제의 농도를 IMP가 소량 용출되는 용출 초기, 대부분의 IMP가 용출되는 용출 중기 및 나머지 소량의 IMP가 용출되는 용출 분기에 따라 변경하여 사용하고 2차 이온교환수지 용출에 있어서 1차 용출에서 분리된 초기 용리액과 말기 용리액을 적의 혼합하여 초기 순환용리제로 사용하므로써 상기 목적을 달성할 수 있음을 발견하고 본 발명을 완성하기에 이르렀다.Under these circumstances, the present inventors have diligently studied, in the first ion exchange resin elution, the concentration of the eluent in the early stage of the elution of the small amount of IMP, the middle of the elution in which most IMP is eluted, and the elution branch in which the remaining small amount of IMP is eluted. In accordance with the present invention, the present invention can be accomplished by mixing the initial eluate and the final eluate separated from the primary elution and using the initial eluent as the initial circulation eluent in the secondary ion exchange resin elution. Reached.

즉, 본 발명의 목적을 통상의 발효법에 의하여 생산한 IMP 발효액으로부터 Cl형 음이온 이온교환수지를 이용하여 IMP를 고농도로 회수하는 새로운 방법을 제공하는 것이다.That is, an object of the present invention is to provide a new method for recovering IMP at a high concentration using Cl type anion ion exchange resin from IMP fermentation broth produced by a conventional fermentation method.

본 발명의 또다른 목적은 IMP 발효액을 원심분리하여 얻은 상징액을 Cl형 음이온 교환수지에 통액시킨 후 용출을 행함에 있어서, 1차 용출후에 소량의 IMP를 함유하는 초기 및 말기 용출 분획을 2차 이온교환수지의 용리액(순환 용리액)으로 재사용함을 특징으로 하는 IMP 회수방법을 제공하는 것이다.Another object of the present invention is to pass the supernatant obtained by centrifuging the IMP fermentation broth to Cl-type anion exchange resin, and then eluting. The initial and final elution fractions containing a small amount of IMP after primary elution are subjected to secondary ions. It is to provide an IMP recovery method characterized by reusing as an eluent (circulating eluent) of the exchange resin.

본 발명의 또다른 목적은 IMP 발효액을 원심분리하여 얻은 상징액을 Cl형 음이온 교환수지에 통액시킨 후 용출을 행함에 있어서, IMP 농도가 0.00%인 초기 용리액(Pre Cut) 1, IMP 농도가 약 0.15%인 초기 용리액 2, IMP 농도가 약 2~2.5%인 중기 용리액(Reach Cut) 1, IMP 농도가 6.5%이상인 중기 용리액2, IMP 농도가 약 2%인 중기 용리액 3, IMP 농도가 약 0.2-0.4%인 말기 용리액(Tail Cut) 1 및 IMP농도가 약 0.05%인 말기 용리액 2를 얻고 이중 중기 용리액 2는 그 다음의 IMP 정제단계에 따라 분리 및 정제하고, 초기 용리액 2와 말기 용리액 2를 합한 순환 용리제1, 중기 용리액 1과 중기 용리액 3을 합하고 여기에 35% 염산을 일정량 첨가하여 제조한 순환 용리제 2 및 말기 용리액 1인 순환 용리제 3을 순환 용리제 시스템으로 사용하여 2차 이온교환수지 컬럼으로루터 IMP를 용출시킴을 특징으로 하는 IMP 회수방법을 제공하는 것이다.Another object of the present invention is to pass the supernatant obtained by centrifuging the IMP fermentation broth to Cl-type anion exchange resin, and then eluting, the initial eluent (Pre Cut) 1 having an IMP concentration of 0.00% and an IMP concentration of about 0.15 % Initial eluent 2, IMP concentration 2 ~ 2.5% medium eluent (Reach Cut) 1, IMP concentration 6.5% or more medium eluent 2, IMP concentration 2% medium eluent 3, IMP concentration about 0.2- A tail cut 1 of 0.4% and a terminal eluent 2 having an IMP concentration of about 0.05% are obtained, and the middle medium eluent 2 is separated and purified according to the following IMP purification step, and the initial eluent 2 and the final eluent 2 are combined. Secondary ion exchange using circulating eluent 1 and circulating eluent 3, the final eluent 1, prepared by combining circulating eluent 1, medium eluent 1, and medium eluent 3, and adding a certain amount of 35% hydrochloric acid. Characterizes eluting Luther IMP into resin columns To provide a method for recovering IMP to.

이하 본 발명을 보다 상세히 설명한다.Hereinafter, the present invention will be described in more detail.

제1도를 참고로 하여 종래 당업계에서 행해지고 있던 순환 용리제를 이용한 이온교환 수지탑의 운전법은 다음과 같다.Referring to FIG. 1, an operation method of an ion exchange resin tower using a circulating eluent conventionally performed in the art is as follows.

제1도에서 보듯이, 2.0% HCI을 이용한 1차 음이온교환수지탑의 용출에 의해 초기 용리액 1 2850m1, 초기 용리액 2 480ml, IMP를 대부분 함유하는 중기 용리액 2650m1, 말기 용리액 1 3850m1, 및 말기 용리액 2 2080ml를 얻는다. 이어, 음이온 교환수지에 2.5-7%의 가성소다를 사용하여 음이온 OH형으로 만든 후 다시 염산 2.5-7% 용액을 사용하여 Cl형으로 재생한다. IMP 발효액을 공간 속도 0.7 또는 1.0으로 수지에 통액한 후, 초기 용리액 2과 말기 용리액 2로 이루어진 순환 용리제(1)과 말기 용리액 1인 순환 용리제(2)를 차례로 이용하여 용출을 행하고 이어 본 용리제로서 2.0% HCI을 이용하여 용출시키고 최종적으로 중류수를 이용하여 용출시킨다. 상기 공지 기술에 따르면, IMP가 용리되기전 공극수를 포함하여 초기용리 1(Pfe Cut 1)의 액량과 폐수 발생량이 많고, 다량의 유틸리티(물)를 사용하여야 하고, 수지탑 운전시간이 길고, 수지의 보충 비용부담이 클 뿐만 아니라 IMP 회수율이 현저히 떨어지며, 다량의 용리제 사용에 의한 운전비용이 많은 단점을 갖고 있다.As shown in FIG. 1, the initial eluent 1 2850m1, the initial eluent 2 480ml, the middle eluent 2650m1, the final eluent 1 3850m1, and the final eluent 2 were eluted by elution of the primary anion exchange resin tower using 2.0% HCI. Obtain 2080 ml. Next, an anion exchange resin was used to make anionic OH using 2.5-7% caustic soda, and then regenerated to Cl using 2.5-7% hydrochloric acid solution. After passing the IMP fermentation broth to the resin at a space velocity of 0.7 or 1.0, the elution was carried out using a circulating eluent (1) consisting of an initial eluent 2 and a late eluent 2, followed by a circulating eluent (2) having a final eluent 1 Eluting with 2.0% HCI as eluent and finally eluting with midstream water. According to the above-mentioned known technology, before the IMP is eluted, the amount of liquid and wastewater generated in the initial elution 1 (Pfe Cut 1) including the pore water is large, a large amount of utility (water) must be used, and the resin tower operation time is long, and the resin In addition to the large burden of replenishment costs, the recovery rate of IMP is remarkably decreased, and the operating cost by the use of a large amount of eluent is disadvantageous.

반면 제2도를 참고로 하여 본 발명의 방법을 설명하면, 1차 용출에서 IMP 농도가 0.00%인 초기 용리액 1, IMP 농도가 0.15%인 초기 용리액 2, IMP 농도가 2.15%인 중기 용리액 1, IMP 농도가 6.5% 이상인 중기 용리액 2, IMP 농도가 1.90%인 중기 용리액 3, IMP 농도가 0.36%인 말기 용리액 1 및 IMP농도가 0.049%인 말기 용리액 2를 얻고 이중 중기 용리액 2는 그 다음의 IMP 정제단계에 따라 분리 및 정제하고, 초기 용리액 1은 폐수로 폐기한다. 초기 용리액 2와 말기 용리액 2를 합한 순환 용리제 1, 중기 용리액 1과 중기 용리액 3을 합하고 여기서 35% 염산을 일정량 첨가하여 제조한 순환 용리제 2 및 말기용리액 1인 순환 용리제 3을 순환 용리제 시스템으로 사용하여 2차 이온교환수지 컬럼으로부터 IMP를 용출시킨다. 그리고 일차 이온교환수지 컬럼에서 얻어진 용리액들은 다시 위와 같은 방법으로 다음번 컬럼에 재순환시킨다.On the other hand, the method of the present invention is described with reference to FIG. 2. In the first elution, the initial eluent 1 having an IMP concentration of 0.00%, the initial eluent 2 having an IMP concentration of 0.15%, and the intermediate eluent 1 having an IMP concentration of 2.15%, Medium eluent 2 with an IMP concentration of at least 6.5%, medium eluent 3 with an IMP concentration of 1.90%, terminal eluent 1 with an IMP concentration of 0.36% and terminal eluent 2 with an IMP concentration of 0.049%, of which intermediate Eluent 2 is the next IMP. Separation and purification are carried out according to the purification step, and the initial eluent 1 is disposed of in waste water. Circulating eluent 1 and circulating eluent 3 prepared by combining the initial eluent 2 and the final eluent 2, the intermediate eluent 1 and the intermediate eluent 3, and adding a certain amount of 35% hydrochloric acid, and the final eluent 1 circulation eluent 3 IMP is used to elute the IMP from the secondary ion exchange resin column. The eluents obtained from the primary ion exchange resin column are recycled to the next column in the same manner as above.

소망하는 결과를 얻기 위해서는 순환 용리제의 Cl-농도 및 공간 속도가 중요하다. 즉, 순환용리제 1은The Cl concentration and space velocity of the circulating eluent are important for obtaining the desired results. That is, circulating eluent 1

그 Cl-농도를 0.9~1.1%로 조절하여야 한다. 순환 용리제 2는 그 Cl-농도가 0.9~1.1%이고, IMP 농도가 2.0∼2.5%인데, 35% 염산 0.8~1 2RV(Resin Volume)를 가하여 그 Cl-농도를 3.0~3.5%로 조정하여 사용하여야 한다. 0.8~1.2RV의 염산은 약 70m1의 부피에 해당한다. 순환 용리제 3으로는 말기 용리액1을 그냥 사용하면 된다. 순환 용리제의 용출 속도는 공간 속도 0.7∼1.0이다.The Cl - concentration should be adjusted to 0.9-1.1%. The circulating eluent 2 has a Cl - concentration of 0.9-1.1% and an IMP concentration of 2.0-2.5%, and the Cl - concentration is adjusted to 3.0-3.5% by adding 0.8-1 2 RV (Resin Volume) of 35% hydrochloric acid. Should be used. Hydrochloric acid at 0.8-1.2 RV corresponds to a volume of about 70 m1. As the circulating eluent 3, the terminal eluent 1 can be used. The elution rate of the circulating eluent is a space velocity of 0.7 to 1.0.

상기와 같은 순환 용리제를 사용하여 초기 용출을 행하고, 이어 2.0% HCI을 이용하여 본 용출을 행하고 마지막으로 증류수를 이용하여 용출을 시킨다.The initial elution is performed using the above circulating eluent, followed by the main elution using 2.0% HCI, and finally the elution using distilled water.

본 발명에 사용할 수 있는 음이온 교환수지는 통상의 것을 사용할 수 있으며, 예를 들면 DIAI0N PA 412, DUOLITE A 162, PUROLITE A 510.S 및 DOWEX MSA 2같은 시판되는 것을 사용할 수 있다.The anion exchange resin which can be used for this invention can use a conventional thing, For example, commercially available things, such as DIAI0N PA 412, DUOLITE A 162, PUROLITE A 510.S and DOWEX MSA 2, can be used.

상기 설명한 본 발명의 용출 방법에 따르면, 기존의 방법에 비해 IMP 농도를 40%이상 상승시켜 다음 용이공정의 부하량이 현저히 감소되고, 둘째 용리제중 염산 사용량이 15∼30% 감소되어 원부재가 절감되고 셋째, 유틸리티(물) 사용량이 대폭 줄어 폐수발생량이 감소되고, 그에 따른 순환과정에서의 저농도 IMP용기액을 폐액으로 버리지 않고 다음공정의 용리제로 사용하여 IMP의 회수율 99.5%이상, 1회 용리시간을 20-30% 감소시키는 효과가 얻어진다.According to the elution method of the present invention described above, the load of the next easy process is significantly reduced by increasing the IMP concentration by more than 40% compared to the conventional method, and the amount of hydrochloric acid in the eluent is reduced by 15 to 30%, thereby reducing raw materials. Third, the amount of utility (water) is drastically reduced, and the amount of wastewater generated is reduced, and the recovery rate of IMP is not less than 99.5% and one time elution time is used as the eluent of the next process without discarding the low concentration IMP solution in the circulating process. The effect of reducing 20-30% is obtained.

본 발명을 실시예에 따라 상세히 설명하면 다음과 같다.Hereinafter, the present invention will be described in detail with reference to Examples.

실시예 1Example 1

통상의 방법으로 수득한 IMP 발효액(IMP : 4.08%. 하이포크산틴(HX) : 0.16%, 이노신(INO) : 0.07%) 3,200m1를 다아이온(DIAI0N) PA 412 수지를 충전한 컬럼에 통액하고 통상의 방법으로 1차 용출하여 IMP 농도가 0.00%인 초기 용리액 1, IMP 농도가 0.15%인 초기용리액 2, IMP 농도가 2.15%인 중기 용리액 1, IMP 농도가 6.5% 이상인 중기 용리액 2, IMP 농도가 1.90%인 중기 용리액 3, IMP 농도가 0.36%인 말기 용리액 1 및 IMP 농도가 0.049%인 말기 용리액 2를 얻었다. 이중 초기 용리액 1은 폐수로 버리고 중기 용리액 2는 그 다음의 IMP 정제단계에 따라 분리 및 정제하였다. 이어, 음이온 교환수지에 2.5∼7%의 가성소다를 사용하여 음이온 OH형으로 만든후 다시 염산 2.5-7% 용액을 사용하여 Cl형으로 재생하였다.IMP fermentation broth obtained by a conventional method (IMP: 4.08%. Hypoxanthin (HX): 0.16%, Inosine (INO): 0.07%) 3,200 m1 was passed through a column packed with DIA0N PA 412 resin and usually The primary eluate 1 was eluted with an IMP concentration of 0.00%, the initial eluate 2 with an IMP concentration of 0.15%, the intermediate eluent 1 with an IMP concentration of 2.15%, and the intermediate eluent 2 with an IMP concentration of 6.5% or more, and the IMP concentration. A middle eluent 3 having 1.90%, a terminal eluent 1 having an IMP concentration of 0.36%, and a final eluent 2 having an IMP concentration of 0.049% were obtained. The initial eluent 1 was discarded into the wastewater and the intermediate eluent 2 was separated and purified according to the next IMP purification step. Subsequently, 2.5 to 7% of caustic soda was used as the anion exchange resin to form anionic OH, followed by regeneration to Cl using 2.5-7% hydrochloric acid solution.

다시, 통상의 방법으로 수득한 IMP 발효액(IMP : 4.08%, 하이포크산틴(HX) : 0.16%, 이노신(INO): 0.07%) 3,200m1를 상기 재생된 수지탑에 공간속도(S.V).0.5 또는 0.7로 통액하고, 하기 표 1에 나타낸 HCl% 농도를 갖는 순환용리제 2,560ml 및 본 용리제 2.0% HC1을 이용하여 용출을 행하고 순환 용리제중의 Cl 이온농도 및 공간 속도가 IMP 회수율 및 폐수로 버려지는 초기 용리액 1의 부피에 미치는 영향을 조사하였다 결과는 표 1과 같다.Again, 3,200 ml of IMP fermentation broth (IMP: 4.08%, hypoxanthine (HX): 0.16%, inosine (INO): 0.07%) obtained by a conventional method was added to the regenerated resin tower. Elution was carried out using 2,560 ml of a circulating eluent having a concentration of HCl% shown in Table 1 and 2.0% HC1 of the present eluent, and the concentration of Cl ions and the space velocity in the circulating eluent were converted into IMP recovery rate and wastewater. The effect on the volume of the initial eluent 1 discarded was investigated.

실시예 2Example 2

IMP 발효액(IMP : 4.08%,하이포크산틴(HX) : 0.16%,이노신(INO : 0.07%) 3,200m1를 실시예 1과 같이 재생한 수지탑에 S.V : 0.7로 통액하였다.IMP fermentation broth (IMP: 4.08%, hypoxanthine (HX): 0.16%, inosine (INO: 0.07%) 3,200 m1 was passed through the resin tower regenerated as in Example 1 as S.V: 0.7.

1차 용출에서 IMP농도가 0.00%인 초기 용리액 1, IMP 농도가 0.15%인 초기 용리액 2, IMP 농도가 2.15%인 중기 용리액 1, IMP 농도가 6.5% 이상인 중기 용리액 2, IMP 농도가 1.90%인 중기 용리액 3,IMP 농도가 0.36%인 말기 용리액 1 및 IMP 농도가 0.049%인 말기 용리액 2를 얻고 이중 중기 용리액 2는 그 다음의 IMP 정제단계에 따라 분리 및 정제하고, 초기 용리액 1은 폐수로 폐기한다. 초기 용리액 2와 말기 용리액 2를 합하고Cl 농도를 0.9∼1.1%로 조절하여 순환 용리제 1을 준비하고, 중기 용리액 1과 중기 용리액 3을 합하고 여기애 35% 염산을 약 70ml 첨가하여 Cl 농도를 3.0~3.5%로 조정한 순환 용리제2를 제조하고 말기 용리액 1을 그대로 순환 용리제 3으로 만든다. 이들 순환 용리제 1∼3으로 이루어진 순환 용리제 시스템을 초기 용리제로 사용하여 공간 속도 0.7로 용출하고, 이어 본 용리제로서 2.0% HCI 3,600m1를 사용하여 S.V.0.7로 용리를 행하였다.In the first elution, the initial eluent 1 with an IMP concentration of 0.00%, the initial eluent 2 with an IMP concentration of 0.15%, the medium eluent 1 with an IMP concentration of 2.15%, the medium eluent 2 with an IMP concentration of 6.5% or more, and the IMP concentration 1.90% Medium Eluent 3, Terminal Eluent 1 with IMP concentration of 0.36% and Terminal Eluent 2 with IMP concentration of 0.049% were obtained, of which Middle Eluent 2 was separated and purified according to the next IMP purification step, and Initial Eluent 1 was disposed of in wastewater. do. Combine the initial eluent 2 and the late eluent 2 with Cl The concentration was adjusted to 0.9 to 1.1% to prepare a circulating eluent 1, the medium eluent 1 and the medium eluent 3 were combined, and about 70 ml of 35% hydrochloric acid was added thereto to give Cl. Circulating eluent 2 was prepared by adjusting the concentration to 3.0 to 3.5%, and the final eluent 1 was changed to circulating eluent 3 as it is. The circulating eluent system consisting of these circulating eluents 1-3 was used as an initial eluent to elute at a space velocity of 0.7, followed by elution with S.V.0.7 using 2.0% HCI 3,600 m1 as the eluent.

초기 용리액 1 2,240ml(IMP : 0.00%)는 폐기하고, 초기 용리액 2 480m1(IMP ·0.15%)와 중기 용리액1 640ml(IMP.2.15%), 중기 용리액 800m1(IMP : 1.90%), 말기 용리액 1 1,840m1(IMP : 0.36%), 말기 용리액 2 2,080rn1(IMP 0.049%)로 이루어진 총 5,680m1는 다음번 수지의 순환용리제로 사용하였다. 중기용리액 2 1,920m1(IMP : 6.75%)를 얻어 통상의 방법으로 탈색 수지 또는 활성탄을 사용하여 탈색한 후 중화 및 농축과 정을 거쳐 IMP 결정 120.2g을 얻었고, 이때의 회수율은 92.06%(IMP 발효액 대비)이었다.Discard the initial eluent 1 2,240 ml (IMP: 0.00%), remove the initial eluent 2 480 m1 (IMP0.15%), medium eluent 1 640 ml (IMP.2.15%), medium eluent 800 m1 (IMP: 1.90%), final eluent 1 A total of 5,680m1 consisting of 1,840m1 (IMP: 0.36%) and the final eluent 2 2,080rn1 (IMP 0.049%) were used as the next circulating eluent. Mid-term eluent 2 1,920m1 (IMP: 6.75%) was obtained and decolorized using a decolorizing resin or activated carbon in a conventional manner, and then neutralized and concentrated to obtain 120.2 g of IMP crystals. The recovery rate was 92.06% (IMP fermentation broth). Contrast).

비교예Comparative example

통상의 방법으로 얻은 IMP 발효액 (IMP : 4.08%,하이포크산틴(HX).0.16%,이노신(INO): 0.07%) 3,200ml를 실시예 1과 같이 재생한 강염기성 음이온 교환수지(Cl형)탑에 S.V : 0.7로 통액하고 이어, 초기 용리액 1과 말기 용리액 2를 단순히 혼합하기만 한 순환용리제 1 및 말기 용리액 1인 순환 용리제 2를 초기 용리제로 사용하여 용출을 행하고 이어 2.0% HCl 본 용리제를 사용하여 S.V : 0.7로 IMP를 용리하였다.Strong base anion exchange resin (Cl type) column regenerated 3,200 ml of IMP fermentation broth (IMP: 4.08%, hypoxanthine (HX), 0.16%, inosine (INO): 0.07%) obtained in the same manner as in Example 1. The solution was passed through SV: 0.7, followed by elution using circulating eluent 1 and final eluent 1, which were simply mixed with the initial eluent 1 and the final eluent 2, as the initial eluent, followed by 2.0% HCl. IMP eluted with SV: 0.7 using Lize.

상기 실시예 2와 비교예에서 얻은 결과를 표 2에 나타내었다.Table 2 shows the results obtained in Example 2 and Comparative Example.

상기 표의 결과에서 알 수 있듯이, 본 발명에 따른 방법을 사용하면 회수된 IMP농도가 40%이상 상숭되고. 용리제인 염산 사용량이 15-30% 감소되어 원부재가 절감되고 유틸리티(물) 사용량이 대폭 줄어 폐수 발생량이 감소되며 IMP의 회수율 99.5%이상, 1회 용리시간을 20-30% 감소시키는 효과가 얻어진다.As can be seen from the results of the above table, using the method according to the invention the recovered IMP concentration is at least 40%. The use of hydrochloric acid as eluent is reduced by 15-30%, which reduces raw materials, drastically reduces utility (water) use, and reduces wastewater generation, and reduces the recovery rate of IMP by more than 99.5% and the one-time elution time by 20-30%. Lose.

Claims (2)

통상의 방법으로 얻은 IMP 발효액으로부터 균체를 제거한 강염기성 음이온 교환수지(Cl형)탑에 흡착시켜 IMP를 회수하는 방법에 있어서, 1차 용출 후에 얻어진 IMP 농도가 약 0.15%인 초기 용리액 2와 IMP 농도가 약 0.05%인 말기 용리액 2를 합한 용액의 Cl-농도가 0.9-1.1%인 순환 용리제 1, 1차 용출 후에 얻어진 IMP 농도가 약 2~2.5%인 중기 용리액 1과 IMP 높도가 약 2.0~2.5%인 중기 용리액 3을 합하고 HC1을 가하여 Cl-농도를 3.0~3.5%로 조정한 순환 용리제 2 및 1차 용출후에 얻어진 IMP 농도가 약 0.2-0.4%인 말기 용리액 1을 순환 용리제로 사용하여 초기 및 중기 용리를 행하고, 이어 2.0% HC1을 사용하여 말기 용리를 행함을 특징으로 하는 고농도 IMP를 회수하는 방법.A method of recovering IMP by adsorbing to a strong basic anion exchange resin (Cl type) column from which cells are removed from an IMP fermentation broth obtained by a conventional method, wherein the initial eluate 2 and IMP concentration having an IMP concentration obtained after the first elution is about 0.15%. Circulating eluent 1 with a Cl concentration of 0.9-1.1%, medium eluent 1 with an IMP concentration of about 2% to 2.5% and an IMP level of about 2.0 Circulating eluent 3 with ~ 2.5% medium eluent 3 added and HC1 added to adjust Cl concentration to 3.0-3.5%, and eluent 1 with a final IMP concentration of about 0.2-0.4% obtained as the primary eluent Recovering the high concentration of IMP, characterized in that the initial and intermediate elution is carried out, followed by the final elution using 2.0% HC1. 제1항에 있어서, 순환 용리제 2의 제조에 있어서, HCI을 0.8-1.2 RV(RESIN VOLUME)의 양으로 사용하여 Cl-농도를 3.0~3.5%로 조정함을 특징으로 하는 방법.The method of claim 1, wherein in the preparation of circulating eluent 2, the Cl concentration is adjusted to 3.0-3.5% using HCI in an amount of 0.8-1.2 RV (RESIN VOLUME).
KR1019920024171A 1992-12-14 1992-12-14 Recovery process of 5'-inosinic acid by using ion exchange resin KR960007617B1 (en)

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