JPS6364200B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention utilizes a fixed bed filled with a strongly acidic cation exchange resin in the form of an alkali metal to produce maltose,
The purpose is to separate maltose from a mixture of oligosaccharides and glucose with high purity and high yield. Maltose is made up of two molecules of D-glucose
It is a 4-linked disaccharide that has a unique flavor and has been widely used in foods as the main component of malt starch syrup since ancient times. However, in recent years, maltose has a higher calorie content than glucose, so its demand as a clinical drug has increased significantly. When producing maltose industrially, starch is generally used as a raw material, and an enzymatic method is used in which starch is first treated with α-amylase to liquefy the starch, and then β-amylase and isoamylase or pullulanase are applied. It will be done. however,
This production method uses three types of enzymes with different functions and requires a very long time to produce a maltose solution with a high maltose content, so the production conditions must be strictly controlled. There must be. Moreover, the produced maltose solution contains oligosaccharides and glucose in addition to maltose, although this differs depending on the two types of enzymes. Furthermore, production is relatively easy when the maltose content is 82 to 83%, but it is extremely difficult when the maltose content is 88% or more. On the other hand, when maltose is used for medical purposes, maltose of extremely high purity is required. Against this background, the present inventors conducted intensive research using a chromatographic separation method to separate high-purity maltose from a maltose solution containing impurities. We have discovered that maltose of high purity can be separated by chromatographic separation. That is, the present invention involves passing a mixed stock solution of maltose, oligosaccharides, and glucose through a fixed bed packed with an alkali metal-type strongly acidic cation exchange resin in a downward or upward flow to separate maltose and other sugars. An oligosaccharide adsorption zone formed while moving in the packed bed by passing a certain amount of stock solution and a specified amount of water in the order of stock solution and water, and circulating the effluent again through the packed bed in the order of flow for at least two or more cycles. A preliminary step is carried out to amplify the distance between the maltose adsorption zone and the glucose adsorption zone, and after the circulation of the preceding step, the dilute oligosaccharide solution,
Of the effluents that flow out with a chromatographic concentration distribution in the order of oligosaccharide solution, oligosaccharide/maltose mixture, maltose solution, maltose/glucose mixture, glucose solution, and dilute glucose solution, a specified amount of oligosaccharide solution and a specified amount are specified. A specified amount of maltose solution and a specified amount of glucose solution are taken out of the system in spots, while other liquids are circulated again in the order of outflow. Oligosaccharide solution, maltose solution, and glucose are injected into the system, and a specified amount of water is injected spot-wise after the inflow of the dilute glucose solution in the circulating fluid, and the total flow rate of the injected stock solution and water is taken out of the system. Circulate the liquid so that it is equal to the total liquid volume. The present invention relates to a method for separating maltose, which is characterized in that a steady step is performed and thereafter the steady step is repeated to sequentially take out an oligosaccharide solution, a maltose solution, and a glucose solution. Hereinafter, an example of an embodiment of the present invention will be described in detail using the drawings. FIG. 1 is an explanatory diagram showing the flow of a maltose separation device, in which one end of an inflow pipe 3 is connected to the upper end of an adsorption tower 2 filled with an alkali metal type strongly acidic cation exchange resin 1, and the other end is It communicates with pump 4. Further, one end of the outflow pipe 5 is connected to the lower end of the adsorption tower 2, and the other end is connected to the outflow branch pipes 5a, 5b, 5c, 5d, 5.
e, 5f, and 5h, respectively. Oligosaccharide thin material tank 6, oligosaccharide liquid tank 7, oligosaccharide and maltose mixed liquid tank 8, maltose liquid tank 9, maltose/glucose mixed liquid tank 10, glucose liquid tank 11, glucose thin material tank each having a weir 24 inside the end. 12, the effluent from the outflow branch pipe 5a is sent to the oligosaccharide thin substance tank 6, the outflow from the outflow branch pipe 5b is sent to the oligosaccharide liquid tank 7, and the outflow from the outflow branch pipe 5c is sent to the oligosaccharide/maltose tank. The effluent from the outflow branch pipe 5d is transferred to the mixed liquid tank 8 and the maltose liquid tank 9.
The effluent from the outflow branch pipe 5e contains maltose.
The configuration is such that the effluent from the outflow branch pipe 5f flows into the glucose liquid tank 10, the outflow from the outflow branch pipe 5h flows into the glucose thin substance tank 12, and the outflow from the outflow branch pipe 5h flows into the glucose liquid tank 12, respectively. On the other hand, one end of the suction pipe 13a is connected to the lower part of the oligosaccharide liquid tank 6, and one end of the suction pipe 13b is connected to the lower part of the oligosaccharide liquid tank 7.
One end of the suction pipe 13c is placed at the bottom of the maltose mixed liquid tank 8, one end of the suction pipe 13d is placed at the bottom of the maltose liquid tank 9, one end of the suction pipe 13e is placed at the bottom of the maltose/glucose mixed liquid tank 10, and one end of the suction pipe 13e is placed at the bottom of the maltose/glucose mixed liquid tank 10. One end of the suction pipe 13f is connected to the lower part of the pump 4, and one end of the suction pipe 13h is connected to the lower part of the glucose thin material tank 12, and the other ends of all the suction pipes are connected to the suction main pipe 14, and the suction main pipe is connected to the pump 4. communicate with. Further, a stock solution tank 15 and a water tank 16 are installed, and one end of the introduction pipe 17a is connected to the bottom of the stock solution tank 15, and one end of the introduction pipe 17b is connected to the bottom of the water tank 16, and the other end of these introduction pipes is connected to the bottom of the stock solution tank 15. are connected to pumps 18 and 19, respectively. In addition, the pump 18,
19, one end of the discharge pipes 20a, 20b is communicated with each other, and the other end is connected to the middle of the inflow pipe 3. In addition, an oligosaccharide liquid extraction tube 21 is branched and attached to the suction tube 13b, a maltose liquid extraction tube 22 is branched and attached to the suction tube 13d, and the suction tube 13f is also attached.
A glucose liquid take-out pipe 23 is branched and attached to the pipe. When separating the oligosaccharide solution, maltose solution, and glucose solution from the stock solution using the maltose separation device configured as described above, in the present invention, first, a specified amount of the stock solution and a specified amount of water are passed through in the order of the stock solution and the water. A preliminary step is performed in which the liquid is drained and the effluent is circulated again through the packed bed in the order in which it flows out for at least two cycles or more. That is, a specified amount of the stock solution is poured into the stock solution tank 1 from the top of the adsorption tower 2, in which some water layer is formed above the packed bed of the strongly acidic cation exchange resin 1.
5 using the pump 18, and then, when the prescribed amount of the stock solution has finished flowing in, a prescribed amount of water is successively introduced from the water tank 16 using the pump 19. On the other hand, at the same time as the raw solution and water flow into the adsorption tower 2,
Let the effluent flow out from the bottom of the. As shown in FIG. 2, the effluent is separated to some extent into the oligosaccharide liquid tank O, the maltose liquid part M, and the glucose liquid part G, and the effluent is sent to the outflow branch pipes 5a, 5b, and 5c, respectively. , 5d, 5e, 5f, and 5h, the liquid flowing out is sent to the oligosaccharide thin tank 6,
It is received in an oligosaccharide liquid tank 7, an oligosaccharide/maltose mixed liquid tank 8, a maltose liquid tank 9, a maltose/glucose mixed liquid tank 10, a glucose liquid tank 11, and a thin glucose tank 12. After receiving the liquid of each portion in each tank in this manner, the liquid of each of these portions is then allowed to flow back into the adsorption tower 2 in the order in which it was discharged, and is circulated. That is, the liquid in the thin oligosaccharide tank 6 is first introduced from the upper part of the adsorption tower 2 through the suction pipe 13a using the pump 4, and then the liquid in the oligosaccharide liquid tank 7,
The liquids in the oligosaccharide/maltose mixed liquid tank 8, the maltose liquid tank 9, the maltose/glucose mixed liquid tank 10, the glucose liquid tank 11, and the glucose thin substance tank 12 are passed through suction pipes 13b, 13c, 13d, 13e, and 1 in the order in which they flow out.
3f and 13h, respectively, and the adsorption tower 2 is made to flow in from the upper part one after another, and the effluent is again received into each tank in the order of outflow. If the above-mentioned circulation is repeated, for example, for three cycles, the concentration distribution of the effluent from the first cycle shown in Figure 2 will change to the concentration distribution of the effluent from the third cycle shown in Figure 3. The concentration curves of liquid part O, maltose liquid part M, and glucose liquid part G gradually diverge. This indicates that the distances between the oligosaccharide adsorption zone, the maltose adsorption zone, and the glucose adsorption zone, which are formed while moving in the packed bed, are amplified. In the present invention, the distances between the oligosaccharide adsorption zone, the maltose adsorption zone, and the glucose adsorption zone can be amplified by performing the preceding step of circulating the effluent as described above, so that high purity can be achieved in the subsequent regular step. It is possible to separate maltose. For example, when obtaining an oligosaccharide solution, a maltose solution, and a glucose solution from the effluent in the third cycle after circulation, the oligosaccharide solution, maltose solution, and glucose solution are sequentially taken out by performing the steady process described below. The outflow of each liquid in the third cycle is as shown in Figure 3. Part A of the oligosaccharide thin liquid that flows out first is transferred to the oligosaccharide thin liquid tank 6, and the next relatively high concentration oligosaccharide is transferred to the oligosaccharide thin liquid tank 6. Part A of the sugar solution is sent to the oligosaccharide solution tank 7.
Part C of the next oligosaccharide/maltose mixture goes to oligosaccharide/maltose mixture tank 8, part E of the next maltose solution goes to maltose tank 9, part E of the next maltose/glucose mixture goes to maltose/glucose. A portion of the next glucose solution is transferred to the mixed liquid tank 10, a portion of the next glucose liquid is transferred to the glucose liquid tank 11, and a portion of the glucose thin substance that is finally discharged is separated into the glucose thin substance tank 12.
Take out part A of the oligosaccharide liquid in the oligosaccharide liquid tank 7 out of the system using the Also, use the glucose liquid extraction pipe 23 to remove the glucose from the glucose liquid tank 1.
A portion of the glucose solution in 1 is taken out of the system. On the other hand, the effluents from the other tanks are again circulated in the order of their outflow using the pump 4, and at this time, a specified amount of the stock solution and a specified amount of water are injected spot-wise into the portions described below. That is, the circulation in the steady process is to flow the effluent from the oligosaccharide thin material tank 6, then the effluent from the oligosaccharide/maltose mixture tank 8 into the adsorption tower 2 using the pump 4, and then add a specified amount of the stock solution. It is made to flow in from the stock solution tank 15 using the pump 18. After injecting a specified amount of the stock solution, the effluent in the maltose/glucose mixed liquid tank 10 and then the effluent in the glucose thin material tank 12 are injected in this order, and then a specified amount of water is injected from the water tank 16 using the pump 19. . on the other hand,
The effluent flowing out from the adsorption tower 2 is divided into parts A, A, C, E, O, F, and G as shown in FIG.
It is received in each tank 7, 8, 9, 10, 11, and 12 in order. Thereafter, the oligosaccharide solution, maltose solution, and glucose solution are sequentially taken out through a regular process in which the removal of the oligosaccharide solution, maltose solution, and glucose solution, and the injection of the stock solution and water are repeated in the same manner. The present invention involves first carrying out the preceding step as described above, and then sequentially repeating the steady-state steps to sequentially extract a high-purity maltose solution, a relatively high-purity oligosaccharide solution, and a glucose solution at a relatively high yield. is possible. In this case, it is necessary not to disturb the outflow curves of maltose, oligosaccharides, and glucose during the circulation in the preceding process, and it is also necessary that maltose, oligosaccharides, and glucose, as formed in the steady process, for example, It is necessary that the concentration distribution of sugar and glucose be reproduced in order from now on. In the present invention, this is achieved by three technical means as explained below. First, when circulating the effluent from the adsorption tower 2 through the preceding process and the steady process, the effluent should flow into the adsorption tower 2 in the order in which the effluents are discharged. In order to re-introduce the effluent into the adsorption tower 2 in the order in which it flows out as it circulates, as many small tanks as possible are provided and the effluent is sequentially received in these large number of tanks.
Once the reception is complete, the effluent from each tank can be flowed into the adsorption tower in the order in which it was received, but this method requires too many tanks and must be changed frequently. Therefore, it is unsuitable for industrial equipment. Therefore, in the present invention, as shown in FIG.
This is achieved by providing a large number of weirs 24 at 7, 8, 9, 10, 11, and 12. For example, regarding the thin oligosaccharide tank 6, if four weirs 24 with their other ends open alternately are provided at intervals in the tank 6, the inside of the tank will be A, B, C, and A, B, C, etc. from top to bottom.
The five chambers D and E can be divided, and the liquid flows in from the top and flows into the chambers E, D, C, B, and in the order of inflow.
Since the liquid remains in each chamber of A, by taking out the liquid from the lower part of the tank, it is possible to drain the liquid in the order in which it entered. Therefore, by providing four of these weirs 24, one tank has the separation capacity of five tanks.
Although there are seven tanks in FIG. 1, if four weirs 24 are provided in each tank, the same effect as dividing into 35 tanks can actually be obtained. Second, the injection of the stock solution in the steady-state process is carried out in spots after the injection of the oligosaccharide/maltose mixture in the circulating fluid. Thirdly, the total amount of stock solution and water injected in the steady-state step must match the total amount of maltose solution, oligosaccharide solution, and glucose solution taken out of the system. Only by making full use of the three technical means described above will it be possible to obtain a highly pure maltose solution with a maltose content of 95% or more in a high yield. To explain the circulation in the preceding step of the present invention, as the circulation continues, the concentration curves of the three types of sugar change as shown in Figures 2 and 3, and the glucose part O and the glucose part G change. Although they are separating, the concentration of each of the three types of sugar in the effluent is also decreasing at the same time. Therefore, if the circulation is repeated too many times, the sugar concentration will drop too much, which is not preferable, and the number of circulation should usually be about 3 to 5 cycles. Next, the inflow amounts of the stock solution and water during the preceding process and the water balance during the steady process in the present invention will be explained. First, the flow rate of the stock solution in the preceding process is 0.1~
It is preferable to use 0.5/- resin (ion exchange resin), and within this range, the more the stock solution is allowed to flow in, the higher the concentration of maltose obtained in the regular process can be. However, if the amount of resin exceeds 0.5/-, the separation efficiency will decrease, which is not preferable. Next, the amount of water flowing in should be approximately proportional to the amount of the stock solution flowing in, and should be 0.1 to 0.7/-resin; if more water flows in, maltose will be diluted more than necessary, which is not preferable. The water balance during the steady process is 0.03 to 0.1/-resin, preferably 0.05.
A portion of the oligosaccharide liquid is received as 0.1 to 0.18/- resin, preferably 0.14/- resin to oligosaccharide thin tank 6 as before and after resin, and a portion of the oligosaccharide solution is received as 0.1 to 0.18/- resin, preferably 0.14/- as before and after resin, to oligosaccharide liquid tank 7. Part C is 0.2 to 0.4/- resin, preferably
0.29/- Received as before and after resin into oligosaccharide/maltose mixed liquid tank 8, and part of maltose liquid is
0.08-0.15/-resin, preferably 0.12/-
- Receive maltose into the maltose liquid tank 9 as before and after the resin, and the partial o of the maltose/glucose mixture is 0.05 to 0.2
/-resin, preferably around 0.1/-resin, into the maltose/glucose mixed liquid tank 10; a partial force of the glucose solution is 0.03-0.08/-resin, preferably around 0.05/-rein, into the glucose liquid tank 11; The partial value of glucose dilute solution is 0.05 to 0.15/- resin, preferably 0.1/
- It is preferable to receive the resin before and after the glucose in the thin liquid tank 12, respectively. Also, the amount of stock solution to be injected is
0.05-0.2/- resin, preferably 0.1/-
Inject resin before and after. The amount of water is 0.15-0.3
/-resin, preferably around 0.2/-resin. The strongly acidic cation exchange resin used in the present invention is preferably porous, and the finer the particle size, the better from the viewpoint of separability. However, if it is too fine, in industrial equipment, the pressure loss will be large and cause the liquid to be unable to pass or flow in one direction.
Use 200 mesh (wet). In the present invention, this strongly acidic cation exchange resin is used in the form of an alkali metal adsorbed thereon, and is usually regenerated with sodium chloride and used in the sodium form. The stock solution usually has a maltose content of around 80% and a sugar solution concentration of 35 to 60%. The liquid passage temperature is preferably 60 to 80°C from the viewpoint of pressure loss during liquid passage and prevention of microbial contamination. Examples of the present invention will be described below. Example 2400 ml of strongly acidic cation exchange resin XT-1022 (manufactured by Tokyo Organic Chemical Industry Co., Ltd.) with a particle size of 40 to 70 mesh was packed into a column with a diameter of 23 mm and a height of 6000 mm, and 1N sodium chloride solution 3 was added. /- The resin was passed through the solution to completely regenerate it into the sodium form. Next, 720 ml of a sugar solution with a stock concentration of 40%, a maltose content of 88.3%, an oligosaccharide content of 7.9%, and a glucose content of 3.7% was passed through this resin at a temperature of 60°C.
The liquid was passed in a downward flow at SV0.2, and 960 ml of water was further passed through. Next, the detected effluent was collected in fractions of 0.02/-resin and sequentially passed through the top of the column in the order in which they were collected, repeating 3 cycles. FIG. 4 shows the state of separation of maltose, oligosaccharides, and glucose in the effluent of the third cycle. In short, each fraction consists of dilute oligosaccharide solution, oligosaccharide solution, oligosaccharide/maltose mixture, maltose solution, maltose/glucose mixture, glucose solution,
Divided into dilute glucose solution. Next, 7 of the fraction numbers No. 25 to 31 in Figure 4
Fraction oligosaccharide solution 336ml (oligosaccharide 10.4
g, maltose 3.4 g) and fraction number No. 46
~51 6 fraction maltose solution 288ml (maltose 108.6g, oligosaccharide 2.8g, glucose 1.1
g) and 4 fractions of glucose solution No. 57 to 60 (120 ml of glucose solution (maltose 2.0 g, glucose
2.2g) out of the system and use 240ml of the stock solution instead.
(maltose 116.7g, oligosaccharide 10.4g, glucose
4.8 g) and 504 ml of water were injected and the circulation was continued again. In other words, injection is performed in the order of dilute oligosaccharide solution, oligosaccharide/maltose mixture, stock solution, maltose/glucose mixture, dilute glucose solution, and water, and from the next cycle onwards, the sugar solution is extracted every cycle and injection is continued. Maltose was separated by the following steps. Table 1 shows the average sugar composition of the maltose solution, oligosaccharide solution, and glucose solution taken out of the system. 【table】

[Brief explanation of drawings]

Each of the drawings shows an example of the embodiment of the present invention, and Fig. 1 is an explanatory diagram showing the flow of the method for separating maltose, and Figs. 2 and 3 show maltose and oligosaccharides in the effluent. and glucose concentration distribution, where the vertical axis represents the concentration of three types of sugar, the horizontal axis represents the amount of effluent, and the second
The figure is a graph of the concentration distribution in the first cycle of circulation, and FIG. 3 is a graph of the concentration distribution in the third cycle of circulation. Moreover, FIG. 4 is a graph showing the concentration distribution of maltose, oligosaccharides, and glucose in the effluent of the third cycle of circulation in the example, where the vertical axis represents the concentration of each sugar, and the horizontal axis represents the concentration of each fraction number. It represents. DESCRIPTION OF SYMBOLS 1... Strongly acidic cation exchange resin, 2... Adsorption tower, 3... Inflow pipe, 4... Pump, 5... Outflow pipe, 6... Oligosaccharide thin substance tank, 7... Oligosaccharide liquid tank, 8... ...Oligosaccharide/maltose mixed liquid tank, 9...
... Maltose liquid tank, 10 ... Maltose/glucose mixed liquid tank, 11 ... Glucose liquid tank, 12 ... Glucose thin liquid tank, 13 ... Inhalation tube, 14 ... Inhalation main tube, 15 ... Stock solution tank, 16 ...Aquarium, 17...
Inlet pipe, 18, 19...pump, 20...discharge pipe, 21...oligosaccharide liquid take-out pipe, 22...maltose liquid take-out pipe, 24...weir.

Claims (1)

[Claims] 1. When passing a mixed stock solution of maltose, oligosaccharides, and glucose in a downward or upward flow through a fixed bed packed with an alkali metal type strongly acidic cation exchange resin to separate maltose and other sugars, a specified amount of the stock solution is An oligosaccharide adsorption zone and a maltose adsorption zone are formed while moving in the packed bed by passing a specified amount of water through the stock solution and water in that order, and circulating the effluent through the packed bed again in the order of outflow for at least two cycles or more. Then, a preceding step is performed to amplify the distance of the glucose adsorption zone, and after the circulation of the preceding step, a dilute oligosaccharide solution, an oligosaccharide solution,
Among the effluents that exhibit a chromatographic concentration distribution in the order of oligosaccharide/maltose mixture, maltose solution, maltose/glucose mixture, glucose solution, and dilute glucose solution, a specified amount of oligosaccharide solution and a specified amount of maltose solution are Then, a specified amount of the glucose solution is taken out of the system in spots, while other liquids are circulated again in the order in which they flow out.At this time, a specified amount of the stock solution is injected in spots after the oligosaccharide/maltose mixture has flowed in. At the same time, a specified amount of water is injected spot-wise after the dilute glucose solution in the circulating fluid has entered.
Also, carry out a steady process in which the total volume of the stock solution and water to be injected is equal to the total volume of the oligosaccharide solution, maltose solution, and glucose solution to be taken out of the system, and then repeat this steady process thereafter. A maltose separation method characterized by sequentially extracting an oligosaccharide solution, a maltose solution, and a glucose solution.