KR20020085699A - Rotational plate-styled membrane separation method and apparatus of protein - Google Patents

Abstract

The present invention is to separate the protein by using a membrane and is a membrane separation technology applying a rotary separation method to improve the conventional plate-type gravity separation method. In general, a technique using a flat separator should be improved by preventing formation of a gel layer which causes a decrease in permeation flux in the direction of gravity because membrane contamination occurs in the membrane surface. The present invention envisions a module in which the transmission direction is transmitted in all directions of "up, down, left and right" rather than the gravity direction by the shear force by rotation to the disk-type flat membrane module. Existing separation methods do not fundamentally reduce membrane fouling, which has made it difficult for you to use them extensively. However, the present invention is not only to reduce the gel layer on the surface of the membrane by the centrifugal force, but also to devise a method that can provide a simple and stable permeation flow rate for the present invention to apply for the present invention.

In the present invention, the disk membrane can be easily separated and can be used to remove membrane contamination by simple operation, as well as using a cheap and easily available membrane such as nylon or polypropylene membrane, so that the selection of the membrane material is artificially selected according to the type of separation material. Has the advantage of selecting.

Description

Rotary plate-styled membrane separation method and apparatus of protein

The present invention relates to a separation device for separating proteins. In order to reduce the fouling phenomenon that blocks the membrane surface, it is important to improve the flow rate of the inflow separation material from the laminar flow to the turbulent flow in the membrane and thus, The present invention relates to a device for increasing the flow rate by maintaining a separation form such as a screen.

Existing disk-type module is a method of preventing the formation of a gel layer that is attached to the top of the membrane to reduce the permeation flux in the gravity direction (when the present invention is a disk-type module in the transmission direction is not the gravity direction by the shear force) The module was conceived to transmit in "up, down, left, right" directions. The former method does not fundamentally reduce membrane fouling, and so far the problem has been difficult for you to use widely.

However, the unique feature of the present invention is that ① the solids collected inside the reactor by centrifugal force are not automatically introduced into the system. ② not only can drastically reduce the formation of the gel layer on the membrane surface, but ③ the operation is simple and stable. The permeation flux is provided, and the dispersion level of the gel layer can be determined by adjusting the speed of the disc. In particular, ⑤ disk membrane can be easily disassembled to remove membrane contamination with simple operation. ⑥ In addition, because it uses inexpensive and easily available membranes such as nylon and pp membranes, the cost is low and the choice of membrane material can be artificially selected according to the type of separation material.

More specifically, the application of membrane module plate membranes with high shear stress due to rotational force to separate proteins, first, the surface area is wide, second, the solvent permeability is increased, and third, the mechanical strength of the membrane is reduced. Fifth, the narrower distribution has a number of advantages that can be easily separated due to the low head (水頭). In selecting materials, high hydrophobic material selection, low protein adsorption, internal structure uniformity, and pore size distribution can be narrowed, and chemical resistance and physical resistance are reduced. Can be increased accordingly.

Accordingly, the present invention was devised to solve the conventional problems of membrane fouling as described above, and its purpose is to increase the flow rate of laminar to turbulent flow in the membrane to separate proteins. There is a need for designs to enhance flow rates by maintaining separations such as screens between membranes. To this end, the rotary microfiltration membrane structural characteristics of the present invention are firstly, the unit structure is composed of a flat plate membrane and the plate membrane is mounted to face each other in the membrane module. Second, the flow rate to the disk membrane in the module is to allow the flow rate of the upper part and the lower part to flow without intersecting with each other. Third, the rotary module is rotated as well as artificially so that artificial turbulence occurs according to the flow rate of the flow. The device structure is shown to control the speed.

Figure 1 is a schematic diagram showing the overall configuration of the apparatus of the present invention

Figure 2 is an enlarged view of the main part of the membrane module of the present invention

* Explanation of symbols for main parts of the drawings

1: penetration pressure gauge

2: exclusion pressure gauge

3: exclusion pressure gauge

4: heat exchanger

5: discharge valve

6: raw water inlet pump

7: Raw water inflow pipe

8: Separator Body

9: permeate

10: permeate pump

11: enemy tank

12: upper layer separator

13: Permeate discharge hole

14: lower layer separator

15: Shaft

16: support

17: O-ring

18: connection support

Hereinafter, the protein separation device according to the present invention will be described in detail.

The present invention is a microfiltration membrane pore size is attached to the membrane material 1.0㎛ or more above and below the membrane module to configure the membrane module device so that the protein is filtered through the filter medium inside.

The membrane module disk is installed in the membrane device, and the flat membrane is placed in the center and tightened with a ball and operated in a sandwich type. The thickness of the upper and lower layer membranes is 20 mm, and the pump, back pressure valve, and raw water are controlled to maintain the multilayer structure. . The internal temperature of the system was maintained at 18 ℃, and the separator was made of Ultipor Nylon membrane having a diameter of 150mm or CA membrane manufactured by Japanese Toyo.

The membrane was washed with deionized water using NaOH mixed water. All experiments were performed using Bovine serum albumin, Hemoglobin, etc. purchased from Sigma Chemical. The pressure difference in the device depends on the feed rate of raw water and protein concentration. Found to be small in general.

Membrane used in the present invention has a micropore size of 0.8 ~ 3㎛ and the pore size of 0.8㎛ or less because the pressure of the membrane device or the head is very high, so the pump is replaced by the pressure to work to properly adjust the size of the pores This very important and suitable pore size is possible from 1.0 to 2.0 mu m.

1 is an important isoelectric point control in the case of protein separation, so that it is possible to automatically adjust the acid and alkali for the control of pH, and if necessary, the heat exchanger (4) of the membrane separation device built-in membrane separation device for temperature control Temperature control was made possible on the body (8). The flow rate control (raw water inflow pump (6), permeate pump (10), raw water inflow (7)) and pH control were automatically controlled. The separation of protein in the permeate (9) was carried out by the Folin reagent method. Was determined using a UV-Spectrophotometer (Shimatzu UV1601-PC). The rotor speed of the disk is shown in the digital tachometer and the exclusion pressure ((2), (3)) and permeation pressure (1) are shown in the pressure gauge.

FIG. 2 is a membrane module in which a rotating disk membrane is mounted for protein permeation by rotor speed, and is a module configuration diagram with a distance of 20 mm between the upper and lower layers of the separation membrane, and the shape of the separation membrane is connected to the connection support 16. It is held by the O-ring 17. The inflow water 7 injected into the inlet of the apparatus using a centrifugal pump to the membrane module equipped with a flat separator is permeated through the upper and lower membranes, and the permeated water discharge hole 13 is along the axis 15. The permeate liquid 9 is permeated using the permeate pump 10. In particular, in order to check whether the protein is separated, UV-Spectrophotometer and recorder can be installed at the outlet of the device to check the protein separation.

In this case, it is important to install two to four layers as a separation membrane layer, that is, if the membrane type is positioned differently by pore, the separation of fractional molecular weight as well as the amount of protein to be separated is possible. In particular, it is possible to change the separation of proteins by different technology types for each application mentioned above.

The standard curve using the UV spectrophotometer (Shimatzu 1601) for each concentration of hemoglobin penetrating the plate membrane is shown in Figure 1 below.

Based on the above standard curve, it was used as a data to measure the concentration of permeate solute.

<Example 1>

In FIG. 1, the protein solution is introduced at a rate of 2 liters per minute through the raw water inlet pump (1). In order to separate proteins using a rotating plate module, the present invention reduces the concentration polarization by forming turbulent flow with high shear stress by rotating the disk in a solution instead of applying pressure to the influent. to be.

This method does not overcome the limitations of membrane fouling in order to separate proteins, but it is two to five times higher than general filtration through separation and concentration experiments of fermenting enzymes or proteins to separate the concentration of microorganisms and products in the fermentation process. It has been shown that the above transmittance can be obtained.

Figure 2 shows the permeation flux for the rotational speed in the steady state on a rotating membrane disk (RMD).

As shown in the figure, as the disk rotation speed increases, the permeation flux for each particle size increases more than twice.The larger the permeate flux in the fluid, the larger the permeate flow rate, and the smaller the permeate flow rate due to membrane contamination on the membrane surface. The phenomenon of more than doubled is shown. The particle separation material used here was tested by purchasing the product of Sigma Chemical Co., Ltd. as polystyrene particles. These experimental results indicate that in the case of rotary filtration, it is important to sufficiently remove the suspended solids from the permeate fluid by pretreatment.

<Example 2>

As a result of applying the separation method of the present invention to a test site of 50ℓ per day, the results of experiments on the eddy-type disk membrane using hemoglobin are shown in Figure 2 below.

The radius-of-gyration of hemoglobin is 140 nm, and the transmission rate in the CA membrane of 1.2 ㎛ used in the present experiment can be expressed as shown in Figure 3. When the rotation speed of the flat panel module is increased from 400rpm to 900rpm, the permeation rate is increased by more than 200%. Also, when the speed is increased from 400rpm to 1500rpm, the permeation rate is increased by about 300%. This result was obtained from the experiments that the permeation rate and permeation rate increased significantly compared to the case where the eddy plate disk membrane had no rotational shear force.

In addition, the membrane permeation rate according to the input amount of the bovine serum albumin purified from Sigma Chemical, the protein used in this experiment, is shown in Figure 4 below. As a result, when the initial protein concentration was low, the permeation rate was changed, but as the input protein concentration was increased, the permeation rate was shown to be without significant change. However, as the input concentration increases, the decrease in the penetration rate tends to decrease.

As described above, the present invention attaches a separator having a pore size of 0.8 to 3.0 μm to the upper and lower surfaces of the disc to separate the fractions by fractional molecular weight while preventing and rotating the gel layer by using a rotor speed outside the membrane. It is a method of manipulating the protein inflow by shear force into the disk module. Experimental results show that membrane fouling of rotating disk membrane module (RMD) can reduce membrane fouling while increasing permeate flow rate if rotation speed is maintained above 1500rpm. In addition, when the protein separation rate is 70% in the existing UF membrane, the separation efficiency is obtained up to 97% using the product, and the collection of cells generated in the fermentation process can be collected more than three times.

Claims (1)

Separation membrane (12), (12), which is composed of a flat membrane and is mounted in a membrane module so that the flat membranes face each other, and the thickness between the upper and lower layer membranes is 20 mm and the separation pore size is 0.8-3.0 μm. 14)) protein membrane separation method characterized in that the permeation rate is increased by three times or more while having a separation efficiency of 95% or more at the speed of the rotary rotor 15 at a speed of 400-1600 rpm.