DD141977A3 - PROCESS FOR PREPARING WATER-SOLUBLE ENZYME POLYSTYRENE COMPLEXES - Google Patents

Abstract

The present invention includes a method for Preparation of water-insoluble enzyme-polystyrene complexes that are , for use in the chemical, pharmaceutical and Food industry and in medicine. With the Processes become economically insoluble in water Enzyme-polystyrene complexes by appropriate functionalization synthetic polystyrenes and subsequent reaction with enzymes won. The preparation of said complexes is carried out by Coupling chloromethylated polystyrenes with amines, preferably with di- or polyamines or amines, the SH or OH groups contained, in suiastic or in solution at temperatures between 0 and 150 ° C and subsequent reaction with enzymes in - aqueous solution at pH values between 3.5 to 8, at temperatures from 0 ° C to the stability limit of the enzymes and at. one Reaction time from less than one hour to 24 h.

Description

187206 -ι-

b) Field of application of the invention

The water-insoluble enzyme-polystyrene complexes are suitable for use in the chemical, pharmaceutical and food industries as well as in medicine.

c) Characteristic of the known technical solutions

The binding of substances to insoluble polymeric compounds has become very important in recent years. Particularly advantageous is the preparation of insoluble enzyme preparations, because these compounds can be easily removed from a reaction mixture and reused several times. Their application can therefore bring about technological advantages in medical diagnostics and therapy, in certain areas of the chemical and pharmaceutical industry and in the food and beverage industry and lead to cost savings. [0003] Up until now, mainly natural based polymers (z * B * cellulose derivatives) have been used as support materials Proteins), synthetic organi-

see polymers (ζ, B # polyacrylic acid derivatives, maleic anhydride copolymers) and inorganic supports (z, B, porous glass, metal oxides) (G. MAHECKE, Chimia 28 (9) , 467 / T9747, G. P, ROYER J, P. AWDREWS, R. UY; Enzymes Techno1 Dig, YoI I / Sarch.7, 99-138 / T9727, M. HELBOECK, D. YAWOREK; Chimia 29 (-3) , 109-9757). ...

Cellulose derivatives and proteins can only be used to a limited extent because they are microbially susceptible to attack and also have poor flow properties in packed reactors. The use of supported enzymes is often limited by the high cost of preparing and functionalizing support materials.

Polystyrene was also used several times as the support material for enzyme binding (H. FILIPPUSSON, WEGEHORLTBY, Biochem., J. , 120 , 215 (1970), WE HORNBY, H. FILIPPUSSON, PEBS Letters, J, 8 / T9707) this purpose is usually nitrated polystyrene, the nitro groups to amino groups is reduced and • then either diazotized to further Punktionalisierung or ζ "as with Chlortriazinderivaten, glutaraldehyde and Suceinanhydrid reacted * the introduction of the functional groups is relatively expense and Plystyrole have often a highly hydrophobic character that an unfavorable influence on the properties of the enzyme resins (see equ., Baum, Biotechn Bioeng ·, / 17 (2), 253, 1975, USP 3,860 486) exerts _e '

d) Object of the invention

The aim of the invention is to present insoluble and stable protein-carrier complexes in a simple, economical manner »

e) Presentation of the essence of the invention

It is an object of the present invention to suitably synthesize styrene-based synthetic polymers and subsequently combine them with an enzyme to form a stable complex *

According to the invention, this is achieved in that chloromethylated polystyrenes with di- or polyamines or amines, which still. contain other functional groups, such as SH or OH, reacted and coupled to the resulting products without additional components enzymes,

The enzyme-polystyrene complexes are very well suited for use in packed reactors, since their particle sizes are variable within wide limits and thus do not have a limiting effect on the flow rate of a reactor. When used in a batch process, the enzyme resin can pass through Filtration quickly and easily separated and then used again Yiferden, without causing a significant loss of activity occurs »

f) embodiment

The presentation of the enzyme-polystyrene complexes is described below using an example:

5 g of chloromethylated polystyrene (8.5% Cl) were washed in dioxane with 5 ml tetraethylenepentamine 12 St closing and dried

stirred with 5 ml of tetraethylenepentamine for 12 hours at 70 ⁰ O, an100 mg of the aminated polystyrene are 1 hour at room temperature? stirred with 4 ml Glucoamylaselösung (acetate buffer pH 4.5) and washed for 2 hours with acetate buffer pH 4 _S 5.

Activity used: · 50 U

Extinguishing activity: 12 U specific activity: > 0.15 U / mg polymer

relative activity: 39 % _f

Claims (3)

Process for the preparation of water-insoluble enzyme-polystyrene complexes, characterized in that chloromethylated polystyrenes are reacted with amines, preferably with di- or polyamines or amines containing SH or OH groups, and coupled with enzymes » 1 872 Claims 2 _# procedure after. Claim. 1, characterized in that the reaction of chloromethylated polystyrenes is carried out with amines in bulk or in solution at temperatures between 0 and 150C · 3 · Method according to claim 1, characterized in that the coupling of the enzymes to the carriers in aqueous solution at pH values between 3.5 to 8, at temperatures of O ⁰ C to sur stability limit of the enzymes and at a reaction time of less takes one hour to 24 hours «