CS221284B2 - Method of decreasing the contents of phenylalanine in the proteinous hydrolysates - Google Patents

Abstract

Protein hydrolysates or meat- aroma concentrates are subjected to chromatography on a chromatographical column filled with a beta -cyclodextrin polymer to reduce their phenylalanine content.

Description

^Zp USO reduction ^{b y} alanine methyl ester in females of protein ^LHY drol zátech ^y, wherein ЬПkovinový hydrolyzate or the concentrate thereof produced and mass-roma about ^0.5 gA to 3 fenylalanmu the total amino acids ^{of 10%} -to ^{30 h} mot. ^; especially ^E Na ² 0 weight%. ^{; 20} to ⁵⁰ wt. ^; in particular ⁴⁰ % by weight. ^, Chromatogr · ^f and U is a column of polymer / J-cyclodextrin in the ratio by weight ^{of 100} dM ^ ^ avovaráho Mlkovinov the hydrolyzate to 45 parts by weight of a polymer of β-cyclodextrin. Polymer / J-cyclodextrin is water-swellable but water-insoluble polymer obtained from the cyclodextrin or cyclodextrin-containing carbohydrate mixture in the presence of polyvinyl alcohol, polyvinyl acetate, vinyl acetates no ^b ^ of the co-interpolymer zesíťovárnm ol with ^{p r} M ^YF unctions crosslinker. reacting with cyclodextrin and vinyl polymer.

Protein hydrolysates, or concentrates produced therefrom, treated according to the process of the invention are important in supplying patients with phenylketonuria with protein.

The invention relates to a method of reducing the phenylalanine content of protein hydrolysates or, optionally, of the same. ^to meat aroma concentrates by chromatographic method.

Phenylalanine-poor protein hydrolyzates and hydrolyzate products, such as meat flavor concentrates, are of vital importance in supplying protein ketones to patients suffering from phenylketonuria.

The literature describes several experiments aimed at reducing the phenylalanine content of protein hydrolysates. Partridge, SM and Brinley RC (Biochem. J., 51, 628, 1951) removed phenylalanine and tyrosine from protein hydrolysates by ion exchange-displacement chromatography (Zeo-Carb 215, Dowey 2 and sulfonated polystyrene resin) and eluted with 0.075 N sodium hydroxide and 0.15 N ammonia.

The preparative ion-exchange method [Zeo-Carb 225 resin] was further developed by Campbell, P. N., Jacobs S. and Werk T. S. (Chem. And Ind. 117, 1975).

Celretti P. and Montesi G. and Silipardi N. (Arch. Scienze Biol., 11, 554, 1957) isolated phenylalanine from the hydrolyzate by preparative method on Arnberlit IRC-50 resin. A shortcoming of known methods for producing 'phenylalanine-poor' protein hydrolysates is that they are very expensive and the taste of the products obtained is unpleasant.

The unpleasant taste of the phenylalanine-free protein hydrolysates prepared by the ion exchange chromatography method results from the hydrolysis process. Ion exchange chromatography requires a protein hydrolyzate with a low salt content, therefore the hydrolysis is not carried out with hydrochloric acid, sulfuric acid, but which can be of the SNA ^D no removed as the calcium salt.

It is further known that protein hydrolysates prepared with sulfuric acid produce unpleasant taste-causing components (German Patent No. 127,693, Predegast, K., Food Trude 44, 14-21, 1974).

British Patent No. 1,091,637 discloses that it is possible. .Beta.-cyclodextrin with various bifunctional compounds (e.g., chlorohydrin, dichlorohydrin, dirpoxypropyl ether) to obtain polymers of .beta.-cyclodextrin.

It is further known that [beta] -betaine polymers form complexes of cyclodextrin with suitable compounds, for example amino acids.

According to Wiedenhof (Stark, 21, 163, 1969), the complexing ability of cyclodextrin polymers served as the basis for the analytical chromatographic method for gel inclusion chromatography.

Zadadon et al. (Stark, 31, 11, 1979) disclose an analytical method according to which it is possible to separate the tryptophan biphenyls and the amino acids simultaneously.

^R ozdělovací metráy for détente aliphatic and aromatic amino acids have not been used for preparative purposes.

Hitherto known analytical methods relate ^{é a de} Lenii aliphatic and aromatic amino acids from amino acid mixtures with polymers of cyclodextrin. Therefore, the aim was not to produce protein hydrolysates with a low phenylalanine content.

The aim of the present invention was to develop a technically usable method for removing ^and rn fenytálaninu of Mkovinových hydrolyzate here ^and their REA--MCH products.

^UK ol & VYN cut is to develop a methodology whereby ^d y can be about ^^^ ^ t ^ rao it from protein hydrolysates with a total content of 10-30 why. ^h mot amino acids and ¹⁰ to ^30% by weight. NaCl phenylalanine, whose solution has long been pushed. ^You n and ^l ez is based on the knowledge that in urmtých circumstances can also be performed isolation of phenylalanine vysocekoncentrovaných solutions.

The above object is solved according to the invention in such a ^way that h ^yd Mkovinový by hydrolysis or flavor concentrate the mass. ^0.5 to 3 g / 1 fenylatamnu with ^Cel containing ¹⁰ ew. up to ³⁰ wt.%, especially ^{20% é} 'weight and the amino-ehy .sušin ^{of 2} 0 ^{and 50%} by weight of a particularly ^E in ^40% chromatographed ^' _ polymer column (S-cklodextri-nu = 100 parts by weight, modified ^{é é} kovmov ^beat him he ^ u · hydrolysis: ^{45 says} lum weight, polymer / R-cyclodextrin.

The polymer jjScyklodextrinu is water-swellable but water-insoluble polymer obtained from the cyclodextrin or mixture of hydrocarbons ^hydroxy stearate ^U contains cyklodtextrin the horizontal being · presence of polyvinyl alcohol polyvinyl acetate or a polyvinyl acetate copolymer, crosslinked ^and MM with polyfunkčmm of a crosslinking ^{dl em-reactive} cyclodextrin and vinyl polymers.

The separation should be carried out under circumstances in which the polymer / cyclodeetrma used exhibits a high complexing activity against phenylalanine.

Numerous ^p us of ^{the y} '^ to take off from the ligament and ^{f b y} en alanine methyl ester of tyrosine and polymer β-cyclodextrin depend on concentration of both amino acid in the protein hydrolyzate.

To reduce the content of NaCl in the used bilkovmovrn h ^y dro ^ly from ^and Tu, zfctarém ^'hy árolýzou with hydrochloric acid, may be used to desalinate it by. No. 127,693, or U.S. Pat.

The amount of phenylalanine bound to 1 g of polymer depends on the diameter of the polymer-filled column, the filling height and the concentration of phenylalanine.

What is important is the fact that the divider ^{to the} apacita column, that is, the amount of phenylalanine absorbed. ^{1 g} poker hundredth ^P and T ^EZ increasing concentrations of phenylalanine.

The measures required for .oddělování nlnu phenylalanine, for example pH, temperature, column size, sec selected as hereinafter described in ^Preparation · ^p Roveda. Equals ^to y division performed are summarized in .tabulce I.

Examples of P mot ^h days ^{the poly} mer (c ^cy lodextřinu ^prepared by DOS · · No. 2927733, was swelled in water and filled into chromatograHckého column (70 x ^2.5 CMJ. The gel bed is applied to 100 parts by weight, casein · h ^yd rol ^y carbazate (solids: ^{40% H} mot., .aminokyselin content: ²⁰ wt.%, the content of NaCl · L: 20 wt% _M contained phenylalanine ^h: 0Д5% wt., pH = 6) . the first volume of the effluent from the column was discarded. by using pressure or vacuum may be ^hydroxy by hydrolysis ⁱⁿ whole or push simply ^let the column. Solvent Composition KATP Whose wipe from the column:

Dry matter: 35 wt.

The content of amino ^alkyl Selin ^17% tooU NaCl content: 18% and the ^b · ^h-phenyl atanrnu: ^0.02% l · ^h mot

Hydrolyzate with a low content of phenylalanine ^of e ^saith to NaCl contained ^h! 1.5% by weight and then has a pleasant taste of meat broth.

The polymer L-cyclodextrin polymer obtained by the previously listed eným způsotem ^d (DCS no. 2927733).

Table · I

Formation of a complex of cyctodextrinphiphenylalanine polymer in aqueous medium as a function of phenylalanine concentration (pH = 7, column diameter: 16 mm)

^Number Qty .beta.-cyclodextrin polymer: ^0.5 g

Column filling height: 2 cm

Phenyl Concentration Amount of Carried Amount of Phenylalanine Amount of Phenylalanine (mg / 100ml) of Phenylalanine mg% Nine Bound 1 g (mg) Cyclodextrin Polymer mg

50 15 mg (in 30 ml) 1,25 8.3 2.5 100 ALIGN! 30 mg (in 30 ml) 1.70 5.7 3.5 150 45. mg (in 30 ml) 2.00 4.4 4.0 Quantity of polymer 1-cyclodextrin: 1.0 g Column filling height: 4 cm 50 15 mg (in 30 ml) 1.75 11,7 ' 1.7 100 ALIGN! 30 mg (in 30 ml) 2.95 9.8 3.0 150 45 mg (in 30 ml) 4.00 8.9 4.0

Quantity polyme.ru / 3cycle · odextrm: 2.0 g

Column filling height: 7 cm

50 15 mg (in 30 ml) 3.25 21.7 1.6 100 ALIGN! 30 mg (in 30 ml) 4.70 15.7 2.4 150 45 mg (in 30 ml) 7.75 17.2 3.9 * Amount of polymer, ₁₁ % cyclodeetrin: 3.0 g Column filling height: 10 cm J * 50 15 mg (in 30 ml) 4.80 32.0 1.6 100 ALIGN! 30 mg (in 30 ml) 7.00 23.3 2.3 150 45 mg (in 30 ml) 11.25 25.0 3.8

Claims (2)

1. A process for reducing the content of protein hydrolysates phenylalanine or concentrates made thereof aroma mass chromatography, characterized in that a protein hydrolyzate or a flavor concentrate mass of 0.5 to 3 g / 1 ^{F y} en lalanmu, having a total ^{of 10} to ^30% especially ^E in ^{20% h} mot. the amino ^y Selin and the dry matter ^{of 20 and 50%} by weight of a mainly ^é to 4 °% ^H mot., chroma ^g rim on skoupci ^P copolymer &, β-cyclodextrin at a ratio of 100 parts by weight , the protein lysate hydroformylation: 45 parts by weight of polymer / cyclodextin. The second ^{are the methods} after le bo ^{d d} u 1 v ^ načujmí in that a polymer / 3cyklodextrinu which is water-swellable but water-insoluble polymer obtained from the cyclodextrin or cyclodextrin-containing carbohydrate mixture in the presence of. polyvinyl alcohol, polyvinyl acetate or polyvinyl acetate copolymer by crosslinking with a polyfunctional crosslinker reactive with a cyclodextrin and a vinyl polymer. sevarography, n. plant 7 Mott