SU1393847A1 - Method of producing immobilized cells displaying glucosoisomerase activity - Google Patents

Description

(21) 4076304 / 31-13

(22) 04/04/86

(46) 05/07/88. Bul. No. 17

(71) Institute of Organoelement Compounds to them. A.N.Nesme new

(72) I.A. Yamskov, L.L.HinKies, V.A.Davankov, M.S. Mosichev

and I.M. Gracheva

(53) 577.5 (088.8)

(56) USSR author's certificate No. 1060675, cl. C 12 N 11/04, 1982.

USSR Author's Certificate No. 1070163, cl. C 12 N 11/04, 1982.

(54) METHOD FOR OBTAINING IMMOBILIZED CELLS WITH GLUCOSE-ISIZED DIFFERENT ACTIVITY

(57) The invention relates to micro. biological industry, in particular, to methods of immobilization of microbial cells with glucose-isomerically active. The aim of the invention is to increase the yield of enzyme activity during immobilization and increase the stability of immobilized cells. The method consists in carrying out the incorporation of Streptorayces cells Al5o5gl3eo1i8 28-3 into an insoluble sorbent by mixing the cells with. aqueous solution of cobalt salts, followed by precipitation with aqueous ammonia, separation and drying of the precipitate. The ratio of dry cells and cobalt acetate is set to 1: (O, 1-10.0), and sedimentation is carried out at pH 6.5-10.0. The method makes it possible to increase the stability, and increase the yield of enzyme activity by 19-22%. 1 tab.

00 with co

00 Niiik

The invention relates to the microbiological industry, in particular to methods for immobilizing microbial cells with enzymatic activity, and to be used in the food industry in the production of hl: aocoso fructose syrups from starch containing raw materials,

The purpose of the invention is to increase the yield of enzymatic activity during immobilization and increase the stability of the target product.

The method consists in the fact that cells with glucose-isomerase activity are included in the insoluble sorbent, for which their suspendable Bi solution is a cobalt salt solution, with a dry weight ratio of dry cells; cobalt salt 1: (O, 1-10.0) and precipitation is carried out by adjusting the pH to 6.5-10.0. The precipitate is filtered off, washed with water and phosphate buffer pH 7.0, dried and crushed.

Pp and mera 1. 5 g of wet cells (humidity 90%) of Streptomyces albogriseolus 28-3, obtained after centrifuging the culture media. and having an activity of 46.2 U / Gz are suspended in 5 ml of the 1 st solution of cobalt acetate (the ratio of dry cells to cobalt acetate l; 0,) j is added dropwise a concentrated solution of ammonia to a pH of 655, the precipitate is filtered, Squeeze with water, phosphate buffer and chop.

The activity of the wet immobilized preparation is 93.2 units / g, the yield of ak-T1-STATE 86%, stability 30 days.

The amount of enzyme catalyzing the isomerization of one is taken as a unit of glucose isomerase activity; micromol glucose in fructose at pH 7.0 and temperature 60 ° С for 1 min. A 2 M glucose solution with the addition of 10 M ions is used as a substrate. The stability of the immobilized preparation is determined as the time of semi-inactivation of the glucose isomerase at 60 ° C and pH 7.0.

Example 2. The immobilization of the cells is carried out analogously to Example I. The ratio of cells to cobalt acetate is 1:10, pH 10.0. The activity of the drug is 71.0 units / g, the yield of activity is 81%, stability is 41 days.

Example 3: Cell immobilization is carried out analogously to example 1.

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J5 0

5 Oh, g

Q

45-0

five

The ratio of cells and cobalt acetate is 1lOsSjpH 8.0. The drug has an activity of 133 units / g, activity is 96%, and stability is 42 sug

Example 4. Cell immobilization is carried out aHajjorH4Ho example 1. The ratio of cells and cobalt acetate 1: 5, pH 9jO. The activity of the drug is 65.1 units / g, the yield of activity is 99%, stability is 45 days.

Example 5. Cell immobilization was carried out analogously to example 1. The ratio of cells and cobalt acetate was 50.04 j pH 6.4. The activity of the drug is 32.0 units / g, the yield of activity is 22%, the stability is 12 days.

Example 6: Immobilization of the cells is carried out analogously to Example 1. The ratio of cells and cobalt acetate 1:11, rP 10.1. Preparation activity. 15.1 U / G, activity yield 40%. stability 6 days.

Example 7. Cells with an activity of 38J6 units / g are used for immobilization. Immobilization is carried out analogously to example 1, the ratio of dry cells and cobalt acetate, 1: 1, pH 8.0. Active drug: 86.2-units / g, activity yield 92%, stability 36 days.

Example 8. Immobilization np o. water as in Example 1, but using 1% cobalt chloride solution, Str, albogriseolus and CoCe ratio 1: 1 pH 8.0. Preparation activity 77.6 u / g, activity yield 88%, stability 28 days .

The use of the proposed method allows one to obtain an immobilized preparation of cells with a high yield of glucose isomerase activity and a rather high stability,

The results of cell immobilization are shown in the table.

The proposed method, in comparison with the known method, allows an increase; to reduce the activity of the immobilized drug in 1.5 - 3.2 times, the yield of activity - by 19 - 22%, stability - more than 20 times.

Claims (1)

Invention Formula The method of obtaining immobilized cells with glucose isomerase activity, by kx mixing, with gel-forming compounds and the subsequent inclusion of cells in ne 31 3938 7 soluble sorbent, differ-. y and so that, in order to increase the yield of enzyme activity during immobilization and increase the stability of the target product, the cells are mixed with an aqueous solution of salt 1: 5 9.0 3.5 65.1 Famous 3 4.2 cobalt at a dry cells: cobalt salt mass ratio of 1: 0.1-10.0, the pH of the medium is adjusted to 6.5-10.0, and the sediment of the cells included in the insoluble sorbent is separated and dried. 99.0 45 41.5 77.0 2