FR2607151A1 - PROCESS FOR THE PREPARATION OF GLUCOSE / FRUCTOSE / SUCROSE MIXTURES FROM SUCROSE USING IMMOBILIZED INVERTASE - Google Patents

Abstract

THE PRESENT INVENTION CONCERNS A PROCESS FOR HYDROLYSIS OF SACCHAROSE BY MEANS OF IMMOBILIZED INVERTASE IN WHICH A SACCHAROSE SOLUTION OF 50 TO 70 GOES THROUGH THE DECOLORATION COLUMN WITH A SPECIFIC LOAD OF 0.5 TO 2.0 H, AN OPERATION IN WHICH THE PH IS SET BETWEEN 4.0 AND 7.0, THEN IS HYDROLYZED.

Description

The present invention relates to a method for the preparation of

  glucose / fructose / sucrose mixtures

  from sucrose using immobilized invertase

  Lisée. These glucose / fructose / sucrose mixtures are usually commercially available in the form of invert liquid sugar or as the case may be

  of invert sugar syrup and are

  important intermediaries in the food, pharmaceutical and chemical industries. They are used for the production of desserts and bakery products, drinks, antibiotics, enzymes, hormones and others. A large number of processes are known

  hydrolysis of sucrose with immature invertase

  are differentiated by the process of immovable

bilization and support.

  By adsorption the invertase is for example

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  linked to an ion exchanger. macroporous polystyrene (Amberlite IRA 93) (French patent

  N 2 233 334) or bone charcoal (British patent

N 564 270).

  In addition, there are known methods of immobilization by covalent bonding. As examples, mention may be made of the binding on shot (P. Monsan, D. Combes and I. Aiemzadeh, Biotechnol Bioeng 26, 658 (1984), P. Monsan and D. Combes, Biotechnol.

  Bioeng. 26, 347 (1984)) and a poly (aminomethyl)

  styrene) (Wofatit Y 58) (J. Mansfeld and A. Schellen

  Shepherd, Acta Biotechnol. 6, 89 (1986)).

  The inclusion of invertase in matrices

  polymers, p. ex. of gelatin (A. I. Krauze, Zesh.

  i5 Nauk Ser. 1 88, 39 (1981)) or calcium alginate (Y.Y. Linko, L. Weckstrom and P. Linko, Food Process Eng Ind., 1979, 81) is another procedure.

  of preparing the immobilized enzyme.

  According to the immobilization method and the support appear with the majority of the invertase-carrier complexes known for the hydrolysis of sucrose a series of disadvantages, so that the use of immobilized invertase spreads only slightly to little on an industrial scale.> These disadvantages can appear in terms of! price, mechanical stability as well as

  of operation stability of the invertase complex

  support, the latter depending to a large extent on the purity and concentration of the substrate and the temperature of the reaction. The importance of the influence of pH on the stability of the operation

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  also depends on the concentration of the substrate.

  The catalytic efficiency of the enzymes also depends on the pH. For invertase an optimum pH of an order of magnitude between 4.0 and 5.5 is indicated (W. Hartmeier, Gordian Int.Z., Lebensmittel und Lebensmitteltechnologie 78, 320 (1978)), the pH optimum is shifted during immobilization according to the process

immobilization and support.

  For the adjustment of the optimum pH is therefore provided in the known methods of enzymatic hydrolysis

  sucrose containers which present

  However, there are disadvantages for the continuity as well as the cost-effectiveness of the process.

  manpower and the importance of investment

  ments. Similar disadvantages occur in the pH adjustment of sweet solutions with

  ion exchangers, which are used in the in-

  sugar industry for the removal of lime

or salts.

  The known methods also indicate that they work at the pH optimum of invertase

  corresponding immobilization, although under the conditions

  known industrial processes this pH is difficult

  to regulate and maintain constant.

  The object of the invention is to achieve a continuous, economical way, while maintaining at a low level the expenses of apparatus and manual,

  avoiding the use of foreign bodies, hydro-

  lysis of sucrose with immobilized invertase

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  for the preparation of glucose / fructose / sac-

  charrose of high concentration and purity.

  The invention responds to the need to develop a method, simple as to the apparatus, to carry out continuously, hydrolysis of the sucrose with immobilized invertase, with which one can prepare the invert liquid sugar or according to

  cases the invert sugar syrup with a concentration

high purity and purity.

  This need is satisfied by the use of a saccharose solution of 50 to 70%, preferably

from 60 to 70%.

  The sucrose solution flows with a specific charge of 0.5 to 2.0 hF along a bleaching column, which is filled with a strongly basic bleaching ion exchanger which is usually found in commercially, for example Wofatit EA 60, in order to prepare for the hydrolysis the substrate with a purity such that it avoids that the unsweetened bodies influence the residence time of the immobilized invertase. The direction

  movement can here be rising or falling.

  From the point of view of economic use

  the capacity of the resin is chosen in decolourism

  ration of clear liquids or thick juices the highest specific loads known,

  which are in color between 2 and 6 h-1.

  Normally there is only a negligible drop in pH which is caused by the exchange adsorption, in which the anions of the salts of acidic compounds

2607151

  present at low concentrations are exchanged for chloride ions, and which is an order of magnitude

  0.3 pH units (I. Stamberg and V. Valter, Ent-

  fârbungsharze, Akademie-Verlag, Berlin, 1970).

  It is surprising that in the discoloration of usual commercial sucrose solutions

  with a specific charge of the decolorizing resin

  From 0.5 to 2.0 h -1, a pH drop of 0.5 to 2.0 pH units is observed which is sufficient for this to lead to hydrolysis in a pH range of 4. 0 to 7.0 corresponding to the optimum of the immobilized invertase including its upper limits. In addition to the adsorption already known exchange, also play a role of other effects, for example the thermal decomposition of unsweetened bodies, following which the decline appears surprisingly

high pH.

  In contrast to the indications of the known technical solutions, it is doubtful whether the hydrolysis of sucrose can be carried out under industrial conditions only at the optimum pH of the immobilized invertase with a degree of

  satisfactory transformation. It should rather ad-

  to put or according to the cases it was found that the hydrolysis

  takes place in an interval that corresponds to the op-

  timum, including its upper limits, of the inver-

  considered, without showing any essential declines in the degree of transformation. We must start from the fact that the stability of the operation

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  corresponding immobilized invertase is compa-

  on the pH range mentioned in the

it has the optimum pH.

  In order to arrive as close as possible to the pH optimum of the immobilized invertase, the process according to the invention is preferably carried out with a specific

  discoloration between 0.5 and 1.0 h-1.

  In addition, the solution of decolorized sucrose and pH adjusted in a known manner traverses a column of product causing an inversion, which is filled with immobilized invertase, for example a complex InvertaseWofatit-Y 58, where the glucose mixtures are formed / fructose / sucrose whose invert sugar fraction depends on the specific charge of the column

  product causing an inversion, the concentration

  temperature and temperature of the saccha-

  pink and can be adjusted correspondingly

at the wishes of the customer.

  The direction of movement here can be up or down. It is important for the simplicity of the process according to the invention that no additional containers or chemicals are needed to adjust the pH, and that a completely continuous operation of the receipt of the sucrose solution is provided. 'to the preparation

of the final product.

  The following examples of

specify the invention.

EXAMPLES OF REALIZATION

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  As an immobilized enzyme, a

  invertase-Y complex 58 whose operability stability

  in a 62% sucrose solution at 70 ° C., expressed as the half-reaction time, is 131 d and the pH optimum at 4.5 (J. Mansfeld et al.

  A. Schellenberger, Acta Biotechnol. 6, 97 (1986)).

  Regardless of the immobilization process and the

  support one can use invertase- complexes

  support whose catalytic efficiency and operating stability above the pH range included

  between 4.0 and 7.0 are comparable to those

mum of pH considered.

EXAMPLE 1

  A filtered 66% sucrose solution having a pH of 8.0 runs for a period of hours a decolorization column which is filled with 50 liters of the discoloration ion exchanger.

  strongly basic Wofatit EA 60 in chloride form.

  The working temperature is 80 C, the specific charge at 1.0 h -1 the color of the solution on average at 50 to 100 ICUMSA (Int Comm Uniform

  Methods Sugar Anal., Units ICUMSA-8420x 103).

  After discoloration, a residual color of 14 to 20% remains. The pH of the sugar solution averages 6.8, and a pH drop of 1.2 pH units is achieved. With this pH, the decolorized sucrose solution then runs through a column of product causing an inversion,

  which is filled with 50 liters of the invertase-

  Y 58. The working temperature is 40 C,

  the specific charge at 1.0 h-1. The degree of trans-

  achieved training is here between 89 and 94%.

  Comparatively, the specific charge of the inverting product column and the temperature and concentration of the solution are achieved with a concordant value.

  of sucrose at the pH optimum of the invertase

  a degree of transformation of 89 to 95%.

  Determination of the degree of transformation

  is carried out according to the volumetric method of W.H.

  Lane-Eynon (Official Methods of Analysis, 494, Asso-

  ciation of Official Agricultural Chemists, Washington

  ton, D.C. (1965) for the determination of sugars

  reducers. The pH is determined by potentiometry.

EXAMPLE 2

  Discoloration and inversion occur

as it is said in Example 1.

  The starting material is a solution of

  67% sucrose having a pH of 6.9. The specific charge

  The amount of discoloration and inversion is 0.5 hl, the residual color remaining after decolourisation at 14 to 20%. A pH drop of 1.7 pH units is obtained, so that the hydrolysis is

  at pH 5.2. Here we reach a degree of trans-

  formation between 95 and 98%, which corresponds to that obtained at the pH optimum of invertase immobilized at the same specific charge of the inverting product column and at the same temperature and concentration of the solution

of sucrose.

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Claims (3)

  1. Process for the hydrolysis of sucrose by means of invert-   immobilized system, in which a solution of saccha-   first runs through a column of bleaching product, which is filled with a strongly basic bleaching ion exchanger, and then hydrolysed, characterized in that a 50 to 70% sucrose solution flows through the bleaching column with a specific charge of 0.5 to 2.0 h 1, an operation in which the pH is set between 4.0 and 7.0, then is hydrolysed.   2. Method according to claim 1, characterized in that a solution of sucrose from 60 to 70%.   3. Process according to claims 1 and 2, characterized   in that the specific charge of the discoloration column is preferably between 0.5 and 1.0 h