CA1185551A - PROCESS FOR THE PREPARATION OF ISOMALTULOSE (6-0-.alpha.- D-GLUCOPYRANOSIDO-D-FRUCTOSE) BY THE USE OF IMMOBILIZED BACTERIAL CELLS - Google Patents
PROCESS FOR THE PREPARATION OF ISOMALTULOSE (6-0-.alpha.- D-GLUCOPYRANOSIDO-D-FRUCTOSE) BY THE USE OF IMMOBILIZED BACTERIAL CELLSInfo
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- CA1185551A CA1185551A CA000424503A CA424503A CA1185551A CA 1185551 A CA1185551 A CA 1185551A CA 000424503 A CA000424503 A CA 000424503A CA 424503 A CA424503 A CA 424503A CA 1185551 A CA1185551 A CA 1185551A
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- isomaltulose
- mother liquor
- sucrose
- sucrose solution
- fructose
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/24—Preparation of compounds containing saccharide radicals produced by the action of an isomerase, e.g. fructose
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- Chemical Kinetics & Catalysis (AREA)
- Biomedical Technology (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Saccharide Compounds (AREA)
- Steroid Compounds (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
ABSTRACT
A process for the preparation of isomaltulose through enzymatic transformation from sucrose with the use of microorganisms forming isomaltulose from sucrose is described. In this process, a pure sucrose solution is mixed with the mother liquor of a preceding isomaltulose crystallization, and the resulting mixture is brought into contact with dead immobilized cells of such microorganisms.
A process for the preparation of isomaltulose through enzymatic transformation from sucrose with the use of microorganisms forming isomaltulose from sucrose is described. In this process, a pure sucrose solution is mixed with the mother liquor of a preceding isomaltulose crystallization, and the resulting mixture is brought into contact with dead immobilized cells of such microorganisms.
Description
TITLE OF THE INVENTION
Process for the preparation of isomaltulose l6-O-a-D-glucopyranosido-D-fructose) by the use ~f immobili~ed bacterial cells.
TECHNICAL FIELD OF ~HE INVENTION
This invention relates to the preparation of isomaltulose (6-O-~-D-glycopyranosido-D-fructose by the use of immobilized bacterial cells.
BRIEF DESCRITPION OF THE PRIOR ART
~ p~ng Canadian patent application 386,779 filed Sep~ber 28, ~81 ~scloses thatsucrcse may be enzymatically co ~ ~
to isomaltulose, by bringing pure sucrose solutions into contact with dead immobili7ed cells of micro-organisms capable of forming isomaltulose from sucrose.
Thus, sucrose solutions having a concentration of 95% by weiyht to 75% by weight, preferably 65% by weight to 75% by weight, are passed at a temperature of 40C to 65~C continuously through a reactor filled with the immobilized cells. The isomaltulose which forms is obtained in crystalline form by methods known per se.
The microorganisms used are Protaminobac~er rubrum tCBS 574 77)r Serratia plvmuthica ~ATCC 15928~, Serratia marcescens (NCIB 8285) and Leuconostoc mesenteroides (NRRL B-512.F (ATCC 10830a)),preferably Protaminobacter rubrum (CsS 574.77).
When carrying out this process, it has been found that often a high proportion (up to about one third3 of the sucrose used is not converted into the desired compound (isomaltulose), but is converted into by-products and/or remains unconverted in the reaction solution. As these by-products inhibit the crystalli-zation of the resultant isomaltulose and thus reduce the crystalline yield even further, it is the object of the present invention to devise a method whlch fully or at least partly prevents the formation of undesirable and disturbing by-products.
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SUMMARY OF THE INVENTION
According to the present invention, there i~
provided~ in a process for preparing isomaltulose ~6-O-~-D-glucopyranosido-D-fructose) wherein sucrose solution is treated with or brought into contact with dead immobilized cells of a microorganism capable of converting sucrose into isomaltulose, the improve-ment which comprises:
(a) mixing said sucrose solution with the mother liquor of a preceding isomaltulose crystallization ~ to form a mixture;
; tb) bringing said mixture into contact with, or tre-ating said mixture with, dead immobilized cells of said microorganism at a temperature of 25 to 4~C
to form.isomaltulose; and lc) recovering said isomaltulose formed in step Ib~
in crystalline form.
~ he present invention also provides a process for obtaining 1 O-~-glucopyranosido-D-fruct~se ~ro~
a mother liquor from which isomaltulose has been separated, which process comprises:
(a) separating isomaltulose from said mother liquor by crystallization from methanol;
(b) removing fermentable sugar from said mother liguor by treatment with free or immobilised yeast;
(c) recovering 1-O-~-glucopyranosido-D-fructose ira~
said mother liquor by chromatographic separation by i means of an ion exchanger or other separating material;
and (d) transforming said l~O-~-glucopyranosido-D-fructose to dry form.
Surprisingly, it has been found that it is possible to obtain isomaltulose of high purity and with a high yield from sucrose by blending pure sucrose solu~o~s having a preferred concentration of 40% by weight to 75% by weight, more preferably 45% by weight to 60~ by weight, with the mo her liquor of a preceding , .
isomaltulose crystallization, by treating this solution or bringing it into contact with immobilized dead cells of microorganisms forming isomaltulose from sucrose at a temperature of 25C to 40C, preferably 30C to 40C, and by obtaining the resultant iso-maltulose in crystalline form by a method known per se.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
An advantageous embodiment of the process consists in dividing the sucrose solution having a concentration of 403 by weight to 60% by weight, into two parts, in passing the first part via a reactor, which is filled with immobilized dead cells of microorganisms forming isomaltulose from sucrose, at ~5C to 40C, preferably 30C to 40C, so that 80% to 90% of the sucrose is converted, in mixing the second part of the sucrose solution ~ith the mother liquor from a preceding isomaltulose crystallization, in passing this solution via a second reactor, which is filled with immobilized dead cells of microorganisms forming isomaltulose from sucrose, at 25C to 40C, preferab~y 30C to 40C, so that the sucrose is almost completely converted, in obtaining the resultant isomaltulose from this solution in crystalline form by methods Xnown per se, in dissolving and crystallizing the crystals in the converted first part of the reaction ; solution, in separating the resultant isomaltulose crystals and in using the mother liquor for the ad-mixture to the second part of the sucrose solution.
The average contact time of the sucrose solution in the reactor depends on the specific activity, present in each case, of the immobilized cells. The specific activity is defined as ~mol of converted sucrose per gram of immobilized cells (dry) per minute. For example, a reactor having a diameter of 100 mm and a bed hei~ht of 450 mm contains 950 g of immobilized cells (dry~
with a specific activity of 60 units/g. A quantity of 1900 ml of sucrose solution (50~ by weight) rnust be made to Elow through this reactor per hour in order to convert 90% of the sucrose. As the activity continuously decreases over the working time, the velocity of flow must be readjusted constantly in order to obtain a product with constant composition.
BRIEF DESCRIPTION OF THE DRAWING
The single Figure of the drawing is a flow she~
illustrating the process described in Example 3 below.
The specific advantages of the process according to the invention are explained in greater detail with the aid of the following examples. Example 1 describes the procedure and results of the known process of Germany Offenlegungsschrift No. P 30 38 219.
(a) Cells from an inoculum of Protaminobacter rubrum (CBS 574.77) are transferred with 10 ml of a sterile nutrient substrate consisting of 8 kg of thick juice from a sugar plant (dry substance content = 65%), 2 kg of corn steep liquor, 0.1 kg of (NH4)2HPO4 and 89.9 kg of distilled water (adjusted, if needed, to pH of 7.2). This suspension is used as the inoculating material for the shaker-machine pre-culture in 1-liter flasks with 200 ml each of the nutrient 25 solution of the above compositionD Twenty of such flasks (total contents 4 liters) are inoculated at 29C for 30 hours.
16 Liters of nutrient solution of the above composition are inoculated in a 30 liter small fermen-ter 30 with the contents of the above twenty flasks (4 liters) and fermented at 29C at an aeration rate of 20 liters/
j minute and a stirring rate of 350 rpm. The increasing cell count is determined with a microscope. When ; a concentration oi 5 x 109 cells~ml is reached, the 35 contents of the fermenter are transferred in-to another container and reacted therein with a cationic flocculant (for instance, PRIMAFLO~ C7 by Rohm & Haas, c 4-*trade mark , Philadelphia, Pa., ~S~). The flocculated cells ~re allowed to settle, decanted, washed ~ith ~.~ M
phosphate buffer of a pH of 7.0i and dewatered in the centrifuse. The substance i~ then extruded into strands, air-dried and ground.
tb) The 0 3 to 0.8 mm sifted fraction from the above-obtained preparation is stirred in a 0.~
glutaraldehyde solution for 10 minutes, washed with phosphate buffer (O.lM, pH=7.0~ and filled under water into a heatable colu~n. The col~mn is then he~ted to 50C and a 70% by ~eight suc~ose solution ~s pumped continuously through this column. The flow rate is adjusted in such a manner that no sucrose can be detected anymore at the end of the column. The isomaltulose solution so obtained has the folla~ing average composition ~as determined ~y ~PLC, ~ e_ high-pressure liquid chromatography):
fructose .. .. .. 7.4 9~OO g TS (total sugar) glucose .. .. .. .. 0.3 g~100 g TS
sucrose .. .. .. .. 0.1 g/100 g TS
isomaltulose .. .. .. 62 6 9~100 g TS
l-O-a-glucopyranosido-D-fructose .. .. .. 16.~ g~100 g TS
oligosaccharide .. .. 13.0 g/100 g TS.
This solution is fed to a cooling crystallizer, cooled to 20C and the crystallized isomaltulose is separated from the mother liquor in wire-basket centrifuges. The mother liquor is concentrated by evaporation to 78% to 82% TS content, cooled in a second cooling crystallization to 20C, and a second quantity of crystallized isomaltulose is separated from the mother liquor in wire-basket centrifuges.
From 100 kg of a sucrose solution containing 70 kg of sucrose there are obtained 43.8 kg of isomaltulose (62.6% of TS content). From this there is obtained in the first crystallization 15.9 kg of pure isomaltulose and in the second crystallization .~ ,~ . .
I
_ 13.4 kg of isomaltulose of 98~ purity. This i~
equivalent to a total yield of 29 kg of pure iso-maltulose from 70 kg of sucrose or 41.4% from the raw material. (In this example and in the following the calculations are based on anhydrous isomaltulose).
The 0.3 to 0.8 mm sifted fraction from ~he preparation prepared according to example l~a) is : stirred in a 0.1% glutaraldehyde solution for 10 minutes, washed with phosphate buffer ~9.1M, pH=7.03 and fil].ed under water into a heatable column. The colu~n is then heated to 30C and a 50~ by weight sucrose solution is pumped through continuously.
The velocity of flow is adjusted so that no sucrose can be detected anymore at the end of the column.
The isomaltulose solution so obtained has the -following average composition ~HPLC):
fructose .. .. .. 3.6 g/100 g TS
glucose .. .. .. .. 1.8 g/lD0 g TS
sucrose .. .. .. .. 0 isomaltulose .. .. .. 78.4 9/100 g TS
l-o-a-D-glucopyranosido-D-fructose .. .. .. 12.6 g/100 g TS
oligosaccharide .. .. 3.6 y/100 9 TS.
.~ 25 This solution is concentrated by evaporation to 78% to 82% TS content, fed to a cooling crystal-lizer a`nd.cooled to 20~C, and the crystallize~
isomaltulose is separated from the mother liquor in wire-basket centrifuges. The mother liquor is again , 30 conecentrated by evaporation to 78~ to 82~ TS content, sub~ected to a second cooling crystallization to 20~C, ; and a second quantity of crystallized isomaltulose is separated from the mother liquor in wire-basket '- centrifuges.
From 1~0 kg of a sucrose solution containing 50 kg of sucrose, there are obtained 39.2 ]cg of isomaltulose (78.4% of TS content). Fro.m this there .. .. ...... .
is obtained in the first crystallization 25.0 kg of pure isomaltulose and in the second crystallization 9.6 kg of isomaltulose with 98~ purity. This is equivalent to a total yield of 3g.4 kg of pure isomaltulose from 50 kq saccharose or 68.8~ from the raw material.
The 0.3 to 0.8 mm sifted fraction from the pre-paration prepared according to example ~(a) is stirred in a 0.1~ glutaraldehyde solution to 10 minutes, washed with phosphate buffer ~O.lM, pH=7.0) and filled under water into two he~table columns heated to 30C.
The procedure is explained ~ith reference to the flow sheet in Figure 1.
A 50% by weight sucrose solution is made to flow through one of the colu~ns 1 conti~uously The velocity of flow is adiusted so t~at only approximately 90% of the sucrose i 5 converted ~hen the~e is flow through the column. The reaction~ solution so obtained ~ 20 has the following exempl~ry compGsition ~EIPLC):
'~ fructose ......... .. .. 2.~ g~100 g TS
glucose .. .. .. .. 1.6 g~lOO g TS
sucrose .. .. .. .. 10.~ g/lOC g TS
isomaltulose .. .. .. 75.2 gJlOO g TS
l-O-a-D-glucopyranosido-D-fructose ....... .. .. 6.~ g/100 g TS
, oligosaccharide ....... .... .. 3.l g/100 g TS.
i Parallel to the first column, the second colomn
Process for the preparation of isomaltulose l6-O-a-D-glucopyranosido-D-fructose) by the use ~f immobili~ed bacterial cells.
TECHNICAL FIELD OF ~HE INVENTION
This invention relates to the preparation of isomaltulose (6-O-~-D-glycopyranosido-D-fructose by the use of immobilized bacterial cells.
BRIEF DESCRITPION OF THE PRIOR ART
~ p~ng Canadian patent application 386,779 filed Sep~ber 28, ~81 ~scloses thatsucrcse may be enzymatically co ~ ~
to isomaltulose, by bringing pure sucrose solutions into contact with dead immobili7ed cells of micro-organisms capable of forming isomaltulose from sucrose.
Thus, sucrose solutions having a concentration of 95% by weiyht to 75% by weight, preferably 65% by weight to 75% by weight, are passed at a temperature of 40C to 65~C continuously through a reactor filled with the immobilized cells. The isomaltulose which forms is obtained in crystalline form by methods known per se.
The microorganisms used are Protaminobac~er rubrum tCBS 574 77)r Serratia plvmuthica ~ATCC 15928~, Serratia marcescens (NCIB 8285) and Leuconostoc mesenteroides (NRRL B-512.F (ATCC 10830a)),preferably Protaminobacter rubrum (CsS 574.77).
When carrying out this process, it has been found that often a high proportion (up to about one third3 of the sucrose used is not converted into the desired compound (isomaltulose), but is converted into by-products and/or remains unconverted in the reaction solution. As these by-products inhibit the crystalli-zation of the resultant isomaltulose and thus reduce the crystalline yield even further, it is the object of the present invention to devise a method whlch fully or at least partly prevents the formation of undesirable and disturbing by-products.
.
~ .
, _ ~ J ' ~
~ " ' ~1'.
.
J~
~.
1 .
SUMMARY OF THE INVENTION
According to the present invention, there i~
provided~ in a process for preparing isomaltulose ~6-O-~-D-glucopyranosido-D-fructose) wherein sucrose solution is treated with or brought into contact with dead immobilized cells of a microorganism capable of converting sucrose into isomaltulose, the improve-ment which comprises:
(a) mixing said sucrose solution with the mother liquor of a preceding isomaltulose crystallization ~ to form a mixture;
; tb) bringing said mixture into contact with, or tre-ating said mixture with, dead immobilized cells of said microorganism at a temperature of 25 to 4~C
to form.isomaltulose; and lc) recovering said isomaltulose formed in step Ib~
in crystalline form.
~ he present invention also provides a process for obtaining 1 O-~-glucopyranosido-D-fruct~se ~ro~
a mother liquor from which isomaltulose has been separated, which process comprises:
(a) separating isomaltulose from said mother liquor by crystallization from methanol;
(b) removing fermentable sugar from said mother liguor by treatment with free or immobilised yeast;
(c) recovering 1-O-~-glucopyranosido-D-fructose ira~
said mother liquor by chromatographic separation by i means of an ion exchanger or other separating material;
and (d) transforming said l~O-~-glucopyranosido-D-fructose to dry form.
Surprisingly, it has been found that it is possible to obtain isomaltulose of high purity and with a high yield from sucrose by blending pure sucrose solu~o~s having a preferred concentration of 40% by weight to 75% by weight, more preferably 45% by weight to 60~ by weight, with the mo her liquor of a preceding , .
isomaltulose crystallization, by treating this solution or bringing it into contact with immobilized dead cells of microorganisms forming isomaltulose from sucrose at a temperature of 25C to 40C, preferably 30C to 40C, and by obtaining the resultant iso-maltulose in crystalline form by a method known per se.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
An advantageous embodiment of the process consists in dividing the sucrose solution having a concentration of 403 by weight to 60% by weight, into two parts, in passing the first part via a reactor, which is filled with immobilized dead cells of microorganisms forming isomaltulose from sucrose, at ~5C to 40C, preferably 30C to 40C, so that 80% to 90% of the sucrose is converted, in mixing the second part of the sucrose solution ~ith the mother liquor from a preceding isomaltulose crystallization, in passing this solution via a second reactor, which is filled with immobilized dead cells of microorganisms forming isomaltulose from sucrose, at 25C to 40C, preferab~y 30C to 40C, so that the sucrose is almost completely converted, in obtaining the resultant isomaltulose from this solution in crystalline form by methods Xnown per se, in dissolving and crystallizing the crystals in the converted first part of the reaction ; solution, in separating the resultant isomaltulose crystals and in using the mother liquor for the ad-mixture to the second part of the sucrose solution.
The average contact time of the sucrose solution in the reactor depends on the specific activity, present in each case, of the immobilized cells. The specific activity is defined as ~mol of converted sucrose per gram of immobilized cells (dry) per minute. For example, a reactor having a diameter of 100 mm and a bed hei~ht of 450 mm contains 950 g of immobilized cells (dry~
with a specific activity of 60 units/g. A quantity of 1900 ml of sucrose solution (50~ by weight) rnust be made to Elow through this reactor per hour in order to convert 90% of the sucrose. As the activity continuously decreases over the working time, the velocity of flow must be readjusted constantly in order to obtain a product with constant composition.
BRIEF DESCRIPTION OF THE DRAWING
The single Figure of the drawing is a flow she~
illustrating the process described in Example 3 below.
The specific advantages of the process according to the invention are explained in greater detail with the aid of the following examples. Example 1 describes the procedure and results of the known process of Germany Offenlegungsschrift No. P 30 38 219.
(a) Cells from an inoculum of Protaminobacter rubrum (CBS 574.77) are transferred with 10 ml of a sterile nutrient substrate consisting of 8 kg of thick juice from a sugar plant (dry substance content = 65%), 2 kg of corn steep liquor, 0.1 kg of (NH4)2HPO4 and 89.9 kg of distilled water (adjusted, if needed, to pH of 7.2). This suspension is used as the inoculating material for the shaker-machine pre-culture in 1-liter flasks with 200 ml each of the nutrient 25 solution of the above compositionD Twenty of such flasks (total contents 4 liters) are inoculated at 29C for 30 hours.
16 Liters of nutrient solution of the above composition are inoculated in a 30 liter small fermen-ter 30 with the contents of the above twenty flasks (4 liters) and fermented at 29C at an aeration rate of 20 liters/
j minute and a stirring rate of 350 rpm. The increasing cell count is determined with a microscope. When ; a concentration oi 5 x 109 cells~ml is reached, the 35 contents of the fermenter are transferred in-to another container and reacted therein with a cationic flocculant (for instance, PRIMAFLO~ C7 by Rohm & Haas, c 4-*trade mark , Philadelphia, Pa., ~S~). The flocculated cells ~re allowed to settle, decanted, washed ~ith ~.~ M
phosphate buffer of a pH of 7.0i and dewatered in the centrifuse. The substance i~ then extruded into strands, air-dried and ground.
tb) The 0 3 to 0.8 mm sifted fraction from the above-obtained preparation is stirred in a 0.~
glutaraldehyde solution for 10 minutes, washed with phosphate buffer (O.lM, pH=7.0~ and filled under water into a heatable colu~n. The col~mn is then he~ted to 50C and a 70% by ~eight suc~ose solution ~s pumped continuously through this column. The flow rate is adjusted in such a manner that no sucrose can be detected anymore at the end of the column. The isomaltulose solution so obtained has the folla~ing average composition ~as determined ~y ~PLC, ~ e_ high-pressure liquid chromatography):
fructose .. .. .. 7.4 9~OO g TS (total sugar) glucose .. .. .. .. 0.3 g~100 g TS
sucrose .. .. .. .. 0.1 g/100 g TS
isomaltulose .. .. .. 62 6 9~100 g TS
l-O-a-glucopyranosido-D-fructose .. .. .. 16.~ g~100 g TS
oligosaccharide .. .. 13.0 g/100 g TS.
This solution is fed to a cooling crystallizer, cooled to 20C and the crystallized isomaltulose is separated from the mother liquor in wire-basket centrifuges. The mother liquor is concentrated by evaporation to 78% to 82% TS content, cooled in a second cooling crystallization to 20C, and a second quantity of crystallized isomaltulose is separated from the mother liquor in wire-basket centrifuges.
From 100 kg of a sucrose solution containing 70 kg of sucrose there are obtained 43.8 kg of isomaltulose (62.6% of TS content). From this there is obtained in the first crystallization 15.9 kg of pure isomaltulose and in the second crystallization .~ ,~ . .
I
_ 13.4 kg of isomaltulose of 98~ purity. This i~
equivalent to a total yield of 29 kg of pure iso-maltulose from 70 kg of sucrose or 41.4% from the raw material. (In this example and in the following the calculations are based on anhydrous isomaltulose).
The 0.3 to 0.8 mm sifted fraction from ~he preparation prepared according to example l~a) is : stirred in a 0.1% glutaraldehyde solution for 10 minutes, washed with phosphate buffer ~9.1M, pH=7.03 and fil].ed under water into a heatable column. The colu~n is then heated to 30C and a 50~ by weight sucrose solution is pumped through continuously.
The velocity of flow is adjusted so that no sucrose can be detected anymore at the end of the column.
The isomaltulose solution so obtained has the -following average composition ~HPLC):
fructose .. .. .. 3.6 g/100 g TS
glucose .. .. .. .. 1.8 g/lD0 g TS
sucrose .. .. .. .. 0 isomaltulose .. .. .. 78.4 9/100 g TS
l-o-a-D-glucopyranosido-D-fructose .. .. .. 12.6 g/100 g TS
oligosaccharide .. .. 3.6 y/100 9 TS.
.~ 25 This solution is concentrated by evaporation to 78% to 82% TS content, fed to a cooling crystal-lizer a`nd.cooled to 20~C, and the crystallize~
isomaltulose is separated from the mother liquor in wire-basket centrifuges. The mother liquor is again , 30 conecentrated by evaporation to 78~ to 82~ TS content, sub~ected to a second cooling crystallization to 20~C, ; and a second quantity of crystallized isomaltulose is separated from the mother liquor in wire-basket '- centrifuges.
From 1~0 kg of a sucrose solution containing 50 kg of sucrose, there are obtained 39.2 ]cg of isomaltulose (78.4% of TS content). Fro.m this there .. .. ...... .
is obtained in the first crystallization 25.0 kg of pure isomaltulose and in the second crystallization 9.6 kg of isomaltulose with 98~ purity. This is equivalent to a total yield of 3g.4 kg of pure isomaltulose from 50 kq saccharose or 68.8~ from the raw material.
The 0.3 to 0.8 mm sifted fraction from the pre-paration prepared according to example ~(a) is stirred in a 0.1~ glutaraldehyde solution to 10 minutes, washed with phosphate buffer ~O.lM, pH=7.0) and filled under water into two he~table columns heated to 30C.
The procedure is explained ~ith reference to the flow sheet in Figure 1.
A 50% by weight sucrose solution is made to flow through one of the colu~ns 1 conti~uously The velocity of flow is adiusted so t~at only approximately 90% of the sucrose i 5 converted ~hen the~e is flow through the column. The reaction~ solution so obtained ~ 20 has the following exempl~ry compGsition ~EIPLC):
'~ fructose ......... .. .. 2.~ g~100 g TS
glucose .. .. .. .. 1.6 g~lOO g TS
sucrose .. .. .. .. 10.~ g/lOC g TS
isomaltulose .. .. .. 75.2 gJlOO g TS
l-O-a-D-glucopyranosido-D-fructose ....... .. .. 6.~ g/100 g TS
, oligosaccharide ....... .... .. 3.l g/100 g TS.
i Parallel to the first column, the second colomn
2 is charged with a substrate which consists of a mixture of sucrose and mother liquor of a preceding isomaltulose crystallization. An exemplary composition (HPLC) of this substrate is:
TS-content .. .. .. 50% by weight fructose .. ... .. 2.0 g/100 g TS
, 35 glucose ..... .. .. .. 1.0 g/100 g TS
sucrose ..... .. .. .. 76.2 g/100 g TS
isomaltulose .'. .. .. 10.8 9/100 g TS
~' .. . .
.'.
~' .. i .
1-0-~-~-glucopyranosido-D-fructose .. .. .. 8.2 9/100 g TS
oligosaccharide .. .. 1.8 g/100 g TS.
The flow rate through this columns is adjusted so that the solution at the end of the column contains at the most 1 g o~ sucrose per 100 g dry substance.
The reaction solution so obtained has the following exemplary composition (HPLC):
fructose ................ .. 5.4 g/100 g TS
glucose ....... .~ ....... .. 2.2 g/100 g TS
; suc~ose ....... .. .. 1 0 9/lOO g TS
isomaltulose ............ .. 72.3 g/100 g TS
1-O-~-D-glucopyranosido-; D-fructose ....... .. .. 16.6 g/100 g TS
oligosaccharide ....... .. 2.5 g/100 g TS.
This solutio~ is concentrated by evaporation to 78~ to 82~ ~S content, subjected to a cooling crystallization to 20C, and the crystallized isomaltulose is separated from this second mother ; 20 liquor in wire-backet centrifuges.
The isomaltulose obtained at this point is dissolved in the reaction solution obtained from the first column, concentrated by evaporation to 78% to 82~ TS content, cooled in a cooling crystallizer to ; 25 20C, and the crystallized pure isomaltulose is separated from the mother liquor in wire-bac~et centrifuges~ The mother liquor obtained at this point is used for preparing the substrate for the second column The second mother liquor, which can also be called molasses,-contains approximately 30~ to 40~, based on TS, of 1-O-a-glucopyranosido-D-fructose ana can be used as raw material for obtaining this sugar.
This sugar can be obtained, for example, by chromatographic separation, by ion exchange or by the use of other suitable separating procedures, after , a prior enrichment by crystallizing out a further , ' ' ,.
.
..... ... ...
part of the isomaltulose from methanol and by removing - the sugars,.by means of fermentation through treatment with free or immobili~ed yeast.
With the procedure outlined in this example, 80.0 kg of pure isomaltulose are obtained per lQ0 kg of sucrose.
Although the invention has been described in conjunction with certain preferred embodiments, it is not intended to be limited thereto, but instead includes all those embodiments within the spirit and scope of the appended claims.
; 25 .
, . .
.
.; .
_ g _ ; '
TS-content .. .. .. 50% by weight fructose .. ... .. 2.0 g/100 g TS
, 35 glucose ..... .. .. .. 1.0 g/100 g TS
sucrose ..... .. .. .. 76.2 g/100 g TS
isomaltulose .'. .. .. 10.8 9/100 g TS
~' .. . .
.'.
~' .. i .
1-0-~-~-glucopyranosido-D-fructose .. .. .. 8.2 9/100 g TS
oligosaccharide .. .. 1.8 g/100 g TS.
The flow rate through this columns is adjusted so that the solution at the end of the column contains at the most 1 g o~ sucrose per 100 g dry substance.
The reaction solution so obtained has the following exemplary composition (HPLC):
fructose ................ .. 5.4 g/100 g TS
glucose ....... .~ ....... .. 2.2 g/100 g TS
; suc~ose ....... .. .. 1 0 9/lOO g TS
isomaltulose ............ .. 72.3 g/100 g TS
1-O-~-D-glucopyranosido-; D-fructose ....... .. .. 16.6 g/100 g TS
oligosaccharide ....... .. 2.5 g/100 g TS.
This solutio~ is concentrated by evaporation to 78~ to 82~ ~S content, subjected to a cooling crystallization to 20C, and the crystallized isomaltulose is separated from this second mother ; 20 liquor in wire-backet centrifuges.
The isomaltulose obtained at this point is dissolved in the reaction solution obtained from the first column, concentrated by evaporation to 78% to 82~ TS content, cooled in a cooling crystallizer to ; 25 20C, and the crystallized pure isomaltulose is separated from the mother liquor in wire-bac~et centrifuges~ The mother liquor obtained at this point is used for preparing the substrate for the second column The second mother liquor, which can also be called molasses,-contains approximately 30~ to 40~, based on TS, of 1-O-a-glucopyranosido-D-fructose ana can be used as raw material for obtaining this sugar.
This sugar can be obtained, for example, by chromatographic separation, by ion exchange or by the use of other suitable separating procedures, after , a prior enrichment by crystallizing out a further , ' ' ,.
.
..... ... ...
part of the isomaltulose from methanol and by removing - the sugars,.by means of fermentation through treatment with free or immobili~ed yeast.
With the procedure outlined in this example, 80.0 kg of pure isomaltulose are obtained per lQ0 kg of sucrose.
Although the invention has been described in conjunction with certain preferred embodiments, it is not intended to be limited thereto, but instead includes all those embodiments within the spirit and scope of the appended claims.
; 25 .
, . .
.
.; .
_ g _ ; '
Claims (9)
1. In a process for preparing isomaltulose (6-O-.alpha.-D-glucopyranosido-D-fructose) wherein sucrose solution is treated with or brought into contact with dead immobilized cells of a microorganism capable of converting sucrose into isomaltulose, the improve-ment which comprises:
(a) mixing said sucrose solution with the mother liquor of a preceding isomaltulose crystallization to form a mixture;
(b) bringing said mixture into contact with, or tre-ating said mixture with, dead immobilized cells of said microorganism at a temperature of 25 to 40°C
to form isomaltulose; and (c) recovering said isomaltulose formed in step (b) in crystalline form.
(a) mixing said sucrose solution with the mother liquor of a preceding isomaltulose crystallization to form a mixture;
(b) bringing said mixture into contact with, or tre-ating said mixture with, dead immobilized cells of said microorganism at a temperature of 25 to 40°C
to form isomaltulose; and (c) recovering said isomaltulose formed in step (b) in crystalline form.
2. The process of claim 1, wherein said temperature in step (b) is 30 to 40°C.
3. The process of claim 1, which comprises:
(a) using a sucrose solution having a concentration of 40 to 75% by weight;
(b) dividing said sucrose solution into a first sucrose solution and a second sucrose solution;
(c) bringing said first sucrose solution into contact with, or treating said first sucrose solution with, dead immobilized cells of said microorganism at a temperature of 25 to 40°C
to convert 80 to 90% of the sucrose to isomaltulose and thereby to form a solution containing isomaltulose:
(d) mixing said second sucrose solution with the mother liquor of a preceding isomaltulose crystallization to form a mixture;
(e) bringing said mixture formed in step (d) into contact with, or treating said mixture formed in step (d) with, dead immobilized cells of said microorganism at a temperature of 25 to 40°C so that the sucrose of said mixture is almost completely converted into isomaltulose;
(f) recovering said isomaltulose formed in step (e) in crystalline form;
(g) dissolving said crystals recovered in step (f) in said solution formed in step (c) and effecting crystallization to obtain isomaltulose crystals and a mother liquor; and (h) using the mother liquor obtained in step (g) as the mother liquor in step (d).
(a) using a sucrose solution having a concentration of 40 to 75% by weight;
(b) dividing said sucrose solution into a first sucrose solution and a second sucrose solution;
(c) bringing said first sucrose solution into contact with, or treating said first sucrose solution with, dead immobilized cells of said microorganism at a temperature of 25 to 40°C
to convert 80 to 90% of the sucrose to isomaltulose and thereby to form a solution containing isomaltulose:
(d) mixing said second sucrose solution with the mother liquor of a preceding isomaltulose crystallization to form a mixture;
(e) bringing said mixture formed in step (d) into contact with, or treating said mixture formed in step (d) with, dead immobilized cells of said microorganism at a temperature of 25 to 40°C so that the sucrose of said mixture is almost completely converted into isomaltulose;
(f) recovering said isomaltulose formed in step (e) in crystalline form;
(g) dissolving said crystals recovered in step (f) in said solution formed in step (c) and effecting crystallization to obtain isomaltulose crystals and a mother liquor; and (h) using the mother liquor obtained in step (g) as the mother liquor in step (d).
4. The process of claim 3, wherein said sucrose solution has a concentration of 45 to 60% by weight.
5. The process of claim 3, wherein said temperature in steps (c) and (e) is 30 to 40°C.
6. The process of claim 1 to 3, wherein said microorganism is selected from the group consisting of Protaminobacter rubrum (CBS 574.77), Serratia plymuthica (ATCC 15928), Serratia marcescens (NCIB 8285) and Leuconostoc mesenteroides (NRRL
B-512 F (ATCC 10830a)).
B-512 F (ATCC 10830a)).
7. The process of claim 1 to 3, wherein said microorganism is Protaminobacter rubrum (CBS 574.77).
8. A process for obtaining 1-0-.alpha.-glucopyra-nosido-D-fructose from a mother liquor from which isomaltulose has been separated, which process com-prises:
(a) separating isomaltulose from said mother liquor by crystallization from methanol;
(b) removing fermentable sugar from said mother liquor by treatment with free or immobilised yeast;
(c) recovering 1-0-.alpha.-glucopyranosido-D-fructose from said mother liquor by chromatographic separation by means of an ion exchanger or other separating material;
and (d) transforming said 1-0-.alpha.-glucopyranosido-D-fructose to dry form.
(a) separating isomaltulose from said mother liquor by crystallization from methanol;
(b) removing fermentable sugar from said mother liquor by treatment with free or immobilised yeast;
(c) recovering 1-0-.alpha.-glucopyranosido-D-fructose from said mother liquor by chromatographic separation by means of an ion exchanger or other separating material;
and (d) transforming said 1-0-.alpha.-glucopyranosido-D-fructose to dry form.
9. The process of claim 8, wherein said mother liquor is that obtained by the process of claim or 3.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DEP3213107.0-42 | 1982-04-07 | ||
| DE19823213107 DE3213107A1 (en) | 1982-04-07 | 1982-04-07 | METHOD FOR PRODUCING ISOMALTULOSE (6-O- (ALPHA) -D-GLUCOPYRANOSIDO-D-FRUCTOSE) WITH THE AID OF IMMOBILIZED BACTERIA CELLS |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1185551A true CA1185551A (en) | 1985-04-16 |
Family
ID=6160532
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA000424503A Expired CA1185551A (en) | 1982-04-07 | 1983-03-25 | PROCESS FOR THE PREPARATION OF ISOMALTULOSE (6-0-.alpha.- D-GLUCOPYRANOSIDO-D-FRUCTOSE) BY THE USE OF IMMOBILIZED BACTERIAL CELLS |
Country Status (9)
| Country | Link |
|---|---|
| EP (1) | EP0091063B1 (en) |
| JP (1) | JPH0661274B2 (en) |
| AT (1) | ATE28084T1 (en) |
| CA (1) | CA1185551A (en) |
| DE (2) | DE3213107A1 (en) |
| DK (2) | DK167366B1 (en) |
| ES (1) | ES8401985A1 (en) |
| FI (1) | FI72345C (en) |
| IE (1) | IE54643B1 (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4898820A (en) * | 1987-07-27 | 1990-02-06 | Showa Denko Kabushiki Kaisha | Process for production of isomaltulose |
| US5578339A (en) * | 1993-05-06 | 1996-11-26 | Sudzucker Aktiengesellschaft Mannheim/Ochsenfurt | Sweetener, process for its preparation and the use thereof |
| US6146856A (en) * | 1997-05-22 | 2000-11-14 | Xyrofin Oy | Process for the production of isomatulose and other products |
Families Citing this family (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH02273192A (en) * | 1989-04-13 | 1990-11-07 | Meito Sangyo Kk | Production of isomaltulose |
| DE4209653A1 (en) * | 1992-03-25 | 1993-10-07 | Herbert Prof Dr Daniel | Gas-filled heat-insulating layer suppressing convection - utilises density gradients determined by temperature gradients in gas |
| DE19747642B4 (en) * | 1997-10-29 | 2004-12-23 | Südzucker AG Mannheim/Ochsenfurt | Process for the production of sweeteners containing isomelecitosis, isomelecitosis and isomaltulose |
| US20100267658A1 (en) | 2009-04-15 | 2010-10-21 | Sudzucker Aktiengesellschaft Mannheim/Ochsenfurt | Trehalulose-containing composition, its preparation and use |
| BR122019001300B1 (en) | 2009-12-23 | 2020-03-03 | Evonik Degussa Gmbh | PROCESS FOR THE PRODUCTION OF SWEETENERS |
| DE102011100772A1 (en) | 2011-05-05 | 2012-11-08 | Evonik Degussa Gmbh | Process for the preparation of isomaltulose from plant juices |
| JP5483482B2 (en) * | 2011-05-23 | 2014-05-07 | 三井製糖株式会社 | Method for producing a solid from a sugar solution and the solid |
| DE102011083030A1 (en) | 2011-09-20 | 2013-03-21 | Evonik Degussa Gmbh | Mixture composition and its use as a sweetener |
| DE102013011977A1 (en) | 2013-07-18 | 2015-01-22 | Südzucker Aktiengesellschaft Mannheim/Ochsenfurt | Optimized process for preparing an isomaltulose-containing composition |
| EP3363909A1 (en) | 2017-02-15 | 2018-08-22 | Evonik Degussa GmbH | Process for production of a solid material containing isomaltulose crystals and trehalulose |
| EP3653708A1 (en) | 2018-11-14 | 2020-05-20 | Evonik Operations GmbH | Isomaltulose production |
| EP3892730A1 (en) | 2020-04-07 | 2021-10-13 | Evonik Operations GmbH | In situ production of isomaltulose |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CH592141A5 (en) * | 1972-04-12 | 1977-10-14 | Sueddeutsche Zucker Ag | |
| DE3066516D1 (en) * | 1979-11-07 | 1984-03-15 | Tate & Lyle Plc | Production of isomaltulose |
| DE3038219A1 (en) * | 1980-10-09 | 1982-04-15 | Süddeutsche Zucker AG, 6800 Mannheim | METHOD FOR PRODUCING ISOMALTULOSE (6-O- (ALPHA) -D-GLUCOPYRANOSIDO-D-FRUCTOSE) WITH THE AID OF IMMOBILIZED BACTERIA CELLS |
| DE3241788A1 (en) * | 1982-11-11 | 1984-05-17 | Süddeutsche Zucker AG, 6800 Mannheim | METHOD FOR PRODUCING 1-0- (ALPHA) -D-GLUCOPYRANOSIDO-D-FRUCTOSE AND USE AS A SWEETENER |
-
1982
- 1982-04-07 DE DE19823213107 patent/DE3213107A1/en not_active Withdrawn
-
1983
- 1983-03-17 DK DK122983A patent/DK167366B1/en not_active IP Right Cessation
- 1983-03-24 IE IE642/83A patent/IE54643B1/en not_active IP Right Cessation
- 1983-03-25 CA CA000424503A patent/CA1185551A/en not_active Expired
- 1983-03-28 EP EP83103078A patent/EP0091063B1/en not_active Expired
- 1983-03-28 DE DE8383103078T patent/DE3372294D1/en not_active Expired
- 1983-03-28 AT AT83103078T patent/ATE28084T1/en not_active IP Right Cessation
- 1983-03-30 FI FI831105A patent/FI72345C/en not_active IP Right Cessation
- 1983-04-05 ES ES521240A patent/ES8401985A1/en not_active Expired
- 1983-04-06 JP JP58059381A patent/JPH0661274B2/en not_active Expired - Lifetime
-
1992
- 1992-01-27 DK DK199200096A patent/DK172544B1/en not_active IP Right Cessation
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4898820A (en) * | 1987-07-27 | 1990-02-06 | Showa Denko Kabushiki Kaisha | Process for production of isomaltulose |
| US5578339A (en) * | 1993-05-06 | 1996-11-26 | Sudzucker Aktiengesellschaft Mannheim/Ochsenfurt | Sweetener, process for its preparation and the use thereof |
| US6146856A (en) * | 1997-05-22 | 2000-11-14 | Xyrofin Oy | Process for the production of isomatulose and other products |
Also Published As
| Publication number | Publication date |
|---|---|
| DE3372294D1 (en) | 1987-08-06 |
| DK122983A (en) | 1983-10-08 |
| DK122983D0 (en) | 1983-03-17 |
| ATE28084T1 (en) | 1987-07-15 |
| DK9692D0 (en) | 1992-01-27 |
| IE54643B1 (en) | 1989-12-20 |
| FI72345B (en) | 1987-01-30 |
| FI831105L (en) | 1983-10-08 |
| DK167366B1 (en) | 1993-10-18 |
| FI831105A0 (en) | 1983-03-30 |
| ES521240A0 (en) | 1984-01-01 |
| DK9692A (en) | 1992-01-27 |
| JPS592695A (en) | 1984-01-09 |
| JPH0661274B2 (en) | 1994-08-17 |
| IE830642L (en) | 1983-10-07 |
| EP0091063A3 (en) | 1984-09-26 |
| EP0091063B1 (en) | 1987-07-01 |
| FI72345C (en) | 1987-05-11 |
| ES8401985A1 (en) | 1984-01-01 |
| DK172544B1 (en) | 1998-12-14 |
| EP0091063A2 (en) | 1983-10-12 |
| DE3213107A1 (en) | 1983-10-13 |
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