JP2002503098A - Process for producing isomaltulose and other products - Google Patents

Abstract

(57) Summary The present invention relates to a method for simultaneously producing converted and unconverted saccharide and / or non-saccharide products. The method is particularly adapted for the simultaneous production of isomaltulose and / or trehalulose and betaine or invert sugar from liquids derived from plants. The sucrose in the liquid is converted to isomaltulose and trehalulose by the enzyme and the desired product is individually recovered from the resulting solution. The isomaltulose can be further converted to isomalt.

Description

DETAILED DESCRIPTION OF THE INVENTION                Process for producing isomaltulose and other products   The present invention relates to inverted and unconverted saccharide and / or non-saccharide The present invention relates to a method for obtaining a product simultaneously. In particular, the present invention relates to isomaltulose e) and / or trehalulose and betaine or converted It relates to the simultaneous production of sugars.   Isomaltulose (or palatinose) is a reductase produced from sucrose. Sugars. Food industry suggests using isomaltulose as sweetener And it is converted to a sweetener isomalt (Palatini) by hydrogenation. (Pala-tinit)). Isomalt is essentially homogeneous α-D-glycopyranosyl- (1,6) -sorbitol and α-D-glucopi It is a mixture of lanosyl- (1,1) -mannitol.   Trehalulose is another type of reducing sugar obtained from sucrose. Trehal Α-D-glucopyranosyl- (1,1) -sorbitol and And α-D-glucopyranosyl- (1,1) -mannitol are obtained, It also has a sweet taste.   Isomerization of sucrose to isomaltulose and / or trehalulose Alternatively, various methods have been reported for transglucosylation. For example, a proto Aminobacter rubrum, Serratica primrumica (S erratica plymuthica), Erwinia rhapontici, Pseu In microorganisms such as Pseudomonas mesoacidophile Isomerization occurs with transglucosylase enzymes that have been found to be present. Is Known isomerization techniques include living microbial cells or dead microbial cells. Includes isomerization using vesicles or using enzymes in an extracted state . The enzyme has been used both in immobilized and free form.   Prior art for the production of isomaltulose and / or trehalulose The method is described, for example, in European Patent EP 0 288 900, US Pat. National patent US4640894, German patent DE1049800, European patent EP1 No. 099, European Patent EP77971, European Patent EP049801, European Patent EP200069, European Patent EP160253, European Patent EP483755 And publications such as EP 625578. Skull A review of transglycosylation of glycine is described in the publication, Munia, M. et al. (Munir, M ), Etc., in Carbohydrate Res., 164 (1987) 477-485. It is listed. Technical background of the method of producing isomaltulose from molasses by enzyme For the scenery, see the Finnish Application 96 filed on May 17, 1996 by the applicant. No. 2095. The specifications of the above patents, patent applications and publications are in the text Has been incorporated as a reference.   Betaine has the formula (CH_Three)_ThreeN⁺CH_TwoCOO^-With a quaternary ammonium compound having is there. It is known that betaine accumulates in sugar beets, and this Various kinds such as ion exchange, crystallization, extraction from sugar beet molasses or distilled meal Collected by technique or by chromatography. Betaine is animal feed And for pharmaceutical and cosmetic applications.   Prior art methods for the recovery of betaine are described, for example, in EP 54544. And European Patent EP 345511, the specification of which is incorporated herein by reference. Has been adopted as.   Sugar cane juice does not contain betaine, but instead they contain It supplies a large amount of converted saccharide, which is a mixture of glucose and fructose. Conversion Can be used directly as a sweetener, but after purification It is also possible to separate into fructose and fructose.   Prior art methods for obtaining converted sugars from sugarcane are described, for example, in the publications, Saska, M. (Saska, M.) and other international sugar journals (Int Sugar Jnl.), 1994, 96, 1150, pp. 403-410; ( Hongisto, H.) et al., Sugarcane by fin sugar chromatography Desuarization of cane molasses by the Finnsugar chromato-gr aphic separation process), Finish Sugar Corporation Limited Ted (Finnish Sugar Co. Ltd.), Kantovik, Finland, Dated March 4, 1977; these specifications are incorporated herein by reference. Has been inserted. In the prior art described above, a batch or continuous process of sugarcane molasses is used. Desaccharification by continuous chromatography has been studied. This method allows you to Separation and recovery of sugars and converted saccharides are possible.   Further, U.S. Pat. No. 3,214,293 discloses a combination of ion exchange and ion exclusion. The separation of non-saccharides from saccharides in aqueous saccharide solutions .   British Patent No. 1 448 524 discloses liquid partitioning chromatography on cation exchangers. For the separation of sugars from non-sugars of sugar beet or sugar cane molasses by chromatography The law is described. Converted saccharides and sugars, including betaines, and non-saccharides Fractions can be collected.   To recover betaine or converted sugars associated with isomaltulose production No prior art method has been supplied for this purpose.   An object of the present invention is to produce valuable sugars and non-saccharides from aqueous solutions derived from plants. To provide a way to get things.   Another object of the present invention is a new method for producing saccharides and non-saccharides from molasses Is to supply.   A special object of the present invention is to provide aqueous solutions containing saccharides, in particular molasses from betaine or Recover converted sugars and isomaltulose and / or trehalulose To provide a method that allows   A further object of the present invention is to provide isomaltulose and / or trehalulose. Providing a method for obtaining the hydrogenation product of the catalyst.   It is now possible to dissolve sucrose in aqueous solutions derived from sugar beets or sugar cane. Has a substantial effect on each of the betaines or converted sugars present in the liquid Be converted to isomaltulose and / or trehalulose without It was discovered that the individual desired products could be recovered in good yield from the liquid obtained in Was. In addition, the sugars in molasses are immediately converted to isomaltulose and / or trehalulose. As well as betaine or conversion from the converted molasses described above. It was discovered that the disaccharides produced can be effectively recovered while recovering the recovered saccharides. Was.   The present invention is based on the above findings. Therefore, the present invention relates to isomalturo A method for simultaneously obtaining a sugar and other saccharide and / or non-saccharide products, comprising: Lower stage a) Derived from plants containing sucrose and the sugar or non-saccharide compound of interest. Supplying the used aqueous solution; b) transglucosylating the sucrose in the solution; c) isomaltulose and the desired sugar from the transglucosylated solution Recovering class and / or non-saccharide products; Provide a method consisting of combining   According to a preferred embodiment of the present invention, the method of recovery is carried out as individual fractions The chromatography consists of recovering the product.   If the liquid derived from the plant contains a large amount of sucrose, e.g. Or, if it is a concentrated solution, the method according to the invention requires a solution derived from plants. Crystallizing sucrose to provide molasses can be included. Or above Total sucrose in a solution derived from said plant isomaltulose and / or It can also be used for conversion to trehalulose. Isoma in the above solution If the ratio of tulose is large, the isomalt is allowed to crystallize before the recovery of other products. Preferably, at least a portion of the loin is separated.   In a preferred embodiment of the present invention, such solutions derived from plants include molasses Is included. If the solution derived from plants is derived from sugar beets, Pllose, Trehalulose and Betaine are mainly recovered in step c) of the process Product. If the solution is derived from sugarcane, isomalturo The source, trehalulose and converted sugars are recovered in step c).   In another embodiment according to the present invention, betaine is transglucosyl of sucrose It is recovered from the above solution before conversion.   The conversion of sucrose to isomaltulose and / or trehalulose is fine. Sucrose isomallose and / or trehalulose in living organisms A transglucosylating enzyme capable of inverting (sucrose-6 glycosyl) Mutase). Preferred microorganisms for the production of isomaltulose are Protaminobacter rubrum (CBS 574.7) 7) or other inverting microorganisms can be used. Immobilized or Using free microbial cells or using enzymes isolated from the cells. Any of the methods of the prior art can be used. In fact, it has enzyme conversion activity Need only be performed, and the manner in which this enzymatic activity is provided is related to the performance of the present invention. Not serious. For example, an increase in activity can be provided by recombination techniques.   An aqueous solution derived from plants containing sucrose and forming the raw material of the method What is used is, for example, a dilute or thick liquid of sugar radish or sugarcane crusher. Such a plant liquid may be used. On the other hand, one or several Use concentrated liquid in a conventional manner to produce sucrose by crystallization in the crystallization stage. Can be used, that is, it is suitable as a solution derived from the plants mentioned above .   In the context of the present invention, the expression “plant derived solution” refers to a large amount of sucrose. And solutions containing compounds derived from other plants. Should be suitable as a raw material for the present invention.   It is easily available in large quantities, inexpensive, and has a residual sucrose component of isomalturo. Molasses is a preferred raw material of the present invention because it is a good starting material for You. Molasses contains sucrose and proteins and mycelium that can be consumed for bacterial growth. Contains other nutrients such as Neral. These nutrient compounds increase the growth of microorganisms. Consumed by micro-organisms to aid the growth of inverting micro-organisms and It is possible to keep the conversion of the source at the same time.   Sugar radish molasses has about 50-70% of sucrose based on dry solids (DS). About 3 to 8% betaine and some other based on DS Nitrogen-containing compounds are included. For sugar cane molasses, about 35 or less based on DS 55% sucrose and about 10-35% invert sugar based on DS Kinds are included. In the prior art, molasses is used for the production of cell suspensions, sucrose. Has been described as a nutrient for microbial growth to convert Was. However, in pure sucrose solution or in dilute or concentrated plant solution Actually produced isomaltulose. Molasses is a good nutrient medium for the above bacteria. And molasses are practical for isomaltulose and / or trehalulose. It has now been discovered to provide a suitable source of sucrose for the production of sucrose. Present in molasses Betaine or the converted sugars, respectively, undergo material changes during the conversion process. And from the resulting reaction mixture isomaltulose and other sugars and Betaine and converted sugars can be recovered separately from Was also discovered.   An advantage of the present invention is that it is possible to use molasses sucrose for the conversion, To produce both turose and betaine or converted sugar fractions The existing molasses chromatography separation and grinding machine can be used.   The invention will be described in more detail with reference to the accompanying figures in the text.   FIG. 1 shows a block diagram of the method according to the present invention.   In the following detailed description, the special properties induced by using molasses For this purpose molasses is generally used as a raw material in the process according to the invention. However, Transglucose of sucrose contained in a solution derived from another plant It is clear that it can be used for saccharification and that described for molasses In the same manner as described above, isomaltulose and other sugars and / or It is also clear that non-saccharide products can be separated.   Sugar-containing solutions are generally prepared in a conventional manner to provide a dilute liquid with sugar beets or radish. It is provided by processing sugar-containing plants such as sugarcane. Dilute liquid Converts sucrose contained in the liquid as isomaltulose and / or It can be used to convert to trehalulose and can be processed into a thick liquid. Then the thick liquid can be crystallized to recover most of the sucrose . The crystallization may be a three-step crystallization as is customary, or it may involve reducing sugars in solution. It is preferable to crystallize the sucrose once or twice to leave more No.   According to the present invention, sucrose is isomaltulose and / or trehalulose The saccharide-containing solution is brought into contact with the transglucosylating enzyme activity that can be converted to glucose. Good In a better way. Product of P. rubrum (CBS 574.77) Growth media for cell suspensions of inverting microorganisms such as molasses or molasses, corn Corn steep solution, and mineral nutrients such as (NH_Four)_TwoHPO_Fourof It is a mixture. By fermentation with good aeration at about 30 ° C. and pH 7, 10⁹/ Ml or more of the desired counts of microbial cells You. The fermentation broth is added to the saccharide-containing solution by a batch method, and To perform the conversion.   As is known in the art, live or dead in a carrier or Can also immobilize microbial cells on a carrier. Alternatively, the enzyme Extraction from the cells and use it as is or in an immobilized state to perform the conversion. Both are possible. For example, referring to the references mentioned in the introductory part of the present specification, These are all incorporated by reference into the text.   Transglucosylation of sucrose is either batch or continuous, immobilized In a tank or column containing immobilized or unimmobilized enzyme system Can be implemented. Batch processes provide an additional source of carbon to the solution during conversion to Can be implemented as a fed-batch method to assist in the growth of the yeast. Optimal temperature of 25-35 ° C The conversion is preferably carried out at about 30 ° C. with suitable aeration. Conversion When molasses is used, it is preferably consumed in the liquid to prevent excessive foaming. Add foam.   The conversion process is performed as a continuous process in a column packed with a carrier with immobilized enzyme. For example, receiving regenerated material in one column while others are operating Several colors that can be connected sequentially and / or in parallel as System exists. Pass through one or several columns to increase conversion yield To recirculate the solution.   It is generally preferred that transglucosylation be continued for a sufficient Convert 80% or more. The converted solution has the main reaction product Contains somaltulose, and-in general, some trehalulose, betaine In addition, it contains fructose and glucose and some unreacted sucrose. Converted or other converted sugars in the solution originally supplied or Not including compounds.   Once the desired conversion has been achieved, live microbial cells need to be killed. This is an example For example, by raising the temperature to about 80 ° C for 30 minutes or longer Is done. Before the solution is separated by chromatography, dead cells and other Must be removed by centrifugation and / or filtration.   If the conversion is carried out in a solution in which sucrose is not crystallized, isomaltulose Due to the large volume, the solution may be removed from the solution in a conventional manner before separation by chromatography. It is preferable to crystallize a major part of somaltulose.   Isomaltulose / trehalulose and betaine or converted sugars Separation by chromatography removes sucrose and betaine or converted sugars. Uses the same methods and equipment used in the prior art to separate each Can be done.   Remove any insoluble impurities from the converted solution, for example by filtration, then It is softened by ion exchange to remove divalent ions. Then, for example, Na, K Solution in a chromatographic separation column containing a monovalent cation exchange resin such as Supply.   In an exemplary method according to the present invention, a transglucosylated solution is Sterilization followed by centrifugation and / or filtration to remove microbial cells. Also remove them. For sugar cane molasses, phosphate precipitation is used to obtain a clear solution. It is necessary to combine the filtration and centrifugation steps. For example, by ion exchange The filtrate is softened and evaporated to reduce volume. Filtrate (preferably pH 5 . (At a pH of 5 or higher) into a separation column and develop with water to Various fractions of the class and non-saccharide products are provided. The actual fraction The composition depends on the initial feed solution used. -In general, individual fractions Evaporate and increase the concentration for the next processing step.   According to the method of the present invention, the fraction is isomaltulose, And betaine or invert sugars, respectively. One of the fractions The portion may be regenerated to provide a more pure product. Iso obtained from separation Crystallize maltulose and supply isomaltulose crystals. Supply isomalt Isomaltulose, which is commercially available as a sweetener. Is used. Isomalt is made from isomaltulose. Isomaltulose fraction after crystallization or from chromatography column It is produced by hydrogenation directly from hydrogen. Hydrogenation of trehalulose is also sweet Feed mixture, so that isomaltulose and trehalulose combine It may be quenched. It can be purified by ion exchange before hydrogenation. Likewise For example, as described in EP 54544 or EP 345511. Betaine can be recovered from the solution as described in Has been incorporated as a reference. The converted sugar fraction is in liquid form Retained or evaporated to a syrup. Invert the converted sugars before evaporation For further purification, for example, ion exchange can be used.   The following examples illustrate the invention, but are not limited to any method. There is no.   Example 1 Molasses as a feedstock for isomaltulose / trehalulose production and sucrose   Protaminobacter rubrum (CBS 574. 77) Production of fermented broth:   P. Material culture of P. rubrum is kept on nutrient agar slant at + 4 ° C. Was. Cells from the culture were diluted with 2 ml of saline. 0.4 to 0.4 of the obtained suspension Use a sterile shielded 500 ml shaking flask using a 0.6 ml aliquot. Co was inoculated with a 9 × 300 ml aliquot of growth medium. The inoculated flask Shake at 230 rpm, 30 ° C. for 1 day. The growth medium contains 0.01 M Na_Two HPO_Four5% sucrose, 1% peptone, 0.5% yeast extract in buffer, pH 7 And 0.3% beef extract. Concentration is 10 × 10⁹Cells / ml reached One from sugar beet molasses and the other from pure sucrose solution, Fermented broth was used for transglucosylation in two separate experiments.   Transglucosylation:   Sugar beet molasses containing 60.7% sucrose based on DS (Cultor Ltd.) was used for the molasses test. The P. described above. Lubrum increase 2600 ml of culture medium together with 7 l of sterile molasses-water solution in an invert fermenter (Bio (Biostat), 10 l). To start the conversion, the sucrose concentration is 200 g / l, the pre-dried solids concentration is 325 g / l, and the pH is adjusted to pH 7. Saved. The conversion is carried out for 13 hours under sterile conditions at 30 ° C. and aeration at 10 l / min. And while stirring at 350 rpm, most of the sucrose is converted. Continued until   Foam formation was very problematic when molasses was used for the conversion. Continuous foam formation Detection and the addition of an antifoaming agent was required. Conversion complete and sucrose detected If no more, raise the temperature of the solution to 80 ° C for 30 minutes to kill the microorganisms. The conversion was stopped. 0.1% Kenite (Ke) as body feed nite) 300 to filter cells and other solid residues by size pressure filtration .   In another experiment, pure sucrose was prepared under conditions similar to those of sugar beet molasses. The solution was inverted, except that the concentration of sucrose was 325 g / l.   The composition of both converted solutions was analyzed by HPLC (Ion exchange resin was Pb²⁺By type) Analyzed and the results are shown in the table below. Example 2 P. Transglucosylation of molasses sucrose by rubrum cells   fermentation   Protoaminobacterium bulum strain (CBS574.77) from an agar slope The cells were diluted with 2 ml of saline. Aliquots of 0.4 to 0.6 ml of the resulting suspension Using a coat, place the growth medium in a sterile shielded 500 ml shake flask. 300 ml were inoculated. The inoculated flask was shaken at 230 rpm and 30 ° C. for 1 day. I'm sorry 0.01M KH in growth medium_TwoPO_Four60 g / in buffer, pH 7 1 sucrose, 10 g / l peptone, 6 g / l beef extract were included. One day old 1,200 ml of a growth medium was added to 8.5 l of a nutrient medium (sucrose 70 g / l, corn Steep solution 40 g / l (NH_Four)_TwoHPO_Four  0.9g / l, pH7) Transferred to fermenter (Biostat, 10 l).   Temperature 30 ° C., pH 6-7, mixing at 350 rpm, and 12 l / Cell density of 10 × 10⁹Fermentation up to count / ml Continued. In a 1500 l fermenter, 1100 l of a sterile nutrient medium of the above composition The whole fermented broth was used for inoculation. 30 ° C, 1100 l / min ventilation rate The fermentation was performed at a stirring speed of 350 rpm.   Concentration is 10 to 20 × 10⁹Sucrose in molasses when reaching cells / ml Whole fermentation to transglucosylate isomaltulose and other sugar products Yeast broth was used.   Transglucosylation   Sugar beet molasses containing 60% sucrose based on DS (Kurta Limite (Cultor Ltd.) (1800 kgDS) into a reaction vessel (10 m^Three) In the 370g Diluted to a concentration of DS / l, sterilized at 120 ° C for 20 minutes and cooled to 30 ° C. pH to H_TwoSO_FourAdjust to pH 6 with. 1100 l of p. Rubrum fermented broth (10 to 20 × 10⁹Cells / ml) into a sugar beet molasses solution (4800 kg) Introduce, aerated at 1500 l / min at 30 ° C. and mixed at 200 rpm Act on sucrose. Defoamer Mazu 100 (PPG Indus Using TREES Inc. (PPG Industries Inc., UK) Avoid significant foam formation during conversion. Sucrose in molasses practically within 25 hours The total amount of sugar was converted and the saccharide composition of the converted solution is shown below. Isomaltulose 82% based on sugars 41% based on DS Trehalulose 8.8% 5.3% Fructose 3.3% 2% Glucose 2% 1% Oligosaccharide 3.9% 2%   After the conversion, p. Rubrum cell material is separated from the solution via a separator, Then the bacterial milk was reused for the new reaction. Is it a separator The solutions were pasteurized at 80 ° C. for 30 minutes to kill any remaining bacteria. The solution was cooled and filtered.   Example 3 Chromatographic separation of isomaltulose, trehalulose and betaine   The transglucosylated solution of Example 2 was chromatographed Separation by chromatography in a system.   Separation is performed in a batch chromatography column It is. All equipment includes supply tanks, supply pumps, heat exchangers, columns, discharge pumps, Pipelines for the introduction and discharge of product containers, feed solutions and dilution water It consisted of a device and a valve for controlling the liquid drained.   A cation exchange resin (Finex Oy, Finnish) and the height of the resin bed was about 5.3 m. Crosslinking of resin The degree of binding was 5.5% DVB and the average particle size was about 0.35 mm. Natori Um (Na⁺) Regenerate the resin into the mold and then assemble the feeder on top of the resin bed I was The temperature of the column as well as the feed solution and developing water was about 80 ° C. column Was adjusted to 630 l / h.   The first pretreatment step of the feed solution is filtration, which is diatomaceous earth as a filter aid. (The temperature of the solution was about 80 ° C, and the concentration was 100 g. About 30 g of dry solid in). After this, ions are removed to remove calcium ions. The solution was softened by exchange (temperature was about 55 ° C., concentration was the same as filtration). Then separation The solution was filtered again before.   Chromatographic separation was performed as follows.   Step 1: Determine the dry solids content of the feed solution and determine the refractive index (RI) of the solution Therefore (adjusted if necessary) to a concentration of about 30 g of dry solids in a 100 g solution. .   Step 2: 120 kg of dry solids (DS) of the feed solution are heat exchanged (through the feeder) It was pushed to the top of the resin bed through the exchanger.   Step 3: Supply ion-exchanged water to the top of the column (again through the heat exchanger) This caused the feed solution to develop in the column in the downward direction.   Step 4: Measure and observe the density and conductivity of the discharged solution with a recorder And collect the discharged liquid according to this information. Fraction; residue fraction number 1 (including salt), regenerated fraction Number 1 (including small amounts of salt and isomaltulose), isomaltulose fura Fraction No. 2 (a small amount of isomaltulose and trehal Lulose), residue fraction number 2 (eg glucose and full And solid infusion:   Amount of dry solids and isomalts in feed solution and product fractions Table 2 shows the content of loin, betaine and trehalulose. The concentration of each component is Expressed as a percentage of total dry solids in the fraction. In the product fraction The yields of isomaltulose, betaine and trehalulose are also shown (discharged Component amount of a particular component in relation to the total amount of a particular component in the whole fraction).   The following separations were performed in a similar manner. Then the regenerated fraction is also separated, And Trehal, regenerated fraction number 2, concentrated to some extent The loin is recovered as a separated fraction immediately after the isomaltulose fraction. Was. The trehalulose content of this fraction was about 30% based on DS.   Example 4 Chromatographic separation of isomaltulose and trehalulose   The regenerated fraction after Example 3 (isomaltulose and second regenerated fraction Trehalulose was concentrated to some extent. To further separate isomaltulose and also trehalulose, The fractions were combined, evaporated and separated again.   This separation was performed in the same manner using the same apparatus as in Example 3. Liner The flow speed was 0.5 m / hour and the feed size was about 160 kgDS. The separation conditions were the same as in Example 3. Isomaltz in a manner similar to that described above. Collect the loin fraction and then treat immediately after the isomaltulose fraction. The Rehalulose fraction was collected. Product components and feeds and products Table 3 shows the composition of the fractions. Example 5 Crystallization of isomaltulose   The isomaltulose fraction from the chromatographic separation of Example 3 was combined. Crystallized. The volume of the syrup is 600 l and the DS content is about 42% there were. A portion of the supplied syrup is placed in a 400 l boiling crystallizer. The mixture was concentrated to a DS content of 77.6 g / 100 g. The crystallization test described above 147 g of the pulverized dry crystalline isomaltulose was added to the above substance (about 150 l in volume). Was inoculated. Evaporative crystallization is continued for about 2 hours until the entire feed syrup is used. Was. The DS content of the material remained approximately the same during crystallization.   Prior to inoculation, the temperature and pressure were 62 and 65 ° C, respectively, and 190 and And 170 mbar. After inoculation, at 65 and 67 ° C respectively, Between 160 and 150 mbar.   After evaporative crystallization, the volume of the material was about 300 l. 400 l of the above substance In a crystallizing crystallizer. Up to 67.5-30 ° C in 41 hours A linear cooling program was run. After about 27 hours, the program was stopped and 41 It was set to a constant temperature of ° C. If water was mixed into the above material. 16 hours mixing Later the centrifugation was started.   The above substance was centrifuged using a Heine Zentrifuge (Heine Zentrifuge). This was performed at 00 rpm for 5 minutes. The crystals were washed with water.   Table 4 shows the results of the centrifugation.   Here, Q is the isomaltulose purity (g isomaltulose / 100 gDS). Represent.   The material was centrifuged and the crystals were dried (60 ° C.) in a rotary drum dryer. Dry The average crystal size was determined to be 200 μm (standard deviation 24%) by sieving analysis of the dried crystals. I found out.   Table 5 shows the analysis results of the samples. ¹Explanation of sample name:       Inoculation: Substance sample immediately before inoculation       Start of cooling ・ ・ ・ ・ ・ ・ ・ ・ ・ Material sample at the start of cooling       End: Substance sample at the start of centrifugation       Cake ... Crystal cake sample centrifuged       Effluent (Run-off) ・ ・ ・ ・ ・ ・ ・ Effluent sample from centrifugation       Dried cakes ... Dried cakes combined into one   ^TwoDS = Dried substance by Karl Fischer method^ThreeApply the ICUMSA method at ICUMSA color = pH5 and filter the solution ( (Filter membrane: 0.45 μm), the color analyzed by measuring at 420 nm.   ^FourPb carbohydrates⁺²Analyzed by HPLC using a type ion exchange column.       Oligo = oligosaccharide       Isom. = Isomaltulose       Gluc. = Glucose       Treh. = Trejarroth   Example 6 Hydrogenation of isomaltulose   From the isomaltulose fraction in the chromatographic separation of Example 3 Isomalulose is subjected to ion exchange and hydrogenation to give glucopyranosyl mannite. And glucopyranosyl sorbitol.   For ion exchange, the following resins are used in the order listed.   1. H⁺Type strongly acidic cation exchange resin (Dow 88)   2. OH^-Type weakly basic anion exchange resin (Dow 66)   3. Adsorption resin regenerated by NaOH (Dowex Optipore  Optipore))   In the ion exchange step, the amount of resin in each column was 0.217. Flow The speed was 1.0 l / h and the temperature of the solution was 40 ° C. All in one cycle An amount of 0.5 kg of dry solid was processed.   The ion-exchanged disaccharide is mixed with a batch type high pressure oil having a volume of 5 l equipped with a mixer. Hydrogenation in an autoclave (Medimex). Hydrogenation pressure is 40 bar And the mixing was kept at about 800 rpm. PH at the beginning with NaOH The pH was adjusted to a value of 6.0. Raney Nickel (Chemcat J10GS ) As hydrogenation catalyst, 10% wet calculated based on the weight of dry solids of the batch It was used at the catalyst loading.   Table 6 shows the analytical values before, after and after hydrogen exchange.RDS = dry solid by refractive index measurement GPM = α-D-glucopyranosyl- (1,1) -mannitol GPS = α-D-glucopyranosyl- (1,6) -sorbitol (Α-glucopyranosyl- (1,1) -sorbitol was not measured separately . )   Example 7 Hydrogenation of trehalulose   In a similar manner as described in Example 6, using Raney nickel The trehalulose fraction obtained in 4 is hydrogenated.   The hydrogenated product contains about 34% of 1,1 GPM based on DS and 1,6 GPS Approximately 19% is included on a DS basis, and about 1% GPS includes about 16% on a DS basis.   Example 8 Betaine crystallization   The betaine from the chromatographic separation of Example 3 crystallizes. DS standard A diluted betaine solution containing about 55% betaine until a concentration of about 76% by weight Allow to evaporate. The resulting solution is inoculated with pure betaine monohydrate crystals. Betaine The monohydrate is treated under a vacuum of 130 to 180 mbar for about 4 hours at 80 to 85 Crystallize at ° C. The crystals are separated from the mother liquor by centrifugation and dried.   Example 9 Recovery of isomaltulose and converted sugars   Sugarcane molasses instead of sugar beet molasses (Cultor Ltd. )) Is used to repeat the transglucosylation of Example 2. Sugar cane molasses Contains 50% sucrose based on DS and dilutes it to about 350 g DS / l . P. produced according to the first part of Example 2. Add rubrum fermented broth to molasses solution And run at about 30 ° C with aeration until most of the sucrose is converted. Continue the sglucosylation reaction. Defoamer (Mazu 100, PPG I Industries (PPG Industries Inc.), UK) To control excessive foam formation.   The transglucosylation product contains most of the isomaltulose, some trace Contains harulose and converted sugars (glucose and fructose).   In the same manner as in Example 3, the transglucosylation solution was purified by filtration. Perform on-exchange. The solution is then separated by chromatography, Supply individual fractions containing loin, trehalulose and invert sugar You.   Combine the isomaltulose and trehalulose fractions into one Hydrogenate as described in Example 6 to provide a sweetener mixture. Inverted sugar Evaporate such fractions and supply syrup.   The invention has been described herein by means of several specific examples. But these examples are simply It is merely illustrative in nature, and the invention departs from the scope of the appended claims. It will be apparent to those skilled in the art that it can be varied in many ways.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) C12P 19/12 C12P 19/12 // C12N 9/10 C12N 9/10 (81) Designated country EP (AT, BE, CH, CY, DE, DK, ES, FI, FR, GB, GR, IE, IT, LU, MC, NL, PT, SE), OA (BF, BJ, CF, CG, CI, CM, GA, GN, ML, MR, NE, SN, TD, TG), AP (GH, GM, KE, LS, MW, SD, SZ, UG, ZW), EA (AM, AZ, BY, KG, KZ) , MD, RU, TJ, TM), AL, AM, AT, AU, AZ, BA, BB, BG, BR, BY, CA, CH, CN, CU, CZ, DE, DK, EE, ES, FI, GB, GE, GH, GM, GW, HU, ID, IL, IS, JP, KE, KG, KP, KR, KZ, LC, LK, LR, LS, LT, LU, LV, MD , MG, MK, MN, MW, MX, NO, NZ, PL, PT, RO, RU, SD, SE, SG, SI, SK, SL, TJ, TM, TR, TT, UA, UG, US, U Z, VN, YU, ZW (72) Inventor Mirja, Lindroos Finland, FFIN-02400 Kirkonumi, Pojoispiha 8 (72) Inventor Paivi, Ojara Finland, FFIIN-02460 Kantvik, Harju 1 Sea (72) Inventor Villi , Lavanco Finland, F.I.N. 02150 Espoo, Jamerantivar 10 B. 26 (72) Inventor Matti, Chile Finland, F.I.N. Vic, squirrel Tikal Rion tea 19

Claims (1)

[Claims] 1. Obtain converted and unconverted saccharide or non-saccharide products simultaneously The method, comprising the following steps: a) sugar radish or saccharose containing sucrose and other saccharide or non-saccharide compounds Providing an aqueous solution derived from cane; b) transglucosylating the sucrose in the solution; c) The above transglucosidic method is carried out by a method including recovery by separation chromatography Isomaltulose and / or Trehalulose and Conversion Recovering unreacted saccharide and / or non-saccharide products; A method consisting of combining. 2. The method comprises, during steps a) and b), from the sugar beets or sugar cane Recovering sucrose from the derived solution, preferably by crystallization, and supplying molasses 3. The method of claim 1, comprising the additional step of: 3. The aqueous solution derived from the sugar beets or sugar cane is sugar cane or sand. The method according to claim 1, which is molasses molasses. 4. The aqueous solution is derived from sugar beets and in step c) isomaltulose and 4. The method of claim 1, 2 or 3, wherein betaine and betaine are recovered. 5. The aqueous solution is derived from sugarcane and, in step c), isomaltulose 4. The method according to claim 1, 2 or 3, wherein the converted saccharide is recovered. 6. In step c), the fraction containing trehalulose and isomaltulose is recovered. 6. A method according to claim 4 or claim 5, wherein the method is performed. 7. The isomaltulose is recovered by crystallization. The method described in one. 8. The method of claim 1 wherein trehalulose is recovered in step c). 9. The transglucosylation is caused by sucrose glucosylmutase enzyme activity The method of claim 1 provided. 10. Free or responsible for dead or living cells of microorganisms. 10. The method of claim 9, wherein said enzyme is immobilized on the body. 11. The microorganism is Protaminobacter rubrum (Protaminobacter rubrum) CBS 574.77). 12. The method according to claim 4, wherein the betaine is recovered by ion exchange. 13. The method according to claim 4, wherein the isomaltulose is recovered by crystallization. 14． Before the transglucosylation, betaine was recovered by ion exchange. The method according to claim 1. 15. The isomaltulose is hydrogenated and α-D-glucopyranosyl- (1, 1) -mannitol and α-D-glucopyranosyl- (1,6) -sorbitol 2. The method of claim 1 wherein the mixture is a mixture of 16. The isomaltulose and / or trehalulose are hydrogenated, 9. The method according to claim 8, wherein the hydrogenation product is recovered. 17． The trehalulose and isomaltulose fractions are hydrogenated, α-D-glucopyranosyl- (1,1) -mannitol, α-D-glucopyrano Sil- (1,6) -sorbitol and α-D-glucopyranosyl- (1,1) 7. The method of claim 6, which results in a mixture comprising sorbitol.