TW466244B - Crystalline 1-kestose and process for preparing the same - Google Patents

Abstract

A process for preparing crystalline 1-kestose which enables crystalline 1-kestose in large fine granulated sugar form to be prepared with high recovery. The process comprises concentrating a high-purity 1-kestose solution to a Brix of not less than 75, depositing crystallites serving as a mother nucleus by adding seed crystals or by concentration under reduced pressure, and repeating at least twice the step of growing crystals by concentration under reduced pressure and the step of redissolving crystallites produced in the concentrate. Alternatively, the process comprises concentrating a high-purity 1-kestose solution to a Brix of not less than 80, growing crystals after addition of seed crystals or induction of crystallization, and repeating at least twice the step of lowering the temperature by 5 to 20 DEG C below the temperature in the previous step and the step of growing crystals growth by allowing the concentrate to stand at the above temperature. An enzyme capable of efficiently converting sucrose to 1-kestose is also disclosed.

Description

466 24 4-V. Description of the invention (1) The present invention relates to a method for crystallizing 1-cane fructose. More specifically, large crystal grains can be obtained by vacuum crystallization or cooling crystallization at a high recovery rate. Crystal 1-cane fructose Crystal 1-Manufacturing method of cane fructose. In addition, the present invention also relates to an enzyme capable of efficiently producing 1_cane triose and a method for producing crystalline 1-cane triose using the same. 1-cane-fructotriose is a fructose at the 1-position of the sucrose molecule. The fructo-oligosaccharides are hard to be tooth-dented and cannot be digested by digestive enzymes in vivo. The growth-promoting effect that is specific to schizophyllum in the intestine has been disclosed by the present invention in Japanese Patent Publication No. 5-9 _ 5 3 8 3, but according to one of the components of this oligosaccharide measured 1_ sucrose triose has the same effect as this fruit oligosaccharide "printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs (please read the note on the back first) Please fill in this page again for item f)) In the past, the crystalline oligosaccharides sold in the market only had beet sugar. This is a small crystal diameter with a long diameter of 0.1 ~ 0., 3mm. At present, the most famous crystalline sugar is granulated sugar, granulated sugar with a large crystal size of about 0.25 to 1 mm as the main component, and sugar production of a mixture of large crystals and medium crystals of about 0.1 to 0.5 mm accounts for its large amount of sugar. Partial production. The related oligosaccharides also have a higher utilization value for these large particles, requiring large crystals. In particular, although 1 cane fructose has the same columnar crystals as granulated sugar, the granulated sugar has a nearly cubic shape, while the 1-caned fructoose has a nearly slender cuboid shape, which has physical properties close to granulated sugar, so it must be It has a larger crystalline form than sugar. In the past, the method for making crystal 1 _ cane fructose was particularly fair. The purity disclosed in the communique No. 6 — 7 0 0 7 5 is more than 70%. Brix 85 This paper size applies Chinese National Standard (CNS) A4 specification (2 丨0X297 mm), _ 4 — 466 24 4 A7 B7 Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs. 5. Description of the invention (2) above 1-Cane triose solution is raised to a temperature above 80 ° C, and then added The method of recovering the precipitated crystal 1-cane triose while crystallizing the suspension while reducing the temperature to 6 5 ~ 7 5 ° C, and then maintaining the temperature at 60 ~ 80 ° C. However, with the crystallization of this method, the concentration and purity of the crystallization solution will gradually decrease, so the recovery rate of crystallization is only about 30 ~ 40%. In addition, the precipitation of microcrystals cannot be avoided in this method, and the viscosity of the crystallizing solution will increase. Therefore, it is necessary to carry out heating while crystallizing, and there are inconveniences. In addition, the particle size of the obtained crystals is not uniform, and the crystal particle size is often small. "On the other hand, the known method of reacting fructose transferase with sucrose to produce 1-cane fructose is to use P. spp. Or a method for producing an enzyme having fructose transfer activity produced by a microorganism of the genus Fusarium, which is adjusted to a sucrose solution having a concentration of 50% or more at pH 4.0 to 7.0 (Japanese Patent Publication No. 63-62184), or Method for causing an enzyme having fructose transfer activity produced by microorganisms of the genus Pythium to act on a sucrose concentration of 20 to 70% at pH 4.0 to 7.0 'temperature 2 5 to 6 5 ° C (Japanese Patent Application Laid-Open No. 61-268190 Bulletin). However, these methods are all a mixture of trisaccharides (1-cane fructose), tetrasaccharides (cane fructose Nystose) and pentasaccharides (fructosyl cane tetraose) obtained from sucrose. For the main purpose, the percentage of conversion from sucrose to 1 cane fructose is only a maximum of 36 to 41% (refer to Table 1 3 in JP 63-62184 and JP 61-268190) example). In addition, when following these methods (please read the precautions on the back before filling in this page)

This paper size applies the Chinese National Standard (CNS) A4 Regulation (210X297 mm) 5 4 6 6 24 4 A7 B7 V. Description of the invention (3), will produce 11 ~ 2 3% cane together with 1_cane fructose Fructose, remaining 1 1 ~ 2 3% unreacted sucrose. In addition, monosaccharides such as fructose or glucose, which are produced as by-products in an amount of 1 to 3 to 2% (refer to the foregoing specification). Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs (please read the note on the back #Wh item to fill the page). Containing these monosaccharides, sucrose, and sucralose is extremely disadvantageous from the following viewpoints. The two-component quasi-mobile layer chromatography separation method is used for the general production of isomerized sugars, etc., and the mixed solution containing two or more sugars of different molecular weights is fractionated by the difference of molecular weights. When using this method to obtain a high-purity 1-cane fructose solution from a mixed solution containing 1-cane fructose, (1) fractionation must be used to separate sugar fractions with a lower molecular weight than 1-cane fructose. And 2 components containing 1-cane triose, with a molecular weight larger than 1-cane triose, or (2) fractionation into sugar fractions with a larger molecular weight than 1-cane triose, and 1_Cane triose, two components of a sugar fraction with a molecular weight smaller than 1 -cane triose, either method. For example, in the former method, a sugar having a molecular weight larger than that of 1-cane fructose will exist in the mixed solution, and the purity of 1-cane fructose after fractionation will inevitably be reduced depending on the amount of the sugar. Contains a large number of sugars with a larger molecular weight than 1 cane-fructotriose, such as Ni stose, and the two-component pseudo mobile 餍 chromatography separation method is used to obtain the high purity (eg 80% or more) 1- In the case of cane fructose solution, the fractionated liquid must be placed in the chromatographic separation device again, and the operation is repeated twice or more, which is extremely disadvantageous in practice. In addition, the latter method is also disadvantageous if the content of monosaccharides and sucrose is large. "For this reason, it is extremely necessary to use an enzyme reaction to obtain these monosaccharides, sucrose, and sucralose. Methods that can selectively produce 1-cane triose require reaction conditions or novel enzymes suitable for these methods. This paper size applies the Chinese National Standard (CNS) Λ4 specification (210X297 mm) 4 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 66 24 4 A7 ____B7_ V. Description of the invention ί 4) Also, there are microorganisms of the genus Qinghui The report indicates that an enzyme with fructose transfer activity exists in penicillium frequentans (Japanese Patent Laid-Open No. 4-29 29494). This report only describes that the above-mentioned microorganisms can produce fructose transferase, and there is no description about the conversion rate of 1-cane-triose. In addition, a method for selectively producing 1-cane fructose has been reported to produce 1-cane fructose in the presence of sucrose in the genus Puccinia, and cultivating glucose-capable microorganisms in a sucrose-containing medium to produce 1 One cane-fructose, method for harvesting 1-cane-fructose from the culture (Japanese Patent Publication No. 5-471 97, Japanese Patent Publication No. 4-41 600). According to this gazette, the conversion rate from sucrose to 1-canetriose can reach 60%. However, the method described in this publication directly uses bacterial cells, so the total sugar concentration in the culture solution, that is, the sucrose concentration at the beginning of the culture, must be reduced to about 15%. In addition, a large amount of impurities such as protein must be removed during purification. Therefore, it is extremely unfavorable. Therefore, I still hope to develop a method that can more selectively produce 1-cane triose, especially to develop new enzymes or new reaction conditions. The present inventors have found that based on the so-called vacuum crystallization or cooling crystallization today, through controlled concentration operation and temperature adjustment, when the viscosity of the separation liquid decreases and the solid-liquid separation crystallizes, it can be used without heating. Separated at room temperature. In addition, it was found that crystalline 1-canetriose can be obtained with a high recovery rate, and large crystal grains can be obtained. In addition, the inventors have discovered that a certain microorganism can produce an enzyme that efficiently produces 1-cane triose. The present invention is based on these findings. The paper size is applicable to the Chinese National Standard (CNS) A4 Specifications (210X297 mm) (Please read the notes on the back before filling out this page) 7 * 66 24 4 B; 5. Description of the invention 丨 5). Therefore, the present invention is to provide high recovery and solid-liquid separation. The purpose is to produce a crystallized 1-cane triose at room temperature. It is also an object of the present invention to provide a method for producing a crystalline 1-sucratotriose which can obtain large crystalline particles, preferably granulated sugar. In addition, the present invention aims to provide sugarcane triose in the form of granulated granules with large crystal grains. Here, the method for producing crystalline 1-canetriose according to the first embodiment of the present invention is performed by the following steps (a '), (b), and (C). (a) Concentrated 1 a high-purity 1-cane fructose solution with a purity of 80% or more, so that the Brix of the solution is 7 5 or more, after adding seeds, the temperature of the concentrated solution is 6 Steps to grow crystals above 0 ° C; and (b) Steps to grow crystals by concentrating the concentrated solution under reduced pressure to grow crystals; Printed with the Shellfish Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs (please read the notes on the back first) (Please fill in this page again for details) (c) After the above crystal growth step, warm up the temperature of the liquid to re-dissolve the microcrystals generated in the above concentrated solution. If all the crystals cannot be dissolved, add water to dissolve the microcrystals. Step; and then repeating the above-mentioned (b) crystal growth step and the (c) microcrystal dissolution step at least once more, and then performing the step of recovering crystalline 1-sucralose. In addition, as a correction method of the method of the first embodiment, instead of the above steps (a) and (b), (a >) concentration 1 _ high purity of sucralose is 80% or more This paper size applies to Chinese National Standard (CNS) A4 (210X297 mm) 8 4 6 6 24 4 A7 B7 Duplicate work and consumption cooperation of the Central Standards Bureau of the Ministry of Economic Affairs. 5. Description of the invention (6) Degree 1-Cane fruit The sugar solution makes the Brix of the solution above 75, and then. The step of warming the concentrated liquid to a temperature above 60 ° C; and then (b /) reducing the concentration of the concentrated liquid under reduced pressure to reduce the temperature of the liquid to start The step of crystallizing and growing the crystal: The above-mentioned (c) microcrystal dissolving step is then performed, and then the above (b) crystal growing step and the (c) microcrystal dissolving step are repeated at least once more, and then the crystal 1 is recovered. Steps of a cane-fructotriose》 Also, according to the second embodiment of the present invention, Crystal 1-A method for producing cane-fructoose is (α) concentrated 1 _ Cane-fructosan has a purity of 80% or more 1 One cane triose high-purity solution, so that the Brix of the solution is more than 80, and then add Warm the concentrated solution at 70 ~ 95 ° C, add seed crystals, and make the concentrated solution grow and crystallize in the above temperature range; and (cold), then reduce the temperature of the concentrated solution to a temperature lower than the temperature of the above step 5 ~ 20 ° C cooling step, then (r) place the concentrated liquid at the temperature lowered by the above cooling step to crystal grow; and then repeat the above (/ 5) cooling step and at least once more In the above (r) crystal growth step, a step of recovering crystal 1-cane fructose is performed after the temperature is lowered to 20-60 ° C. In addition, another correction method of the second embodiment may be to perform (α —) concentration instead of the above step (α) to obtain a 1-sugar triose high-purity solution having a purity of 80% or more, so that Brix of this solution is (please read the precautions on the back before filling this page)

This paper size is applicable to China National Standard (CNS) A4 specification (210X297 mm) 1-9-4 24 4 Economy, Qiu Central Standard Bureau employee consumer cooperative printed A7 B7, V. Invention description (7) 80 or more, and then Set the concentrate at a temperature ranging from 50 to 60 ° C. Start crystallization, and after the step of growing the crystal at 70 to 95 ° C, repeat the above (j8) cooling step and the above one more time. (R) Crystal growth step, the method of recovering the crystal 1_cane fructose after reducing the temperature to 20 ~ 60 ° T, and the long diameter of the crystal 1_cane fructose of the present invention is 0.3 mm or more (Preferably 0.3 to 2 mm), crystal 1-cane fructose of columnar crystals having a purity of 95% or more. In addition, according to a third embodiment of the present invention, a method for producing crystalline 1-cane fructose is provided, which includes (i) (1) an enzyme having fructose transfer activity from a microorganism derived from the genus Corydalis, which acts on sucrose Solution, the step of converting 43% by weight or more of sugar produced into 1-cane fructose, (2) allowing an enzyme having a fructose transfer activity from a microorganism belonging to the genus Penicillium to act on the sucrose solution, converting more than 46% by weight to produce The step of sugar being 1-cane fructose, or (3) enabling an enzyme from a microorganism belonging to the genus Lichenella having fructose transfer activity to act on a sucrose solution to convert 53 to 3% by weight of sugar to 1-cane fructose Any step of the sugar step, and (ii) the step of fractionating (i) obtained by chromatographic separation method (1) cane fructose is a step with a purity of 80% or more, and then (iii) the crystallization step (ii_) The obtained 1-cane triose to obtain a columnar crystal with a diameter of 0.3 mm or more and a purity of more than 9 5%. The step of 1-cane fructose, the paper size applies the Chinese National Standard (CNS) A4 specification (210X 297mm) (Please read the note on the back first Please fill out this page again

-10-4 Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 6 6 24 4 A7 _B7_ V. Method of Invention (8). The method of the first and second embodiments of the present invention. Crystallized stock solution The methods of the first and second embodiments of the present invention all use the crystallized stock solution containing high-purity 1-cane triose as a starting material. 1 The purity of cane-fructose is as high as possible, but in the method of the present invention, it is preferably 80% or more, and more preferably 90% or more. The purity of 1-cane-triose refers to the weight ratio of 1-cane-triose in the sugar component of the crystallized stock solution. The 1-cane-triose crystallized stock solution is prepared or contains 1- Sugar source of cane fructose. Refining this sugar source can improve the purity of 1 -cane fructose. Modulation of the sugar source containing 1 -cane fructose can use, for example, plants, microorganisms and other fructose transferases to act on sucrose. The substrate can be obtained, and the microorganism can be obtained by culturing the microorganism on a sucrose-containing medium. In addition, various fructo-oligosaccharide mixtures are commercially available, and these mixtures can be purified by conventional methods such as chromatographic separation, membrane separation, or the Stephen method using lime to precipitate impurities. According to the comparative embodiment 1 of the present invention, it is preferred that the crystallized raw solution of cane-fructose is one that contains less cane-fructose among other sugar components. According to the experience of the present inventors, in order to obtain a larger 1-sucrafructose crystal, it is advantageous to minimize the sucrose-tetrasaccharide in the crystallization stock solution as much as possible. More specifically, if it is known that sucralose has hindered crystallization of 1-sucralose, the amount of sucralose is preferably 10% by weight or less of 1-sucralose. (Please read the notes on the back before filling out this page) Equipment · I: Order

The size of this paper is in accordance with the Chinese National Standard (CNS) A4 specification (210X297 mm) -11-4 66 24 4 Printed by the Buyers and Consumers Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs A7 B7 V. Description of the invention (9) In the methods of the first and second embodiments of the present invention, the conventionally known method for producing cane-fructose also has the above phenomenon. Method of the First Embodiment The method of the first embodiment of the present invention is to first concentrate the above-mentioned crystallization solution to B.rix 75 or more, preferably 80 to 85. It can be concentrated by heating or depressurizing. Thereafter, a seed crystal was added to the concentrate. In this series, it is advisable to add 1% by weight or less of the solid content of the concentrated liquid to the seed crystals. 9 After adding the seed crystals, use this as the mother nucleus to grow the crystal. The concentrated liquid system is best maintained at a temperature above 60 eC of about 0.5 ~ The temperature is preferably about 6 hours, and more preferably maintained at 70 ~ 95 ° C for 1 ~ 3 hours. According to another preferred embodiment of the present invention, the mother liquor can be formed without adding the above-mentioned seed crystals. Specifically, the concentrated solution is concentrated under reduced pressure to a temperature above 60 ° C, and the microcrystals are precipitated. The mother nucleus grows crystals under the above conditions. The formation of the microcrystals is preferably performed at an absolute pressure of 10 to 160 mmHg and a temperature of 30 to 70 ° C, and more preferably at an absolute pressure of 40 to 12 OmmHg and a temperature of 50 to 60 ° C. Then, the concentrated solution which was crystal-grown as described above was concentrated under reduced pressure. The reduced-pressure concentration system is preferably performed for 10 to 60 minutes at an absolute pressure of 40 to 20 OmmHg, and more preferably for 30 to 60 minutes at an absolute pressure of 60 to 1 2 OmmHg. The temperature of the liquid is reduced to 40 ~ 70 ° C to promote crystal growth (crystallization growth steps) "(Please read the precautions on the back before filling this page) — ^ 丨 Γ I.: ― This paper size applies Chinese National Standard (CNS) A4 specification (210X297 mm) 12-Economy. Which is printed by the Consumer Standards Cooperative of the Central Bureau of Standards 4 66 24 4 A7 B7 V. Description of the invention (10 ) This crystal growth step is followed by applying a second microcrystalline dissolution step to the concentrated solution. Although the crystal growth of 1-sucratotriose can be seen through the above-mentioned reduced-pressure concentration operation, micro-crystallization of 1-sucratotriose is usually generated due to the concentration operation. The concentrated solution is preferably heated at 70 ° C to 95 ° C, more preferably at 75 ° C to 85 ° C for 0.5 to 6 hours, preferably 1 to 2 hours, and the microcrystals are dissolved again while maintaining the temperature. If it is impossible to completely re-dissolve the microcrystals at this temperature, water may be added, and it is preferable to re-dissolve all the microcrystals. In the method of this first embodiment, after the microcrystal dissolution step, the same crystal growth step and the microcrystal dissolution step as described above are repeated at least once more. That is, it is preferable to place the concentrated solution at an absolute pressure of 40 to 2 OmmHg • for 10 to 60 minutes to allow the crystals to grow, and it is preferable to warm the concentrated solution at 70 ° C to 9 5eC and maintain 0.5 to 6 The formed microcrystals are re-dissolved at this temperature for one hour. If it is impossible to dissolve all the microcrystals by heating, it is best to add water to the microcrystals to re-dissolve them. By the above steps, crystallized 1-cane triose can be sufficiently precipitated to have a desired size. The resulting crystals can be recovered individually by conventional methods (e.g., centrifugation). According to the preferred embodiment of the present invention, it contains sufficient growth and becomes a low-viscosity liquid with a low viscosity after the crystallization of granulated sugar 1-sugar triose has been crystallized, so the crystals can be recovered without heating. For its best advantage. After recovery, the crystals can be dried to obtain crystal 1-cane fructose. According to the method of the first embodiment, i-cane fructose can be obtained with a high yield of 50 to 60%. In addition, according to a preferred embodiment of the present invention, it can be obtained with a long diameter of 0.3 mm or more (preferably 0.3 to 2 mm, and more preferably 0 5). This paper uses the Chinese National Standard (CNS) A4. Specifications (210X297 mm) 1._l · ——Γ ^ I — (Please read the notes on the back before filling in this page) Order _ -13-4 66 24 4 A7 B7 V. Description of the invention (11) ~ 1. 2mm), the knot of columnar crystals with a purity of more than 9 5% (please read the precautions on the back before filling this page) Crystal 1 _ Cane Triose. Specifically, in the above crystal growth step, controlling the Brix of the concentrated liquid to be about 75 to 85, preferably about 79 to 83, can control the generation of microcrystals, promote crystal growth, and efficiently obtain larger crystals as described above. 1 -Sucrose triose》 Also, in this embodiment, the volume reduced by the crystallization step may be performed after the crystal growth step but before the microcrystal dissolution step or before the crystal growth step after the microcrystal dissolution step. Replenishing the crystallized stock solution • This supplement can increase the processing capacity per military operation, and at the same time can suppress the decrease in the purity of cane fructose caused by the precipitation of crystals, which can be maintained at a high level of 1-cane fruit three Crystallization was performed at sugar purity. Printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs. In addition, for the sugar solution containing 1 1 cane fructose after recovering and crystallizing 1_cane fructose, the same operation can be performed again or by the second implementation described later. The method of morphology, successively made crystal 1-cane fructose. In addition, after the crystallized 1-cane triose is recovered, the 1_cane triose-containing sugar solution can be re-added, and then the crystallization stock solution can be re-added, and then the same operation as described above or the method of the second embodiment described later can be performed again. . By continuously performing the crystallization operation as described above, the recovery rate of 1-cane triose crystals can be further improved, and sometimes a high recovery rate of 80% or more can be achieved. Method according to the second application form The method according to the second embodiment of the present invention is to first concentrate the above crystallization solution to Brix 80 or more, preferably 83 to 88. It can be concentrated by heating or depressurizing. After that, the concentrated solution was added at 70 ~ 9 5 ° C, and seed crystals were added to the thick paper. The Chinese national standard (CNS) A4 specification (2〗 0X297 mm) was applied. -14-4 66 24 4 A7 B7 V. Description of the invention (12) Shrinkage. In this case, a seed crystal of 1% by weight or less is preferably added to the solid content of the concentrated solution.-According to another preferred embodiment of the present invention, the mother core can be formed without adding the seed crystal described above. The inventors have found that it is very interesting to find that the concentrated solution that has been heated and concentrated to Brix 80 or above can be efficiently obtained at room temperature (about 25 ° C) to 50 ~ 60 ° C. It is the micro-crystal of the mother nucleus, and the heating rate can be controlled to control its quantity and size. Specifically, the heating rate from room temperature to the temperature in the above range can be accelerated, that is, the number of microcrystals used as the mother core can be reduced to obtain larger crystals. On the other hand, the heating rate can be slowed down and increased as The number of microcrystals in the mother nucleus formed into smaller crystals. After adding seed crystals or forming microcrystals as the mother core as described above, the heat-retaining concentrated liquid is kept at 70 ~ 95 ° C, preferably 75 ~ 85X, and the crystals are grown in 30 minutes ~ 6 hours *. Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs (please read the precautions on the back before filling this page) Secondly, this method is used as a cooling step to make the temperature of the concentrated liquid 5 ~ 20 ° C lower than the above step, preferably Cool at about 10 ° C. Thereafter, the concentrated solution is maintained at the temperature for 30 minutes to 6 hours, and a crystal growth step is performed for crystal growth * In this method, the cooling step and the crystal growth step are repeated at least once after the cooling step and the crystal growth step. That is, the temperature of the concentrated solution is lowered by 5 to 20 ° C, preferably about 10 ° C, from the previous stage, and then the concentrated solution is kept at the temperature for 30 minutes to 6 hours to grow the crystals. If a large amount of microcrystals are precipitated during this crystallization, the temperature will be raised to make it smaller than the micro paper size. It will be applicable to Chinese national standards (CNS > A4 specification (210X297)). 4 6 6; 24 4 A7 B7 V. Description of the invention (l3). The liquid of crystal precipitation is about 10 ~ 20 ° C high, and then dissolve the microcrystals, and then repeat the cooling step and crystal growth step. The cooling step and the crystal growth step are alternately and regularly combined, and are implemented until the temperature of the final crystallization solution can be 20 ~ 60 ° C, preferably about 40 ~ 50 ° C. With the above The operation can fully crystallize 1-cane triose as the target. The obtained crystal can be recovered separately by ordinary methods (such as centrifugation). According to a preferred embodiment of the present invention, it is the same as the first embodiment described above. The form contains sufficient growth, and it becomes granulated crystal 1-sucrose triose, after the crystallization of the concentrated liquid is low, so it can be recovered without heating, which is also the most advantageous. Dry it after recycling Crystallization can obtain crystal 1-sucrose triose. According to the method of the second embodiment, a recovery rate of more than 40% can be obtained. 1 In a preferred embodiment, sucralose can obtain 4 3 to 5 5%. High recovery rate. According to a preferred embodiment of the present invention, the long diameter is 0.3 mm or more (preferably 0.3 to 2 mm, and more preferably 0.5 to 1.2 mm), and the purity is 9 5% or more of the crystals of the columnar crystal 1 sucralose. Also, as in the method of the first embodiment described above, the 1_sucralose-containing The sugar solution can perform the same operation again or use the method of the first embodiment described below to continuously produce crystalline 1-cane triose. In addition, the sugar solution can be recovered after the crystalline 1-cane triose is recovered. 1-Cane fructose is added again to the sugar solution. The paper size applies the Chinese National Standard (CNS) A4 specification (210X297 mm) —Ι1--Ϊ- ^^ 1 In i ^ i ^^ 1 H. I ( (Please read the notes on the back before filling out this page)

P -16-4 6 6 24 4 A7 B7 printed by Zhengong Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of the invention 丨 14) After crystallization of the dope, perform the same operation as above or the first implementation described later. Method of form. By continuously performing the crystallization operation as described above, the recovery rate of 1-cane triose crystals can be further improved. Method of the third application form of the present invention According to the third embodiment of the present invention, a method for producing 1-cane triose from sucrose using an enzyme derived from a certain microorganism can be provided. In addition, one unit of the enzyme having fructose transfer activity according to the third embodiment of the present invention is: When it comes to the origin of the microorganism of the genus Pleurotus, it means at 40 ° C 'pH 5.0, the matrix (sucrose ) At a concentration of 10% by weight, an enzyme that can produce 1 millimolar 1-cane triose within one minute; when it is an enzyme derived from a microorganism belonging to the genus Cymbidium, it means at 50 ° C 'pH 7.0. The substrate ( Sucrose) concentration at 10 reset% can produce 1 millimolar 1-cane triose enzyme in one minute; the enzyme derived from microorganisms belonging to the genus Gliocladium refers to 40 ° C, pH 7. 〇, at a concentration of 10% by weight of the substrate (sucrose), 1 millimolar 1-canetotriose enzyme can be produced in one minute; step (i): 1-canetotriose is produced from sucrose-the first aspect of the present invention In the three embodiments, the sucrose is first converted into 1-cane-triose, and the steps are implemented by any one of (1), (2) or (3). (i) (1) Fructose transfer is caused by microorganisms belonging to the genus Pycnomyces m. ^^ 1 In m (please read the precautions on the back before filling out this page) 1 ”Order — The paper size of the paper is applicable to the country National Standard (CNS) A4 specification (210X297 gong) -17-466244 V. Description of the invention (15) The active enzyme acts on the sucrose solution to make 4 3% of the sugar produced by the child. Converted to 1-cane fruit three The step of sugar, (2) a step of causing an enzyme having fructose transfer activity from a microorganism belonging to penicillium spp. To act on a sucrose solution, and converting 46% by weight or more of the sugar produced into 1-sucralose, (3) Step (i) (1) the step of causing an enzyme having fructose transfer activity derived from a microorganism belonging to the genus Brassica to act on a sucrose solution to convert 53 to 3% by weight of the sugar produced into 1-sucralose. (I) The enzyme used in (1) can be obtained from microorganisms of the genus Pleurotus and has fructose transfer activity. The enzyme is preferably from Aspergillus niger, more specifically from the enzyme ATCC 2 0 6 1 1. The above enzymes can be used in appropriate media (for example, containing 3.0 ~ 10. 0% sucrose, 3.0 ~ 10 · 0% yeast extract, printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs (please read the precautions on the back first) Fill in this page again) 0.3 ~ 1.0% CMC medium), culture the microorganisms at an initial pH of 6.0 to 7.0 and a temperature of 25 to 30% for 48 to 96 hours, and recover from the culture solution. The characteristic of this enzyme is that it can act on a sucrose solution with a concentration of 50% by weight or more under the conditions of PH5 · 0 ~ 10.0, temperature 50 ~ 65 ° C in an amount of 0.5 to 100 units per 1g of sucrose, More than 43% by weight of the generated sugar is converted into 1-canetriose. It also acts on sucrose solution under the same conditions, and converts less than 13% by weight of the sugar produced. The paper size applies the Chinese standard (CNS > A4 size (210 X 297 mm)), 0-18-4 66 24 4 Α7 Β7 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs. 5. Description of Invention (16). Those who produce cane and fructose. The above-mentioned Special Publication No. 6 3 — 6 2 1 84 and Special Publication No. 6 1 — 2 6 8 In the method described in 190, 'the conversion rate from sucrose to 1-cane fructotriose is only limited to 36 to 41%. In addition, when using these methods, the At the same time, 1 1 ~ 2 3% of sucrose tetraose is produced. Therefore, the method of the present invention is extremely suitable for the selective production of 1-sucrose triose compared with the prior art described in these publications. The above enzyme acts on sucrose to produce 1_cane triose. According to a preferred embodiment of the present invention, the enzyme is at pH 5.0 ~ 10. 0, preferably at 5.5 ~ 10. 0, and the temperature is 50 ~ 65 °. C, preferably 50 to 60 ° (: reducing the concentration of 50% by weight or more, preferably 50% to 60% by weight of sucrose solution. Also, according to the present invention In a preferred embodiment, an enzyme having fructose transfer activity is added to the sucrose solution at 0.5 to 100 units per lg of sucrose, preferably at 3 to 100 troops, at 50 to 6 at 5 ° C, preferably at 5 0 ~ 6 0 ° C reaction for 2 ~ 100 hours, preferably 2 ~ 40 hours. It is best to deactivate the enzyme after the transfer reaction. 0.1 to 1.0% by weight of activated carbon is added to each solid component. (For example, Taige Activated Carbon S-W 50), it is suitable to heat the enzyme reaction solution at 90 ~ 95 ° C for 20 minutes to 60 minutes to decolorize and deactivate it. According to a preferred embodiment of the present invention, sugar is generated. Composition 1-Cane fructotriose is 43% by weight or more, and cane fructose is 13% by weight or less. When performed under preferred conditions, 'may be 4 to 4% by weight or more. And cane-fructose below 7% by weight. --- C ------- ί.ν / —v-(Please read the precautions on the back before filling this page) IIU. Thread_

0T This paper size applies Chinese National Standard (CNS) A4 specification (210X297 mm) -19-46624 ^ A7 __B7 V. Description of the invention (π) Step (i) (2) The above steps (i) (2) Enzymes are derived from Penicillium and have the ability to act on sucrose and convert more than 46% by weight of sugar to 1-sucralose. The enzyme having a fructose transfer activity suitable for use in the present invention may be a novel enzyme described below. This enzyme can be obtained from a microorganism of the genus Penicillium, preferably from Penicillium Loudi, more specifically, from Penicillium loui (Penici 1 1 i u m r 〇 q u e f rt i)-IAM7254 strain. The above enzyme system uses an appropriate medium (for example, containing 5 ~ 30% sucrose, 1 ~ 10% corn pulp, 0.05 ~ 0 3% urea, 0.2 ~ 3.0% potassium trihydrogen phosphate, 0.01 ~ 0.1% magnesium sulfate. Heptahydrate salt medium), cultured at an initial pH value of 6.5 ~ 7.5, a temperature of 25 ~ 30 ° C for 2 ~ 5 days, and recovered from the culture solution. Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs (please read the precautions on the back before filling this page) Enzymes are best refined by known refining methods. The preferred purification methods include salting out by ammonium sulfate, precipitation by organic solvents (such as methanol, ethanol, propanol, etc.), adsorption by raw starch, ultrafiltration, gel filtration chromatography, and ions. Exchange chromatography, and other various chromatography. Reviewing the properties of this novel enzyme, the results are shown below. <Function> It can cut sucrose 1 _ sucrose triose, beet sugar and other 4 _ D _ fructofuranosyl bonds with Θ-D — fructofuranoside bonds, and specifically transfer the generated fructosyl groups to saccharides The terminal fructosyl group is at the 0: _1 (hydroxyl) position. This paper size applies the Chinese National Standard (CNS) A4 specification (210 × 297 mm) _ 20 &quot; 466244 A7 _______B7 _____ 5. Description of the invention (18) In addition, 'This enzyme can be 0.5 to 1 gram per 1 g of sucrose. Unit quantity, at PH6. 0 ~ 9. 0, temperature 35 ~ 55 ° C, 'react with sucrose solution with a concentration of 50% by weight or more for 2 to 100 hours' can convert 4 to 6% by weight of sugar to 1 — cane fruit triose. In addition, under the same conditions, the sucrose solution was converted to less than 7% by weight of saccharides to sucralose. Therefore, it can be seen that this enzyme is very suitable for the selective production of 1-cane triose. <Matrix Specificity> It can interact with sucrose, 1-cane fructose, beet sugar, etc., but not with sucrose, maltose, etc. effect. <Optimum temperature> Optimum temperature is 40 ~ 50 ° C «<Temperature stability> Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs (please read the precautions on the back before filling this page) at PH7. 0, 30 Under the condition of minutes, relative activity above 60% can be maintained below 55Ϊ. <Optimal pp value> The optimum pH value is 6.0 to 7_0. 〈Stability p Η value〉 Extremely stable in the pH range of 4.0 ~ 8. 0, 40 ° C, this paper size applies Chinese National Standard (CNS) A4 specification (210 X 297 mm) n, -Δι-4 6 6 24 4-A7 B7 printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of the Invention (19) Relative activity of more than 90% can be maintained under the condition of 30 minutes. <Molecular weight> The molecular weight obtained by the gel filtration method is 3 15.0. <Inhibition of Activity> This enzyme is inhibited by glucose, which is a by-product produced during the fructose transfer reaction. <Enzyme kinetics> The km (rice soil constant) of this enzyme is 1.1M. In step (i) (2) of the present invention, the above-mentioned enzyme is acted on sucrose to generate 1-canetriose. According to a preferred embodiment of the present invention, it is at pH 6 · 0 to 9 0, preferably 6 · 0 to 8. 0, a temperature of 35 to 5 5 ° C, and preferably a concentration of 5 at 4 to 5 0 ° C. 0% by weight or more, preferably 55% to 65% by weight of sucrose solution. In addition, according to a preferred embodiment of the present invention, the sucrose solution is added to 0.5 to 100 units per gram of sucrose, Preferably, 2 to 50 units of an enzyme having fructose transfer activity is reacted at a temperature of 35 to 55 ° C, preferably 40 to 50 ° C, for 2 to 100 hours, preferably 4 to 100 hours. It is best to deactivate the enzyme after the transfer reaction. Add 0.1 to 1.0% by weight of activated carbon (such as Taiko activated carbon S — W 50) to each solid component, and heat at 40 to 70 ° C for 20 minutes to 60 It ’s better to decolorize the enzyme reaction solution in minutes, and to deactivate it ”(please read the precautions on the back before filling in the form page). Binding. The paper size is applicable to Chinese national standards (CNS &gt; A4 specifications (210X297) (Centi) -22-4 printed by the Central Bureau of Standards of the Ministry of Economic Affairs and Consumer Cooperatives 6 6 24 4 A7 ___B7 V. Description of the invention (20) According to a preferred embodiment of the present invention, the sugar composition system 1-cane fruit is produced. The sugar is more than 46% by weight, and the sucrose tetraose is not more than 7% by weight. "Step (i) (3) The enzyme used in the above step (i) (3) is from the genus Tetraomyces, which can act on sucrose Those capable of converting 53% by weight or more of sugar to 1-cane fructose. The enzyme having fructose transfer activity that is more suitable for use in the present invention may be a novel enzyme as described below. This enzyme may be obtained from the genus Euphorbia The microorganism is preferably a short-handled broom emblem, more specifically, the short-handled emblem opsis brevicairlis) was obtained from IF04834 strain. The above enzyme system was prepared using an appropriate culture medium (for example, containing 3.0 to 10.0% sucrose, 5 to 15% corn slurry, 0.05 to 0.3% urea, and 0.2 to 3 0% potassium trihydrogen phosphate, 0.01 ~ 0.1% medium of magnesium sulfate and heptahydrate), cultured at an initial pH of 6.5 ~ 7.5, and a temperature of 3 ~ 8 at a temperature of 20 ~ 30 ° C It is recovered from the culture liquid. It is preferable to recover the bacterial cells by using a centrifugal separator such as a basket type centrifuge, and then suspend the dried bacterial cells in a buffer solution of pH 7.0, and then apply ultrasonic treatment and membrane treatment to obtain a crude product. The enzyme solution can be obtained by subjecting the crude enzyme solution to anion, exchange, gel filtration, chromatofocusing, etc. After reviewing the properties of this novel enzyme, the results are shown below. Paper size is applicable to China National Standard (CNS) A4 (210X297 mm) (Please read the notes on the back before filling out this page) -23-4 Printed by the Staff Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 66 24 4 A7 ___ + —__ B7_ V. Description of the invention '(21) Can cut off sucrose 1 sucrose triose, beet sugar Having JS - D - -D-fructopyranose for stone bond of sugars - -fructofuranosidic linkage, the resulting anisotropically transfer of fructose Kit fructosyl saccharide at the end of the C-1 (hydroxy) bit on.

In addition, this enzyme can be at pH 6.0 ~ 10.0, preferably 7. 0 ~ 9_8, temperature 35 ~ 50eC, preferably 35 ~ 40 ° C, and a concentration of 50% by weight or more, preferably 50 ~ 5. The 5% sucrose solution is reacted for 2 to 100 hours, and then 53% by weight or more of sugar can be converted into sucralose. In addition, the enzyme is at a pH value of 6.0 to 10 0, more than 7.0 to 9.8, a temperature of 35 to 50 ° C, preferably 35 to 40 ° C and a concentration of 50% by weight or more. 5-50% by weight of the sucrose solution can be converted to more than 53% by weight of sugar to become 1-cane fructose. More preferably, sucrose per ig is 0.5 to 1 0. 0 units, preferably in an amount of 2 to 20 units, causes the enzyme to act. In addition, the enzyme of the present invention acts on a sucrose solution under the above-mentioned conditions, and converts less than 5% by weight of sugar to sucrose tetraose. Therefore, it can be seen that this enzyme system is extremely suitable for selectively producing 1-canetriose. <Matrix specificity> It can interact with sucrose, 1-canetriose, beetose, etc., but not with sucrose, maltose, etc. 〈Optimum temperature〉 This paper size is in accordance with Chinese National Standard (CNS) A4 (210X297mm) (Please read the precautions on the back before filling in this page) -24-466244 A7 B7 V. Description of the invention (22) Optimum temperature 40 ° C. <Optimal PP value> The optimum pH value was 7.0. 〈Stability P Η value〉 It is extremely stable in the pH range of 60 ~ 10 · 〇, and can maintain a relative activity of more than 80% under the conditions of 40 ° C and 30 minutes. <Molecular weight> The molecular weight obtained by the gel filtration method is 3 6 0 to 3 8 0 kD a ο. When analyzed by SD S PAGE, it is confirmed that the molecular weight is about 5 4 / c D a. The molecular weight after sugar bond removal is Approx. 5 1 «D a subunit structure. <Isoelectric Point> Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs (please read the note f on the back before filling out this page). The secondary electrophoresis and hydrophobic chromatography were found to be approximately 3. 8 ~ 3.9. <Inhibition of Activity> This enzyme is inhibited by glucose, which is a by-product produced during the fructose transfer reaction. 〈Enzyme reaction rate theory〉 The paper size is applicable to the Chinese National Standard (CNS) Λ4 specification (210X297 mm) -25-4 6 6 2 4 4 A7 B7 V. Description of the invention (23) The km of this enzyme is 0.75M . In addition, it will be subject to the non-competitive inhibition of glucose, and its inhibition constant (Ki) is 0 _ 125M »(Please read the precautions on the back before filling this page) Step (i) (3) in the present invention is the above Enzymes act on sucrose to produce 1 cane fructose. According to a preferred embodiment of the present invention, it is at ρΗ 6. 0 ~ 10. 0, preferably at 7. 0 ~ 9.8, temperature 3 5 ~ 50 0 ° C, preferably 3 5 ~ 4 0 ° C. The effect of sucrose solution at a concentration of 50% by weight or more, preferably 50% to 55% by weight. Also, according to a preferred embodiment of the present invention, the sucrose solution is added in an amount of 0.5 to 100 for each 1g of sucrose. Units, preferably 2 to 20 units of enzymes with fructose transfer activity, for 2 to 100 hours, preferably 9 to 60 hours, at a temperature of 3 5 to 5 0 ° C, preferably 3 5 to 4 01 * Transfer reaction After that, it is best to deactivate the enzyme. Add 0.1 · 1.0% by weight of activated carbon (such as Taiko activated carbon s_w 50) to each solid component, and heat at 90 ~ 95 ° C for 20 to 60 minutes to make It is desirable that the enzyme reaction solution is decolorized and deactivated at the same time. Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs According to a preferred embodiment of the present invention, the sugar composition is 1-cane fructose is 53% by weight or more and cane-fructose is 5% by weight or less. When carried out under preferred conditions, it may be 1 to 5 wt% of cane fructose, and 4 to 4 wt% of cane fructose. According to a preferred embodiment of the present invention, it is best to obtain a solution containing 1 cane-fructose from any one of the steps (i) (1) to (3) before performing the next step (i). It is advisable to remove bacteria and the like by filtration, and then perform decoloring and deodorizing operations of activated carbon. The decolorization and deodorization of activated carbon are based on the addition of 0.1 to 1.0% by weight of solid components containing 1 cane fructose. The paper size is in accordance with China National Standard (CNS) A4 (210X297 mm) -26 -4 66 24 4 Α7 Β7 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of the invention (24). Activated carbon is stirred at 40 ~ 70 ° C for 20 ~ 60 minutes. In addition, desalting treatment is performed by cation and anion exchange resin, and filtering again to make it colorless and transparent 1-cane fructose-containing liquid β step (ϋ): Chromatographic separation In order to obtain crystalline 1-canetotriose from the enzyme reaction solution of cane triose, a purification operation is performed by chromatography. The chromatographic separation method is best to use pseudo-mobile layer chromatography usually used in the production of isomerized sugars. By this chromatographic separation operation, 1 cane triose has a purity of more than 80%. It is preferably a chromatographic fractionation solution with a content of more than 90%. "According to the method of the present invention, the relative content of sugars having a larger molecular weight than 1-canetotriose, such as mangotetraose, can be reduced as described above. Therefore, a two-component pseudo-fluid-layer decantation separation method is used. If the reaction conditions of the enzyme shown in the steps are used, those who have to repeat the operation more than twice in the past can effectively improve the 1-cane triose content by only one operation. purity. According to a preferred embodiment of the present invention, a 1_cane triose having a purity of 80 to 95% can be obtained by this operation. In this step, a fractionated liquid having a purity of 1-cane triose of S 0%, preferably 90% or more, can be obtained. Further, it is preferable that such a high-purity solution of 1-cane fructose is prepared as a solution containing a small amount of cane-fructose as another sugar component by a fractionation operation. The smaller the amount of cane-tetraose, the larger the crystalline 1-cane-triose can be obtained. The amount of cane fructose is preferably 10% by weight or less of 1-cane triose. This paper size applies Chinese National Standard (CNS) A4 specification (210 X 297 mm) DL ------ u /! .Iv— ^ n 1 (Please read the precautions on the back before filling this page) Order- 27-4 6 6 244 A7 B7 V. Description of the invention (25) According to the preferred embodiment of the present invention, it is best to apply the activated carbon treatment and desalination treatment to the obtained fractionation liquid before implementing the next step (iii). Better. Crystallization in Step (Twisting) In the third embodiment of the present invention, step (iii) is a crystallization step. It is preferable that the method of directly implementing the first or second embodiment of the present invention is used as this step. In addition, the preferred embodiments of the first and second embodiments of the present invention can also be implemented as the better of the third embodiment. Crystal 1-Printed by the Central Standard of the Ministry of Economics of Cane, Fruit, and Sugar, printed by employee consumer cooperatives (please read the note on the back, and then fill out this page). According to the above-mentioned preferred embodiment of the present invention, 0.3 ~ 2 can be obtained. mm, preferably granulated crystalline 1-cane fructose of granulated sugar with a size of 0 · 5 ~ 1 ~ 2mm. This kind of large crystal 1-cane fructose is different from crystals of fine powder, and it is difficult for the crystals to co-aggregate with each other when dried. So you can get the particles of uniform sentence. In addition, the crystalline 1-canetriose system of the present invention is also excellent in fluidity, and fine powder flying or coagulation rarely occurs. Therefore, it can be filled accurately on the table when it is filled with small bags such as sugar, and it can also be mixed very evenly when mixed with granulated sugar. Hereinafter, the present invention will be described in more detail through examples and the like, but the present invention is not limited by these examples. In addition, the following percentages are% by weight unless otherwise specified. In addition, the crystallization stock solution used in the following examples is a oligosaccharide mixture Meioligo G or Meioligo p sold from the market by chromatographic separation method (Meiji Paper size applies Chinese National Standard (CNS) A4 specification (210X297 mm)- 28-4 6 6 24 4 Printed by the Central Standards Bureau of the Ministry of Economic Affairs, Consumer Cooperatives, A7, B7 V. Description of the invention (26) Made by 杲 Company) Fractionation and purification, or fructose, as in the examples described below. Transferase acts on sucrose The subsequent solution is prepared by methods such as fractionation and purification by chromatography. The sugar composition of the crystallization stock solution used in the following examples is slightly different due to the different separation conditions used.

Example A Example A 1 The crystallized stock solution obtained from Example C 1 described later and having a sugar composition of 9 2% 1-sucrose triose, 5% sucrose, and 3% sucrose tetraose was concentrated. Brix up to 80. In addition, after heating the concentrated solution to a temperature of 75 ° C, a seed crystal having a solid content of 1% of the concentrated solution was added, and the crystal was grown in 0.5 hours. The solution was then concentrated under reduced pressure at 30 mmHg for 30 minutes under reduced pressure. As a result, the temperature of the concentrated operating solution was reduced to 5 5 ° C, and Brix was 83. As a result of this concentration operation, 1-cane triose microcrystals are formed, so the concentrated solution is heated at 7 5 eC and maintained at this temperature for 1 hour to dissolve the generated microcrystals. As a result, the Brix series was about 80 in which the solution portion of the crystalline portion in the concentrate was removed. The crystal growth step and the microcrystal dissolution step were repeated 4 times, so that the crystal 1 and cane-triose were sufficiently precipitated. The crystals were then recovered at room temperature using a centrifugal separator, and dried to obtain 1-sucraticose crystals. The purity of the obtained 1-cane fructose crystals is 99%, and the crystal recovery rate is about 60%. * Moreover, most of the results are observed by microscopy, and most of them have a major diameter of 0.3 ~ This paper size is applicable to Chinese national standards (CNS ) A4 size (210X297mm) (Please read the notes on the back before filling this page)

-29-B 24 4 A7 B7 V. Description of the invention (27). Crystals in the range of 2mm will not cause the cohesion of the crystals to each other. The crystals obtained by the electric sieve with the sieve openings of 20, 40 and 60 will be screened 5 The particle size distribution is shown in the following table in minutes. It can be seen that most of the obtained crystals have a long diameter of 0.5mm ~ 12mm, and 1-cane fructose is similar to the shape of an elongated cuboid, so the length of the remaining crystals for screening should be larger than that of the sieve holes. Large corners. Table 1 printed by the Employees' Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs Table 1 Dimensions of one side of the Mesh mesh hole—diagonal size screening results 0.5% 2 0 0.84 _ ---- 1.19 7 5.9% 4 0 0.37 0.52 18.1% 6 0 0.25 0 3 5 5.5% Example A 2 Example A 2 This paper size applies the Chinese National Standard (CNS) A4 specification (210X297 male feet) ---: ------ Γ / ίν ^-(please first Read the note on the back of the page and fill in this page)-Order 4 66 24 4 A7 B7 V. Description of the invention 丨 28) Result obtained from Example D 2 described later, with 95% 1-cane fructose, 4% sucrose, Crystallized stock solution composed of 1% cane-fructose sugar (please read the note on the back before filling the page). Concentrate the crystallized stock solution to Brix 80. In addition, after heating the concentrated solution to a temperature of 80 ° C, a seed crystal of 1-cane triose having a solid content of 0.5% of the concentrated solution was added, and the crystals were grown in 0.5 hours. Then, the solution was concentrated under reduced pressure for 30 minutes at an absolute pressure of 160 mmHg. The temperature of the concentrated operation solution was lowered to 70 ° C, and Brix was 8 3. »This concentration operation produced 1-cane triose microcrystals, so the concentrated solution was heated to 9 5 ° C and maintained at this temperature for 1 hour. , Dissolve the resulting microcrystals. The crystal growth step and the microcrystal dissolution step were repeated 4 times. After the fourth dissolution step, the microcrystals could not be completely dissolved, so water with a solid content of about 2% of the concentrated solution was added to dissolve the microcrystals. It repeats the same crystal growth step and microcrystal dissolution step as described above. The precipitated crystals are recovered at room temperature using a centrifugal separator * and dried to obtain 1-cane triose crystals. Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 1 The purity of the cane-fructose crystals is 99%, and the crystal recovery rate is about 57%. As a result of microscopic observation, most of them were crystals with a major diameter of about 1 mm. Example A 3 Crystallized dope obtained from Example D1 described below and having a sugar composition of 9 5% 1-sucrose triose, 4% sucrose, and 1% sucrose tetraose was concentrated. This crystalline dope was concentrated to Brix 7 5 . In addition, after heating the concentrated solution to 70 ° C, it is concentrated for 15 minutes at an absolute pressure of 80mmHg. The paper size is applicable to Chinese National Standard (CNS) A4 (210X297 mm) -31-4 6 6 2- «^ A7 B7 V. Description of the invention ί: 29) Microcrystals are produced. With this microcrystal as the mother nucleus, the crystals were grown at 6.0 hours at 70 ° C. Then, the solution was concentrated under reduced pressure for 30 minutes at an absolute pressure of 8 mmHg. As a result, the temperature of the concentrated operating liquid is reduced to 5 5 ° C. This concentration operation generates 1-cane triose microcrystals, so the concentrated solution is heated at 75 ° C for 1 hour to dissolve the generated microcrystals. The crystal growth step and the microcrystal dissolution step were repeated 4 times, so that the crystal 1 and cane-triose were sufficiently precipitated. Then, the obtained crystals were recovered using a centrifugal separator at room temperature, and dried to obtain 1-cane fructose crystals. The purity of the obtained 1-cane fructose crystals was 99%, and the crystal recovery rate was about 59%. Moreover, most of them were crystals with a major diameter of about 1 mm when observed under a microscope. The obtained crystals were sieved through sieves with 20, 24 '32, and 40 sieve openings, and their particle size distributions are shown in the following table. It can be seen that most of the obtained crystals are crystals having a major diameter of 0.7 mm to 1.2 mm. And 1_ cane fructose is very similar to the shape of an elongated cuboid, so the length of the remaining crystals of the screening should be larger than the diagonal of the sieve. 11_-L__rrv ^ .II (Please read the back Note: Please fill in this page again.) Order] J. Printed by the Consumer Standards Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs. The paper size is applicable to China National Standard (CNS) A4 (210 × 297 mm) _ 32 ― 4 6 6 24 4- A7 B7 5 Description of the invention: 3〇) Table 2 Table

Mesh sieve mesh size diagonal size screen. Selection result 6.5% 2 0 0.84 1.19 3 5.8% 2 4 0.71 1.00 4 1.7 3 2 0.50 0.70 13-8% 4 2 0.35 0.49 2.2% (Please read the back first (Please note that this page is to be completed before filling in this page.)-Example · Printed by the Central Bureau of Standards, Ministry of Economic Affairs, Consumer Cooperative, Example A 4 Chromatographic separation using a two-component pseudo-mobile layer chromatography separation device with Me ioli go P as the raw material By operation, a crystallized stock solution having a sugar composition of 9 2% 1-cane fructose, 2% sucrose, and 6% cane fructose was obtained. This crystallization stock solution was concentrated to Brix 80. In addition, the concentrated solution was heated to 70 ° C and then concentrated at an absolute pressure of 8 OmmHg for 10 minutes. The paper size is -33-466244 of China National Standard (CNS) A4 specification (210X297 mm) in the Ministry of Economic Affairs * · Printed by the Consumer Bureau of Standards Bureau A7 B7 V. Invention Description (31) Microcrystals are produced. This microcrystal was used as the mother nucleus, and the crystal was grown at 70 ° C for 3 hours. Then, the solution was concentrated under reduced pressure at a pressure of 80 mmHg for 30 minutes under reduced pressure. As a result, the temperature of the concentrated operating liquid is reduced to 5 5 ° C. This concentration operation generates 1-cane triose microcrystals, so the concentrated solution is heated at 75 ° C for 1 hour to dissolve the generated microcrystals. . Repeat the crystal growth step and the microcrystal dissolution step three times, and then add the crystallization stock solution that has been reduced by the concentration operation, and repeat the secondary crystal growth step and the microcrystal dissolution step. The obtained crystals were recovered at room temperature using a centrifugal separator, and dried to obtain 1-cane fructose crystals. The purity of the obtained 1-sucratotriose crystals was 99%, and the crystal recovery was about 60%. In addition, most of them were crystals with a major diameter of 0.3 to 1.0 mm as a result of observation with a microscope. Example A 5 A crystallized raw solution composed of sugars obtained from Example D2 described later and having a sugar content of 9 5% 1-sucralose, 4% sucrose, and 1% sucrose tetraose was used. This crystallization stock solution was concentrated to Brix 8.5. In addition, after heating the concentrated solution to a temperature of 80 ° C, a seed crystal of 1-sugar triose having a solid content of 0.5% of the concentrated solution was added, and the crystal was grown in 30 minutes. The liquid was then cooled to 60 ° C over a period of 40 minutes, and crystals were grown at 60 ° C over a period of 30 minutes. The liquid was then cooled to 50 ° C in 60 minutes, and crystals were grown at 50 ° C in 30 minutes. The obtained crystals were recovered using a centrifugal separator at room temperature, and dried to obtain 1_ sucralose crystals. (Please read the precautions on the reverse side before filling out this page) Binding _ Order 〇 This paper size applies the Chinese National Standard (CMS) A4 specification (210X297 mm)-34 _ Printed by the Central Consumers Bureau of the Ministry of Economic Affairs and Consumer Cooperatives 4 β 6 24 厶 Α7 B7 V. Description of the invention (32) The purity of the 1-sugar triose crystals is 98%. The crystal recovery rate is about 45%. The results of microscopic observation revealed that most of them were crystals with a major diameter of 0.3 to 2 mm. Example A 6 A crystallized dope composed of 9 5% 1-sucrose triose, 4% sucrose, and 1% sucrose tetraose sugar was obtained using Example D2 described later. This crystallization stock solution was concentrated to Brix 8.5. In addition, the concentrated solution was slowly warmed from 20 ° C to 80 ° C, and 50 to 6 Ot: microcrystals were generated. "With the microcrystals as the mother core, crystals were grown at 80 ° C for 30 minutes. The liquid was then cooled to 70 Ot over 30 minutes; the crystals were grown at 70 ° C for 30 minutes. The liquid was then cooled to 60 ° C over 40 minutes, and the crystals were grown at 60 ° C for 30 minutes. The liquid was then cooled to 50 ° C in 60 minutes, and the crystals were grown at 50eC for 30 minutes. The obtained crystals were recovered using a centrifugal separator at room temperature, and the 1-sucratotriose crystal β obtained by drying was 99% pure and the crystal recovery rate was about 43%. The results of microscopic observation revealed that most of them were crystals with a major diameter of about 0.3 to 2 mm. Example A 7 Crystallized dope obtained from Example D2 described below and having a sugar composition of 9 5% 1-cane fructose, 4% sucrose, and 1% sucrose tetraose was concentrated. This crystallization dope was concentrated to Brix 8 5 . Add the solid content of the concentrated liquid to the concentrated liquid as 0.1% of 1—seeds of cane fructose. The paper size applies to Chinese national standards (CNS &gt; Α4 size (210 ×: 297 mm)) (please first (Read the notes on the back and fill out this page)

-35-A7 B7 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 6 6 24 4 V. Description of the invention 丨 33), the crystals are grown at 80 ° C for 30 minutes +. Then, the solution was cooled to 60 eC in 40 minutes, and crystals were grown at 30 minutes' 60 ° C. Then, the liquid was cooled to 50 ° C in 60 minutes, and the crystals were grown in 30 minutes at 50 ° C. Then, it was cooled to 30 ° C in 120 minutes, and crystals were grown at 30 ° C for 6 hours. Then, the obtained crystal was recovered by using a centrifugal separator at room temperature, and dried to obtain 1 cane-fructotriose crystal. The purity of the obtained 1-deertotriose crystal was 99%, and the crystal recovery rate was about 5 1%. The purity of a sucrose triose system was 92%, so it was concentrated to Brix 8 4 and crystallized by the same operation. The purity of the obtained 1-cane fructose crystal was 9 7%, and the crystal recovery rate was about 4 4%. So far the total 1-cane fructose recovery rate is 72%. Since the 1-cane triose purity of this honey-sweetening liquid was 89%, it was concentrated to Brix 84 and crystallized in the same manner. The purity of the obtained 1-sucrose triose crystals was 9 5%, and the crystal recovery was about 5 5%. So far, the total recovery of cane fructose is 85%. At this time, the purity of the honey syrup 1_cane fructose is 79%, so it is concentrated to Brix 8 4 and crystallized in the same operation, but a large amount of fine crystals are precipitated, which is difficult to separate at room temperature. Finishing operation Example A 8 The crystallized stock solution obtained from Example D2 described later and having a sugar composition of 9 5% 1-maltotriose, 4% sucrose, and 1% sucrose tetraose was used. This paper size applies to Chinese National Standard (CNS) A4 (210X297 mm) (please read the notes on the back before filling in this page)

-36 — 4 66 24 4 A7 A7 B7 Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of the invention (34) Concentrate this crystallization stock solution to Brix 8 5. In addition, warm up the concentrate. After the temperature is 80 ° C, add 1 to 0.5% of the solid content of the concentrate—seeds of cane fructose, and grow the crystals at 80 ° C for 15 hours. . The liquid was then cooled to 60 ° C over 2 hours, and crystals were grown at 60 ° C for 5 hours. Then the solution was cooled to 40 eC for 4 hours, and the crystals were grown at 40 ° C for 4 8 hours. The obtained crystals were recovered using a centrifugal separator at room temperature, and dried to obtain 1-cane fructose crystals. The purity of sugar crystals is 99%, and the crystal recovery rate is about 50%. Most of the crystals with a major diameter of about 0.3 to 2 mm were obtained through microscope observation. The crystals obtained were screened with sieves of 20, 24, 32, and 40 sieve. The particle size distribution is shown in the following table. The obtained crystals were crystals having a major diameter of 1.0 mm to 12 mm. And 1- cane fructose is very similar to the shape of an elongated cuboid, so the size of the major axis of the remaining crystals for screening should be larger than the diagonal of the sieve openings. (Please read the note on the back before filling in this page) -Packing. I. Order ": The size of thread paper is applicable to Chinese National Standard (CMS) A4 specification (210X297mm) -37-466244 A7 B7 V. Description of the invention (35 Table 3 Table Mesh Mesh Dimensions One Side Diagonal Size Screening Results 7.3% 2 0 0.84 1.19 6 8.5% 2 4 0.71 1.00 16.0% 3 2 0.50 0.70 6-4% 4 2 0.35 0.49 1.8% (Please first Read the notes on the back and fill in this page again.] The Central Consumers Bureau of the Ministry of Economic Affairs prints a comparative test example. The mixed sugar was crystallized in the above-mentioned Example A. 1-cane fructose. Investigate the characteristics of the carcass. Example 1 2 1-Cane fructose and granulated sugar HA (manufactured by Japan Beet Sugar Co.), 80% granulated sugar, 20% 1 cane fructotriose ratio mixed with 40 or more sieve holes as a large crystal type (large This paper size applies Chinese National Standard (CNS) A4 (210 X 297 mm) 〇〇 ™ 〇〇4 6 6 24 4 A7 B7 V. Description of the invention (36) in 40 sieve) Granulated sugar 1_ cane fructose The mixture was pulverized with a desktop grinder, respectively, and the above-mentioned implementation was screened. Crystal 1 obtained from A 2 1 cane fructose and granulated sugar HA (manufactured by Japan Beet Sugar Co., Ltd.), mixed with 80% granulated sugar and 20% 1-cane fructose ratio less than 1 0 0 sieve as powder sugar Granulated sugar 1-cane-fructotriose mixture. The items in the following table were measured by a conventional method using a powder tester (manufactured by Hosokawa Micron Co., Ltd.). The results are shown in the following table. Large crystal indium fluidity repose angle 4 9 3 9 Scraping angle 6 5 4 0 Compression degree 4 9 13 Dispersion degree 2 7 11 Poor haikuness during filling is good Good printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs It can be seen that the large crystalline type has the most fluidity, the unevenness when filling is the least, and the powder type will coagulate into a solid when compressed.

Example B Example B 1 (1) Put 15 liters of 5% sucrose in a 30 liter fermentation tank. The paper size is applicable to the Chinese National Standard (CNS) A4 specification (210X297 mm) &quot; &quot; &quot; ~ ~ ~~ _ jy-4 6 6 24 4 A7 B7 Printed by the Consumers 'Cooperatives of the Bureau of Missing Standards, Ministry of Economic Affairs, 5. Description of the Invention (37), 6.5% yeast extract' 〇. 5% CMC medium, adjusted to pH 6.5, sterilize at 120 ° C for 30 minutes. On the other hand, in a 1-liter Erlenmeyer flask containing 5.0% sucrose, 20% powder gravy, and 0.5% CMC medium, inoculate the black badger ATCC 2 0 6 1 1 and incubate at 2 8 ° C for 2 hours . Inoculation of this medium is equivalent to the above-mentioned medium, and cultured at 28 ° C for 96 hours. After the cultivation, use a basket-type centrifugal separator to recover bacterial cells from the culture solution, and then freeze-dried to recover the dried bacterial cells, and use them as the following reaction. The fructose transfer activity of the dried bacterial cells is about 14,000 units (2) per 1 g of dried bacterial cells, followed by putting 10 liters in the reaction tank to prepare a concentration of 60% by weight and a pH value of 10.0 ' The sucrose solution at a temperature of 60 ° C was added with 60 units of sucrose per 1 g of the enzyme prepared in the above method (1), and reacted at 60 ° C, pHIO · 0 for 40 hours. Then add activated carbon (Taigo activated carbon S-W 50) with a solid content ratio of 10%, and apply enzyme deactivation treatment and decolorization treatment at 95 ° C for 30 minutes. Sugar composition of this inactivated solution Line 4 4% 1-cane fructose, 3 2% sucrose, 17% monosaccharides, 7% cane fructose. This inactivated liquid was filtered to remove impurities such as bacterial cells and activated carbon. Then add activated carbon (Taiwan activated carbon S — W 50) with a solid content of 0.5% to the filter solution, and stir at 60 ° C for 20 minutes, and then apply decoloring treatment again. After filtering and removing the activated carbon in the same way as above, the cation and anion exchange resin are used for desalination treatment, and the filtering is performed again to obtain colorless and transparent 1 --------- factory-installed f Note $ item and fill out this page}.

Lr Γ First-line-This paper size applies to China National Standard (CNS) A4 (210X297 mm) -40-4 66 24 4 A7 B7 ----------------- V. Invention Explanation (38) _ Cane triose liquid is a chromatographic stock solution. In addition, during the decolorization and desalting treatment, no decomposition of sugarcane triose was observed at all. In order to further improve the purity of the 1-sucratotriose of the obtained chromatographic stock solution, a two-component moving bed chromatography separation device was used to perform chromatographic separation, and the operation was obtained. The chromatographic separation and fractionation liquid composed of sugar of cane and fructose "is then decolorized, and the chromatographic and fractionation liquid is desalted to obtain a colorless and transparent 1-cane fructose fractionation liquid, which is a crystalline raw liquid. Example B 2 Put 10 liters of sucrose solution at a concentration of 50% by weight, pH 9 · 0, and a temperature of 50 ° C into a reaction tank, and add 100 units per sucrose to the above Example Bl (1) In this method, the enzyme prepared was reacted at 50 ° C, pH 9.0 for 2 hours. Then add activated carbon (Taige activated carbon S-W50) with a solid content ratio of 1.0%, and apply enzyme deactivation treatment and decolorization treatment at 95 ° C for 30 minutes. Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs (please read the notes on the reverse page). The sugar composition of this inactivating solution is 4 3% 1—cane fructose, 30% sucrose, 18% Monosaccharides, 9% cane-tetraose. This inactivated liquid was filtered to remove impurities such as bacterial cells and activated carbon. Then add activated carbon (Taiko activated carbon S-W50) with a solid content of 0,5% to the filter solution, and stir at 60 ° C for 20 minutes, and then apply decolorization treatment again. After filtering and removing the activated carbon in the same manner as above, the cation and anion exchange resin were used for desalination treatment, and then filtering was performed again to obtain a colorless and transparent 1-sugar triose-containing liquid, that is, a chromatographic raw liquid. In addition, the decolorization and desalting processes are completed when the paper size is in accordance with the Chinese National Standard (CNS) A4 specification (210X297 mm) &quot; &quot; &quot; —41-4 6 G 24 4 · Beijing Consumer Cooperative, Central Standards Bureau, Ministry of Economic Affairs Printing A7 __B7_ V. Description of the Invention (39) None of the 1-cane fructose decomposition was seen. In order to further improve the purity of 1-canetotriose of the obtained chromatographic stock solution, a two-component moving bed chromatography separation device was used to perform chromatographic separation. The operation yielded 90% 1-canetriose, 3% sucrose, and 7%. Chromatographic separation and fractionation of sucrose tetrasaccharides. Then it is decolorized, and the chromatographic fractionation solution is desalted to obtain a colorless and transparent sucralose triose fractionation solution, which is a crystallization stock solution. Example B 3 To a solution of 10 liters of sucrose at a concentration of 60%, pH 5.5, 60t: was added Example Bl (3.0) per gram of sucrose in the same manner, at 60 ° C. , PH 5.5 reaction for 4 hours. Then add activated carbon (Taige activated carbon S — W 50) at a ratio of 0.3% to solid content, and apply enzyme deactivation treatment and decolorization treatment at 95 ° C for 30 minutes. The sugar composition of this / first living fluid is 4 3% 1_matrotriose, 2 2% sucrose, 22% monosaccharides, and 13% sucralose. This inactivated liquid was filtered to remove impurities such as bacterial cells and activated carbon. Then add activated carbon (Taige activated carbon S-W50) with a solid content of 0.2% to the filtrate, and stir at 60 ° C for 20 minutes, and then apply decolorization treatment again. After filtering and removing the activated carbon in the same manner as above, the cation and anion exchange resin were used for desalting treatment, and filtering was performed again to obtain a colorless and transparent liquid containing sucralose, that is, a chromatographic stock solution. In addition, during the decolorization and desalting treatment, 1-sugar triose was not decomposed at all. (Please read the precautions on the back before filling this page]-Binding and binding. Γ, _line · This paper size applies to China National Standard (CNS) A4 (210X 297 mm) —42 — 4 66 24 4 Liji A7 ____B7 printed by the Shell Standard Consumer Cooperative of the Ministry of Standards of the People's Republic of China. 5. Description of the invention (40) In order to further improve the purity of the 1_ cane fructose of the obtained chromatographic stock solution, the chromatographic separation device using a 20% fen mobile bed decantation was applied After separation and operation, 80% 1 sucrose triose, 7% sucrose, and 13% sucrose tetraose chromatographic separation and fractionation solution are obtained. Then, the chromatographic fractionation solution is decolorized and desalted to obtain colorless and transparent No. 1_Sucralose triose fractionation solution is the crystallization stock solution. Example B 4 The sugar obtained from Example B 2 described later is used, which has 90% 1-sucralose, 3% sucrose, and 7% sucrose tetraose sugar. Constitute # 化 原液. Concentrate this crystallization stock solution to Brix 80. In addition, after heating this concentrated solution to a temperature of 70 ° C, concentrate it at an absolute pressure of 8 OmmHg for 10 minutes to produce microcrystals β. Nuclei, crystal growth at 3 ° C, 70 ° C. Then the absolute pressure 8 OmmHg minus in 30 minutes Concentrate this solution. The temperature of the concentration operation solution is lowered to 5 5 ° C. This concentration operation generates 1_cane fructose microcrystals, so the concentrated solution is heated at 7 5 ° C and maintained at this temperature for 1 hour. Dissolve the generated microcrystals. Repeat the crystal growth step three times and the microcrystal dissolution step, and then replenish the crystallization stock solution reduced by the concentration operation, and then repeat the secondary crystallization or long step and the microcrystal dissolution step. The recovered crystals were recovered at room temperature using a centrifugal separator, and dried to obtain 1 cane fructose crystals. 'The purity of the obtained 1 cane fructose crystals was 99%, and the crystal recovery rate was about 60%. Microscopic observation results, most of which are long diameters 0.3 ~ HH H-----r./L·._ Packing I (Please read the precautions on the back before filling this page) Order ', -line_ This paper The scale is applicable to China National Standard (CNS) 8-4 specification (210X297 mm) _ 43-4 Printed by the Shellfish Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 66 24 4 A7 ____B7_ V. Description of the invention (41) 1. Crystal of about 0mm Example B 5 Result obtained using Example B 1 and having 9 2% 1 _ cane fructose, 3% Sugar, a crystallization solution consisting of 5% maltotetraose sugar. Concentrate this crystallization solution to Brix 8 5 "To this concentrate, add 1-cane fructose to the concentrated solution with a solid content of 0.1% The seed crystal 'allowed the crystal to grow for 30 minutes at 80 ° C. Then the liquid was cooled to 70 ° C for 30 minutes, and the crystal was grown for 30 minutes at 70 ° C. The liquid was then cooled to 60 ° for 40 minutes. C, crystals were grown at 60 ° C for 30 minutes. The liquid was then cooled to 50 ° C over 60 minutes, and the crystals were grown at 50 ° C for 30 minutes. The liquid was then cooled to 30 ° C over 120 minutes and allowed to crystallize at 30 ° C over 6 hours. The obtained crystals were recovered at room temperature using a centrifugal separator, and after drying, 1 cane fructose crystals were obtained. The purity of the obtained .1-cane-triose crystals was 99%, and the crystal recovery was about 51,%. The 1-cane fructose purity of this honey fraction is 8 5%, so it is concentrated to Brix 8 4 and crystallized in the same manner. The purity of the obtained 1-cane fructose crystals was 9 5%, and the crystal recovery rate was about 5 5%. So far, the total recovery of cane fructose is 78%. At this time, the purity of cane-fructose, 1 of the honey solution at this time, was 74%, so it was concentrated to Br i X 8 4 and crystallized by the same operation, but a large amount of fine crystals precipitated, which is difficult to separate at room temperature. Therefore, the operation of this paper is finished. The Chinese standard (CNS) A4 specification (21〇 &gt; &lt; 297 mm) is applicable. -44-4 6624 4 A7 B7 V. Description of the invention (42)

Example C (Please read the precautions on the back before filling this page) Example C 1 (1) Put 30 liters of fermentor in 15 liters containing 2 2.5% sucrose, 1.5% yeast extract Material, 3.0% corn slurry culture medium, adjusted to pH 6.8, sterilized at 120 ° C for 30 minutes. On the other hand, in a 1-liter Erlenmeyer flask containing 15.0% sucrose, 1.0% yeast extract, and 2.0% corn slurry culture medium, Penicillium laudenii (I AM7 2 54 strain) was placed at 2 8 ° C was cultured for 3 days. Inoculate this medium as described above and incubate at 28 ° C for 4 days. After the cultivation, the bacterial cells were recovered from the culture solution using a basket-type centrifugal separator, and the dried bacterial cells were recovered by freeze-drying, and used as the following reaction enzyme. The fructose transfer activity of the dried bacterial cells is 9 5 ～ 1 3 5 units per 1 g of dried bacterial cells. Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs (2) Secondly, put 10 liters in the reaction tank to prepare 5 5 A sucrose solution having a concentration of% by weight, pH 7.0, and a temperature of 50 ° C, was added with the enzyme prepared in the above method at 10 units per sucrose, and reacted at 50 ° C and pH 7.0 for 4 hours. Then add activated carbon (Taige activated carbon S_W 50) with a solid content of 0.3%, and apply enzyme deactivation treatment and decolorization treatment at 95 ° C for 30 minutes. The sugar composition of this inactivated solution is 4 6% 1-sucrose triose, 18% sucrose, 30% monosaccharides, 6% sucralose. This inactivated liquid was filtered to remove impurities such as bacteria, and then decolorized and desalted with activated carbon and ion exchange resin, and filtered again to obtain a colorless and transparent 1_cane triose liquid. This paper size applies to Chinese National Standard (CNS) A4 specifications (210 × 297 mm) -45-Α7 Β7 Printed by the Shellfish Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs β 6 24 4 V. Description of the invention (43) In order to improve the obtained chromatography 1 of the original liquid, the purity of cane-fructose was applied to a chromatographic separation using a two-component moving bed chromatography separation device, and operation was performed to obtain 9 2% 1-cane-fructose, 5% sucrose, and 3% cane-fructan The composition of the chromatographic separation and fractionation liquid 'was then decolorized, and the chromatographic fractionation liquid was desalted to obtain a colorless and transparent 1-cane fructose fractionation liquid, that is, a crystallization stock solution. Example C 2 An enzyme prepared in the same manner as in Example C 1 (1) with 50 units per gram of sucrose was added to a 10 liter solution of sucrose bran prepared at a concentration of 65%, pH 8. 0, 40 ° C. , And reacted at 40 ° C, pH 8.0 for 5 hours. Then add activated carbon with a solid content of 1.0% (Taiko Activated Carbon S-W50), and apply enzyme deactivation treatment and decolorization treatment at 95 ° C for 30 minutes * The sugar composition of this solution is 4 7% 1 -Sucrose triose, 18% sucrose, 28% monosaccharides, 7% sucrose tetraose》 After filtering this inactivated liquid to remove impurities such as bacteria, it is decolorized and desalted with activated carbon and ion exchange resin, again Filter to obtain a colorless and transparent 1-cane-fruit-three-bran liquid. "To further improve the purity of the 1-cane-fructose of the obtained chromatographic stock solution, use a two-component moving bed chromatography separation device to perform chromatographic separation / operation to obtain 9 0.% 1 Chromatographic separation and grading solution consisting of 1 cane fructose, 3% sucrose, and 7% sucrose tetrasaccharides. "Then decolorize and desalinize the chromatographic fractionation solution to obtain colorless and transparent. National Standard (CNS) A4 specification (210X297 mm) (Please read the notes on the back before filling in this tile)

-46-4 Printed by Shellfish Consumer Cooperative, Central Bureau of Standards, Ministry of Economic Affairs

0 6 24 4 A7 ____B7 V. Description of the Invention (44) 1—Sucralose triose fractionation solution is the crystallization stock solution. Using this stock solution in the same manner as in Example B 4 described above, about the same crystalline 1-canetotriose was obtained. Example C 3 An enzyme prepared in the same manner as in Example C1 (1) with 2 units per lg of sucrose was added to a solution of 10 liters of hempose prepared at a concentration of 65%, pH 6.0 and 40 ° C at 40 °. C, pH 6.0 reaction for 100 hours. Then add activated carbon with a solid content of 1.0% (Taiko activated carbon S-W50), inactivate the enzyme and decolorize it at 95 ° C for 30 minutes. The sugar composition of this solution is 46% 1-cane Fructose, 19% sucrose, 28% monosaccharides, 7% sucrose tetraose. After filtering the inactivated liquid to remove impurities such as bacteria, it is decolorized and desalted with activated carbon and ion exchange resin, and filtered again to obtain colorless Transparent liquid containing 1 cane fructose. In order to further improve the purity of 1-sucratotriose of the obtained chromatographic raw solution, a two-component moving bed chromatography separation device was used to perform chromatographic separation. The operation yielded 80% 1 monocanuctotriose, 7% sucrose, 1 3 Chromatographic separation and fractionation of sucrose tetraose sugar. Then decolorize and desalinate the chromatographic fractionation solution to obtain a colorless and transparent 1-cane-fructotriose fractionation solution, which is a crystallization stock solution. Example D (Please read the precautions on the back before filling this page) Line-This paper size applies to Chinese National Standard (CNS) A4 (210X297 mm) -47-Printed by Male Workers Consumer Cooperatives, Central Bureau of Standards, Ministry of Economic Affairs 4 66 24 4 A7 ______B7 ____ 5. Description of the Invention (45) Example D 1 (1) Purification of enzymes Put 15 liters of medium containing 5.0% sucrose, 0.5% corn pulp, 0.1% urea in a 30 liter fermentation tank, adjust to pH 7.0, at 120 ° C for 30 minutes. In addition, a 1-liter Erlenmeyer flask containing 5.0% sucrose, 1.0 yeast extract, and 2.0 corn slurry was inoculated with 3 strains of F. brevis I F04 8 4 and cultured at 28 ° C for 29 hours. , 7 days at 28 ° C. After cultivation, use a basket-type centrifugal separator to recover the bacterial cells from the culture solution, and then freeze-dried to recover the dried bacterial cells. The fructose shifting activity of the dried bacterial cells is 5.2 to 8.3 per 1 g of dried bacterial cells unit. The freeze-dried powder of the bacterial cells obtained as described above was suspended in Mcllvaine buffer (PP7.0), and the cells were crushed by ultrasonic treatment. The crushed product obtained by membrane fractionation using an ultrafiltration membrane (molecular weight of 100,000 fractions) was used as the crude enzyme solution. An ion exchange column (Q-Sephalocolumn 4 0/1 0 0, manufactured by Pharmacia) equilibrated with Tris-HCl buffer (P Η 7.3) was fractionated with a concentration gradient of 0 to 1 μM N a C 1 This crude enzyme solution was purified by gel filtration column chromatography (Superdex 2 0 1 0/3 0 Pharmacia), and then two fractions of enzyme activity were observed. In the obtained fraction, about a single band fraction was observed on electrophoresis, and this was used as a partially purified enzyme. Then use 0. 0 2 5 M bis-Tris * HCJ? (PH6-3) as the starting buffer, and a 10-fold dilution of the multiple buffer solution 7 4 · This paper size applies the Chinese National Standard (CNS) Α4 Specifications (210X297mm) (Please read the notes on the back before filling this page)

-48-4 ㈣ 24 4 Yingan A7 B7, Shellfish Consumer Cooperative, Central Bureau of Standards, Ministry of Economic Affairs 5. Description of the invention (46) HC 1 (pH 3. 8) is used for column isoelectric point chromatography of the eluent (Mono P 5 / 20 column, Pharmacia) Refined this part of the purified enzyme * The outline of the enzyme purification is shown below. Table 5 Amount of total enzyme solution (ml) Pieces (single) Recovery rate (¾) Freeze-dried extract 7700 219,000 100 Ultrafiltration membrane 1200 228, 6 00 104 Anion exchange chromatography 130 214,200 98 Gel filtration chromatography 15 657 0. 3 isostatic point decantation 8 32 B 0.15 (2) The molecular weight and isoelectric point of the whole enzyme obtained by gel filtration method are about 3 60 to 3 8 0 / c D a. The molecular weight of SDS-PAGE was about 54 / cDa, and it was confirmed that the subunit structure was present. In addition, the method of Muramatsu et al. (Methods in Enzymology, 50'555 (1987)) was changed as follows to remove sugar bonds. That is, 1 to 2 times (w / w) SDS was added per leg enzyme, and then modified in a hot water bath. Add 1 for every 1 modified protein. This paper size is applicable to Chinese National Standard (CNS) A4 (210X297 mm) (Please read the notes on the back before filling this page)

4 6 6 24 4 A7 __B7 ____ V. Description of the invention (47) mm Endoglycosidase unit-point-点 'at 50 mM orthoacid (please read the note on the back before filling this page) buffer (P Η 6 · 0), incubate at 37 ° C for 12 hours or more. As a result, the molecular weight after removing the sugar bond was about 5 1 «D a. It was confirmed that a subunit structure was present. The isoelectric point of the enzyme was estimated from the results of two-dimensional electrophoresis and hydrophobic chromatography from 3.8 to 3.9. Example D 2 In 10 liters of a sucrose solution adjusted to a concentration of 55% by weight and at a pH of 9.8 'and a temperature of 40 ° C, the purified enzyme obtained in Example D 1 with 20 units per sucrose was added. 0 eC, P Η 9.8 for 9 hours. Then add activated carbon with a solid content of 1.0% (Taiko Activated Carbon S-W50 0) at 95. (: Enzyme inactivation treatment and decolorization treatment for 30 minutes. The sugar composition of this treatment solution is 5 5% 1-sucrose triose, 16.6% sucrose, 25% monosaccharides, 4% cane fructose. Economy After printing and filtering this inactivated liquid to remove impurities such as activated carbon, it was printed and filtered by activated carbon and ion exchange resin, and filtered again to obtain a colorless and transparent liquid containing 1 cane-fructose. In order to further improve the purity of 1-cane-fructotriose of the 1-cane-fructotriose solution, a two-component pseudo-moving bed chromatography separation device was used for chromatographic separation operation to obtain 95% A chromatographic fractionation solution composed of sucrose and 1% sucrose tetraose sugar. Then the chromatographic fractionation solution was decolorized and desalted to obtain colorless and transparent. The paper size applies the Chinese painting national standard (CNS) A4 specification (210 X 297). (Mm) -50-466244 Printed by A7 _____ ^ _ B7__ of the Central Laboratories of the Ministry of Economic Affairs, Shellfish Consumer Cooperative. V. Description of the Invention (48) 1-A cane-fructotriose fractionation solution, that is, a crystallization stock solution. Example D 3 at 10 Liter adjusted to 50% by weight, pH 7.0, temperature 3 5 ° C To the sucrose solution, 2 units of sucrose per lg of the purified enzyme obtained in Example D 1 was added, and the reaction was performed at 35 ° C, p Η 7.0 for 60 hours. Then, 0.3% of activated carbon with solid content was added ( Taiko activated carbon S-W50), enzyme deactivation treatment and decolorization treatment were performed at 95 ° C for 30 minutes. The sugar composition of this treatment solution was 5 3% 1-sucrose triose, 1 7% sucrose, 25% single Sugars, 5% cane and fructose "After filtering this inactivated liquid to remove impurities such as activated carbon, decolorization and desalting treatment with activated carbon and ion exchange resin, and filtering again to obtain a colorless and transparent liquid containing 1 cane fructose trisaccharide" In order to further improve the purity of the 1-cane-fructotriose solution, the decantation separation operation using a two-component pseudo-moving bed chromatography separation device was performed to obtain 90% 1_cane-fructoose, 3%. A chromatographic fractionation solution composed of sucrose, 7% sucrose tetrasaccharide sugar. Then, the chromatographic fractionation solution is decolorized and desalted to obtain a colorless and transparent 1-cane fructose fractionation liquid, which is a crystallization stock solution. The stock solution was treated in the same manner as in Example B 4 above to obtain approximately the same crystalline 1-canetriose. (Please read the note f on the back before filling out this page). Packing--Ordering-Thread · This paper size adopts China National Standard (CNS) A4 specification (210X297 male sauce)

Claims (1)

Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs. I. ABCD Lee Scope No. 85115394 Patent Application Chinese Patent Scope Amendment June 1990 Revision of Seed Crystal 1 _ Cane Fructose Manufacturing Method, Its Features In order to perform concentrated 1-high-purity 61 with a purity of more than 80% of cane-fructose: 丨 the parent is more than 75, the step of adding crystals above 60 ° C to make the crystal grow 1-the solution of cane-fructose makes the After the solution is seeded, the temperature of the concentrated solution is warmed; and (b) E of the concentrated solution is concentrated under reduced pressure at an absolute pressure of 40 to 200 ° C, so that the crystal grows (c) the crystal growth step When the temperature of ° C dissolves the production part in the concentrated solution, add water to dissolve it as appropriate and then repeat the above (c) microcrystalline dissolution step at least once more. The rix is controlled in the range of 7 5 ~ 85 mmHg, and the temperature is 40 ~ 70 crystal growth steps; and after the step, the solution is heated to 70 ~ 95 5 microcrystals and then dissolved. If the microcrystals cannot be completely dissolved, the microcrystalline dissolution step is described above. (B) The crystal growth step is the same as above. After that, a step of recovering crystalline 1_cane triose is performed. 2 · — Seed Crystal 1-Method for making cane fructose, characterized by application (please read the precautions on the back before filling this page) J ---- I——Order --------- Line) Concentrated 1-Brix of high-purity liquid with a purity of 80% or more of fructose is above 75, followed by a step of ° C or higher; then a 1-sugar triose solution is used to warm the solution and warm the concentrate The temperature is 60. This paper size is in accordance with the National Standard (CNS) A4 specification (210 X 297 mm). 4 Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economy 66244 Jaw C8.! D8 VI. Application scope of patents &gt;) The above concentrated liquid is concentrated under reduced pressure at an absolute pressure of 10 to 160 mmHg at a temperature of 30 to 70 ° C, a step of lowering the temperature of the liquid to start crystallization and growing the crystal; Thereafter, perform the (c) microcrystal dissolution step as described in item 1 of the patent application scope, and then repeat the above (b) crystal growth step and (c) 'microcrystal dissolution step as described above in the patent application item 1 at least once more, Thereafter, the step of crystallizing 1-cane triose is recovered. 3. A method for producing crystal 1 monocane triose, which is characterized in that (α) concentration is performed. 1-cane triose has a purity of more than 80%. 1-cane triose high purity solution, so that Brix is 80 or more, and then the concentrated solution is heated to 70 to 95 ° C, and a seed crystal is added to make the concentrated solution grow and crystallize in the above temperature range; and (Lu) then reduces the temperature of the above concentrated solution The cooling step is 5 ~ 20 ° C lower than the temperature of the above step, and then (7 ·) the concentrated liquid is placed at the temperature lowered by the above cooling step, so that the crystal grows in the crystal growing step; then, the above is repeated at least once more (Million) cooling step and the above (r) crystal growth step, lowering the temperature to 20 ~ 60 ° C and then recovering the crystal 1-the step of cane fructose "4. A kind of crystallization 1 the production of cane fructose The method is characterized by performing (α &gt;) concentration of a high-purity solution of cane-fructose with a purity of 80% or more, so that the Brix of the solution is more than 80, following the paper standard applicable to China National Standard (CNS) A4 (210 X 297 mm) (please first Read the note on the back of the page and fill in this page again) -2-Α8 Β8 C8 D8 4 6 6 244 6. Apply for a patent and set the concentrate at a temperature ranging from 50 to 60 ° C to start crystallization 'and at 7 0 ~ 9 After the step of growing the crystal at 5 ° C, repeat the process more than once, such as (β) cooling step and the above (γ) crystal growth step in the scope of the patent application, and reduce the temperature to 20 ~ 60 ° C. The step of recovering crystalline 1-cane fructose. 5. For example, the method for producing crystalline 1-cane fructose of item 3 or 4 of the patent application group, wherein the (α) step or (α &gt; 1) High purity in the step 1-1 of cane fructose solution-the purity of cane fructose is more than 90%. 6. For example, the crystalline 1_cane fructose in any one of the items 1 to 4 of the scope of patent application. The manufacturing method, wherein in the steps (a), (a〃), (α) or in the step, as a high-purity 1_ sucrose triose solution, the amount of sucralose is 1 to 1 Less than 10% of the trisaccharide. &Quot; '...--........ 7' If the crystal of any one of items 1 to 4 of the scope of patent application 1-cane fruit three The manufacturing method of 'wherein the recovery step of crystallized 1-cane triose at room temperature is performed. 8—a continuous production method of crystallized 1-cane triose, which is characterized by The method of any one of 4 items recovers the 1-cane triose sugar solution after crystallization, and then applies the method of any one of claims 1 to 4 to recover the crystal 1-cane fructose. If necessary, for the sugar solution containing 1-canetotriose after being recovered and crystallized, the method according to any one of claims 1 to 4 of the scope of patent application is applied to recover the crystalline 1-canetotriose. This paper size applies to Chinese National Standard (CNS) A4 ^ (210 X 297 public love) (Please read the precautions on the back before filling out this page) ... V "Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs ~ 3 -4 Printed clothing for employees' cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs 66244 i ____ ^ _ S VI. Application for patent scope 9. If the manufacturing method of any one of the patent scope 1 to 4 is applied, the purity can be obtained as 9 5% or more columnar crystals with a major diameter of 0 3 to 2 mm. 1 0 · —A kind of crystal 1 A method for producing cane fructose, which comprises (i) an enzyme having fructose transfer activity derived from the genus Pleurotus niger, at pH 5.0 to 10.0.0, and temperature 50 Under the condition of ~ 65 ° C, it can convert more than 43% by weight of sugar to 1 to 3% by weight of sucrose solution with a concentration of 50% by weight or more. An enzyme that converts to sucralose, acts on a sucrose solution, and converts 43% by weight or more of the generated sugars into 1-sucralose, and (ii) chromatographic separation step (i) The step of obtaining 1-cane fructose to a purity of 80% or more. For (twisting) the fraction of purity 80% or more obtained in step (ii), the first to the scope of the patent application is applicable. The method according to any one of item 4, a step of crystallizing 1_cane triose to obtain a columnar crystal having a major diameter of 0.3 mm or more, and a crystalline 1-cane triose having a purity of 9 5% or more . 1 1. The method for producing 1-cane fructotriose according to item 10 of the scope of application for patent, wherein the genus P. niger is ATCC20611. 12. The method for producing a 1-cane fructose according to item 10 or 11 of the scope of patent application, wherein the sucrose concentration of the sucrose solution in step (i) is 50% by weight or more for 1 g of sucrose. Adding Fructose Transfer Active Enzyme 0 _ 5 to 100 units * and the pH is 5 _ 0 to 10.0. 〇 This paper size is applicable to China National Standard (CNS) A4 (210 X 297 mm) (Please read the back (Please fill in this page again for attention) &quot; ------ 丨 Order --------4-466244 A8 R8 C8 D8 Printed clothing by employees' cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs It is carried out under the conditions of 50 to 6 ° C and a reaction time of 2 to 100 hours. 1 3. A kind of crystal 1 _ A method for producing sucrose triose, which comprises (1) an enzyme having fructose transfer activity derived from Penicillium spp. Penicillium spp. At pH 6-〇 ~ 9 · 0, temperature 35 ~ Acting at 55 ° C on a sucrose solution with a concentration of more than 50% by weight, an enzyme that can convert more than 46% by weight of sugar to 1 _ sucralose is used in the sucrose solution to make 4 A step of converting 6% or more by weight to 1_sutotriose, (ii) a step of fractionating the 1_sutotriose obtained in step (i) by a chromatographic separation method to a step of 80% or more in purity, and ( ΰΐ) For the distillate having a purity of 80% or more obtained in step (ii), the method of any one of the items 1 to 4 of the scope of patent application is applied to crystallize 1-cane triose to obtain a long diameter A step of crystallizing 1-cane triose with a columnar crystal body of 0.3 mm or more and a purity of 95% or more. 1 4 _ As for the manufacturing method of item 13 in the scope of patent application, wherein Penicillium spp. Loudi penicillium is I AM7 2 54. 15. The method for producing crystal 1-cane fructose according to item 13 or 14 of the scope of the patent application, wherein in step (i), the sucrose concentration of the sucrose solution is 50% by weight or more, and 1 g of sucrose Specifically, 0.5 to 100 units of an enzyme containing fructose transfer activity is added, and the reaction is carried out under conditions of pH 6.0 to 9.0, temperature 35 to 55 ° C, and reaction time 2 to 100 hours. This paper size applies to China National Standard (CNS) A4 specification (210 X 297 mm) (Please read the precautions on the back before filling this page) l ·;. · Clothing --------, 0. -5-6 6 244 A8 B8 C8 1) 8 VI. Application for patent scope 1 6 .—Crystal 1 Manufacturing method of cane fructose 'It's feature is included (please read the precautions on the back before filling this page) ) (i) Fructose transferase-derived enzymes derived from Gliocladium sphaeroides are used at a pH of 6.0 to 〇_1〇_〇 and a temperature of 35 to 50 ° C to act on sucrose at a concentration of 50% by weight or more A solution that converts more than 53% by weight of sugar to 1-sugarose triose, and acts on the sucrose solution to convert more than 53% by weight of sugar to 1-sugarose, and (ii) The step of fractionating the 1-cane fructose obtained in step (i) by a chromatographic separation method into a fraction having a purity of 80% or more, and (Xun) the purity obtained in step (ii) is 80% or more The fraction 'applies the method of any one of the first to the fourth of the patent application scope, which crystallizes 1-cane triose to obtain a long diameter of 0.3 mm or more. Knot-like crystals, and a purity of more than 95% of the crystal - the step of kestose. 17. The manufacturing method of crystal 1_cane fructose according to the scope of application for patent No. 16, wherein the genus Pleurotus spp. Is IF04843. 1 8 For example, the crystallization of item 16 or 17 in the scope of patent application 1—Manufacturing method of printed fructose by the consumer cooperative of employees of the Intellectual Property Bureau of the Ministry of Cane Economy, wherein in step (i), the sucrose concentration of the sucrose solution is 5 0% by weight or more, 0.5 to 100 units of an enzyme containing fructose transfer activity is added to 1 g of sucrose, at a pH of 6.0 to 10, 0, a temperature of 3 to 5 to 50 ° C, and a reaction time of 2 to 100 hours. Next implementation. 19. If there is a crystalline 1-cane-fructose manufacturing method in the scope of patent application No. 10, 13 or 16 ', in which the national paper standard (CNS) A4 specification is applied to the paper size as in the patent application scope No. 3 (210 X 297 mm) 6 4 6 AS B8 C8 D8 6. The scope of patent application or the method of item 14 is used in (α) step or (α-) step 1-cane fructose purity is 90% The above high-purity 1 cane-fructotriose solution 〇20. For example, a method for producing crystalline 1-cane-fructotriose according to any one of the scope of application patents 10, 13 or 16, wherein the crystallization is performed at normal temperature. 1 _ Recovery steps of cane fructose. 21—A continuous manufacturing method of crystalline 1-cane fructose, characterized in that for crystals containing 1-cane fructose after recovery by crystallization according to any one of the methods of patent application No. 10, 13 or 16 The sugar solution of sugar can be fractionated by chromatographic separation method if necessary. The 1-cane triose in the sugar solution has a purity of 80% or more. One method recovers crystalline 1-cane fructose, and if necessary, for a sugar solution containing 1-cane fructose after the crystals are recovered, if necessary, chromatographic separation is used to fractionate 1- The sucrose triose has a purity of 80% or more, and the method such as any one of claims 1 to 4 of the scope of patent application is applied to recover the crystallized sucralose. 2 2. If the manufacture of the scope of application for the patent No. 10, 13 or 16 ----------- IV ^ -------- order -------! Line „, Y (please read the precautions on the back before filling in this page) Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs 3 The shape of the paper can be the same, m method m square 2 This paper size applies to China National Standard (CNS) A4 (210 X 297 mm)-1- Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs. I. ABCD Lee Scope No. 85115394 Patent Application Chinese Patent Scope Amendment June 1990 Revision of Seed Crystal 1 _ Cane Fructose Manufacturing Method, Its Features In order to perform concentrated 1-high-purity 61 with a purity of more than 80% of cane-fructose: 丨 the parent is more than 75, the step of adding crystals above 60 ° C to make the crystal grow 1-the solution of cane-fructose makes the After the solution is seeded, the temperature of the concentrated solution is warmed; and (b) E of the concentrated solution is concentrated under reduced pressure at an absolute pressure of 40 to 200 ° C, so that the crystal grows (c) the crystal growth step When the temperature of ° C dissolves the production part in the concentrated solution, add water to dissolve it as appropriate and then repeat the above (c) microcrystalline dissolution step at least once more. The rix is controlled in the range of 7 5 ~ 85 mmHg, and the temperature is 40 ~ 70 crystal growth steps; and after the step, the solution is heated to 70 ~ 95 5 microcrystals and then dissolved. If the microcrystals cannot be completely dissolved, the microcrystalline dissolution step is described above. (B) The crystal growth step is the same as above. After that, a step of recovering crystalline 1_cane triose is performed. 2 · — Seed Crystal 1-Method for making cane fructose, characterized by application (please read the precautions on the back before filling this page) J ---- I——Order --------- Line) Concentrated 1-Brix of high-purity liquid with a purity of 80% or more of fructose is above 75, followed by a step of ° C or higher; then a 1-sugar triose solution is used to warm the solution and warm the concentrate The temperature is 60. This paper size applies to the national standard (CNS) A4 specification (210 X 297 mm).