JPH0198601A - Preparation of high-purity isomalto-oligosaccharide - Google Patents

Abstract

PURPOSE:To prepare a high-purity isomalto-oligosaccharide in a high efficiency and in a large amt., by passing an specified high-conversion syrup through a strongly acidic cation exchange resin. CONSTITUTION:A high-conversion syrup, obtd. by the reaction of a starch partially hydrolyzed with an enzyme or an acid with 0.001-1.0% saccharifying alpha-amylase and 0.001-1.0% glucoamylase, is caused to react with a transferase or a debranching enzyme to give a high-conversion syrup contg. 10-60% glucose, 10-60% isomaltose and 10-50% isomalto-oligosaccharide other than isomaltose per unit solid content. The obtd. syrup is passed at a rate (SV) of 0.01-1.0 through a strongly acidic cation exchange resin of alkali (earth) metal type at 30-80 deg.C, thereby separating a fraction contg. at least 90% glucose per unit solid content, to give a high-purity isomalto-oligosaccharide consisting of a fraction contg. at least 80% isomalto-oligosaccharide, including isomaltose, per unit solid content.

Description

[Detailed Description of the Invention] [Industrial Application Fields] The high purity isomalto-oligosaccharide obtained by the present invention can be used in breads, confectionery, gums, powdered foods, soft drinks, dairy drinks, frozen desserts,
Dairy products, canned goods, bottled goods, tubed goods, jams, tsukudani, pickles, paste products, soy sauce, sauces, seasonings, prepared foods, prepared foods, alcoholic beverages, feed, pharmaceuticals, health foods, household sweeteners, etc. 0 [Prior art] In recent years, research and development of oligosaccharides has been actively conducted, and oligosaccharides have various physiological functions such as mild sweetness, good hygroscopicity, and indigestibility. For this reason, high conversion syrups containing isomaltose have been developed. This isomaltose-containing high-conversion syrup is characterized by a high content rate of glucose and isomaltose. Examples of its characteristics include a DE (glucose equivalent) of 60 to 75, and a glucose content (@-type). per) 10% to 60%, isomaltose content (same) 10%
9b to 60%), Inmal other than isomaltose),
T! J djlh minute (same) 1 (1~5096"t'!
D. The total isomalto-oligosaccharide including inmaltose per solid is 30% to 60%. On the contrary,
Efficiently and economically produce large quantities of high-purity isomalto-oligosaccharides containing 60% or more and less than 80% of isomalto-oligosaccharides including diisomaltose in solid form, or more than SOS of isomalto-oligosaccharides including diisomaltose in solid form There is no established method for producing it, and expectations are high for its development for use in food and medicine.

[Problem that the invention seeks to solve]

As explained above, the present invention has an isomalto-oligosaccharide content of 60
96 or more and less than 80% or isomaltooligosaccharide content SO
In view of the current situation where an industrial production method for high-purity isomalto-oligosaccharides containing S or more has not been established, we provide an industrial production method for efficiently and economically producing large quantities of high-purity isomalto-oligosaccharides. Means to solve the problem] The raw materials for obtaining isomalto-oligosaccharides including isomaltose with high purity are: ■Retrosynthetic product produced by the action of glucose-rich glucoamylase〇■ Pullulan, Dexto2 Partially hydrolyzed products such as 0 ■ Starch or partial hydrolysates of starch (liquefied products) k Liquefied type d
- There are reaction products produced by the action of amylase, β-amylase, debranching enzyme, and transferase singly, simultaneously, or in stages. Method A detailed explanation is given in Tabe et al., ``Amylase retrosynthesis reaction and transfer reaction 11 Starch Science, Vol. 25, No. 2 (1978).'' Generally, a high concentration of glucoamylase is applied to a glucose solution obtained by hydrolyzing a starch liquefied solution with glucoamylase, but the reaction takes a long time and there are problems with high concentrations of glucoamylase. I'm leaving it behind. Method (2) uses expensive pullulan and dextran as raw materials, and method (0) is not suitable for the efficient and economical industrial production method that is the objective of the present invention. Although this method uses the enzyme preparations of Tori, the basic reaction mechanism is an enzyme that mainly produces maltose and an enzyme that has a sugar transfer effect (for example, Yatabe Fuyo et al. 1. Manufacture of sugar materials for 93-year-old sake I Journal of the Japan Fermentation Association, Volume 75,
No. 12 (1980) provides a detailed explanation). This method leaves room for investigation of even better enzyme conjugation conditions from the viewpoints of isomalto-oligosaccharide yield, workability, and economic efficiency.

Under these current circumstances, the inventors of the present invention have conducted extensive research into an industrial method for producing high-purity isomalto-oligosaccharides.
The most suitable method for industrial production is to apply high-conversion syrup containing isomaltose as a raw material for chromatographic separation using a strongly acidic cation exchange resin. Alternatively, by spray-drying the syrup to form a powder, an extremely efficient and economical industrial production method was discovered, leading to the present invention.

(Production of high-conversion syrup containing isomaltose) High-conversion syrups other than isomaltose are produced by combining monosaccharification type α-amylase and glucoamylase on starch partial hydrolyzate using enzymes or acids. The high-conversion syrup is then treated with a transferase or a combination of a transferase and a debranching enzyme to cause the reaction sugar solution to contain isomaltose and isomalto-oligosaccharides other than isomaltose. The amount of enzyme is 0.5% of the saccharification agent d-amylase (for example, Novo Industries, Fungamil 800L) per substrate (per solid).
001～toq! I, glucoamylase (e.g.
Manufactured by Novo Industries, AMG300L) 0.001~
3fl of about 0.001-to% of transferase (for example, Ohno Pharmaceutical Co., Ltd., transglucosidase)! [Combination of enzymes or additional debranching enzymes (e.g.
Manufactured by Novo Industries, Promozyme 200L) α00
It is a combination of four enzymes to which about 1 to 0.1% are added. The viscosity reaction conditions are usually a temperature of 40 to 65°C and a pH of
3.0 to 6.5, substrate concentration 10 to 60% (solid), and saccharification time of about 40 hours, but if the amount of enzyme is large, it will be shorter, and if it is less, it will be longer. The sugar composition of glucose, isomaltose, and isomalto-oligosaccharides other than isomaltose in the final saccharification reaction solution can be changed by adjusting the usage ratio of saccharification 1JLA-amylase, glucoamylase, transferase, and excision enzyme. Can be done. Normally, when an enzyme liquefied liquid is used, a saccharification reaction liquid containing high inmaltose and a low glucose value can be obtained compared to an acid liquefied liquid. The starch used may be corn starch, potato starch, sweet potato starch, tapioca starch, etc., and there is no restriction as long as it is a starch commonly used for saccharification. The important thing is that the DE set in advance
(glucose) and the reaction by glucoamylase is stopped when the sugar composition is reached. Don't stop! As the reaction progresses, the amount of glucose increases. To stop the reaction, the saccharification reaction solution is heated at 80°C for about 40 minutes or at 85°C for 5 minutes. Thereafter, the saccharification reaction solution is passed through the mouth, subjected to ion exchange purification, activated carbon treatment, and concentrated to obtain a high conversion syrup containing isomaltose. The ingredients obtained by this are 10% to 60% glucose content and 10% isomaltose content per solid.
% to 60%, isomaltooligosaccharide content other than isomaltose is 10% to 50%, maltooligosaccharide content is 5% to 10%, and the total isomaltooligosaccharide content including isomaltose is 50% to 60%. Here, isomalto-oligosaccharides other than isomaltose include panose, isomaltotriose, and oligosaccharides having a glucose degree of polymerization (DP) of 4 or more having α-1,6 bonds; examples of malto-oligosaccharides include maltose and maltotriose. and α-1
It is an oligomer with a glucose polymerization degree (DP) of 4 or more and has 4 bonds.

(Manufacture of high-purity isomalto-oligosaccharide syrup by chromatographic separation) Chromatographic separation is an excellent method for separating high-conversion syrup containing isomaltose in large quantities at low cost, and strong acid cation exchange resin is used as a filler. is suitable. The strongly acidic cation exchange resin used in the present invention is of the sulfonic acid type, such as Diaion 5KIB.

DIAIO/5K102, DIAION 5K104, DIAION 5K106, DIAIO/5K110, DIAION 5K112, DIAIO/5K116, DIAION FRLI 1 (all product names manufactured by Mitsubishi Kasei) or DOWEX 50WX2, DOWEX 50WX4
, DOWEX 50WX8, DOWEX 88 (all product names manufactured by Dow Chemical) or Amberlite IR-
116, Amber Light IR-118, Amber 2 Light IR-120B.

Amber Light XT1022E, Amber Light XT4
71E.

Amberly) G102 (all product names, manufactured by Organo)
etc. are suitable, and are used as alkali metal type or alkaline earth metal type before use. The separation device consists of a stock solution tank, developing solvent tank, liquid pump, separation tower, detector, separation division fraction mechanism, and control mechanism, and efficiently separates isomalto-oligosaccharides including glucose and inmaltose. The operations for this include: undiluted sugar concentration of 50-80%;
Preferable conditions are a temperature of 30 to 80° C. and a liquid passing rate of SV of 0.01 to to. The separated high-purity isomalto-oligosaccharide (e) is separated by mouth filtration, ion exchange purification, activated carbon treatment, and concentration to obtain high-purity isomalto-oligosaccharide syrup.

(Production of powdered high-purity isomaltooligosaccharides by spray drying) Powdered high-purity isomalto-oligosaccharides can be produced by spray-drying high-purity isomaltooligosaccharide syrup. The spray-drying conditions for high-purity isomalto-oligosaccharide syrup are: (1) The high-purity isomalto-oligosaccharide syrup is pumped or sprayed; and (K) it has sufficient fluidity in a liquid state. This fluidity depends on the sugar solid content and product temperature, but the sugar solid content is 40-80% and the product temperature is 40-80℃.
and maintain the viscosity between 10 and 500 cp. In this case, the high-purity isomalto-oligosaccharide syrup does not need to be in a semi-condensed fluid state in which fine crystals are precipitated, and is characterized in that it can be sprayed in a liquid state. ■It is suitable to instantly produce high-purity isomalto-oligosaccharide syrup using a micro-particle spray method.
The rotation speed is 10.000 to 40.00 Or, p,
It is to maintain m and k. ■Atomized droplets of highly pure isomalto-oligosaccharide syrup come into instant contact with hot air,
The evaporated air is turned into powder, but this hot air is distributed unevenly at the inlets 40 to 9.
The temperature should be kept at 0°C and the outlet temperature between 20 and 60°C. ■The powdered particles are spherical, porous, fluid,
Good solubility and dispersibility.

Examples of the present invention are shown below, but the present invention is not limited to these examples.

〔Example〕

Example 1 1. Preparation of high-conversion syrup containing isomaltose
-Amylase (manufactured by Novo Industries, 'Fa/Gamil 800L) 0.02% (based on solids, the same applies below), glucoaminase (manufactured by Novo Industries, AMG 300)
L) 0.01%, transferase (manufactured by Ohno Pharmaceutical Co., Ltd., transglucosidase) 0.03%, debranching enzyme (manufactured by Novo Industries, prosozyme 200L) 0.02-0. After carrying out the saccharification reaction for an hour,
Hyphenperjotriose containing isomaltose was heated at 85°C for 5 minutes, passed through the mouth, purified by ion exchange, treated with activated carbon, and concentrated. Isomaltooligosaccharides include panose, isomaltotriose and d-1,6
It is an oligosaccharide with a glucose polymerization degree (DP) of 4 or more that has a bond.

Analyzer High performance liquid chromatography [Model] Waters model 590 Shimadzu LCJA model [Kara A J
SCR101NC Island iQt) Zorbax N
H2 (Dubo/) [Solvent] Water
Preparation of high-purity isomalto-oligosaccharide syrup by chromatography using acetonitrile:water = 70:502. The inmaltose-containing high conversion syrup obtained in step 1 was diluted to a predetermined concentration and transferred to a strongly acidic cation exchange resin column. (8) Category containing 90% or more of glucose per inverted form, (8) 60% or more of isomalto-oligosaccharides including diisomaltose per solid.
C) a category containing less than 80% isomalto-oligosaccharides including isomaltose per solid.

(4) is the segment to be removed from the product, and (8) and 0 are the segments to be used as the target product.High purity isomalto-oligosaccharide syrup was prepared by filtration, ion exchange purification, activated carbon treatment, and concentration.

(Separation conditions) Packing agent: Diaion FRO1 (Na type) (manufactured by Mitsubishi Kasei Corporation) Separation column: 40m x 2000mH (resin [2t) Sugar load: 1549/BX65 (1) Separation temperature: 65°C Liquid passing rate t: SVα1 Other components include maltotriose as a maltooligosaccharide, panose, isomaltotriose as an isomaltooligosaccharide, and an oligosaccharide with a glucose degree of polymerization (DP) of 4 or more having a tt -1,6 bond. It is.

Analyzer High performance liquid chromatography [Neizan] Waters type 590 Shimadzu LCdA type [column] 5CR
101N (Shimadzu) Z orbax NH2 (DuPont) [Solvent] Water Acetonitrile:Water = 70:303, l! A: Preparation of powdered high-purity isomalto-oligosaccharide by mist drying The high-purity isomalto-oligosaccharide obtained in step 2 was diluted to a predetermined degree of quality, and the drying chamber diameter was 2,000 m, the height of the drying chamber was 4,
Spray drying was carried out using a 900+ alt spray dryer under the following conditions.

(Sugar liquid conditions) Moisture 62%, solid content 68%, temperature 50°C, stiffness 60-7
0 cp (Drying conditions) Spraying method: High-speed rotating disc Rotation speed: 24.00 Or, = 20 odor/H for p-1 treatment (Powdered sugar produced by spray drying) Isomalto-oligosaccharides including inmaltose A spherical and porous powdery high-purity isomalto-oligosaccharide was prepared by spray-drying a high-purity isomalto-oligosaccharide syrup containing 60% or more and less than 80% and containing inmaltose at 80% or more. .

〔Effect of the invention〕

The present invention uses high-purity isomalto-oligosaccharide containing 60% or more and less than 80% of isomalto-oligosaccharides including isomaltose per solid and high-purity isomalto-oligosaccharides containing 80 or more isomalto-oligosaccharides including inmaltose per solid. Alternatively, it provides an industrial manufacturing method for producing a large iK in powder form.

Conventional high-conversion syrups containing isomaltose contain less than 60 grams of isomalto-oligosaccharides, including diisomaltose, per solid state. By carrying out the process and, if necessary, spray-drying, it becomes possible to efficiently and economically produce high-purity isomalto-oligosaccharide in the form of syrup or powder.

This high-purity isomalto-oligosaccharide contains large amounts of isomaltose, isomaltotriose, panose, isomaltotetraose, etc., which were previously difficult to produce on an industrial scale, and has a mellow sweetness and good taste. Due to its hygroscopicity and indigestibility, it has various physiological functions and can be used in many fields as a flavoring agent and functional sugar.

Claims (2)

[Claims] (1) High conversion syrup containing 10% to 60% glucose, 10% to 60% isomaltose, and 10% to 50% isomaltooligosaccharides other than isomaltose per solid is passed through a strongly acidic cation exchange resin. The segment containing 90% or more of glucose per solid is separated, and the segment containing 60% or more and less than 80% of isomaltooligosaccharide including isomaltose per solid, and the segment containing 80% or more of isomalto-oligosaccharide including isomaltose per solid. % or more. (2) The method for producing high-purity isomaltooligosaccharide according to claim 1, wherein the high-purity isomalto-oligosaccharide is in the form of syrup or powder.