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Method for preparing isomaltulose by polyvinyl alcohol immobilized sucrose isomerase producing bacteria

A technology of sucrose isomerase and isomaltulose is applied in the field of functional sugar production to achieve the effects of reducing pollution, less immobilization operation procedures and high strength

Inactive Publication Date: 2013-04-17
广西科学院生物研究所有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0015] The purpose of the present invention is to: aim at the shortcomings of the current sodium alginate immobilization method, to provide a simple and fast operation, high strength of immobilized bacteria, easy preservation of immobilized bacteria, less loss of enzyme activity and no pollution and waste. The enzyme carrier polyvinyl alcohol immobilizes the bacteria, and provides a method to decompose the residual sucrose in the conversion solution by producing sucrose isomerase, effectively reducing the sucrose content in the isomaltulose obtained after crystallization to prepare high-quality isomaltulose method

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] In a 30-liter fermenter, add 22 liters of fermentation medium, insert 2 liters of strain Protaminobacter rubrum (CBS574.77) after sterilization, medium sucrose 4% (W / V), yeast extract 2% (W / V ) yeast extract 0.4% (W / V), (NH 4 ) 2 HPO 4 0.1% (W / V), the fermentation temperature is 30°C, the pH value is 6.5, and the aeration rate is 0.8V / V.m Stirring for fermentation culture, the stirring speed is 300rpm, after 48 hours of fermentation, filter with a 0.2μm ceramic membrane Collect the cells to obtain about 1 liter of cell suspension with a wet cell concentration of 30%. After boiling 1.6 liters of water, add 0.45 kg of polyvinyl alcohol, stir, continue to heat until the polyvinyl alcohol is fully dissolved, then add 0.23 kg of diatomaceous earth and stir evenly, cool to 30°C, add the cell suspension, stir well, Place in -20°C freezer for 24 hours. When it is time to use, take out the immobilized bacteria and thaw naturally, cut into particles with a diameter of about 3...

Embodiment 2

[0034] In a 30-liter fermenter, use Erwinia rhapontici (NCPPB 1578) as the fermentation strain, obtain the immobilized thalline by the fermentation process and the immobilization process of Example 1, and put the immobilized thallus into 30 liters of batch type In the reactor, add 20 liters of concentration and be 35% (W / V) sucrose solution, at 26.5 DEG C, pH is 5.5 conditions stirring conversion, when the sucrose amount in the conversion solution is less than 2.6%, stop conversion, add new sucrose solution Carry out conversion, collect two batches of conversion liquids and altogether 40 liters, the conversion liquid is poured into the container and is heated up to 45 ℃, adds 0.3 gram of Erwinia rhapontici (NCPPB 1578) enzyme, after reacting for 5 hours, after detection, at this time by Erwinia rhapontici (NCPPB 1578 ) after enzymatic decomposition, the conversion liquid components (W / V) are 77.5% isomaltulose, 16.5% isotrehalose, 2.3% glucose, 2.6% fructose and 0.8% others. A...

Embodiment 3

[0036] In a 30-liter fermenter, use Serratia phymuthica (ATCC 15928) as the fermentation strain, obtain the immobilized thallus by the fermentation process and the immobilization process of Example 1, and put the immobilized thallus into 30 liters of batch type In the reactor, add 20 liters of concentration and be 35% (W / V) sucrose solution, at 30 DEG C, pH is 5.5 conditions stirring conversion, when the sucrose amount in the conversion solution is less than 2.6%, stop conversion, add new sucrose solution Carry out transformation, collect two batches of transformation liquid, a total of 40 liters, pour the transformation liquid into a container and heat up to 55°C, add 0.25 grams of sucrose isomerase, and react for 3.5 hours. After detection, it is decomposed by sucrose isomerase The conversion solution components (W / V) are 77.3% isomaltulose, 14.5% isotrehalose, 2.1% glucose, 3.2% fructose and 0.9% others. After decolorization and filtration, separation, concentration and cry...

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Abstract

The invention discloses a method for preparing isomaltulose by polyvinyl alcohol immobilized sucrose isomerase producing bacteria. The polyvinyl alcohol immobilized sucrose isomerase producing bacteria are obtained by adding 5 to 30 weight percent of sucrose isomerase producing wet bacteria into 4 to 50 percent of water solution pulp of an enzyme carrier which is polyvinyl alcohol or / and kieselguhr at 25 to 30 DEG C and uniformly stirring and hardening at -20 DEG C for later use, wherein the ratio of polyvinyl alcohol to kieselguhr is 2:(0.8-1.21). When used, the immobilized enzyme producing bacteria are taken out, naturally unfrozen at room temperature, cut into grains and added into sucrose solution to perform isomerization conversion; then produced sucrose isomerase is used to decompose sucrose remaining in conversion solution; the conversion solution is purified by decolorizing and ion exchange; and an isomaltulose product is obtained by concentration crystallization and centrifugal separation. The method has the advantages that: the operation is simple and efficiency; the material is cheap and readily available; the grains of immobilized bacteria have high strength and are insusceptible to breakage; the activity of sucrose isomerase is high; the service life of the sucrose isomerase is long; and the like. The isomaltulose has high quality, the purity of the isomaltulose is over 98 percent, the sucrose residue content is less than 0.1, and the isomaltulose is more suitable for preparing high-quality isomalt by hydrogenation.

Description

technical field [0001] The invention belongs to the field of functional sugar production and relates to a method for preparing isomaltulose by immobilizing sucrose isomerase-producing bacteria with polyvinyl alcohol. Background technique [0002] Isomaltulose, also known as Palatinose, is a reducing disaccharide, which was first discovered by Weidenhagen et al. in sugar beet production in 1957. The industrialized production of isomaltulose is based on sucrose as raw material, isomerized by sucrose isomerase (also known as α-glucosyltransferase) to form isomaltulose syrup, then decolorized and purified, concentrated and crystallized, centrifuged and dried. get. Isomaltulose has similar sweetness properties to sucrose, its sweetness is about half that of sucrose, without any off-flavor. Compared with sucrose, the outstanding advantages of isomaltulose are: (1) low cariogenic properties; (2) suitable for diabetics. As a promising functional sweetener, isomaltulose has been w...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N11/14C12R1/01C12R1/18C12R1/425
Inventor 蒋永强卢汉浪唐峰陈业良
Owner 广西科学院生物研究所有限责任公司
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