Method for producing fructooligosaccharide by using transfructosylase

A technology of fructosyltransferase and fructooligosaccharides, applied in the field of fructooligosaccharides production, can solve the problems of unfavorable fructooligosaccharides accumulation, production limitation, low activity, etc.

Inactive Publication Date: 2011-07-20
山东文远生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Most of the enzyme-producing bacteria used in industrial production are molds, such as Aspergillus niger ACTT20611, Aspergillus foetidus NRRL4337, Aspergillus japonicus TITKJ1, etc. Many plants, yeast and molds can produce fructosyltransferase, but the activity of the enzyme produced by plants is low and the yield is low. Restricted by season; and yeast invertase activity is strong but not conducive to the accumulation of fructooligosaccharides

Method used

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  • Method for producing fructooligosaccharide by using transfructosylase
  • Method for producing fructooligosaccharide by using transfructosylase

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Experimental program
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Embodiment 1

[0021] The Penicillium spp. was inserted into a fermentation tank containing 5% sucrose, 3.5% soybean meal, 0.5% corn meal culture solution 50L, culture conditions were 30℃, PH5.20, aeration rate 200ml / min, stirring speed 150r / min, After 20 hours of fermentation, a large amount of mycelium was obtained by centrifugal separation, and the enzyme activity was detected by HPLC. The mycelium was put into the enzyme reaction tank of 45% sucrose solution according to the enzyme amount of 3500u / kg sucrose, and the reaction temperature was controlled at 30℃, PH5. 2±0.2, react for 24 hours, the content of oligofructose reached above 50% by HPLC, the reaction solution is separated by ceramic microfiltration membrane to separate the mycelium to obtain a transparent light yellow oligofructose crude liquid, which is decolorized by macroporous resin and concentrated Fructooligosaccharides with a chromaticity of less than 0.05 are obtained.

Embodiment 2

[0023] The penicillium species was connected to a fermentation tank containing 8% sucrose, 4.0% soybean meal, 1.5% corn meal culture solution 5000L, the culture condition was 32℃, the aeration rate was 500ml / min, the stirring speed was 120r / min, and the fermentation was carried out for 24 hours. , Centrifugal separation to obtain a large number of mycelium, detect the enzyme activity by HPLC, put the mycelium into the enzyme reaction tank of 50% sucrose solution according to the enzyme amount of 4500u / kg sucrose, control the reaction temperature of 31 ℃, pH 6.0 ± 0.2, After reacting for 20 hours, the content of fructo-oligosaccharides reached more than 50% by HPLC. The reaction liquid was separated by ceramic microfiltration membrane to obtain a transparent light yellow fructo-oligosaccharide crude liquid, which was decolorized by macroporous resin and concentrated to obtain a color less than 0.05 fructooligosaccharide.

Embodiment 3

[0025] The Penicillium spp. strain was inserted into a fermenter containing 20% ​​potato, 3% sucrose, 0.2% potassium dihydrogen phosphate, 0.3% magnesium sulfate, 0.3% sodium nitrate, 0.2% agar culture solution 50L, and the culture condition was 27°C. The aeration rate is 300ml / min, the stirring speed is 100r / min, after 50 hours of fermentation, a large amount of mycelium is obtained by centrifugal separation, and the enzyme activity is detected by HPLC. The mycelium is poured into a 40% sucrose solution at an enzyme amount of 3000u / kg. Enzyme reaction tank, control the reaction temperature at 27℃, pH 6.3±0.2, react for 30 hours, detect the oligofructose content by HPLC to reach more than 50%, the reaction liquid is separated by ceramic microfiltration membrane to separate the mycelium, and the transparent light yellow is obtained. The crude polyfructose liquid is decolorized by a macroporous resin and concentrated to obtain fructooligosaccharides with a color less than 0.05.

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Abstract

The invention relates to the technical field of food biotechnology, particularly a method for producing fructooligosaccharide by using transfructosylase, which comprises preparation of transfructosylase and industrial production of fructooligosaccharide by using the enzyme preparation. The method is characterized by comprising the following steps: selecting a good strain containing high-activity transfructosylase, culturing the strain in an appropriate culture medium, separating to obtain large-scale cultured mycelia containing high-activity transfructosylase, and storing the mycelia for later use; and converting sucrose into fructooligosaccharide by a batch process by using the mycelia containing high-activity transfructosylase as a biocatalyst. The method provided by the invention has the advantages of short bioconversion time, and high conversion rate of fructooligosaccharide; the mycelia can be repeatedly used; and the content of fructooligosaccharide (trisaccharide) prepared by using the transfructosylase is up to 57-62%, which is 3-5% higher than that of the fructooligosaccharide prepared by using Aspergillus niger.

Description

(1) Technical field [0001] The invention relates to the field of food biotechnology, in particular to a method for producing fructooligosaccharides by using fructosyl transferase. (2) Background technology [0002] Fructooligosaccharide is a water-soluble dietary fiber. Long-term use can reduce serum cholesterol and improve lipid metabolism. Experiments on animals and humans have confirmed that oligofructose has the following physiological functions: it is used by beneficial bacteria such as bifidobacteria. Bifidobacterium has a two-way regulation effect when it proliferates 10-100 times. After the human body ingests oligofructose, the number of beneficial bacteria Bifidobacterium in the body increases, which can inhibit the growth and reproduction of exogenous pathogenic bacteria and inherent intestinal spoilage bacteria such as Salmonella, reduce the growth and accumulation of intestinal spoilage substances, and promote the intestinal tract Peristalsis to prevent constipation ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/04
Inventor 赵素亮赵洪涛
Owner 山东文远生物技术有限公司
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