Rhubarb horsetails Erwinia sp. and its application in preparation of isomaltulose

A technology of isomaltulose and Erwinia, applied in the fields of fermentation engineering and enzyme engineering

Inactive Publication Date: 2010-09-29
NANJING TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the above-mentioned bacteria can convert sucrose into isomaltulose, the yield is very unstable, and the conversion rate is 8-86%.
Moreover, in the process of catalyzing the conversion of sucrose by these isomaltulose-producing strains, in addition to the main product, trehalulose, isomaltose, isomelezitose, and by-products of glucose and fructose produced by the hydrolysis of sucrose also exist in the enzymatic conversion liquid. product, the product specificity is not high, on the one hand reduces the yield of the target product, on the other hand makes the downstream process very difficult, the production cost increases greatly

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Slant medium: peptone 10g / L, beef extract 3g / L, NaCl 5g / L, agar 20g / L, pH 7.0.

[0037] Shake flask medium: sucrose 50g / L, yeast extract 10g / L, Na 3 PO 4 ·12H 2 O 5g / L, pH7.0.

[0038] Erwinia rhubarb NX-5 CGMCC No.2222 was cultured on a slant medium at 30°C for 24h, then a loop of this bacteria was placed in a shake flask medium, cultured at 30°C for 10h, and the shake flask speed was 200r / min. The bacterial content in the solution is 20g / L, the enzyme production reaches 2.5-5.0U / ml, and it is transformed with free cells. Weigh 10g of cells and add them to 100ml of 450g / L sucrose solution (the amount of cells added is 1L55% per 100g of cells). sucrose solution), under the condition of 30 ℃, shake and transform to produce isomaltulose in a shaker flask, the reaction time is 8h, and the substrate conversion rate and product concentration are measured after the reaction is terminated. The conversion rate of sucrose is 99.5%, and the conversion rate of isomaltulose is ...

Embodiment 2

[0040] With 50g / L maltose as carbon source, 5g / L bean cake flour as nitrogen source, other components of medium and bacterial culture conditions are the same as in Example 1, prepare 1.5% sodium alginate colloidal solution, and the bacterial suspension prepared with physiological saline After mixing evenly, add 8% diatomaceous earth for adsorption, drop 2% CaCl with a syringe 2 It was solidified in the solution to make spherical immobilized cells with a diameter of about 3-4 mm, which were placed in a refrigerator at 4 °C for several hours, then the supernatant was poured out, washed twice with distilled water, and 10 g of wet cells were wrapped in sodium alginate. 20 g of immobilized cells were obtained by burying, and 100 ml of 550 g / L sucrose solution was transformed with this in a shaker flask at 30 °C. The transformation time of each batch was 10 h, and 40 batches were transformed. The average conversion rate of sucrose was 99.5%. The average conversion of maltulose was 8...

Embodiment 3

[0042] 225g of glucose, 45g of corn steep liquor, 22.5g of NaCl, pH 6.0 were made up to 4.5L with water to form a culture medium, put into a 7.5L glass stirring fermenter, and steam sterilized at 121°C for 15min. Preparation of seed medium: sucrose 50g / L, yeast extract 10g / L, Na 2 HPO 4 ·12H 2 O 5g / L, pH 6.0. Connect Erwinia rhubarb NX-5 CGMCC No.2222 to the seed medium, cultivate at 30°C for 8 hours to obtain the seed liquid, and insert the seed liquid into the cooled fermentation medium according to the inoculum of 3% of the volume of the fermentation liquid. , 30 ℃ culture (ventilation volume 1vvm, stirring speed of 600r / min) 12h. The fermentation broth is filtered bacteria in an ultrafilter, and the ultrafiltration membrane cut-off molecular weight is 10 6 Dalton, the operating pressure is 0.2MPa, the control temperature is 50°C, and the membrane surface speed is 4m / s. After the 90g cells obtained by the interception are immobilized according to the method of Example 2...

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Abstract

The invention discloses an Erwiniarhapontici NX-5 and a method of producing sucrose isomerase for the preparation of isomaltulose by the Erwiniarhapontici. The strain CGMCC No.2222 is inoculated in an aseptic culture medium containing carbon source, nitrogen source, inorganic salt and water for the aerobic culture. Cells containing the sucrose isomerase are obtained after the centrifugation or the ultrafiltration. Furthermore, dissociative cells containing the sucrose isomerase are directly adopted to be converted into sucrose to generate the isomaltulose or immobilized cells containing the sucrose isomerase which are then converted into the sucrose isomerase to generate the isomaltulose. With the strain adopted, the conversion rate of the sucrose reaches to 99.5 percent (w / w), the conversion rate of the isomaltulose reaches to 90 percent, and the concentration of the isomaltulose in the conversion solution reaches to 500g / L. no hydrolysis side effects happen. Almost no glucose and fructose are contained in the conversion solution. The product, the isomaltulose, can not be converted into other components with the reaction. The invention is beneficial to the industrial production of the isomaltulose.

Description

technical field [0001] The invention belongs to the technical fields of fermentation engineering and enzyme engineering, and relates to a microbial strain Erwinia rhapontici NX-5 producing sucrose isomerase and its use in the production of isomaltulose. Background technique [0002] Isomaltulose (6-O-α-D-glucopyranosyl-D-fructose, Isomaltulose) is a reducing sugar that exists in small amounts in natural honey and beet juice, and has the most similar physical properties to sucrose and taste. As a substitute for sucrose, it has the following advantages: (1) Since it releases monosaccharides in the blood slower than sucrose and does not stimulate insulin secretion after being eaten by the human body, it can be eaten by diabetics and people who lose weight; The oral streptococcus that causes dental caries metabolizes and does not corrode teeth; (3) In the microflora of the human intestine, it selectively stimulates the growth of bifidobacteria; (4) It has high food safety perfo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P19/12C12R1/18
Inventor 徐虹李莎环民霞林璐蔡恒欧阳平凯
Owner NANJING TECH UNIV
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