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A novel method for directional preparation of endo-inulinase

A technology for endo-inulinase and inulinase is applied in the directions of biochemical equipment and methods, enzymes, hydrolase, etc., to achieve the effects of improving yield, reducing monosaccharide content, and low-cost production methods

Active Publication Date: 2019-08-06
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method is a two-step process to obtain a precipitated protein containing endo-inulinase activity. Although the I / S ratio is high, the precipitated protein still contains exo-inulinase activity

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Add the aqueous solution of 50wt% ethanol in the conical flask, add inulin, make the inulin concentration reach 80g / L; 0.1mol / L hydrochloric acid adjusts the pH value to be 5.0; mL endo-inulinase activity and 10.63U / mL exo-inulinase activity), after standing at 0-5°C, centrifuge, remove the supernatant to obtain a milky white solid, redissolve the milky white solid with distilled water To 6.5mL, the enzyme activity in the reconstituted solution was determined by the DNS method. After measurement, the enzyme activity of endo-inulinase is 8.22U / mL, the enzyme activity of exo-inulinase is 0U / mL, only endo-inulinase remains in the inulinase enzyme system, and the recovery of endo-inulinase rate reached 88%.

Embodiment 2

[0031]Add the aqueous solution of 70wt% ethanol in the conical flask, add inulin, make the inulin concentration reach 20g / L; 0.1mol / L hydrochloric acid adjusts the pH value to be 4.0; mL endo-inulinase activity and 10.63U / mL exo-inulinase activity), after standing at 0-5°C, centrifuge, remove the supernatant to obtain a milky white solid, redissolve the milky white solid with distilled water to 1.6mL, the enzyme activity in the reconstituted solution was determined by the DNS method. After measurement, the enzyme activity of endo-inulinase is 8.04U / mL, the enzyme activity of exo-inulinase is 0U / mL, only endo-inulinase remains in the inulinase enzyme system, and the recovery of endo-inulinase rate reached 86%.

Embodiment 3

[0033] Add the aqueous solution of 60wt% ethanol in the conical flask, add inulin, make the inulin concentration reach 50g / L; 0.1mol / L hydrochloric acid adjusts the pH value to be 4.6; Then add natural inulinase enzyme liquid 4mL (comprising 9.34U / mL Endo-inulinase activity and 10.63U / mL exo-inulinase activity), after standing at 0-5°C, centrifuge, remove the supernatant, and obtain a milky white solid, redissolve the milky white solid with distilled water to 4mL, the enzyme activity in the reconstituted solution was determined by DNS method. After measurement, the enzyme activity of endo-inulinase is 8.51U / mL, the enzyme activity of exo-inulinase is 0U / mL, only endo-inulinase remains in the inulinase enzyme system, and the recovery of endo-inulinase The rate reached 91%.

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Abstract

The invention relates to a novel method for directional preparation of endoinulinase. The method is characterized by: dissolving inulin in an aqueous solution of ethanol, fully mixing the substances evenly, adjusting he pH value of the solution to 4.0-5.0, adding microorganism produced natural inulase, performing standing, then conducting centrifugation to obtain a solid, and adding distilled water to redissolve the solid, thus obtaining an enzyme liquid only containing endoinulinase activity. The method provided by the invention can efficiently split endoinulinase and exoinulinase in natural inulase system, can achieve directional preparation of enzyme liquid only containing endoinulinase activity, and can be used for preparation of fructooligosaccharide by hydrolysis of inulin. The method provided by the invention greatly improves the yield for enzymatic preparation of fructooligosaccharide, reduces the content of monosaccharide without specific physiological activity to human body in the product, and provides a high efficiency and low cost production method for preparation of fructooligosaccharide from microorganism produced natural inulase.

Description

technical field [0001] The invention relates to the field of oligosaccharides produced by microorganisms in bioengineering, in particular to a novel method for directional preparation of endo-inulinase. Background technique [0002] Inulin, also known as inulin, is widely distributed in nature, and mostly exists in plants as sugar storage materials, such as Jerusalem artichoke, chicory, garlic, onion, etc. It is a plant polysaccharide and a polymer of D-fructose. D-fructose is a chain polysaccharide linked by β(2-1) glycosidic bonds, and D-glucose is linked by β(1-2) bonds at the end of the molecule. The molecular formula of inulin has a GFn structure (G: glucosyl, F: fructosyl, n: number of units of fructosyl). The degree of polymerization of inulin ranges from 2 to 60, and the average molecular weight is around 5500. Inulin is slightly soluble in cold water, can dissolve quickly in hot water, does not show color reaction with iodine, has no reduction, has optical activit...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/24C12P19/14
CPCC12N9/2402C12P19/14C12Y302/01007
Inventor 李鑫王伟勇强赖晨欢周瑾
Owner NANJING FORESTRY UNIV