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Aspergillus and application of aspergillus in fructosyltransferase preparing

A fructosyl transferase, Aspergillus technology, applied in the direction of transferase, immobilized on/in organic carrier, microorganism-based method, etc., can solve the narrow application range of fructo-oligosaccharides, and there are not many reports on fructosyl transferase research. , No bacterial species and other problems, to achieve the effect of promoting effective utilization

Active Publication Date: 2014-09-24
广西多得乐生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, restricted by the price of fructo-oligosaccharide and sucrose and the production cost, the scope of application of fructo-oligosaccharide is relatively narrow, the awareness of the public is not high, and the goal of benefiting the public with fructo-oligosaccharide has not yet been realized
[0010] There are many domestic research reports on fructooligosaccharides, but there are not many research reports on fructosyltransferase
In addition to the research on fructosyltransferase by the above-mentioned several units, although Jiangmen Quantum High-Tech, Shandong Baolingbao, Wuhan Shengshi Tianyuan and other units currently produce fructo-oligosaccharides, none of the above-mentioned enterprises have strains, but purchased enzyme preparations
And most of the strains are intracellular enzymes, which increases the cost of enzyme separation and purification

Method used

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  • Aspergillus and application of aspergillus in fructosyltransferase preparing

Examples

Experimental program
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Effect test

preparation example Construction

[0072] Preparation of basic medium: 30g sucrose, 1.5g yeast powder, 1.5g peptone, NaNO 3 3.0g, KH 2 PO 4 1.0g, MgSO 4 ·7H 2 O0.5g, KCl0.5g, FeSO 4 ·7H 2 0.01g, sodium carboxymethyl cellulose 2g, dissolves with 800mL deionized water, then adjusts the pH value to 4.0-6.0 (pH is 6.0 in the following examples) with concentrated hydrochloric acid or NaOH solution, constant volume with deionized water To 1L, sterilize with damp heat at 115°C for 30min.

[0073] Preparation of citric acid-disodium hydrogen phosphate buffer solution at pH 6.0: 7.71 ml of 0.2M Na 2 HPO 4 The aqueous solution was mixed with 12.29 ml of 0.1M aqueous citric acid solution.

[0074] In Example 1 and Example 2, the fructosyltransferase activity was measured using 3,5-dinitrosalicylic acid (3,5-dinitrosalicylic acid, DNS) method to measure the reducing sugar produced (reference "Miller GL. Use of dinitrosalicylic acid reagent for determination of reducing sugar.Anal Chem1959,31:426-428), the specific...

Embodiment 1

[0080] Example 1, the isolation and identification of Aspergillus aculeatus strain M105

[0081] 1. Acquisition of strains

[0082] 1. Isolation and screening of strains producing fructosyltransferase

[0083] (1) Use deionized water to prepare the separation medium (Chas' solid medium); every liter of separation medium contains: sucrose 30g, NaNO 3 3.0g, KH 2 PO 4 1.0g, MgSO 4 ·7H 2 O0.5g, KCl0.5g, FeSO 4 ·7H 2 O0.01g, 2g sodium carboxymethylcellulose, 15g agar, natural pH, 115°C damp heat sterilization for 30min; after sterilization, cool to 50°C and pour plate.

[0084] (2) Take 2 μL of each of the 29 strains of Aspergillus strains stored in the laboratory and spread them on the plate prepared in step (1), and incubate them upside down in a constant temperature incubator at 28°C for 6 days.

[0085] (3) Prepare Chapei culture medium (liquid) with pH 6.0, the method is the same as step (1), without adding agar, adjust the pH to 6.0.

[0086] (4) Inoculate the fungal...

Embodiment 2

[0097] Embodiment 2, utilize Aspergillus strain M105 to prepare fructosyltransferase

[0098] 1. Obtaining culture medium

[0099] 1. Preparation of spore solution

[0100] (1) Sterilize the PDA medium (solid) at 115° C. for 30 minutes.

[0101] (2) the spores of Aspergillus aculeatus M105 obtained by embodiment 1 that were subcultured and activated for 6 days on the PDA plate were washed with sterile water to make spore suspension, and the spore concentration was 1 × 10 8 individual / mL.

[0102] 2. Preparation of crude enzyme solution

[0103] (1) Take 150mL basic culture base in a 500mL Erlenmeyer flask.

[0104] (2) Get the spore liquid of Aspergillus aculeatus M105 and inoculate it into the basic medium according to the inoculum amount (volume percentage) of 1%, and cultivate it for 6 days at 28°C and 180rpm (26-32°C, 180rpm-200rpm, cultivate 5 -7 days), collect the culture.

[0105] (3) Centrifuge the culture, 4°C, centrifugal radius 7cm, 3500rpm, centrifuge the cul...

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Abstract

The invention discloses aspergillus and an application of aspergillus in fructosyltransferase preparing. The preservation number of the aspergillus is CGMCC No.8912. The aspergillus aculeatus bacterial strain is a bacterial strain for producing the high-activity and ectoenzyme fructosyltransferase, the bacterial strain can be applied to producing fructo-oligosaccharide with saccharose or molasses as raw materials, and the aspergillus plays a significant role in promoting deep processing of saccharose and effective utilizing of waste.

Description

technical field [0001] The invention relates to an aspergillus strain and its application in the preparation of fructosyltransferase, which belongs to the field of biotechnology. Background technique [0002] Fructose oligose refers to the fructose oligomers kestose, kestose, and kestose formed by connecting 1-3 fructose molecules on the fructose residue of the sucrose molecule through a β-(2-1) glycosidic bond. Sugar and its mixture (Clevenger M A, Turnbull D, Inone H. Toxicological evaluation of neosugar: genotoxicity, carcinogenicity and chronic toxicity. Int J Toxicol, 1988, 7: 643–662), because fructooligosaccharides have the ability to promote beneficial bacteria in the human gut The proliferation of Bifidobacterium, lowering blood cholesterol, improving blood lipids, promoting calcium absorption, low calorie, low dental caries and other special physiological effects have become the fastest growing health food ingredients in the market in recent years. [0003] Althou...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N9/10C12N11/10C12N11/04C12P19/18C12R1/66
Inventor 冯家勋黄妹平徐强胜伍敏莫德姣
Owner 广西多得乐生物科技有限公司
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