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Aspergillus niger-spores for converting glucose to produce gluconic acid and preparation and application method of aspergillus niger-spores

A technology of Aspergillus niger spores and gluconic acid is applied in biochemical equipment and methods, methods of using spores, methods based on microorganisms, etc., to achieve the effects of simple preparation method, simple composition and simple process

Inactive Publication Date: 2016-04-13
CENTRAL SOUTH UNIVERSITY OF FORESTRY AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to overcome the shortcomings of the existing methods and provide a biocatalyst Aspergillus niger spore that is not easily inactivated and easily separated in the reaction of converting glucose to produce gluconic acid, and its preparation and application method

Method used

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  • Aspergillus niger-spores for converting glucose to produce gluconic acid and preparation and application method of aspergillus niger-spores
  • Aspergillus niger-spores for converting glucose to produce gluconic acid and preparation and application method of aspergillus niger-spores

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Embodiment 1

[0030] The preparation of embodiment 1 relevant culture medium

[0031] 1) PDA medium Wash the potatoes, peel them, cut them into small pieces, then weigh 200g, add water and boil until rotten (boil for 20-30min, it can be pierced by a glass rod), filter with eight layers of gauze, heat, Then add 15g of agar according to the actual experiment needs, continue heating and stirring to mix evenly, after the agar is dissolved, add glucose, stir evenly, and then make up water to 1000mL after cooling down a bit, divide into test tubes or Erlenmeyer flasks, add stoppers, bandage, 121°C After sterilizing for about 20 minutes, take out the test tube and place it on an inclined plane to obtain the PDA medium, cool it and store it for later use.

[0032] 2) Buckwheat seed culture medium Buckwheat seeds (300g) are washed with deionized water, boiled with an equal amount of deionized water, softened in a boiling water bath (100°C, 15min), poured off excess water, and packed in conical In t...

Embodiment 2

[0033] Example 2 Acquisition of Aspergillus niger spores (derived from Aspergillus niger ATCC9029, purchased from the American type culture integration library, but not limited to this strain)

[0034] Aspergillus niger was inoculated on the PDA medium slant, and cultivated for 7 days at 30°C; the Aspergillus niger PDA culture slant was washed with sterilized deionized water containing 0.1% Tween-80 (or Tween-20), to obtain the Aspergillus niger spore suspension, and Nephelometric method adjusted to a concentration of 10 8 spores / mL of the spore suspension, which was inoculated in the buckwheat seed medium (the inoculum size was 10 8 spores / 100g buckwheat seeds), cultivated for 200h. Add sterilized deionized water containing 0.1% Tween-80 (or Tween-20) to the buckwheat seed medium that has cultivated Aspergillus niger for 200h, shake at 180rmp for 1h, filter it with sterile gauze under aseptic conditions, and collect The filtrate was centrifuged at 10,000 g for 10 minutes, t...

Embodiment 3

[0035] The processing of embodiment 3 Aspergillus niger spores

[0036] The Aspergillus niger spore obtained above, because its spore wall is its natural barrier, has hindered the contact of glucose and intracellular glucose oxidase, therefore, to process Aspergillus niger spore, the method is:

[0037]1. Add sterilized deionized water containing 0.1% Tween-80 (or Tween-20) to the Aspergillus niger spore precipitate, centrifuge at 10,000g, 4°C for 10min, discard the supernatant, collect the lower precipitate, repeat the above After the step n times (1≤n≤25), the spores are collected for use.

[0038] 2. Put the obtained Aspergillus niger spore precipitate into -20℃ refrigerator, -80℃ low-temperature refrigerator or liquid nitrogen tank for more than 48 hours, then take it out from the refrigerator, let it dissolve rapidly at room temperature, and add a certain concentration Terpenoids (citral in this example) are treated for a certain period of time (3-18h), and the spores ar...

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Abstract

The invention discloses aspergillus niger-spores for converting glucose to produce gluconic acid and a preparation and application method of the aspergillus niger-spores. The preparation method includes the following steps of firstly, conducting cultivation through a solid fermenting method, wherein aspergillus niger-spores containing high-activity glucose oxidase in cells are obtained; secondly, conducting washing, freeze thawing and terpenoid reagent treatment, wherein aspergillus niger-spores high in permeability and free of germination in the biological conversion process are obtained. The prepared aspergillus niger-spores are added to a conversion container, and the content of glucose and the content of gluconic acid are measured through a binitro salicylic acid method and a gluconic acid kit respectively after the aspergillus niger-spores are converted for 200 hours. It is shown through the result that the prepared spores can be used for converting glucose to produce gluconic acid and have the advantages of being not prone to inactivation, free of fixation, easy to separate and capable of being repeatedly used in the conversion process. The method provides the theoretical foundation and technical support for the actual application of the aspergillus niger-spores for biologically converting glucose to produce gluconic acid.

Description

technical field [0001] The invention relates to an aspergillus niger spore for converting glucose to produce gluconic acid and a preparation and application method thereof. Background technique [0002] Gluconic acid is a multifunctional organic acid widely used in industries such as food, washing, medicine and construction. Gluconic acid can specifically catalyze the formation of β-D-glucose by glucose oxidase in the presence of oxygen. Because microorganisms have the characteristics of fast growth and reproduction and wide sources, they become the main source of glucose oxidase. The main production strain of microorganisms is Aspergillus niger (NRRL3). In industry, Aspergillus niger vegetative cells or synthetic enzymes are often used as biocatalysts in the continuous operation of biocatalytic systems to obtain gluconic acid. Various physical and chemical factors, chemicals and the release of degrading enzymes can easily cause enzyme inactivation. In addition, the bioca...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N3/00C12P7/58C12R1/685
CPCC12N1/14C12N3/00C12P7/58
Inventor 任佳丽周玉庭张慧唐云鹏张紫莺林亲录李忠海
Owner CENTRAL SOUTH UNIVERSITY OF FORESTRY AND TECHNOLOGY
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