Flat plate color developing method for fast detecting fructooligosaccharide synthesizing capacity of filamentous fungi

A filamentous fungus, fructooligosaccharide technology, applied in the field of detection, to achieve the effect of shortening the screening process, rapid method, strong application and promotion value

Active Publication Date: 2016-04-06
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

After searching, there is no report in the relevant patents and literatures about the detection method based on the medium plate and using the double enz

Method used

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  • Flat plate color developing method for fast detecting fructooligosaccharide synthesizing capacity of filamentous fungi
  • Flat plate color developing method for fast detecting fructooligosaccharide synthesizing capacity of filamentous fungi
  • Flat plate color developing method for fast detecting fructooligosaccharide synthesizing capacity of filamentous fungi

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0033] Embodiment 1. The source of fructo-oligosaccharide synthesis filamentous fungus and activated sporulation culture

[0034] The fructo-oligosaccharide-synthesizing filamentous fungi involved in this embodiment include: Aspergillus niger (Aspergillus niger) XG530, Aspergillus niger XG530 ultraviolet mutagenesis strain, Aspergillus tubingensis (Aspergillustubingensis) and Aspergillus aculeatus (Aspergillus aculeatus);

[0035] Wherein: Aspergillus niger XG530 bacterial strain is used as the standard bacterial strain of this detection filamentous fungus synthetic fructooligosaccharide ability plate color development method, and it is the commonly used production strain of fructooligosaccharide industry at present, purchased from Shandong Xingguang Sugar Industry Group Co., Ltd.; Aspergillus tubingensis) and Aspergillus aculeatus are filamentous fungi capable of synthesizing fructo-oligosaccharides isolated from soil samples collected from the sugar factory of Shandong Xinggu...

Example Embodiment

[0041] Embodiment 2. establishes the optimal condition of plate color development of detection standard strain synthetic fructo-oligosaccharide ability

[0042] A. Determination of the best detection time: take fresh spores of the standard strain Aspergillus niger XG530, and dilute them appropriately to a spore concentration of 1×10 7 1 / mL, take 1 μl and plant it on the center of Cha's solid medium plate, inoculate 6-8 plates under the same experimental conditions, culture at 30°C for 2h, 5h, 10h, 24h and 48h, take out and pour into the pre-prepared glucose respectively Detect the reagent and make it evenly cover all the medium surface on the plate. After standing at 40°C for 10 minutes, observe and record the size of the glucose color circle to determine the optimal detection time.

[0043] The experimental results show that the standard strain can produce a red halo with a diameter of 0.5 cm when cultured for 5 hours. As the culture time prolongs, the red halo gradually cove...

Example Embodiment

[0047] Example 3. Evaluation of the effect of the plate color method for rapid detection of the ability of filamentous fungi to synthesize fructo-oligosaccharides

[0048] A. The strains used

[0049] Standard strain - Aspergillus niger XG530, strain to be tested - Aspergillus niger XG530 ultraviolet mutagenesis strain, 16 strains were selected, respectively numbered A1, A2, A3, A4, A5, A6, A7, A8, A9, A10, A11, A12, A13, A14, A15 and A16.

[0050] B. Utilize the plate chromogenic method of the present invention to detect the synthetic ability of 16 strains of mutagenized fructooligosaccharides

[0051] Cultivate and collect the fresh spores of the above-mentioned standard strain XG530 and its 16 mutagenized strains (see Example 1 for the method), prepare the spore suspension count and dilute to 1×10 under the same experimental conditions. 7 1 / mL, take 1 μl and plant on the Chapei solid medium plate, the distance between the spores of different strains should not be less tha...

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Abstract

The invention discloses a flat plate color developing method for fast detecting the fructooligosaccharide synthesizing capacity of filamentous fungi. Fresh spore suspension of filamentous fungi to be detected and standard fungi is spot planted on a flat plate of a Czapek's medium, the spot planting distance between strain spores is not smaller than 2 cm, cultivation is performed for 5 h at 30 DEG C, glucose detection liquid is directly poured on the flat plate where a bacterial colony grows, the whole medium surface of the flat plate is evenly covered with the glucose detection liquid, standing is performed for 10 min at 40 DEG C, red halos formed around the bacterial colony are observed, the size of the red halo formed by the bacterial strains to be detected and the size of the red halo formed by the stand bacterial strains are compared, if the red halo formed by the bacterial strains to be detected is larger than the red halo formed by the stand bacterial strains, it can be judged that the fructooligosaccharide synthesizing capacity of the bacterial strains to be detected is high, and if the red halo formed by the bacterial strains to be detected is smaller than the red halo formed by the stand bacterial strains, it can be judged that the fructooligosaccharide synthesizing capacity of the bacterial strains to be detected is low. The method is easy to operate and low in cost, is a simple, convenient, efficient and reliable method for identifying the fructooligosaccharide synthesizing capacity of filamentous fungi, has industrial application value and is worthy of popularization.

Description

technical field [0001] The invention relates to a method for detecting the ability of filamentous fungi to synthesize fructo-oligosaccharides, and belongs to the technical field of detection. Specifically, it relates to a plate color development method for rapidly detecting the ability of filamentous fungi to synthesize fructo-oligosaccharides. Background technique [0002] Fructo-oligosaccharides (FOS for short), also known as functional fructo-oligosaccharides, is a typical prebiotic substance, which widely exists in natural products such as onions, Jerusalem artichokes, bananas, pears, wheat, and honey. It has been widely used in the development and production of medicines, food, health products, feed and other products at home and abroad. Its molecular formula is G-F-Fn (G is glucose, F is fructose, n=1~3), and it is kestose (GF) formed by combining several D-fructose with β-1,2 glycosidic bonds on the sucrose molecule. 2 ), Kestose (GF 3 ), Kippentose (GF 4 ) and th...

Claims

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Application Information

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IPC IPC(8): C12Q1/54C12Q1/28C12Q1/26
CPCC12Q1/26C12Q1/28C12Q1/54C12Q2326/50C12Q2326/96
Inventor 钟耀华张静李宁谢益佳宁占国周焕霞
Owner SHANDONG UNIV
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