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Method of producing exoinulinase with utilization of solid state fermentation of streptomyces grisepoplanus S501

A technology of exo-inulinase and Streptomyces griseignis, which is applied in the field of microorganisms, can solve problems such as environmental pollution, waste, and unsuitability for industrial production, and achieve the effect of overcoming high cost and improving the activity of exo-inulinase

Active Publication Date: 2015-05-13
DALIAN NATIONALITIES UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most studies have used inulin-containing plants Jerusalem artichoke, chicory, etc. as carbon sources to induce inulinase-producing strains, resulting in high cost of using inulin to produce enzymes, which is not suitable for industrial production. If you can find a price Inulinase with high enzyme activity can be obtained from cheap and easy-to-get carbon sources, which can meet the needs of industrial production
[0004] Food processing plants will produce a large amount of garlic skins in the production process. These garlic skins are used in a very low rate in daily life and are usually disposed of as garbage. The annual discarded garlic skins in the country reach 200,000 tons, resulting in a large amount of waste. polluted the environment

Method used

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  • Method of producing exoinulinase with utilization of solid state fermentation of streptomyces grisepoplanus S501
  • Method of producing exoinulinase with utilization of solid state fermentation of streptomyces grisepoplanus S501
  • Method of producing exoinulinase with utilization of solid state fermentation of streptomyces grisepoplanus S501

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Experimental program
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preparation example Construction

[0038] Preparation of fructose standard curve:

[0039] Take 1mg / mL fructose standard solution 0, 0.2, 0.4, 0.6, 0.8, 1.0, 1.2, 1.4, 1.6mL respectively in test tubes, add distilled water to make up to 2.0mL, add 1.5mL of DNS solution respectively, mix well, put in boiling water bath React for 5 minutes, cool to room temperature, set the volume to 25.0mL, mix well, measure the absorbance value at 540nm, draw the fructose standard curve with the fructose content as the abscissa and the absorbance value (Abs) as the ordinate.

[0040] DNS inulinase activity assay:

[0041] The DNS method was used to determine the average value of three parallel experiments, and the fructose standard curve method was used to calculate the reducing sugar content. At the same time, the solid-state fermentation medium without inoculation was used as a control. The unit of inulinase activity is defined as: at 50°C, the amount of enzyme required to convert inulin into 1 μmol of reducing sugar per min...

Embodiment 1

[0049] 1. Strain culture

[0050] Make the lyophilized powder of strain S501 into a bacterial suspension with sterile water, dip an appropriate amount on the selection medium with a sterile swab, and cultivate it at 28°C for 3-5 days, and gently scrape off the spores on the plate with a sterile spatula , placed in sterile water and diluted to a spore suspension concentration of 10 6 cfu / mL.

[0051] Described selection medium is made up of the component that comprises following content:

[0052]

[0053]

[0054] The selection medium has a pH of 7.2 and is autoclaved at 112° C. for 20 minutes.

[0055] 2. Exo-inulinase Preparation

[0056] The above-mentioned bacterial suspension was inserted into a solid-state fermentation medium with an inoculation amount of 8% (v / w), and cultured statically at 28° C. for 6 days; the components of the solid-state fermentation medium were:

[0057]

[0058] The above-mentioned solid-state fermentation culture was sterilized under...

Embodiment 2

[0060] 1. Strain culture

[0061] Make the lyophilized powder of strain S501 into a bacterial suspension with sterile water, dip an appropriate amount on the selection medium with a sterile swab, and cultivate it at 28°C for 3-5 days, and gently scrape off the spores on the plate with a sterile spatula , placed in sterile water and diluted to a spore suspension concentration of 10 6 cfu / mL.

[0062] Described selection medium is made up of the component that comprises following content:

[0063]

[0064]

[0065] The selection medium has a pH of 7.2 and is autoclaved at 112° C. for 20 minutes.

[0066] 2. Exo-inulinase Preparation

[0067] The above-mentioned bacterial suspension was inserted into a solid-state fermentation medium with an inoculation amount of 8% (v / w), and cultured statically at 28° C. for 6 days; the components of the solid-state fermentation medium were:

[0068]

[0069] The pH of the solid-state fermentation medium is 8.0, and it is autoclave...

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Abstract

The invention provides a method of producing exoinulinase with utilization of solid state fermentation of streptomyces grisepoplanus S501 and belongs to the technical field of microorganisms. The method comprises the following steps: preparing a bacterial suspension by freeze-dried powder of a strain S501 and aseptic water on a selective medium, culturing for 3-5d at the temperature of 28 DEG C, scraping spores, putting the spores into the aseptic water, diluting until the concentration of a spore suspension is 106cfu / mL, inoculating to a solid state culture medium, and carrying out static culture for 3-6d at the temperature of 28 DEG C; and after fermentation, adding deionized water, carrying out oscillating extraction for 1h, filtering, carrying out centrifugation on a filtrate for 20 minutes at the temperature of 4 DEG C and the rotational speed of 8000rpm / min to obtain a supernatant containing the exoinulinase, and treating the supernatant by virtue of the conventional freeze-drying method to prepare an inulinase preparation. According to the method provided by the invention, with wheat bran as a solid fermentation matrix and garlic husk powder which is cheap and easily acquired as a carbon source, the strain S501 is induced to produce the exoinulinase, the enzyme activity of the exoinulinase reaches up to 209.63 + / - 0.96U / g, and the defects of high cost, time and labor consumption in a liquid state fermentation method and non suitability in industrial production in the prior art with inulin as a carbon source are overcome.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a method for producing exo-inulinase by solid-state fermentation of Streptomyces griseoplanus S501. Background technique [0002] Inulin is a kind of natural fructan, which is connected by D-fructofuranose with β-2,1 glycosidic bonds, and its reducing end is connected with a glucose group, showing a straight chain structure. Inulin-rich plants mainly include Jerusalem artichoke, chicory, etc. Inulin has attracted the attention of the food industry in recent years because of its unique functions such as prebiotics, and its application prospects are very broad. Inulinase is a kind of hydrolase that can hydrolyze β-2,1-D-fructan glycosidic bonds, and is mainly divided into endoenzyme and exoenzyme according to the mode of action. Exo-inulinase can hydrolyze inulin to produce high-purity fructose, while endo-inulinase can hydrolyze inulin to produce fructo-oligosa...

Claims

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Application Information

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IPC IPC(8): C12N9/24C12R1/465
CPCC12N9/2402C12Y302/01007
Inventor 于基成陈娇刘秋于东宁
Owner DALIAN NATIONALITIES UNIVERSITY
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