Leuconostoc citreum for producing acid inulase and application

A Leuconostoc lemon, acid inulin technology, applied in the direction of microbial-based methods, enzymes, bacteria, etc., to achieve high-stability effects

Inactive Publication Date: 2020-02-18
GUANGXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] There are many microorganisms that can produce inulinase in nature, such as bacteria, fungi and yeasts. At present, Aspergillius sp., Penicillium sp. and Kluyveromyces are more studied. sp.), but there are few research reports on acid inulinase from food-grade microbial sources

Method used

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  • Leuconostoc citreum for producing acid inulase and application
  • Leuconostoc citreum for producing acid inulase and application
  • Leuconostoc citreum for producing acid inulase and application

Examples

Experimental program
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Effect test

Embodiment 1

[0035] Screening and identification of a strain producing inulinase

[0036] (1) Preliminary screening of lactic acid bacteria producing inulinase

[0037] Leuconostoc 1-1, Lactobacillus plantarum 1-2, Lactobacillus plantarum 1-3, Lactobacillus plantarum 3-2, Lactobacillus plantarum 4- 3. Lactobacillus plantarum 6-3, Lactobacillus fermentum 9-4, and Lactobacillus bulgaricus, Lactobacillus helveticus, Pediococcus pentosaceus, and Lactobacillus casei preserved in the laboratory were streaked onto MRS solid medium, and cultured at 30°C until A single colony appears;

[0038] Streak the single colony normally grown in the above solid medium into the solid fermentation medium with inulin as the sole carbon source, and culture at 30°C until a single colony appears;

[0039] (2) Secondary screening of lactic acid bacteria containing acid inulinase gene

[0040] Pick a single colony that normally grows in the solid fermentation medium and inoculate it into the liquid fermentation m...

Embodiment 2

[0045] Production of Acid Inulinase by Fermentation of Leuconostoc citreum 1-1

[0046] Use Leuconostoc citreum 1-1 for fermentation to produce acid inulinase, inoculate the strain into the liquid fermentation medium with an inoculum size of 1% (v / v), and ferment at 30°C for 1-2 days , Determination of its enzyme activity. Among them, one unit of inulinase activity (I) is defined as the amount of enzyme required to catalyze the conversion of inulin into 1 μmol of fructose per minute, and one unit of invertase activity (S) is defined as the amount of enzyme required to catalyze the conversion of sucrose into 1 μmol of fructose per minute. Enzyme amount.

[0047] Determination of protein content: use Kangwei BCA protein quantification kit to measure the protein content of acid inulinase, and calculate the specific activity of the enzyme. According to the operating instructions of the BCA protein quantification kit, dilute the standard and prepare the BCA working solution. Dil...

Embodiment 3

[0051] Optimum Reaction Temperature and Temperature Stability of Acid Inulinase Produced by Leuconostoc citreum 1-1

[0052] The preparation method of acid inulinase is the same as example one. Take 100μl acid inulinase and add 0.5% 200μL inulin solution prepared with pH 5.0, 0.2M acetic acid buffer solution, respectively at 20℃, 25℃, 30℃, 35℃, 40℃, 45℃, 50℃, 55℃ , 60°C, 65°C, 70°C, 75°C, and 80°C for 30 minutes, and then immediately boiled water bath for 5 minutes to terminate the reaction, and then the enzyme activity was measured by DNS method. The highest enzyme activity of the measurement results was 100%, and the relative value of each measurement group was calculated. enzyme activity. Three parallel samples were set for each temperature.

[0053] Incubate 100 μl acid inulinase at the above temperature for 30 minutes, add 0.5% 200 μL inulin solution prepared with pH 5.0, 0.2M acetate buffer, react at 30°C for 30 minutes, and then measure the enzyme activity by DNS meth...

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Abstract

The invention discloses leuconostoc citreum for producing acid inulase and application. The strain is collected in the China Center for Type Culture Collection (CCTCC) on November 25, 2019 and has thecollection number of CCTCC NO:M 2019971. The leuconostoc citreum 1-1 is a lactobacillus strain 1-1 selected from traditional food raw-pressed rice noodles in a Guangxi Zhuang nationality region and having inulase producing capacity, inulin serving as a carbon source, beef extract serving as a nitrogen source and the like form a fermentation medium for fermentation production of the acid inulase,the strain grows well on the fermentation medium and has high acid resistance, the acid inulase produced by the strain has the survival rate of 96% after 1 h at the pH of 4.0, and has capacity of degrading inulin and fructan substances and preparing high fructose syrup through detection after fermentation.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to the field of development of functional lactic acid bacteria and production and development of products thereof. It specifically relates to a strain of Leuconostoc citrate producing acid inulinase, and the invention also includes the application of the Leuconostoc citrea. Background technique [0002] At present, the main source of inulin in industrial production is Jerusalem artichoke, which has the characteristics of low price, high yield, and non-grain crops, making it the focus of biorefinery. According to the different ways that inulinase acts on the substrate, it can be divided into exo-inulinase and endo-inulinase. Exo-inulinase acts on the fructosidic bond at the non-reducing end of fructan, cutting off fructose one by one, and finally The product is fructose and a small amount of glucose; endo-inulinase randomly acts on the fructosidic bonds inside inulin, and the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/24C12P19/14C12P19/02C12R1/01
CPCC12N1/20C12N9/2402C12P19/02C12P19/14C12Y302/01007
Inventor 王成华黄澄陈睿
Owner GUANGXI UNIV
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