Aspergillus niger strain with high yield of beta-D-fructofuranosidase and liquid-state fermentation and enzyme production method of aspergillus niger strain

A technology of fructofuranosidase and Aspergillus niger strains, applied in the field of microorganisms or enzymes, can solve the problems of undisclosed applicability, heat resistance, and low production costs, and achieve the effects of shortening the fermentation cycle, reducing fermentation costs, and reducing production costs

Active Publication Date: 2017-05-31
SHANDONG LONGKETE ENZYME PREPARATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] However, in summary of the invention patents, the wide range of applicability and heat resistance of its enzyme activity at different pHs and different temperatures have not been disclosed.
Because the enzymatic activity of β-D-fructofuranosidase is easily restricted by the applicable conditions of the enzyme, it is difficult to meet the needs of industrial production in the existing technology FFase. The production cost is low, and the invention and creation that can significantly promote large-scale production

Method used

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  • Aspergillus niger strain with high yield of beta-D-fructofuranosidase and liquid-state fermentation and enzyme production method of aspergillus niger strain
  • Aspergillus niger strain with high yield of beta-D-fructofuranosidase and liquid-state fermentation and enzyme production method of aspergillus niger strain
  • Aspergillus niger strain with high yield of beta-D-fructofuranosidase and liquid-state fermentation and enzyme production method of aspergillus niger strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Embodiment 1 culture medium preparation

[0054] PDA slant medium: sucrose 25g, NaNO3 2g, MgSO 4 0.5g, KCl 0.5g, FeSO 4 0.01g, K 2 HPO 4 1g, 20g agar, 1000mL distilled water, pH 4.5, sterilize at 121°C for 20min;

[0055] Seed medium: 150mL wort juice, 3g agar, adjust pH to 4.5, sterilize at 121°C for 20min;

[0056] The composition of the seed tank medium by mass fraction: glucose 2%, yeast powder 1.5%, KH 2 PO 4 0.15%, CaCl 2 0.005%, prepared with distilled water, adjusted to pH 4.5, sterilized at 121°C for 30 minutes;

[0057] The fermenter culture medium is composed by mass fraction: 4% corn starch, 4% corn flour, 4% bean cake powder, 1% ammonium sulfate, KH 2 PO 4 0.3%, corn steep liquor 0.6%, CaCl 2 0.5%, prepared with distilled water, adjusted to pH 4.5, sterilized at 121°C for 30 minutes;

[0058] The feed medium is composed by mass fraction: cornstarch 10%, ammonium sulfate 1%, KH 2 PO 4 0.1%, corn steep liquor 0.6%, CaCl 2 0.5%, prepared w...

Embodiment 2

[0060] Example 2 Aspergillus niger strain QL-225 fermented to produce enzyme and the extraction and refining method of the produced FFase

[0061] An Aspergillus niger strain with high production of β-D-fructofuranosidase, specifically the Aspergillus niger strain (Aspergillus niger) QL-225, and the preservation number is CGMCC No.13168.

[0062] Aspergillus niger strain QL-225 produces β-fructosidase by liquid fermentation, the optimum pH is 4.0-7.0, and the optimum temperature range is 40°C-60°C.

[0063] The method for producing enzyme by liquid fermentation of Aspergillus niger strain QL-225 mainly comprises the following steps:

[0064] Slant culture: Take a ring of Aspergillus niger strain QL-225 and inoculate it on the PDA slant medium, and cultivate it at a constant temperature at 30°C for 72 hours to obtain first-class seeds;

[0065] Shake flask culture: Pick a ring of first-grade seeds and insert them into the seed medium, and cultivate them for 72 hours at a const...

Embodiment 3

[0075] Example 3 Aspergillus niger strain QL-225 fermented to produce enzyme and the extraction and refining method of the produced FFase

[0076] An Aspergillus niger strain with high production of β-D-fructofuranosidase, specifically the Aspergillus niger strain (Aspergillus niger) QL-225, and the preservation number is CGMCC No.13168.

[0077] Aspergillus niger strain QL-225 produces β-fructosidase by liquid fermentation, the optimum pH is 4.0-7.0, and the optimum temperature range is 40°C-60°C.

[0078] The method for producing enzyme by liquid fermentation of Aspergillus niger strain QL-225 mainly comprises the following steps:

[0079] Slant culture: Take a ring of Aspergillus niger strain QL-225 and inoculate it on the PDA slant medium, and cultivate it at a constant temperature at 30°C for 72 hours to obtain first-class seeds;

[0080] Shake flask culture: Pick a ring of first-grade seeds and insert them into the seed medium, and cultivate them for 72 hours at a const...

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Abstract

The invention discloses an aspergillus niger strain with a high yield of beta-D-fructofuranosidase and a liquid-state fermentation and enzyme production method of the aspergillus niger strain and belongs to the field of microorganisms or enzymes. The aspergillus niger strain is aspergillus niger QL-225 and the preservation number is CGMCC (China General Microbiological Culture Collection Center) No.13168; liquid deep-layer fermentation is carried out through a 50L fermentation tank; meanwhile, materials are supplemented and fermented and cultured for 120h to 140h; finally, the enzyme activity of fermentation liquid can reach 19000U / ml to 20000U / ml. According to the established method for producing the beta-D-fructofuranosidase through liquid fermentation, the most suitable operation pH (Potential of Hydrogen) of the produced beta-D-fructofuranosidase is 4.0 to 7.0, and the most suitable operation temperature range is 40 DEG C to 60 DEG C; the residual enzyme activity is 51.4 percent after heat is preserved at 70 DEG C for 8h; the aspergillus niger strain has good acid resistance and heat resistance and can be widely applied to the industrial fields of foods, medicines, wine brewing and the like.

Description

[0001] Technical field: [0002] The invention belongs to the field of microorganisms or enzymes, and in particular relates to an Aspergillus niger strain with high production of β-D-fructofuranosidase and a method for producing the enzyme by liquid fermentation. [0003] Background technique: [0004] β-D-fructofuranosidase (β-D-fructofuranosidase, EC 3.1.2.26, abbreviated as FFase), also known as sucrase or invertase, widely exists in the biological world, the β-fructofuranosidase from different microbial sources The properties are different, β-D-fructofuranosidase can catalyze the hydrolysis of β-D-fructosidic bonds, and can also catalyze the transglycosylation reaction to generate galactosyl sucrose, that is, lactosucrose (LS). Fructo-oligosaccharides cannot be directly digested and absorbed by the human body, but can only be absorbed and utilized by intestinal bacteria. After ingestion, it will not cause fluctuations in blood sugar levels and insulin levels in the body, an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N9/26C12R1/685
CPCC12N9/2431C12Y302/01026C12N1/145C12R2001/685
Inventor 王兴吉刘文龙郭庆亮贾仁洁
Owner SHANDONG LONGKETE ENZYME PREPARATION
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