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Application of aspergillus niger in preparation of beta-fructofuranosidase

A fructofuranoside, Aspergillus niger technology, applied in the directions of glycosylase, enzyme, hydrolase, etc., can solve the problems of instability, low yield of production strains, low β-enzyme activity, etc. Commercial prospects, the effect of less loss of enzyme activity

Inactive Publication Date: 2016-01-27
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] However, the yield of the currently reported production strains is not high, and the enzyme activity of β- is low and unstable (Ma Geli, Zhang Xuejun, Jin Li. Research on the enzymatic properties of β-fructofuranosidase produced by Aspergillus niger. China Brewing, 2009, 207 (6): 22-24), so screening high-yielding and stable strains and isolating and purifying their transferases is one of the important research contents of this industry at present

Method used

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  • Application of aspergillus niger in preparation of beta-fructofuranosidase
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Effect test

Embodiment 1

[0036] (1) Inoculate the bacterial suspension of Aspergillus niger FOS-0620 in the logarithmic phase growth in the fermentation induction medium with an inoculation amount of 5% by volume, and induce fermentation on a constant temperature shaker at 33°C and 133r / min After cultivating for 48 hours, filter the obtained culture solution with a 400-mesh filter bag, wash the obtained bacteria sludge with sterile water, and then filter it with a 400-mesh filter bag. Repeat the cleaning and filtering operations twice to obtain bacteria cells. , drying under a vacuum of 0.030 psi to obtain Aspergillus niger cells with β-FFase activity. The fermentation induction medium: by weight, 7 parts of sucrose, 2 parts of yeast extract, 1.5 parts of corn flour, NaNO 3 0.3 parts, pH 6.5, dilute to 100 parts with distilled water, mix well, sterilize and cool at 121°C for later use.

[0037] (2) In (1), add trypsin inhibitor containing 2 μg / mL and 1.5 μmol / mL dithiothreitol (abbreviated DTT) enzym...

Embodiment 2

[0053] (1) Inoculate the bacterial suspension of Aspergillus niger FOS-0620 in the logarithmic phase growth in the fermentation induction medium with an inoculation amount of 5% by volume, and after cultivating for 36h at 30°C and 100r / min, the obtained fermentation The liquid was filtered with a 400-mesh filter bag, and the obtained bacteria sludge was washed with sterile water and then centrifuged at 8000r / min for 10 minutes. The cleaning and centrifugation operations were repeated twice to obtain bacteria, which were dried at -40°C and a vacuum of 0.030psi to obtain Aspergillus niger bacteria. The fermentation induction medium: by weight, 2 parts of sucrose, 1 part of yeast extract, 1.0 part of corn flour, NaNO 3 0.1 part, pH 6.0, dilute to 100 parts with distilled water, mix well, sterilize and cool at 121°C for later use.

[0054] (2) In the Aspergillus niger strain in (1), add the enzyme buffer solution containing 1 μg / mL trypsin inhibitor and 0.5 μmol / mL DTT to make th...

Embodiment 3

[0059] (1) Inoculate the bacterial suspension of Aspergillus niger FOS-0620 in the logarithmic phase growth in the fermentation induction medium with an inoculation amount of 10% by volume, and after cultivating for 72h at 40°C and 200r / min, the obtained fermentation The solution was filtered with a 400-mesh filter bag, and the obtained bacteria sludge was washed with sterile water and then filtered with a 400-mesh filter bag. The cleaning and filtering operations were repeated twice to obtain bacteria, which were dried at -40°C and a vacuum of 0.030psi. Obtain Aspergillus niger cells. Described fermentation induction medium: by weight, 10 parts of sucrose, 5 parts of yeast extract, 5 parts of corn flour, NaNO 3 0.5 parts, pH 7.5, dilute to 100 parts with distilled water, mix evenly, sterilize and cool at 121°C for later use.

[0060] (2) In the Aspergillus niger strain in (1), add the enzyme buffer solution containing 3 μg / mL trypsin inhibitor and 2 μmol / mL DTT to make the A...

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Abstract

The invention discloses application of aspergillus niger in preparation of beta-fructofuranosidase. The aspergillus niger FOS-0620 has been deposited at the China General Microbiological Culture Collection Center with the accession number of CGMCC No. 6640. A preparation method comprises the following steps of filtering, washing and performing freeze drying, enzymolysis and cell wall breaking on the aspergillus niger FOS-0620 after the aspergillus niger FOS-0620 is fermented in an inducing fermentation medium to obtain beta-fructofuranosidase crude enzyme; and performing separation purification on the crude enzyme to obtain high-purity beta-fructofuranosidase. The prepared beta-fructofuranosidase is high in enzyme activity, and has a wide temperature application scope; and fructo-oligosaccharide which is catalytically prepared from the beta-fructofuranosidase by using sucrose as a substrate is simple in process and high in content, and reaches national standard.

Description

technical field [0001] The invention relates to the field of biological products, in particular to an application of Aspergillus niger in the preparation of β-fructofuranosyltransferase. Background technique [0002] Fructose oligosaccharide (FOS) is a kestose triose (GF) produced by combining 1 to 3 fructosyl groups with fructosyl groups in sucrose (GF) through β-1,2 glycosidic bonds. 2 ), Kestose (GF 3 ) and fructopentose (GF 4 ) etc., is a new type of functional fructooligosaccharide. FOS is low in calories, has excellent properties such as reducing blood fat, preventing dental caries, and preventing the growth of harmful bacteria. It also has physiological activities such as promoting the growth of human bifidobacteria and other probiotics. Therefore, it is widely used in food, feed, and cosmetics industries. [0003] At present, there are mainly five methods for producing FOS: (1) extracting directly from natural plant materials; (2) obtaining by enzymatic hydrolysis...

Claims

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Application Information

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IPC IPC(8): C12N9/26C12R1/685
CPCC12N9/2431C12Y302/01026
Inventor 刘冬梅周劲松杨丹霞吴晖肖性龙余以刚唐语谦李晓凤
Owner SOUTH CHINA UNIV OF TECH
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