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Cyclodextrin glycosyltransferase compound enzyme preparation

A glucose-based, cyclodextrin technology, applied in the directions of enzyme stabilization, microorganism-based methods, microorganisms, etc., can solve the problems of high cost and loss of enzyme activity, and achieve the effects of low cost, simple compounding method, and abundant raw material sources.

Active Publication Date: 2012-05-30
山东黄三角生物技术产业研究院有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In industrial applications, due to the high cost of solid powder enzyme preparations and the loss of enzyme activity in the process of processing, industrial enzyme preparations tend to use liquid fermentation broth directly as enzyme preparations, but their storage stability is not as high as powder enzyme preparations

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] The preparation of embodiment 1 cyclodextrin glucosyltransferase

[0016] Fermentation medium (1 L) of Bacillus softening bacillus (Paenibacillus macerans, CCTCC M 208063):

[0017] Glycerin: 10g; Peptone: 18g; Yeast extract: 20g; NH 4 Cl: 12g; KH 2 PO 4 : 2.31g; K 2 HPO 4 -3H 2 O: 16.43; MgCl 2 : 5 mmol.

[0018] Fermentation conditions of Bacillus softening bacillus (Paenibacillus macerans, CCTCC M 208063):

[0019] In a 3L fermenter, the volume of the culture medium is 1.5L. Insert the cells into the tank and culture them at 37°C for 72 hours, then put them into the tank, centrifuge the fermentation liquid at 12,000rpm for 15 minutes to remove the cells, and obtain the supernatant, which is the desired fermentation liquid.

Embodiment 2

[0021] The pH of the fermentation broth is adjusted to 6.5, and a cellulose acetate filter membrane with a molecular weight cut-off of 30,000 Daltons is selected for ultrafiltration and concentration until the protein content is 8-10 g / L.

[0022] The content of each component of the compound liquid enzyme preparation is as follows:

[0023] Cyclodextrin Glucosyltransferase Protein content 3g / L

[0024] Glycerin 10%

[0025] CaCl 2 0.1g / L

[0026] PEG400 5%

[0027] Gelatin 0.5%

[0028] The reconstituted liquid enzyme preparations were stored at 40°C for 60 days, and the enzyme activity retention rates were 95%, as shown in Table 1.

Embodiment 3

[0030] The pH of the fermentation broth is adjusted to 6.5, and a cellulose acetate filter membrane with a molecular weight cut-off of 30,000 Daltons is selected for ultrafiltration and concentration until the protein content is 8-10 g / L.

[0031] The content of each component of the compound liquid enzyme preparation is as follows:

[0032] Cyclodextrin Glucosyltransferase Protein content 5g / L

[0033] Glycerin 20%

[0034] CaCl 2 0.2g / L

[0035] PEG400 5%

[0036] Gelatin 0.5%

[0037] The reconstituted liquid enzyme preparations were stored at 40°C for 60 days, and the enzyme activity retention rates were 96%, as shown in Table 1.

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Abstract

The invention discloses a cyclodextrin glycosyltransferase compound enzyme preparation, which has extremely excellent storage stability, can be stored for 60 days at the temperature of 40 DEG C, has retention rate of enzyme activity still reaching over 95 percent, and solves the problem of reduced application performance of products caused by long product transportation or storage time. The liquid enzyme preparation provided by the invention has a simple compounding method, rich raw material sources and low cost, and is particularly suitable for industrial popularization.

Description

technical field [0001] The invention relates to a compound enzyme preparation, in particular to a cyclodextrin glucosyltransferase compound enzyme preparation for cyclodextrin production. Background technique [0002] Cyclodextrin glucosyltransferase (CGTase, EC 2.4.1.19) is a multifunctional enzyme that can catalyze three transglycosylation reactions (disproportionation reaction, cyclization reaction, coupling reaction) and one hydrolysis reaction. Through the intramolecular transglycosylation reaction, CGTase can convert starch and starch analogues into cyclodextrins. Common molecules with 6, 7 and 8 glucose units are called α-, β- and γ-cyclodextrins, respectively. . Cyclodextrin can form inclusion complexes with guest molecules, thereby changing the physical and chemical properties of guest molecules, so it can be widely used in analytical chemistry, agriculture, biotechnology, medicine, food and cosmetics and other fields. [0003] The methods of producing cyclodextri...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/96C12R1/01
Inventor 吴敬吴丹郑贤良陈坚陈晟
Owner 山东黄三角生物技术产业研究院有限公司
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