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Streptomyces coelicolor mutant strain, method of producing beta-agarase by using same, and method of producing neoagaro-oligosaccharides by using same

A technology of Streptomyces coelicolor and agarase, applied in the direction of microorganism-based methods, biochemical equipment and methods, glycosylase, etc.

Active Publication Date: 2021-03-26
DYNEBIO
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far, there are few reports on strains capable of producing DagAβ-agarase at commercially applicable levels

Method used

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  • Streptomyces coelicolor mutant strain, method of producing beta-agarase by using same, and method of producing neoagaro-oligosaccharides by using same
  • Streptomyces coelicolor mutant strain, method of producing beta-agarase by using same, and method of producing neoagaro-oligosaccharides by using same
  • Streptomyces coelicolor mutant strain, method of producing beta-agarase by using same, and method of producing neoagaro-oligosaccharides by using same

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Embodiment Construction

[0029] Hereinafter, the present invention will be described more specifically by way of examples. However, the following examples are only used to clearly illustrate the technical features of the present invention, and are not intended to limit the protection scope of the present invention.

[0030] 1. Enzyme activity measurement method

[0031] (1) Measure the β-agarase (β-agarase) activity of the sample

[0032] The β-agarase activity of the sample was measured by a reducing sugar quantification method (dinitrosalicylic acid method (DNS method)). Specifically, 490 μL of a 20 mM Tris-HCl solution (pH 7) in which agarose (agarose) was dissolved at a concentration of 0.5% (w / v) was mixed with 10 μL of the sample, and reacted at 40° C. for 15 minutes. , and then the same amount of DNS reagent as the reaction solution (prepared by dissolving 6.5 g of dinitrosalicylic acid, 325 ml of 2M NaOH and 45 ml of glycerol in 1 L of distilled water) was added thereto, and After boiling f...

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Abstract

The present invention provides Streptomyces coelicolor strain A3(2)_M22-2C43 obtained by inducing a point mutation in the base sequence of the DagB gene in a wild-type Streptomyces coelicolor strain A3(2) by UV radiation. Since the Streptomyces coelicolor strain A3(2)_M22-2C43 according to the present invention expresses a DagB mutant enzyme expressing little or no DagB beta-agarase or exhibitinglittle or no beta-agarase activity, there is no need for separate isolation and purification of DagA enzymes from culture fluid, and the culture fluid of the Streptomyces coelicolor strain A3(2)_M22-2C43 or supernatant thereof can be used to produce, from agar or agarose, neoagarose oligosaccharides with a higher content of neoagarotetraose or neoagarohexaose than that of neoagarobiose.

Description

technical field [0001] The present invention relates to a mutant strain of Streptomyces coelicolor, a method for producing Beta-agarase using it and a method for preparing Neoagaro-oligosaccharides using it, more specifically , relating to a mutant strain of Streptomyces coelicolor mainly expressing DagA enzyme compared with a parental strain, a method of using it to mass-produce DagA enzyme in β-agarase, and using it to prepare new agarose from agar or agarose (neoagarotetraose) or new agarose (neoagarotetraose) content than new agarobiose (neoagarobiose) higher new agar oligosaccharide method. Background technique [0002] For a long time, agar has been widely used as a representative polysaccharide (derived from seaweed) in food additives, pharmaceuticals, cosmetics, livestock feed, and industrial raw materials. In Korea, its annual production volume reaches about 2000~ One of the richer aquatic resources of 5,000 tons. However, in actual use, only a part of the total p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/38C12P19/04C12P19/14C12P21/00C12R1/465
CPCC12Y302/01081C12P19/04C12R2001/465C12N1/205C12N1/20C12N9/2468C12P19/14C12P21/00
Inventor 李济贤金恩妵李娟嬉
Owner DYNEBIO
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