Arylsulfatase and preparation method and applications thereof

A technology of radical sulfatase and radical sulfate, applied in the fields of botany equipment and methods, biochemical equipment and methods, applications, etc. rate drop etc.

Active Publication Date: 2013-10-16
THIRD INST OF OCEANOGRAPHY STATE OCEANIC ADMINISTATION +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In 2004, Lim JM et al. (Lim JM, Jang YH, Kim HR, Kim JK, Nam SW. Overexpression of arylsulfatase in E.coli and its application to desulfation of agar, J Microbiol Biotechnol.2004,14:777–782.) It was found that the arylsulfatase from Pseudoalteromonas carrageenovora ATCC43555 strain can degrade the substrate p-nitrophenyl sulfate (pNPS), and its gene can degrade SO on agar after expression in E. coli 4 2- The ability to increase the gel strength and decrease the conductivity of agar
At present, there is no patent on arylsulfatase, and there is no patent on sulfatase related to the treatment of agar.

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  • Arylsulfatase and preparation method and applications thereof
  • Arylsulfatase and preparation method and applications thereof
  • Arylsulfatase and preparation method and applications thereof

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Embodiment Construction

[0065] The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental methods without specific conditions in the following examples usually follow conventional conditions such as the experimental conditions described in the Molecular Cloning Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 2001), or according to the reagent or instrument manufacturer’s Suggested conditions.

[0066] The production strain used to prepare the aromatic sulfatase gene Ary423 is Flammeovirga Pacifica H2. This strain has been deposited in the China Type Culture Collection on June 13, 2012, address: Wuhan, China, Wuhan University, and the deposit number is: CCTCC NO: M2012229.

[0067] 1. Preparation of arylsulfatase Ary423

[0068] 1) Extraction of Flammeovirga Pacifica H2 Genomic DNA

[006...

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Abstract

An arylsulfatase and a preparation method and applications thereof, relate to an arylsulfatase with a new gene sequence and preparation thereof. Provided is arylsulfatase Ary423 originated from the Flammeovirga Pacifica H2. Preparation methods of arylsulfatase gene Ary423 and arylsulfatase Ary423 are also provided. Gene Ary423 is cloned, recombinant plasmid carrying with the gene Ary423 is constructed and then converted to the escherichia coli BL21, the positive clones are selected and cultured in a culture medium, then the cultured positive clones are re-suspended in a lysis buffer, after the processes of cracking and centrifugation, the supernatant is collected and homogeneously mixed and purified with Ni-NTA Agarose. Arylsulfatase Ary423 can stably express in escherichia coli, recombinase maintains high catalysis efficiency in a relatively wide temperature range and pH range, so that the shortages of low vigor of arylsulfatase and tedious processes of separating and purifying in the prior art are overcome.

Description

Technical field [0001] The invention relates to an aryl sulfatase with a novel gene sequence, a preparation method thereof, and its application in an agar extraction process. Background technique [0002] Agar is a natural polysaccharide colloid extracted from red algae seaweeds. It has a wide range of applications in food industry, pharmaceutical industry, daily chemical industry, biological engineering and many other aspects. Gel strength is the main indicator to measure the quality of agar products, and the sulfuric acid group is the key factor hindering the formation of gel from agar molecules. In the process of producing agar, the crude agar molecule in the seaweed contains a high content of sulfuric acid groups, and it needs to be treated with alkali (NaOH) to reduce the sulfuric acid group (SO) in the agar molecule. 4 2- ) Content to improve the gel strength of agar products, so alkali treatment is an indispensable process in the agar industrial production process. In the...

Claims

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Application Information

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Patent Type & AuthorityApplications(China)
IPC IPC(8): C12N15/55C12N9/16C12N15/70C08B37/12
Inventor曾润颖高超易志伟产竹华
OwnerTHIRD INST OF OCEANOGRAPHY STATE OCEANIC ADMINISTATION