Purine nucleoside phosphorylase and preparation method thereof

A purine nucleoside phosphorylase and polynucleotide technology, which is applied in the field of genetic engineering, can solve the problems of low PNP yield, inactivation, EcPNP changes sharply at 50-55 ℃, and the effect cannot be achieved.
CN103468656AActive Publication Date: 2013-12-25AILEX TECH GRP CO LTD +1
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
AILEX TECH GRP CO LTD
Publication Date
2013-12-25

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Abstract

The invention relates to purine nucleoside phosphorylase and a preparation method of the purine nucleoside phosphorylase, in particular to a mutational pseudoalteromonas-genus purine nucleoside phosphorylase. Compared with a wild-type amino acid sequence, the amino acid sequence of the mutational purine nucleoside phosphorylase is characterized in that Asp at the 98th bit is mutated into Tyr. The invention further discloses the preparation method of the purine nucleoside phosphorylase. The purine nucleoside phosphorylase and the preparation method of the purine nucleoside phosphorylase have the advantages of being high in yield and good in heat stability. The invention further discloses polynucleotide for encoding the amino acid sequence of the purine nucleoside phosphorylase, a carrier containing the polynucleotide and a host cell containing the polynucleotide.
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Description

technical field

[0001] The invention relates to the field of genetic engineering, in particular to a purine nucleoside phosphorylase and a preparation method thereof. Background technique

[0002] Purine nucleoside phosphorylase (PNP for short), is one of the key enzymes in the purine salvage synthesis pathway, widely exists in mammals, parasites and microorganisms. According to the protein structure of purine nucleoside phosphorylase, it can be divided into two categories: low molecular weight homotrimers and high molecular weight homohexamers. Among them, the purine nucleoside phosphorylases of mammals and some microorganisms (such as Salmonella typhimurium, thermophilic archaea Sulfolobus solfataricus, etc.) belong to the homotrimer class, and its molecular weight is about 80-100kDa. The molecular weight of each subunit is 30-32kDa, and usually this kind of purine nucleoside phosphorylase can only use guanosine and inosine as substrates. The homohexamer purine nucleosid...

Claims

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