Oxalate decarboxylase and recombinant expression method of oxalate decarboxylase

Abstract

The invention discloses an oxalate decarboxylase and a recombinant expression method of the oxalate decarboxylase. The oxalate decarboxylase is subjected to recombinant expression through an Escherichia coli expression system, and is stable and active when the pH value is less than 3.0; and compared with the primary zymoprotein sequence, a signal peptide segment is deleted in the protein sequence of the oxalate decarboxylase in the expression process. The recombinant expression method is suitable for oxalate decarboxylase which is still active when the pH value is lower than 3.0: the DNA segment of the optimized oxalate decarboxylase coding gene and an Escherichia coli expression vector are mixed according to the mole ratio of (3-9):1 to perform connection so as to establish the recombinant expression vector, and the recombinant expression vector is used for transforming DH5a competent cells. PCR (polymerase chain reaction) and sequencing verification are utilized to screen the successfully established recombinant expression vector transformed bacteria, the recombinant expression vector is extracted after culture and is used for transforming the molecular-chaperone-containing Escherichia coli expression strain, and 15-30-DEG C culture and protein expression induction are performed. The method enhances the expression efficiency, simplifies the production steps and lowers the production cost.

Description

technical field [0001] The invention relates to a novel stable and efficient oxalate decarboxylase in a low pH environment and a recombinant expression method of the oxalate decarboxylase. Background technique [0002] Oxalic acid (oxalic acid) is a molecular formula C 2 o 4 h 2 , a dibasic organic acid with a molecular weight of 90.04, which exists in most common foods, and is very high in specific foods (such as spinach or other green vegetables, green tea, cocoa beans, soybean milk, coffee, etc.). Oxalic acid is the final product in the body and cannot be further decomposed. The main way of excretion is to excrete it with urine. Oxalic acid in the human body includes exogenous oxalic acid and endogenous oxalic acid. Exogenous oxalic acid refers to oxalic acid ingested through diet. Endogenous oxalic acid refers to oxalic acid produced by the human liver itself. It is basically the same as the content of endogenous oxalic acid. For some people with high risk of kidney...

AI Technical Summary

Problems solved by technology

[0005] In order to solve the technical problem that low pH (pH<3.0) stable and active OXDCs cannot be expressed in Escherichia coli in the prior art, the present invention obtained multiple OXDCs in the gastrointestinal tract environment (pH1) through high-throughput screening. .5~7.5) have high oxalate degrading enzyme activity, and have high activity oxalate decarboxylase in low pH gastric environment (pH1.5~3.0), and its coding gene has been cloned to achieve its recombinant expression

During the recombinant expression, the DNA sequence of the signal peptide part of these oxalate decarboxylases is removed, so that the transformed gene does not contain the signal peptide coding sequence, and the recombinant expression of E. The problem that the active oxalate decarboxylase cannot be produced by the E. coli expression system reduces the production cost

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