Pectinase artificial sequence and expression method and application thereof

Abstract

The invention relates to a pectinase artificial sequence and an expression method and application thereof; a gene herein at least comprises a DNA fragment of one of nucleotide sequences, including: 1)a nucleotide sequence of SEQ ID NO. 1 in a sequence table; 2) a nucleotide sequence having more than 90% homology with the nucleotide sequence shown in SEQ ID NO. 1, and encoding a same biological functional protein; or 3) a nucleotide sequence hybridized with the nucleotide sequence shown in SEQ ID NO. 1, and encoding the same biological functional protein. The gene sequence according to the invention allows a recombinant vector to be further constructed and yeast to be transformed, the secretion expression of the recombinant pectinase under the induction of methanol can be achieved, the DEAE anion exchange purification can be performed to obtain active recombinant pectin protein with the purity of higher than 95%, and the active zymoprotein here has strong activity of clarifying fruit juice.

Description

technical field [0001] The invention belongs to the technical field of biomolecular cloning, and relates to a preparation method and application of an artificial sequence of a high-activity pectinase gene and a recombinant protein thereof. Background technique [0002] my country is a big fruit producing country in the world. In the past ten years, my country's total fruit output has remained above 45 million tons. Rich fruit resources provide a good foundation for the development of my country's fruit juice industry. At present, the competition in the fruit juice market is fierce, there are many brands, and many processing enterprises are in trouble. How to develop the fruit juice industry has become a topic worthy of attention. A big problem in juice production is that the resulting juice is turbid, which affects the quality and taste of the juice. The use of pectinase to clarify fruit and vegetable juices has a long history and has been used as early as the 1930s. Usua...

AI Technical Summary

Problems solved by technology

Finally, obtaining high-purity proteins often requires multi-step purification operations. The more purification steps, the lower the yield of the protein, and it is more likely to cause the inactivation of the target product. Therefore, this expression system is not suitable for Mass production of recombinant recombinant pectinase

Exogenous gene expression systems such as insect cell expression systems, plant expression systems, and mammalian expression systems have too high requirements for technology, equipment, and technical level, so the products produced are often expensive, and are also not suitable for large quantities of pectinases. low cost production

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