Recombinant pectinase, gene thereof, recombinant vector, preparation method and application

Abstract

The invention relates to recombinant pectinase, a gene thereof, a recombinant vector, a preparation method and application. The gene at least comprises DNA fragment of one of a nucleotide sequence ofSEQ ID NO.1 in a sequence table, a nucleotide sequence having higher than 90% homology with the nucleotide sequence shown as the SEQ ID NO.1 and coding same biological functional protein and a nucleotide sequence hybridized with the nucleotide sequence shown as the SEQ ID NO.1 and coding same biological functional protein. The recombinant vector is further built according to the gene sequence, yeast is transformed, in this way, constitutive secretory expression of the recombinant pectinase in a high-density fermentation condition can be realized; active protein higher than 95% in purity and with high protein concentration can be obtained through simple nickel affinity purification; the active protein has high pectin hydrolyzing activity at low temperature, and a novel and efficient enzymeproduct can be provided for increasing juice yield of fruit juice.

Description

technical field [0001] The invention belongs to the technical field of biomolecular cloning, and relates to a recombinant pectinase and its gene, recombinant vector, preparation method and application. Background technique [0002] my country is a vast country with a land area of ​​about 9.6 million square kilometers. It spans the three major regions of the cold zone, temperate zone and subtropical zone in the north and south. It is suitable for the growth of many kinds of fruit trees. Therefore, my country is a big country in fruit production in the world, such as: hawthorn, apple, Pears, peaches, apricots, grapes, jujubes, seabuckthorns, black currants, kiwis, tangerines, oranges, tangerines, pineapples, bananas, guavas, prickly pears, passion fruit, etc. 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 competit...

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 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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