Construction and application of ganoderma laccase pichia pastoris genetic engineering strain

Abstract

The invention provides construction and application of ganoderma laccase pichia pastoris genetic engineering strain. The construction comprises the following steps: cloning a laccase gene GlLac from ganoderma; carrying out EcoRI and XbaI double-enzyme cleavage; cloning the double-enzyme cleavage product into a pichia pastoris expression plasmid to construct an expression vector; finally, electrically converting the expression vector into a pichia pastoris expression host strain; carrying out cultivation, amplification and screening to obtain a pichia pastoris engineering strain. According to the construction, the yeast secreting type co-expression vector pPICZ<Alpha>A-GlLac of ganoderma laccase (Lac) is constructed, and converted into pichia pastoris X33; recombinant of high Zeocin resistance is selected out to serve as a pichia pastoris engineering strain. The extracellular expression vector pPICZ<Alpha>A of pichia pastoris is a carrier for judging whether a multi-copy gene is inserted into a pichia pastoris genome or not, and can express high-copy genes.

Description

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AI Technical Summary

Benefits of technology

This patented technology allows for screening out high yield cellulosic pulps that have been produced by certain types of microorganisms such as Saccharomyces cerevisiae or Bacillus subtilis without losing their ability to produce specific compounds called lignocystokines. By inserting these enzyme genes together, this process helps create more efficient production processes while reducing costs associated therewith.

Problems solved by technology

This patented technical problem addressed in this patents relates to improving the efficiency and quality of making industrials' worthful sources of lactic acid, particularly those derived from glutamic acid metabolism pathways. Current methods involve expensive processes like chemical synthesis, electroporsis, lysera rodenseidge mutants, and cell culture techniques. There also exist attempts to create more economically viable ways to make these valuable resources available for commercial purposes without compromising their value.

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