Method for producing ligninolytic enzymes

Abstract

The invention discloses a method for producing ligninolytic enzymes, and belongs to the biotechnical field. The method comprises the following steps: 1, respectively cloning lignin peroxidase, manganese peroxidase and laccase genes from microbes; 2, constructing a Pichia pastoris expression vector containing the expression cassettes of the above three genes, and a Saccharomyces cerevisiae expression vector containing the expression cassettes, and converting the above vectors to corresponding host bacteria to obtain engineering bacteria; and 3, respectively fermenting Pichia pastoris expression engineering bacteria and Saccharomyces cerevisiae engineering bacteria to produce recombinant lignin peroxidase, manganese peroxidase and laccase. The method has the following advantages: engineering bacteria are used to substitute natural bacterial strains to produce the lignin peroxidase, manganese peroxidase and laccase, so the enzyme outputs are improved through increasing the copy numbers of the genes; and the engineering bacteria containing three enzyme gene expression cassettes substitute engineering bacteria containing a single enzyme gene expression cassette to produce single enzymes, so the raw material, the energy consumption and the manpower resource are saved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for constructing microbial engineering bacteria to produce recombinant proteins. Specifically, the method uses microbial engineering bacteria to produce recombinant mixed lignin peroxidase, manganese peroxidase and laccase. Background technique [0002] Lignin Peroxidase (LiP), Manganese Peroxidase (MnP) and Laccase have the function of degrading lignin together. [0003] Plant resources are constantly regenerated and are very rich. The basic components of plants are cellulose, hemicellulose and lignin. Among them, the most useful component in the manufacture of biogas and ethanol industry is cellulose, followed by hemicellulose. The basic component of the former is glucose, while the basic component of the latter is pentose. These sugars are converted into ethanol and biogas. Lignin is an amorphous, aromatic polymer containing oxyphenylpropanol or its derivatives in its ...

AI Technical Summary

Problems solved by technology

However, due to the slow growth of natural enzyme-producing strains or (and) high nutritional requirements, the production cost is higher

Currently, there are studies using engineering bacteria to produce these three enzymes separately. Since one strain is used to produce one of the enzymes, three processes are required to produce the three enzymes, which makes the production cost too high.