Optimized high-activity human lysozyme gene as well as expression vectors and applications thereof

Abstract

The invention provides an optimized high-activity human lysozyme gene as well as expression vectors thereof. The gene sequence is optimized according to the codon bias of Pichia pastoris and the human lysozyme gene is artificially synthesized; recombinant plasmids pUC18-T-HZ and pPICZ(alpha)A-HZ are constructed; the recombinant plasmid pPICZ(alpha)A-HZ is linearized and then transfected to Pichiapastoris, and positive transformants are obtained by screening. The study results show that the recombinant Pichia pastoris can secrete and express high-activity human lysozyme, and the expression product has significant bacteriostatic activity. According to the invention, the Pichia pastoris expression system is adopted to produce the human lysozyme with high expression level and high activity, so as to overcome the problems in the prior art, such as high cost, low expression level and low activity. The production and the promotion and application of the high-activity and low-cost lysozyme can bring considerable economic and social benefits for the development of feed and animal husbandry industries in China and huge ecological benefit as well.

Description

technical field [0001] The invention belongs to the technical field of molecular biology and pharmacy, and in particular relates to an optimized human lysozyme gene and its expression vector and application. Background technique [0002] Lysozyme (Lysozyme) is called 1,4-β-N-lysozyme, also known as mucopeptide N-acetylmuramoyl hydrolase. It was first discovered in human saliva, tears and nasal mucus by the British bacteriologist Fleming in 1922. It is named for its ability to dissolve bacterial cell walls and have a bacteriolytic effect. Lysozyme is an important defense factor in human and animal body fluids and tissues. It has strong bactericidal effect, wide bactericidal spectrum, strong stability, and is easily accepted by the body. It can also promote the recovery of immune function and has no toxicity. Side effects and adverse reactions. Therefore, as a biological agent, lysozyme is widely used in food, feed industry and clinical medicine. Human lysozyme participates...

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Problems solved by technology

[0003] The present invention provides an optimized high-activity lysozyme in the prior art in view of the defects of slow growth, low density and unsatisfactory exogenous protein expression and post-translational processing in the production of human lysozyme using the Saccharomyces cerevisiae expression system Human lysozyme gene and its expression vector, the present invention uses Pichia pastoris expression system as the expression system for producing human lysozyme, which has the advantages of low production cost, simple operation, rapid growth, high expression efficiency, and post-transcription processing modification and good fermentation and secretion performance, etc., the human lysozyme produced by the optimized human lysozyme gene of the present invention through the Pichia pastoris expression system has the advantages of high activity and high expression

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