The feasibility of surfactant assisted refolding, which was performed by diluting denatured protein sample into a refolding buffer containing a surfactant of appropriate concentration, was examined by using recombinant human lysozyme (rhLys) and recombinant β-mannanase (rMan) as protein samples.

Recombination of mannanase gene man23 and its expression in Brevibacillus brevis;

Expression of recombinant mannanase gene Man23 in Bacillus sp. WB600;

Construction of a double functional recombinant strainof Pichia pastoris co-expressing phytase and mannanase and the enzymatic analyses;

Study on the immobilized β-mannanase by chitin;

Study on the Production of β-mannanase by Bacillus M-21;

Breeding of β-mannanase Producing Strains;

Optimization of β-mannase production by bacillus subtilis;

Acetone precipitation of β-mannase;

External Secretion of β-Mannase by Transgenic E.coli in Intestine of Swines;

A plasmid ,pMD-18Tman2XcmI is developed,the plasmid is constructed by cloning an Xcm?I cassette with mannanase gene (man 1?101?bp long) between two Xcm?I restriction sites into pMD-18T.