Construction of transgenic Bacillus mucilaginosus strain with improved phytase secretion

Abstract

Aims: To construct a transgenic Bacillus mucilaginosus strain to increase the secretion capability of a wild-type isolate of B. mucilaginosus D 4B1 to hydrolyse phytate phosphorus, which can be used as a microbial fertilizer in field application. Methods and Results: We constructed a phytase secreting expression vector pSP43 with a mini-Tn5 transposon and a Aspergillus fumigatus phytase expression cassette. The vector pSP43 was successfully transferred into the wild-type B. mucilaginosus using the particle bombardment method, and three transgenic strains with a stable copy of phytase expression cassette integrated into the chromosome of the B. mucilaginosus by Tn5 transposition were selected. The phytase activity of the engineered strains increased 36-46-fold when compared with the wild-type strain of D 4B1. Conclusions: The A. fumigatus phytase gene can be expressed under the direction of p43 promoter in B. mucilaginosus. The expression protein is secreted extracellularly and newly constructed strains showed a high phytase activity. Significance and Impact of the Study: A transgenic Bacillus strain by the particle bombardment method was constructed. © 2005 The Society for Applied Microbiology.