Expression and secretion of an acid-stable alpha-amylase gene in Bacillus Subtilis by SacB promoter and signal peptide.
Alpha amylase gene from Bacillus licheniformis was mutated by site-directed mutagenesis to improve its acid stability. The mutant gene was expression in Bacillus subtilis under the control of the promoter of sacB gene which was followed by either the alpha-amylase leader peptide of Bacillus licheniformis or the signal peptide sequence of sacB gene of Bacillus subtilis. Both peptides efficiently directed the secretion of alpha-amylase from the recombinant B. subtilis cells. The extracellular alpha-amylase activities in two recombinants were 1001 and 2012 U ml(-1), respectively. The purity of the recombinant product was confirmed by SDS-PAGE.
Heng
Cai
C
Tianjin Key Lab of Industrial Microbiology, College of Biotechnology, Tianjin University of Science & Technology, 300222, Tianjin, PR China. caihengmail@yahoo.com.cn
Chen
Zhongjun
Z
Du
Lianxiang
L
Lu
Fuping
F
D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D015870:Gene Expression; D006863:Hydrogen-Ion Concentration; D016297:Mutagenesis, Site-Directed; D011401:Promoter Regions, Genetic; D021382:Protein Sorting Signals / Q000502:physiology*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D000516:alpha-Amylases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
D001426:Bacterial Proteins; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D000516:alpha-Amylases