Improvement of alpha-amylase production by modulation of ribosomal component protein S12 in Bacillus subtilis 168.
The capacity of ribosomal modification to improve antibiotic production by Streptomyces spp. has already been demonstrated. Here we show that introduction of mutations that produce streptomycin resistance (str) also enhances alpha-amylase (and protease) production by a strain of Bacillus subtilis as estimated by measuring the enzyme activity. The str mutations are point mutations within rpsL, the gene encoding the ribosomal protein S12. In vivo as well as in vitro poly(U)-directed cell-free translation systems showed that among the various rpsL mutations K56R (which corresponds to position 42 in E. coli) was particularly effective at enhancing alpha-amylase production. Cells harboring the K56R mutant ribosome exhibited enhanced translational activity during the stationary phase of cell growth. In addition, the K56R mutant ribosome exhibited increased 70S complex stability in the presence of low Mg2+ concentrations. We therefore conclude that the observed increase in protein synthesis activity by the K56R mutant ribosome reflects increased stability of the 70S complex and is responsible for the increase in alpha-amylase production seen in the affected strain.
Kurosawa
Kazuhiko
K
National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan.
Hosaka
Takeshi
T
Tamehiro
Norimasa
N
Inaoka
Takashi
T
Ochi
Kozo
K
D001412:Bacillus subtilis / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins; D015964:Gene Expression Regulation, Bacterial; D005818:Genetic Engineering / Q000379:methods; D010447:Peptide Hydrolases / Q000378:metabolism; D017354:Point Mutation; D000091243:Ribosomal Protein S9; D012269:Ribosomal Proteins / Q000235:genetics* / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism; D000516:alpha-Amylases / Q000096:biosynthesis*
D029968:Escherichia coli Proteins; D000091243:Ribosomal Protein S9; D012269:Ribosomal Proteins; C000717190:RpsI protein, E coli; C034212:ribosomal protein S12; D000516:alpha-Amylases; D010447:Peptide Hydrolases