Affiliation:
1. Division of Enzymology, Institute of Applied Microbiology, The University of Tokyo, Bunkyo-ku, Tokyo, Japan
Abstract
Deoxyribonucleic acid (DNA) of
Bacillus subtilis
var.
amylosacchariticus
showed almost the same ability as
B. subtilis
Marburg to induce transfer of several genetic markers in DNA-mediated transformation. DNA-DNA hybridization data also showed an intimate relationship between the two strains. Genetic elements involved in the production of extracellular α-amylase (EC 3.2.1.1.) in
B. subtilis
var.
amylosacchariticus
were studied by using DNA-mediated transformation. Two Marburg derivatives, NA20(
amyR2
) and NA20-22(
amyR1
), produced about 50 and 10 U of α-amylase per mg of cells, respectively, whereas
B. subtilis
var.
amylosacchariticus
produced as much as 150 U of the enzyme per mg of cells. When
B. subtilis
var.
amylosacchariticus
was crossed with strain NA20-22 as recipient, transformants that acquired high α-amylase productivity (about 50 U/mg of cells) were obtained. Genetic analysis revealed that a regulator gene (
amyR
) for α-amylase synthesis was found in
B. subtilis
var.
amylosacchariticus
, as in the case of
B. natto
1212 (
amyR2
) and
B. subtilis
Marburg (
amyR1
). The allele was designated
amyR3;
it is phenotypically indistinguishable from
amyR2
, but is readily distinguishable from
amyR1
. The presence of
amyR3
was not sufficient for an organism to render production of an exceptional amount of α-amylase. Extra-high α-amylase producers could be obtained by crossing
B. subtilis
var.
amylosacchariticus
as donor with strain NA20 as recipient. The transformants produced the same or even greater amounts of the enzyme than the donor strain. Results suggest the presence of another gene that is involved in the production of the exceptional amount of α-amylase.
Publisher
American Society for Microbiology
Subject
Molecular Biology,Microbiology
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