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KMID : 1007519960050040358
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Food Science and Biotechnology
1996 Volume.5 No. 4 p.358 ~ p.362
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Expression of Streptomyces albus KSM - 35 Amylase with a Bacillus Subtilis Secretion Vector
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Kim, Young Bae
Rhim, Seong Lyul/Lee, Jae Woo/Choi, Won Jin
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Abstract
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A gene encoding the Streptomyces albus KSM-35 amylase (Samy) was expressed in Bacillus subtilis with the aid of an α-amylase-based secretion vector. The S. albus KSM-35 amylase structural gene, lacking its own promoter and some of signal sequence, was introduced into secretion vector pUBA240 containing the promoter and the signal sequence of B. subtilis α-amylase. The resulting plasmid pASA240 was transformed into B. subtilis LKS88 and the amylase activity was detected in starch plate assay. Over 90% of Samy J produced by the transformed B. subtilis was found in the culture medium. The enzyme production by B. subtilis started in early log phase of growth, reached to its maximum in the stationary phase. After 24 hr of cultivation, the amount of amylase activity was about 27-fold higher than that of the parental strain, Streptomyces. The amylase expressed by Bacillus seemed to have the same molecular mass and amino-terminal sequence to those of the parental strain. Plasmid pASA240 was stably maintained up to 24 hr.
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