사찰의 된장에서 분리된 Bacillus licheniformis YB-1234의 내열성 α-Amyalse
Thermostable α-Amyalse of Bacillus licheniformis YB-1234 Isolated from the Fermented Soybean of a Korean Buddhist Temple
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초록
국내 사찰에서 제조된 된장으로부터 내열성 α-amylase 생산균으로 분리된 YB-1234는 형태적 특성, 생화학적 성질 및16S rRNA 유전자 염기서열에 근거하여 Bacillus licheniformis 로 동정되었다. B. licheniformis YB-1234의 α-amylase 유전자를 클로닝하여 그 염기서열을 결정하였으며 그로부터유추된 α-amylase의 아미노산 서열은 glycosyl hydrolase family 13에 속하는 B. licheniformis의 내열성 α-amylases 와 매우 높은 상동성을 보였다. α-Aamylase 유전자를 함유한 재조합 대장균과 B. licheniformis에 의해 각각 생산된 α-amylase는 pH 6.0에서 최대활성을 보였으나, 최적 반응온도는 약간의 차이가 있었다. 또한 B. licheniformis로부터 α-amylase는 재조합 대장균에서 생산된 효소보다 열안정성이매우 높았다. 이들 효소에 의한 maltotetraose와 maltohexaose 의 주된 가수분해산물로는 glucose, maltose 및 maltotriose 가 관찰되었다.
Abstract
A bacterial strain was isolated from soybean paste fermented in a Korean Buddhist temple as a producer of the extracellular thermostable α-amylase. The isolate YB-1234 has been identified as Bacillus licheniformis on the basis of its 16S rDNA sequence, morphology and biochemical properties. A gene encoding the thermostable α-amylase of B. licheniformis YB-1234 was cloned into Escherichia coli and its nucleotide sequence was determined. The deduced amino acid sequence of α-amylase was very highly homologous to those of the thermostable α-amylases of B. licheniformis belonging to the glycosyl hydrolase family 13. The α-amylase produced by recombinant E. coli carrying the α-amylase gene exhibited maximal activity at pH 6.0, identical to α-amylase in the culture filtrate of B. licheniformis, while the temperature profile was somewhat different between the two. Particularly, α-amylase produced from B. lcheniformis is much more thermostable than that from recombinant E. coli. The predominant products resulting from the α-amylase hydrolysis were glucose, maltose and maltotriose for maltotetraose and maltohexaose.
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