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국문 초록 (Abstract)

토양시료를 미생물 분리 시료로 이용하여 CM-cellulose를 기질로 함유한 배지상에서 명확한 cellulase 활성환을 형성함과 동시에 액체배양액으로부터 높은 cellulase 활성을 보이는 균주를 단리하였...

토양시료를 미생물 분리 시료로 이용하여 CM-cellulose를 기질로 함유한 배지상에서 명확한 cellulase 활성환을 형성함과 동시에 액체배양액으로부터 높은 cellulase 활성을 보이는 균주를 단리하였으며, 본 균주 유래의 16S rRNA 유전자 분석을 통하여 본 균주를 B. subtilis 로 동정함과 동시에 B. subtilisKB1으로 명명하였다. B. subtilis KB1 유래 cellulase는 pH 5.0과 50°C의 조건하에서 가장 높은 효소활성을 보였으며, 30, 40, 50, 60, 그리고, 70°C 에서의 효소 half-life는 각각 5,404,133, 37, 24, 그리고, 6분을 나타내었다. 그리고, 본 효소의 기질특이성 검토에서 전형적인 endo-type cellulase 기질인 CMcellulose에 대하여 가장 높은 효소활성을 보이는 것으로부터 endo-type cellulase임이 강하게 시사되었다. B. subtilis KB1균주 유래 cellulase 유전자를 cloning하여 유전자 배열을 규명한 결과 B. subtilis KB1 유래 cellulase 유전자는 아미노산499개를 암호화하는 1500 bp의 open reading frame (ORF)으로 이루어져 있었으며, 아미노산 배열로부터 추정되는 분자량은 55, 251 Da이었다. 그리고, 규명된 cellulase 유전자로부터 추정된 아미노산 배열을 이용하여 상동성을 검토한 결과 cellulase는 Glycoside hydrolase family (GH) 1에 속하는 cellulase와 높은 상동성을 나타내었다.

다국어 초록 (Multilingual Abstract)

We isolated a strain producing celluase from environmental soil. The strain was formed activity zone on the LB agar plate containing carboxymethylcellulose (CM-cellulose) stained with trypan blue as substrate with its cellulase activity. In the analysis of the 16S rRNA gene sequence, it was identified as Bacillus subtilis and was named as Bacillus subtilis KB1. The B. subtilis KB1 cellulase showed the maximum activity at pH 5.0 and 50°C. The half-lives of the enzyme at 30, 40, 50, 60 and 70°C were 5,404, 133, 37, 24, and 6 min, respectively. In the investigation of substrate specificity, B. subtilis KB1 cellulase showed the most high enzyme activity for the CM-cellulose substrate. To clone the cellulase gene from B. subtilis KB1, the cellulase gene was amplified from genomic DNA of the B. subtilis KB1 with designed primers based on the cellulase gene from Bacillus subtilis (GenBank: AGN52749) by PCR. In the analysis of the cellulase gene sequence, it was confirmed that the cloned cellulase gene was consisted of a 1,500 bp open reading frame (ORF) encoding a 499 amino acid protein with calculated molecular mass of 55,251 Da. The deduced amino acid sequences from cellulase gene showed high identity with glycosyl hydrolases family (GH) 1.

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