Bacillus licheniformis 분리균 2종의 α-Galactosidase 생산성과 효소특성
Production and Characterization of α-Galactosidases from Two Bacillus licheniformis Isolates
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전통 발효된장으로부터 α-galactosidase를 분비 생산하는 두 종류 Bacillus licheniformis 균주로 YB-1413과 YB-1414를 분리하여 당 이용능을 비롯한 생화학적 특성 및 16S rRNA 염기서열과 중합효소 연쇄반응에 의한 임의증폭 DNA 다형성의 유전학적 특성을 비교한 결과 두 균주는 매우 유사하였지만 동일 균주는 아닌 것으로 확인되었다. 탄소원으로 wheat bran을 사용하였을 때는 두 균주의 α-galactosidase 생산성이 증가하였으나, melibiose, raffinose 또는 sucrose는 효소 생산성이 급격하게 감소하였다. 질소원으로 yeast extract가 첨가된 배지에서 효소 생산성이 높았으며, YB-1413은 1.87 U/ml, YB-1414는 1.69 U/ml의 효소생산성을 보였다. 이들 균주가 생산하는 α-galactosidases는 pH 6.0과 45℃에서 모두 최대활성을 보였으며, 낮은 농도의 ribose와 galalctose에 의해서도 효소활성이 급격하게 저해되었다. 한편 이들 효소는 대두와 콩과식품에 존재하는 항영양인자인 raffinose와 starchyose를 완전히 가수분해 하였는데, 이로 보아 분리균 YB-1413과 YB-1414는 콩 발효식품과 대두 사료의 영양가를 개선하는데 활용할 가치가 있다고 여겨진다.
Abstract
Two bacterial strains, Bacillus licheniformis YB-1413 and YB-1414, producing extracellular α-galactosidase, were obtained from homemade Doenjang. On the basis of their biochemical properties, 16S rRNA sequences and random amplified polymorphic DNA patterns by polymerase chain reaction, they were found to be somewhat different from one another. α-Galactosidase productivities of the two isolates were increased by wheat bran, but drastically decreased by melibiose, raffinose and sucrose which were used as carbon sources. The enzyme productivities were increased by yeast extract as a nitrogen source with maximum levels of 1.87 U/ml for YB-1413 and 1.69 U/ml for YB-1414, respectively. The enzymes of both isolates exhibited maximum activity for hydrolysis of para-nitrophenyl-α-D-galactopyranoside (pNP-αGal) under reaction conditions of pH 6.0 and 45℃. Their hydrolyzing activities for pNP-αGal were drastically decreased by the addition of low concentrations of ribose and galactose. They were capable of hydrolyzing completely α-1,6 linked galactosyl residue in melibiose, raffinose and stachyose, which are known to be anti-nutritional factors in products of soybean and legume. In relation to the latter, the isolates YB-1413 and YB-1414 have potential applicability in improving soybean-fermented foods and the nutritional value of soybean feed.
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