• Journal of Microbiology and Biotechnology
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• 제24권8호
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• pp.1096-1104
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• 2014

$\small{L}$-Asparaginase from gram-positive bacteria has been poorly explored. We conducted recombinant overexpression and purification of $\small{L}$-asparaginase from Staphylococcus sp. OJ82 (SoAsn) isolated from Korean fermented seafood to evaluate its biotechnological potential as an antileukemic agent. SoAsn was expressed in Escherichia coli BL21 (DE3) with an estimated molecular mass of 37.5 kDa, determined using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Consistent with asparaginases in gram-negative bacteria, size-exclusion chromatography determined SoAsn as a homodimer. Interestingly, the optimal temperature of SoAsn was $37^{\circ}C$ and over 90% of activity was retained between $37^{\circ}C$ and $50^{\circ}C$, and its thermal stability range was narrower than that of commercial E. coli $\small{L}$-asparaginase (EcAsn). Both SoAsn and EcAsn were active between pH 9 and 10, although their overall pH-dependent enzyme activities were slightly different. The $K_m$ value of SoAsn was 2.2 mM, which is higher than that of EcAsn. Among eight metals tested for enzyme activity, cobalt and magnesium greatly enhanced the SoAsn and EcAsn activity, respectively. Interestingly, SoAsn retained more than 60% of its activity under 2 M NaCl condition, but the activity of EcAsn was reduced to 48%. Overall, the biochemical characteristics of SoAsn were similar to those of EcAsn, but its kinetics, cofactor requirements, and NaCl tolerance differed from those of EcAsn.

• 한국식품영양학회지
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• 제27권5호
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• pp.897-905
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• 2014

고구마 경엽을 식품소재로 활용하기 위해 신황미, 하얀미, 스이오우를 수확시기(60, 90, 120일) 및 마디별(1~5, 6~10, 11~15마디)로 채취, 동결건조한 후 분쇄하여 L값과 총 폴리페놀 함량, DPPH 라디칼 소거 활성을 측정하였다. 그 결과, 신황미는 하얀미, 스이오우에 비해 높은 폴리페놀 함량과 DPPH 라디칼 소거 활성을 나타냈으며, 이는 재배기간이 길수록 정상의 순에 가까울수록 높았다. 90일 동안 재배한 후 1~10마디의 경엽을 채취, 2단계 열풍건조법으로 제조한 신황미, 하얀미, 스이오우 경엽 분말의 총 아미노산 함량은 각각 2,469, 3,384, 2,531 mg/100 g, 총 무기물 함량은 각 7.23, 6.54, 7.11 g/100 g, 총당은 각 11.6, 12.8, 18.2 g/100 g, 비타민 E 함량은 3.04, 3.18, 1.52 mg/100 g이었다. 또한 총 폴리페놀 함량은 각각 5.03, 3.90, 4.02 mg/g, ${\beta}$-카로티노이드는 75.6, 71.6, 63.1 mg/g이었고, DPPH 라디칼 소거 활성($EC_{50}$)은 각각 0.29, 0.36, 0.33 mg, ABTS 라디칼 소거 활성($EC_{50}$)은 0.12, 0.15, 0.11 mg으로 시금치보다 2배 이상 강한 활성을 나타내었다.