효모 표층 Arylsulfatase에 의해 제조된 Agarose의 특성
Characterization of Agarose Produced by Yeast Cell Surface Displayed-Arylsulfatase
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초록
Agar로부터 agarose 제조 시 유기용매를 이용해서 황을 제거하는 방법이 일반적으로 많이 사용되어진다. 하지만 agar 의 황을 가수 분해하는 효소를 사용할 경우 agarose 제조 시공정과정을 획기적으로 간소화할 수 있다. 따라서 arylsulfatase 로 agaropectin에서 황 제거를 통해 agarose로 바꾸는 공정은 간단하고 높은 수율을 얻을 수 있기 때문에 agarose 생산에 효율적으로 적용할 수 있다. 본 연구에서는 arylsulfatase를세포표면에 발현하는 효모생촉매를 이용하여 제조한 agarose의 황 함량과 gel 강도를 측정하였다. 처리한 효모(효)소의 농도가 증가할수록 증가된 탈황 반응에 의해 황함량이 줄어들었고, 특히 35 unit/mL의 효소 농도로 처리하였을 때 황 함량은 0.2%까지 감소하는 것을 확인할 수 있었다. 황 함량을 가장 낮출 수 있는 최적 조건은 0.6% agar(Junsei)용액에 효모 표층 arylsulfatase 35 unit/mL로 처리하고40oC에서 3시간 반응시켰을 때 였다. 또한 1.0% DNA 전기영동용 agarose의 gel 강도는 효모 표층 arylsulfatase 처리로제조된 agarose의 경우 559.8±0.12로 상업적 agarose의 gel 강도(880.6±0.15 g/cm2) 보다는 낮았다. 따라서 효모 S. cerevisiae의 세포 표면에서 발현된 재조합 arylsulfatase 효소를 이용하여 agar로부터 전기영동용 agarose의 생산 공정에 적용 가능함을 알 수 있었다.
Abstract
Enzymatic hydrolysis of sulfate groups in agaropectin or agar simplifies the production process of high-quality or low sulfate-content agarose. This study was investigated that cell surface displayed arylsulfatase can be applied to desulfatation of agar for production of agarose. Sulfate content of agarose prepared by treatment of yeast surface-displayed arylsulfatase was decreased in a enzyme dosedependent manner. Especially, 35 unit/mL of yeast surface arylsulfatase attenuated sulfate content of agarose up to 0.2%. In the 0.6% agar(Junsei), 35 unit/mL enzyme treated at 40oC for 3 h showed the lowest content of sulfate. Therefore, this result was determined to be the optimal condition to desulfatation of agar for production of agarose. In addition, the gel strength of yeast surface arylsulfatase treated agar and commercial agarose were compared. Agarose prepared by treatment of yeast surface arylsulfatase showed 559.8±0.12 of gel strength, and it is a similar compared to the commercial agarose.
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