Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Enzymatic Extraction of Pilocarpine from Pilocarpus jaborandi

Pilocarpus jaborandi로부터 필로카르핀의 효소반응추출

  • Cho, Jun-Ho (Department of Pharmaceutical Engineering, Sunmoon University) ;
  • Bhattarai, Saurabh (Department of Pharmaceutical Engineering, Sunmoon University) ;
  • Oh, Tae-Jin (Department of Pharmaceutical Engineering, Sunmoon University) ;
  • Jang, Jong Hwa (Department of Dental Hygiene, Hanseo University)
  • Received : 2013.03.25
  • Accepted : 2013.04.25
  • Published : 2013.06.28
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Abstract

Pilocarpine is an imidazole alkaloid, found exclusively in the Pilocarpus genus, with huge pharmaceutical importance. In order to extract pilocarpine from Pilocarpus jaborandi, environmentally friendly enzyme-assisted extraction was applied. Viscozyme$^{(R)}$ L, a commercially available enzyme cocktail, was used for the study. The conditions for extraction were optimized on the basis of substrates, enzymes, temperatures and pHs. Optimum conditions for extraction with the highest yield were 30 h reaction of 100 mg substance at $45^{\circ}C$ in 40 ml of 50 mM acetic acid, pH 4. A 10% enzyme concentration was found to be the best for extraction. Total pilocarpine content after extraction was analyzed by HPLC. The total pilocarpine content ($1.14{\mu}g/mg$) obtained from Viscozyme$^{(R)}$ L treatment was 3.08-fold greater than those of the control treatment ($0.37{\mu}g/mg$).

필로카르핀은 Pilocarpus 속으로부터 유일하게 분리되는 이미다졸계 알칼로이드로서 상당히 제약적으로 중요하다. Pilocarpus jaborandi로부터 필로카르핀을 추출하기 위하여 환경친화적인 효소를 이용한 추출법을 이용하였다. 본 연구에서는 상업적으로 이용할 수 있는 효소칵테일인 Viscozyme$^{(R)}$L을 사용하였다. 추출 조건은 기질, 효소, 온도 및 pH 등에 기초하여 최적화되었다. 가장 높은 수율을 위한 최적화 조건은 pH4인 50 mM 아세트산 40 ml 하에서 $45^{\circ}C$, 100 mg 기질, 30시간 반응이였다. 최적의 추출 효소농도는 10%이였다. Viscozyme$^{(R)}$L 처리로부터 얻어진 전체 필로카르핀 함유량($1.14{\mu}g/mg$) 수준은 기존 처리방법에서 얻어지는 양($0.37{\mu}g/mg$)보다 3.08배 높은 것을 확인하였다.

Keywords

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