KSBB Journal

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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High-Yield Production of Astragalosides from Transgenic Hairy Root Cultures of Astragalus membranaceus

형질전환된 황기 모상근으로부터 Astragalosides의 생산을 위한 연구

  • Hwang, Sung-Jin (Department of Oriental Medicine Materials, Dongshin University)
  • 황성진 (동신대학교 한약재산업학과)
  • Published : 2006.04.28
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Abstract

A transgenic hairy root clone AG-04 of Astragalus membranaceus was obtained following co-cultivation of leaf explants with Agrobacterium rhizogenes ATCC15834. This clone was examined for its growth and production of cyclolanostane-type saponins, astragalosides I, II, and III, under various culture conditions. Among the five basal media tested, Shenk and Hildebrandt(SH)(18) medium was best for roots growth and astragalosides production. The maximum root biomass was obtained at inoculum size of 500 mg FRW per flask, initial sucrose concentration of 3%, and shaking speeds of 90 rpm. The astagalosides production was promoted when the hairy root clone AG-04 was cultured at shaking speeds of 120 rpm and light irradiation of 18 h. Astragaloside contents was also stimulated with high initial sucrose concentration, and the maximum astargalosides contents of 6.21 %/g DRW was obtained at initial sucrose concentration of 6%. The addition of chitosan(100 mg/L) to the culture medium was significantly increased astragalosides production. This was 2.1 times higher than that obtained in a control culture without chitosan.

황기의 형질전환된 모상근으로부터 약리물질인 astragalosides I, II, 그리고 III을 효율적으로 생산하기 위하여 다양한 물리화학적 조건들 즉, 배지, 초기 당농도, 교반속도, 초기 접종농도, 광조사, 그리고 chitosan 처리에 한 elicitation 효과를 조사 하였다. 선발된 AG-04 clone의 성장과 astragalosides의 함량은 4종의 배지들 중 SH배지에서 가장 좋았으며, 배지에 첨가하는 초기 당 농도는 3%와 6%에서 각기 건물중과 astragalosides의 함량이 가장 높게 나타났다. 또한, 초기 접종량은 50 ml 배지에 500 mg(FRW)씩을 주입하는게 가장 효과적이었으며, 18시간 광조사가 되는 Shaking incubator에서 90 rpm과 120으로 교반시켰을 때 모상근의 성장과 astargalosides의 함량이 높게 나타났다. 배양 2주 후 chitosan의 처리는 모상근의 성장에는 크게 영향을 미치지 않았으나, 100 mg/L 처리구에서 대조구의 약 2.1배의 astragalosides 함량 증가를 가져왔다. 이와같은 연구결과는 황기의 대량배양을 통한 saponins 생산 연구에 효율적으로 활용될 수 있을 것으로 본다.

Keywords

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