줄기세포 배양을 통한 들깻잎 로즈마린산 대량 생산 기반 기술 개발
Scaling Up Rosmarinic Acid from Perilla Frutescens Leaves through Stem Cell Culture for Mass Production
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초록
Background: The objective of the present study is to establish a cambium-derived stem cell culture method for stable mass production of rosmarinic acid (RA) from Perilla leaves for industrialization. Methods and Results: Perilla frutescens leaves of the Bora variety were used as starting material. These leaves accumulated the highest levels of RA among the 32 Perilla varieties examined. Callus was successfully induced in solid MS medium containing 2.0 ㎎/ℓ 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.05 ㎎/ℓ benzylaminopurine. Then, white, soft, and crushable stem cells were isolated using a microscope. RA was obtained at a concentration of 40.4 ± 1.4 ㎎/g after culturing callus stem cells in a quarter-strength MS liquid medium containing 0.5 ㎎/ℓ 2,4-D to produce RA in large quantities. To determine the properties of stem cells, we performed the catalase test. When hydrogen peroxide, a strong oxidizing chemical, was applied to stem cells, bubbles appeared almost immediately, demonstrating the antioxidant ability of these cells. In addition, microscopic examination revealed that these stem cells included a relatively large number of tiny vacuoles, whereas callus contained large vacuoles. Conclusions: A cambium-derived stem cell culture method was developed. The method enables the production of RA from P. frutescens leaves in large, industrial-scale quantities.
Abstract
Background: The objective of the present study is to establish a cambium-derived stem cell culture method for stable mass production of rosmarinic acid (RA) from Perilla leaves for industrialization. Methods and Results: Perilla frutescens leaves of the Bora variety were used as starting material. These leaves accumulated the highest levels of RA among the 32 Perilla varieties examined. Callus was successfully induced in solid MS medium containing 2.0 ㎎/ℓ 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.05 ㎎/ℓ benzylaminopurine. Then, white, soft, and crushable stem cells were isolated using a microscope. RA was obtained at a concentration of 40.4 ± 1.4 ㎎/g after culturing callus stem cells in a quarter-strength MS liquid medium containing 0.5 ㎎/ℓ 2,4-D to produce RA in large quantities. To determine the properties of stem cells, we performed the catalase test. When hydrogen peroxide, a strong oxidizing chemical, was applied to stem cells, bubbles appeared almost immediately, demonstrating the antioxidant ability of these cells. In addition, microscopic examination revealed that these stem cells included a relatively large number of tiny vacuoles, whereas callus contained large vacuoles. Conclusions: A cambium-derived stem cell culture method was developed. The method enables the production of RA from P. frutescens leaves in large, industrial-scale quantities.
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