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Azadi, Pejman,Otang, Ntui Valentaine,Chin, Dong Poh,Nakamura, Ikuo,Fujisawa, Masaki,Harada, Hisashi,Misawa, Norihiko,Mii, Masahiro The Korean Society of Plant Biotechnology 2010 Plant biotechnology reports Vol.4 No.4
Lilium ${\times}$ formolongi was genetically engineered by Agrobacterium-mediated transformation with the plasmid pCrtZW-N8idi-crtEBIY, which contains seven enzyme genes under the regulation of the CaMV 35S promoter. In the transformants, ketocarotenoids were detected in both calli and leaves, which showed a strong orange color. In transgenic calli, the total amount of carotenoids [133.3 ${\mu}g/g$ fresh weight (FW)] was 26.1-fold higher than in wild-type calli. The chlorophyll content and photosynthetic efficiency in transgenic orange plantlets were significantly lowered; however, after several months of subculture, they had turned into plantlets with green leaves that showed significant increases in chlorophyll and photosynthetic efficiency. The total carotenoid contents in leaves of transgenic orange and green plantlets were quantified at 102.9 and 135.2 ${\mu}g/g$ FW, respectively, corresponding to 5.6- and 7.4-fold increases over the levels in the wild-type. Ketocarotenoids such as echinenone, canthaxanthin, 3'-hydroxyechinenone, 3-hydroxyechinenone, and astaxanthin were detected in both transgenic calli and orange leaves. A significant change in the type and composition of ketocarotenoids was observed during the transition from orange transgenic plantlets to green plantlets. Although 3'-hydroxyechinenone, 3-hydroxyechinenone, astaxanthin, and adonirubin were absent, and echinenone and canthaxanthin were present at lower levels, interestingly, the upregulation of carotenoid biosynthesis led to an increase in the total carotenoid concentration (+31.4%) in leaves of the transgenic green plantlets.
반응표면분석법을 이용한 Paracoccus sp.의 Astaxanthin 생산배지 최적화
최종일(Jong-il Choi),이희섭(Hee-Sub Lee),최선강(Seon-Kang Choi),김재훈(Jae-Hun Kim),김진규(Jin Kyu Kim),Norihiko Misawa,변명우(Myung-Woo Byun),이주운(Ju-Woon Lee) 한국생물공학회 2009 KSBB Journal Vol.24 No.3
Paracoccus sp.의 astaxanthin 생산성 증가를 위해 반응표면 분석법을 사용하여 최적의 배지조성을 설계하였다. Paracoccus의 성장 배지인 Marine Broth와 modified Blaszczyk 배지를 사용하여 astaxanthin 생산성을 측정한 결과 각각 0.39 mg/L, 0.40mg/L의 astaxnathin 농도를 보였다. Modified Blaszczyk 배지의 조성을 각각 변수로 두어 가장 많은 영향을 주는 성분을 알아본 결과 MgSO₄와 yeast extract로 확인되었다. 중심합성계획법에 따라 MgSO₄ (0.397~4.621 g/L), yeast extract (2.879~7.121 g/L)를 달리하였을 때, astaxanthin 생산성에 대한 회귀식의 R²은 0.894로 나타났고, 이에 따른 최대 생산량은 0.925 mg/L로 예상되었으며 이때의 MgSO₄ 와 yeast extract의 농도는 각각 2.83과 7.02 g/L로 나타났다. 이에 대한 확인실험 결과 2.83 MgSO4 g/L, 7.02 yeast extract g/L에서 1.021 mg/L의 astaxanthin이 생산되었으며, 배지의 최적화에 따라 250% 이상의 생산성 증가가 확인되어졌다. This study was to optimize the medium components for astaxanthin production in Paracoccus sp. through surface response methodology. A screening test was first conducted on 5 medium components using a Plackett-Burman design, from which MgSO₄ and yeast extract were identified as the significant factors affecting astaxanthin production. These significant factors were optimized by central composite design of experiments and response surface methodology, as 2.83 g/L MgSO4 and 7.02 g/L yeast extract, respectively. The expected astaxanthin concentration with these optimized medium compositions were 0.925 mg/L. In flask culture, the experimentally obtained concentration of astaxantin was 1.021 mg/L, where it had been 0.4 mg/L before optimization.