碩士 / 國立中興大學 / 基因體暨生物資訊學研究所 / 103 / The Chlorella sp. DT, an endemic species in Taiwan, was found with astaxanthin producing ability. To maximize productivity of astaxanthin in Chlorella sp. DT, NaNO3, Na2CO3 and CaCl2 were selected as inducers by factors screening. Further, response surface methodology was used to optimize the cultural condition for astaxanthin production. The results showed astaxanthin productivity was 0.749 mg L-1 after 10 days of cultivation and induction by used the optimized the cultural condition, BG11 medium with NaNO3 2779.08 mg L-1, Na2CO3 273.97 mg L-1 and CaCl2 75.34 mg L-1. Furthermore, based on factors screening, the culture conditions for two-stage to produce astaxanthin by Chlorella sp. DT were developed. The PBD-2 medium was used to accumulate biomass in green-stage, and the Pred-medium was used for astaxanthin production in induce-stage. Astaxanthin productivity is 0.805 mg L-1 after 12 days of cultivation and induction in two-stage strategy. We successfully maximize the astaxanthin production in Chlorella sp. DT by one and two-stage of cultivation strategies. To increase the production of astaxanthin, we cloned the Chlorella sp. DT β-carotene ketolase (BKT), the key enzyme of astaxanthin synthesis, and overexpressed the BKT gene in Nicotiana tabacum and N. benthamiana by using agrobacterium-mediated transformation. The transgenic N. tabacum and N. benthamiana plants can produce 604.6 and 385.8 μg astaxanthin per g (dry weightof leaves), respectively. This study successfully used β-carotene ketolase gene form Chlorella sp. DT to build a transgenic plant system for astaxanthin production.
| Identifer | oai:union.ndltd.org:TW/103NCHU5105010 |
| Date | January 2015 |
| Creators | Yi-Ting Kuo, 郭一廷 |
| Contributors | Yu-Ting Chen, 陳玉婷 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 138 |
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