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題 名 | 兩水相萃取技術在生物殺蟲劑蘇力菌素的分離純化應用 |
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作 者 | 曾耀銘; 吳美貌; 徐泰浩; | 書刊名 | 大葉學報 |
卷 期 | 2:1 1993.12[民82.12] |
頁 次 | 頁109-122 |
分類號 | 433.85 |
關鍵詞 | 兩水相系統; 蘇力菌素; 生物檢定; Two-phase aqueous system; Thuringiensin; Bioassay; |
語 文 | 中文(Chinese) |
中文摘要 | 蘇力菌素(thuringiensin),俗稱B-外毒素,是由某些蘇力菌亞種分泌而得的一種有效蠅類防治成分,具有相常高的商品化潛力。本研究主要是利用兩水相系統(two-phase aqueous systcem)進行蘇力菌素的分離純化,並以高效液相層析法(HPLC) 及生物檢定法確認回收蘇力菌素的活性。多次重複試驗的結果顯示,含15% 聚乙烯醇(PEC)8000,和10% 磷酸氫二鉀(K2HPO4)溶液的分配係數最佳,蘇力菌素大都集中到下層液(磷酸鹽相);在500mL樣品處理之平均回收率達85.5%。由HPLC定量和利用家蠅幼蟲進行生物檢定的關聯性探究得知,經兩水相系統處理後的上、下層溶液均有和醱酵上清液相當的殺蟲活性,足見兩水相系統十分適合具活性之蘇力菌素的回收,由此也同時証明所使用HPLC法具有優良精確的定量能力。 |
英文摘要 | Some Bacillus thuringiensis (B. t.) strains secret a heat-stable insecticidal adenine-nucleotide analog, thuringiensin, which is also known as “fly factor” with a high commercial potential in fly control. In this study, we used a series of two-phase aqueous systems to separate the thuringiensin of B. t. subsp. darmstadiensis. Thuringiensin at different processing stages were analyzed with high performance liquid chromatography (HPLC) method and/or bioassay. The results showed that 15% PEG 8000/10% K2HPO4 two-phase aqueous system had highest partition coefficient. In a modest large scale 500mL operation, the averaged recovery of thuringiensin in the lower solution (phosphate phase) was 85.5%. Bioassay results indicated that the additional toxicity from the d-endotoxin and spores existing in the broth was obvious. Comparison of the correlation of HPLC and bioassay methods indicated that the lower solution and the upper solution of the two-phase system had similar effective control rate with supernatant. In conclusion, the 15% PEG 8000/10% K2HPO4 two-phase aqueous system was suitable for recovering active thuringiensin from fermentation broth. In addition, a series of analytical results showed that the HPLC method used in this study is very sensitive and precise in thuringiensin quantitative analysis. |
本系統中英文摘要資訊取自各篇刊載內容。