- 期刊
兩水相萃取技術在生物殺蟲劑蘇力菌素的分離純化應用
Partitioning of Thuringiensin from Bacillus thuringiensis subsp. darmstadiensis Using Polyethylene Glycol and Dipotassium Phosphate Two-phase Aqueous System
摘要
蘇力菌素（thuringiensin），俗稱β-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 system）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。
並列摘要
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 δ-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.
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