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題 名 | Purification of Thuringiensin from Bacillus Thuringiensis Using Calcium Silicate Adsorption and Dibasic Sodium Phosphate Desorption Methods=利用矽酸鈣及磷酸氫二鈉對蘇力菌素的吸附性及去吸附性作為純化的方法 |
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作 者 | 劉正民; 劉炳嵐; 曾耀銘; 童吉士; | 書刊名 | Journal of the Chinese Institute of Chemical Engineers |
卷 期 | 32:6 2001.11[民90.11] |
頁 次 | 頁537-545 |
分類號 | 433.85 |
關鍵詞 | 蘇力菌素; 生物殺蟲劑; Bacillus thuringiensis; Calcium silicate adsorption; Capillary electrophoresis; Insecticide; Thuringiensin; |
語 文 | 英文(English) |
中文摘要 | 蘇力菌素是由蘇力菌所分泌出的代謝產物,它是一種極具潛力取代化學殺蟲劑的 生物殺蟲劑。蘇力菌素由於其分子量小,所以純化不易。本文介紹改良式蘇力菌素回收法以 取代傳統之過濾膜回收法,此一改良之新方法不但簡易而且經濟有效。首先將含有蘇力菌素 之發酵液先加入矽酸鈣經調至微酸性,大約百分之八十九之蘇力菌素可被矽酸鈣所吸收,吸 收後的矽酸鈣經離心分離再以高濃度的磷酸氫二鈉將蘇力菌素從矽酸鈣解離出。本實驗發現 用 0.4 M 之磷酸氫二鈉在 pH 9.2 的條件下可釋放出最多量之蘇力菌素, 此釋放出的蘇力 菌素可經由毛細管電泳作定量分析後,做為配方製劑之依據。如欲進一步純化,則可經由製 備型 HPLC 分離後以電透析去除過多的鹽分,最終產物可由毛細管電泳及質譜儀加以鑑定。 |
英文摘要 | Thuringiensin (β-exotoxin) is a metabolic product derived from Bacillus thuringiensis. This exotoxin is a very effective pesticide and has great potential for replacing synthetic chemical pesticides in the future. In this report, we introduce an improvement in the method of thuringiensin recovery, which is much simpler and more effective than the traditional membrane filtration method. Calcium silicate was used to adsorb thuringiensin from the fermentation broth, with pH adjusted to 5. The conditions for adsorption were optimized to achieve a recovery percentage of up to 89%. After thuringiensin was captured by calcium silicate, the liquid phase was easily removed by means of centrifugation. Subsequently, the thuringiensin was successfully desorbed from calcium silicate by adding a high ionic strength dibasic sodium phosphate solution. We observed that 0.4 M dibasic sodium phosphate (pH 9.2), with a slurry-to-solution ratio of 1: 70 (w/v) , allowed the maximal amount of thuringiensin to be released from the slurry. This preliminary purified thuringiensin could be quantified by means of capillary electrophoresis and used to formulate the final product. Further purification of thuringiensin was achieved by using semi-preparative HPLC and electrodialysis. The purity of the final product was verified by means of capillary electrophoresis and mass spectrometry. |
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