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利用響應(yīng)面法優(yōu)化巴卡亭III生成10-DAB的工藝條件

發(fā)布時間:2019-01-21 17:59
【摘要】:10-脫乙;涂ㄍII(10-DAB)是紫杉烷類抗腫瘤藥物多烯紫杉醇的前體物質(zhì)。采用響應(yīng)面法優(yōu)化成團(tuán)泛菌P2-A-8轉(zhuǎn)化巴卡亭III生成10-DAB的工藝條件。利用Box-Behnken試驗(yàn)和方差分析,從菌體培養(yǎng)時間、菌體濃度OD600、底物終濃度、轉(zhuǎn)化溫度、轉(zhuǎn)化時間5個方面優(yōu)化成團(tuán)泛菌P2-A-8轉(zhuǎn)化巴卡亭III生成10-DAB的工藝條件。結(jié)果表明,獲得最佳轉(zhuǎn)化工藝條件為:菌體經(jīng)過二級培養(yǎng)12 h,轉(zhuǎn)接后調(diào)節(jié)初始OD600為3.0,此時,加入溶解于甲醇的巴卡亭III溶液,使其終濃度為0.007 mg/m L,32℃,200 r/min下轉(zhuǎn)化4 d,此時10-DAB的生成率達(dá)到24.5%,比優(yōu)化前提高了446.8%。實(shí)驗(yàn)結(jié)果表明,響應(yīng)面法優(yōu)化成團(tuán)泛菌P2-A-8轉(zhuǎn)化巴卡亭III生成10-DAB的工藝條件合理可行。該研究成果對于多烯紫杉醇生物合成具有重要的應(yīng)用價值。
[Abstract]:III (10-DAB) is a precursor of docetaxel, a taxane antitumor drug. Response surface method (RSM) was used to optimize the process conditions for the transformation of Vaca III into 10-DAB by panbacterium P2-A-8. Box-Behnken test and variance analysis were used to optimize the process conditions for the transformation of III from P2-A-8 to 10-DAB from five aspects: cell culture time, final concentration of OD600, substrate, transformation temperature and transformation time. The results showed that the optimum transformation conditions were as follows: after 12 hours of secondary culture, the initial OD600 was adjusted to 3.0, and the final concentration of Vaca III dissolved in methanol was 0.007 mg/m L ~ (2 +) at 32 鈩,

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