摘要
蒎烯是一种重要的平台化合物,可以用来合成高密度燃料、精细化学品及材料。将来源于黄花蒿的β-蒎烯合成酶基因与外源杂合甲羟戊酸(MVA)途径一起整合到BL21(DE3)中共表达,通过气相色谱-质谱(GC-MS)和气相色谱(GC)对发酵产物进行了定性及定量检测。通过发酵条件优化,对影响发酵产β-蒎烯的因素(诱导温度、诱导剂浓度和N源)进行了考察。结果表明:来自黄花蒿的蒎烯合酶可以在宿主细胞内高效表达,且能高效催化β-蒎烯的合成。通过对重组菌进行发酵条件优化得到最佳培养条件:诱导温度为28℃,IPTG浓度为0.1 mmol/L,最佳有机N源为MD牛肉粉。在此条件下,检测出β-蒎烯产量达到22.89 mg/L,比优化前(4.60 mg/L)提高了3.98倍。
Pinene is an important natural product that is widely used in high-density renewable fuels and fine chemicals. The Artemisia annua β-pinene synthase gene （OH6） was successfully assembled in Escherichia coli BL21 （DE3） with the heterologous hybrid mevalonate （MVA） pathway. Pinene was quantitatively analyzed and characterized by gas chromatography （GC） and GC-mass spectrometry （MS）. The influences of induction temperature, inducer concentration, and nitrogen on production of β-pinene were studied. After shake-flask fermentation, β-pinene productivity of the engineered strain was 4.60 mg/L. Under the optimum conditions of induction temperature 28℃ ,inducer （IPTG） concentration 0. 1 retool/L, and beef extract powder （ MDbio Inc. ）, theβ-pinene yield was up to 22.89 mg/L, 3.98 times higher than that before optimization.
出处
《生物加工过程》
CAS
2015年第1期28-34,46,共8页
Chinese Journal of Bioprocess Engineering
基金
国家自然科学基金(21376255)
国家高技术研究发展计划(863计划)(SS2013AA050703-2)
青岛科技发展计划(12-1-4-9-(3)-jch)
