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紫穗槐-4,11-二烯合酶催化FPP(farnesyl pyrophosphate,法尼基焦磷酸)生成青蒿素前体紫穗槐-4,11-二烯,是青蒿素生物合成途径中的关键酶。本文对紫穗槐-4,11-二烯合酶的分子生物学和代谢工程研究进行了综述。紫穗槐-4,11-二烯合酶编码基因及其相关核酸序列已经得到了克隆。紫穗槐-4,11-二烯合酶cDNA全长1641bp,编码546aa。紫穗槐-4,11-二烯合酶最适pH范围较宽,但需要二价金属离子作为辅酶才能发挥作用,其产物和底物的特异性不高。在紫穗槐-4,11-二烯合酶作用下,FPP首先进行的是1,6-合环,然后是1,10-合环,形成紫穗槐-4,11-二烯。由于紫穗槐-4,11-二烯合酶在青蒿素生物合成中具有重要的意义,自从其基因被克隆测序后,先后被导入E.coli、S.cereviseae、烟草、拟南芥和A.nidulans,获得了能产生紫穗槐-4,11-二烯的各种工程菌或细胞,研究通过不同方式提高工程菌中紫穗槐-4,11-二烯产量的方法。
Amorpha-4,11-diene synthase catalyzes the formation of the artemisinin precursor amorpha-4,11-diene by farnesyl pyrophosphate (FPP) Key enzyme. In this paper, the molecular biology and metabolic engineering of amorpha-4,11-diene synthase were reviewed. Amorpha-4,11-diene synthase encoding genes and their related nucleic acid sequences have been cloned. Amorpha-4,11-dienol synthase cDNA is 1641bp in length and encodes 546aa. Amorpha-4,11-diene synthase has the widest pH range, but requires divalent metal ions as a coenzyme to play a role, and the specificity of its product and substrate is not high. Under the action of amorpha-4,11-diene synthase, FPP undergoes 1,6-cyclization first and then 1,10-cyclization to form amorpha-4,11-diene. Since amorpha-4,11-diene synthase is of great importance in artemisin biosynthesis, it has been introduced into E.coli, S.cereviseae, tobacco, Arabidopsis and A. nidulans, obtained a variety of engineering bacteria or cells that can produce amorpha-4,11-diene, and studied ways to improve the yield of amorpha-4,11-diene in engineering bacteria by different ways.