Roessner C A, Devagupta R, Hasan M, Williams H J, Scott A I
Chemistry Department, Texas A&M University, College Station 77843-3255.
Protein Expr Purif. 1992 Aug;3(4):295-300. doi: 10.1016/1046-5928(92)90004-g.
The gene for the indole alkaloid biosynthetic enzyme, strictosidine synthase, of Catharanthus roseus has been cloned into an inducible Escherichia coli expression vector using an expression cassette polymerase chain reaction technique. Induction of the gene resulted in overexpression of the enzyme which accumulated mainly as insoluble inclusion bodies. Denaturation and refolding of the insoluble protein resulted in the ability to purify up to 6 mg of active enzyme from a single liter of cell culture. The recombinant enzyme has good activity (approximately 30 nkat/mg).
利用表达盒聚合酶链反应技术，将长春花中吲哚生物碱生物合成酶——异胡豆苷合成酶的基因克隆到一个可诱导的大肠杆菌表达载体中。该基因的诱导表达导致该酶的过量表达，其主要以不溶性包涵体的形式积累。对不溶性蛋白进行变性和复性处理后，能够从一升细胞培养物中纯化出高达6毫克的活性酶。重组酶具有良好的活性（约30纳卡特/毫克）。
