23 Jul A human kallikrein, and a coding gene and an application and preparation method thereof
Abstract
The invention provides a human tissue kallikrein-1 (hK1) coding gene, an expression and purification manner, an activity detecting method, and a modified pichia pastoris fermentation BSM culture medium. The optimized human tissue kallikrein-1 (hK1) coding gene is shown as SEQ ID NO:1. The sequence is obtained by codon optimization for a pichia pastoris expression system. Compared with natural sequences, the sequence is more conductive to hK1 expression in pichia pastoris. The established fluorescence enzyme activity detecting method is simple in operation, and can be used for screening of high-expression monoclonal recombinant engineering bacteria. In addition, the modified fermentation BSM culture medium can promote efficient expression of the hK1 protein and inhibit expression of host endogenous protein under the premise of not influencing production of the pichia pastoris, thus facilitating subsequent separation and purification. Accordingly, the coding gene, the expression and purification manner, the activity detecting method and the fermentation BSM culture medium lay foundations for expression and application of the hK1 protein in the pichia pastoris.
Year: 2015
Country: CN
Doc No: 104745613