Boldenone is a protein-assimilating androgen steroid that can promote protein synthesis,support nitrogen storage,and enhance renal erythropoietin release.The industrial production of boldenone mainly relies on chemica...Boldenone is a protein-assimilating androgen steroid that can promote protein synthesis,support nitrogen storage,and enhance renal erythropoietin release.The industrial production of boldenone mainly relies on chemical synthesis,which has various problems,such as a complex conversion process,excessive byproducts,and serious environmental pollution.There-fore,it is of great significance to explore a new biosynthetic route.Recently,the enzymatic synthesis of steroid compounds has been performed more frequently than in the past.In this work,boldenone was produced from androstenedione(AD)in two steps by a dual-enzyme cascade of 17β-hydroxysteroid dehydrogenase(17β-HSD)and 3-sterone-Δ^(1)-dehydrogenase(KstD).The conversion efficiency of three isoenzymes of 17β-HSD from Mycobacterium sp.LY-1 for substrate AD was first analyzed.After that,the 17β-HSD2 with high selectivity and specificity for AD was screened and co-expressed with KstD3 in Escherichia coli BL21 to construct a dual-enzyme catalytic system.The results showed that the synthesis of boldenone from AD could be achieved by constructing the dual-enzyme expression system of 17β-HSD and KstD,as we determined that the concentration of boldenone reached 24.3 mg/L.To further improve the synthesis efficiency of boldenone,the expression conditions of the dual-enzyme system were optimized,and the concentration of boldenone reached 31.9 mg/L.The explora-tion of this route will provide a foundation for the efficient enzymatic synthesis of boldenone.展开更多
基金supported by National Key Research and Development Program of China(No.2019YFA0905300)the National Natural Science Foundation of China(No.22078126)+1 种基金Qing Lan Project in Jiangsu ProvinceFundamental Research Funds for Central Universities of China(No.JUSRP221025).
文摘Boldenone is a protein-assimilating androgen steroid that can promote protein synthesis,support nitrogen storage,and enhance renal erythropoietin release.The industrial production of boldenone mainly relies on chemical synthesis,which has various problems,such as a complex conversion process,excessive byproducts,and serious environmental pollution.There-fore,it is of great significance to explore a new biosynthetic route.Recently,the enzymatic synthesis of steroid compounds has been performed more frequently than in the past.In this work,boldenone was produced from androstenedione(AD)in two steps by a dual-enzyme cascade of 17β-hydroxysteroid dehydrogenase(17β-HSD)and 3-sterone-Δ^(1)-dehydrogenase(KstD).The conversion efficiency of three isoenzymes of 17β-HSD from Mycobacterium sp.LY-1 for substrate AD was first analyzed.After that,the 17β-HSD2 with high selectivity and specificity for AD was screened and co-expressed with KstD3 in Escherichia coli BL21 to construct a dual-enzyme catalytic system.The results showed that the synthesis of boldenone from AD could be achieved by constructing the dual-enzyme expression system of 17β-HSD and KstD,as we determined that the concentration of boldenone reached 24.3 mg/L.To further improve the synthesis efficiency of boldenone,the expression conditions of the dual-enzyme system were optimized,and the concentration of boldenone reached 31.9 mg/L.The explora-tion of this route will provide a foundation for the efficient enzymatic synthesis of boldenone.
