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
The analytical method developed is based on HPLC-MS/MS to simultanous determine Progesterone,Boldenone and Trenbolone in Chicken,Beef and Pork meats.The sample is freeze drying reduced in powder and a Methanol-Acetate...The analytical method developed is based on HPLC-MS/MS to simultanous determine Progesterone,Boldenone and Trenbolone in Chicken,Beef and Pork meats.The sample is freeze drying reduced in powder and a Methanol-Acetate Buffer (pH=5.2) solution is added in order to perform the extraction and sonicated with a solution prior to deconjugation using β-glucosidase.The sample is purified using C18 and SiOH solid phase extraction (SPE) cartridge prior to analyze on reversed phase with a elution gradient performed on Agilent C18 coupled with a HPLC-MS/MS.The detection Limit is respectively 0.11,0.17 and 0.02 μg/kg for Trenbolone,Boldenone and Progesterone with a ratio Signal/Noise>3 in the 3 different kind of meat.The quantification was based on the peak area and overall recoveries of synthetic growth hormones were 62%-99%.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.
文摘The analytical method developed is based on HPLC-MS/MS to simultanous determine Progesterone,Boldenone and Trenbolone in Chicken,Beef and Pork meats.The sample is freeze drying reduced in powder and a Methanol-Acetate Buffer (pH=5.2) solution is added in order to perform the extraction and sonicated with a solution prior to deconjugation using β-glucosidase.The sample is purified using C18 and SiOH solid phase extraction (SPE) cartridge prior to analyze on reversed phase with a elution gradient performed on Agilent C18 coupled with a HPLC-MS/MS.The detection Limit is respectively 0.11,0.17 and 0.02 μg/kg for Trenbolone,Boldenone and Progesterone with a ratio Signal/Noise>3 in the 3 different kind of meat.The quantification was based on the peak area and overall recoveries of synthetic growth hormones were 62%-99%.
