The invention relates to a construction of a
Comamonas testosteroni 3,17beta-HSD
gene reinforced strain, and belongs to the microbial field. The construction of the
Comamonas testosteroni 3,17beta-HSD
gene reinforced strain is characterized in that a recombinant
plasmid is pK-Tacpromoter-3,17beta-HSD; and the construction comprises the following steps: carrying out PCR technological amplification, and target segment
recovery and purification of a
Comamonas testosteroni
genome which is a template to connect with a pUCm-T carrier; 2, carrying out PCR technological amplification, and target segment
recovery and purification of a
plasmid pAH10 which is a template to connect with the pUCm-T carrier; 3, extracting a recombinant
plasmid 3,17beta-HSD-T through adopting a column
plasmid DNA small-amount extraction kit; and 4, respectively extracting the recombinant plasmid and pK-3,17beta-HSD through adopting the column
plasmid DNA small-amount extraction kit. According to the invention, the
Comamonas testosteroni 3,17beta-HSD
gene reinforced strain is constructed for the first time and is applied, and the
estriol degradation efficiency, the
cholesterol degradation efficiency and the stosterone degradation efficiency of the
Comamonas testosteroni 3,17beta-HSD gene reinforced strain are 30%, 15% and 150% higher than that of a wild
Comamonas testosteroni strain respectively.