Highly efficient synthesis of boldenone from androst-4-ene-3,17-dione by Arthrobacter simplex and Pichia pastoris ordered biotransformation

Yeast as a promising heterologous host for steroid bioproduction

“…The bacterial strain Arthrobacter simplex previously identified to exhibit high ∆ 1 -dehydrogenation activity was used to convert AD into ADD; and P. pastoris GS115 was engineered to express 17βHSD from S. cerevisiae to enhance the conversion of ADD into BD [50]. With the optimized medium condition (5.0 g/L AD with molar ratio 1:1 HP-β-CD and 75-g/L glucose), the conversion of AD into BD achieved 83% [50]. The efficient conversion of AD into BD in the bacteria-yeast coculture later on inspired Mycobacterium neoaurum-P. pastoris GS115 coculture, where M. neoaurum can natively convert phytosterols into ADD (with a small amount of AD supplemented) [51].…”

“…The medium condition of steroid biotransformation in yeast is relatively simple, usually common yeast rich or minimal medium supplemented with organic solvent, such as ethanol and N,N-dimethylformamide (DMF) to help dissolve the steroid substrates [48,56]. Addition of β-cyclodextrin often helps the transport of steroid substrates into the cells [50,57]. Enzymes responsible for steroid hydroxylation and dehydrogenation have been largely identified within the past 20 years, with many characterized in yeast.…”

“…Therefore, the cosolvent 2-hydroxypropyl-β-cyclodextrin (HP-β-CD) was supplemented to improve the solubility of steroids in the medium and enhanced the conversion of AD into BD to 56.76% (with molar ratio 1: 1 of HP-β-CD to AD) [ 49 ]. The conversion was further improved with the bacterial and yeast co-culture strategy in the follow-up investigations [ 50 , 51 ]. The bacterial strain Arthrobacter simplex previously identified to exhibit high ∆ 1 -dehydrogenation activity was used to convert AD into ADD; and P. pastoris GS115 was engineered to express 17βHSD from S. cerevisiae to enhance the conversion of ADD into BD [ 50 ].…”

“…The conversion was further improved with the bacterial and yeast co-culture strategy in the follow-up investigations [ 50 , 51 ]. The bacterial strain Arthrobacter simplex previously identified to exhibit high ∆ 1 -dehydrogenation activity was used to convert AD into ADD; and P. pastoris GS115 was engineered to express 17βHSD from S. cerevisiae to enhance the conversion of ADD into BD [ 50 ]. With the optimized medium condition (5.0 g/L AD with molar ratio 1:1 HP-β-CD and 75-g/L glucose), the conversion of AD into BD achieved 83% [ 50 ].…”

Microorganisms as Biocatalysts and Enzyme Sources

“…Boldenone 124is an important steroid hormone drug which is the derivative of testosterone. Biotransformation of 124 by Arthrobacter simplex and recombinant Pichia pastoris with 17β-hydroxysteroid dehydrogenase from Saccharomyces cerevisiae produces BD (124) from androst-4-ene-3,17-dione (79, AD) efficiently [65]. Many microorganisms such as Mucor racemosus, Nostoc muscorum, and Arthrobacter oxydans can utilize androst-1,4-diene-3,17-dione (80, ADD) as substrate to produce testosterone through 17β-carbonyl reduction reactions ( Table 5).…”