Method for preparing duloxetine chiral intermediate through microbial conversion
A technology for microbial transformation and chiral intermediates, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve problems such as not ideal paths, achieve easy large-scale industrial production, improve transformation efficiency, and be environmentally friendly. Effect
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Embodiment 1
[0039] The CGMCC No. 2266 strain was inoculated into the slant medium, and cultivated at 30 ℃ for 4-6 days to obtain the slant. Use an inoculating needle to take an inoculum loop from the slant surface of the bacteria and inoculate it in a 250 mL Erlenmeyer flask containing 100 mL of liquid culture medium, and incubate at 30 ℃ and 180 r / min for 24 h to obtain a seed solution. Inoculate 10 mL of seed solution (inoculation amount based on the volume fraction of medium 10%) in a 250 mL Erlenmeyer flask containing 100 mL of liquid culture medium, and culture for 24 h at 30 ℃ and 180 r / min to obtain fermentation broth. Centrifuge the fermentation broth to obtain enzyme-containing bacterial cells.
[0040] The dry weight of the bacteria is determined by centrifuging the fermentation broth and taking a small part from the wet bacteria of the enzyme-containing bacteria cells and drying them at 120 ℃ for 48 hours to a constant weight. The weight of the dry cells is measured and the unit c...
Embodiment 2
[0046] The CGMCC No. 2266 strain was inoculated into the slant medium, and cultivated at 30 ℃ for 4-6 days to obtain the slant. Use an inoculating needle to take an inoculum loop from the slant surface of the bacteria and inoculate it in a 250 mL Erlenmeyer flask containing 100 mL of liquid culture medium, and incubate at 30 ℃ and 180 r / min for 24 h to obtain a seed solution. Inoculate 10 mL of seed solution (the inoculum volume is calculated as 10% of the volume of the medium) in a 250 mL Erlenmeyer flask containing 100 mL of liquid culture medium, and culture for 24 h at 30 ℃ and 180 r / min to obtain fermentation broth. The fermentation broth is centrifuged to obtain enzyme-containing bacterial cells, and the dry weight of the enzyme-containing bacterial cells per liter of fermentation broth is 50 grams.
[0047] The wet cells obtained after centrifugation of the above 400 ml fermentation broth were added to ten Erlenmeyer flasks each containing 100 mL pH 7.0 phosphate buffer. T...
Embodiment 3
[0051] The Saccharomyces cerevisiae CGMCC No. 2266 strain was inoculated into the slant medium, and cultivated at 30 ℃ for 4-6 days to obtain the slant. Use an inoculating needle to take an inoculum loop from the slant surface of the bacteria and inoculate it in a 250 mL Erlenmeyer flask containing 100 mL of liquid culture medium, and incubate at 30 ℃ and 180 r / min for 24 h to obtain a seed solution. Inoculate 10 mL of seed solution (inoculation amount based on 10% of the volume of the medium) in a 250 mL Erlenmeyer flask containing 100 mL of liquid medium, and cultivate for 24 h at 30 ℃ and 180 r / min to obtain fermentation broth. The fermentation broth is centrifuged to obtain enzyme-containing bacterial cells, and the dry weight of the enzyme-containing bacterial cells per liter of fermentation broth is 50 grams.
[0052] In five Erlenmeyer flasks each containing 100 mL of pH 7.0 phosphate buffer, add the wet cells obtained after centrifugation of the above 400 mL of fermentati...
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