Method for preparing duloxetine chiral intermediate through microbial conversion
A technology for microbial transformation and chiral intermediates, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve problems such as non-ideal pathways, and achieve easy large-scale industrial production, high yield, and large-scale Cultivate easy effects
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Embodiment 1
[0038] The CGMCC No. 2266 strain was inoculated into the slant medium, and cultured at 30°C for 4-6 days to obtain the slant surface of the bacteria. An inoculation loop was taken from the slant of the bacteria with an inoculation needle and inoculated into a 250 mL Erlenmeyer flask containing 100 mL of liquid medium, and cultured at 30 °C and 180 r / min for 24 h to obtain a seed liquid. Inoculate 10 mL of seed solution (the inoculum volume is calculated as 10% of the medium volume fraction) into a 250 mL Erlenmeyer flask containing 100 mL of liquid medium, and culture it at 30 °C and 180 r / min for 24 h to obtain a fermentation broth. The fermentation broth is centrifuged to obtain enzyme-containing bacterial cells.
[0039] The determination of the dry weight of the bacteria is to take a small part of the wet bacteria containing the enzyme cells after centrifuging the fermentation broth, and dry it at 120 °C for 48 hours to a constant weight, measure the weight of the dry cell...
Embodiment 2
[0044] The CGMCC No. 2266 strain was inoculated into the slant medium, and cultured at 30°C for 4-6 days to obtain the slant surface of the bacteria. An inoculation loop was taken from the slant of the bacteria with an inoculation needle and inoculated into a 250 mL Erlenmeyer flask containing 100 mL of liquid medium, and cultured at 30 °C and 180 r / min for 24 h to obtain a seed liquid. Inoculate 10 mL of seed liquid (the inoculum amount is calculated as 10% of the medium volume) into a 250 mL Erlenmeyer flask containing 100 mL of liquid medium, and culture at 30 °C and 180 r / min for 24 h to obtain a fermentation broth. The fermented liquid is centrifuged to obtain enzyme-containing bacterium cells, and the dry weight of the enzyme-containing bacterium cells in every liter of fermented liquid is 50 grams.
[0045] Add the wet cells obtained after centrifugation of the above-mentioned 400 milliliters of fermented liquid into ten portions of Erlenmeyer flasks each containing 100...
Embodiment 3
[0049] Saccharomyces cerevisiae CGMCC No. 2266 was inoculated into the slant medium, and cultured at 30°C for 4-6 days to obtain the slant. An inoculation loop was taken from the slant of the bacteria with an inoculation needle and inoculated into a 250 mL Erlenmeyer flask containing 100 mL of liquid medium, and cultured at 30 °C and 180 r / min for 24 h to obtain a seed solution. Inoculate 10 mL of seed solution (the inoculum amount is calculated as 10% of the medium volume) into a 250 mL Erlenmeyer flask containing 100 mL of liquid medium, and culture it at 30 °C and 180 r / min for 24 h to obtain a fermentation broth. The fermented liquid is centrifuged to obtain enzyme-containing bacterium cells, and the dry weight of the enzyme-containing bacterium cells in every liter of fermented liquid is 50 grams.
[0050] Add the wet cells obtained after centrifugation of the above 400 ml fermentation broth to five parts of Erlenmeyer flasks each containing 100 mL of pH7.0 phosphate buff...
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