Method for preparing (3S)-3-(tertbutyloxycarbonyl)amino-1-chlorin-4-phenyl-(2R)-butanol by microbial transformation
A technology for the conversion of tert-butoxycarbonyl and microorganisms, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of large environmental pollution, cumbersome preparation process, and high price, and achieve high molar conversion rate, Environmentally friendly and low cost effect
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[0029] Example 1
[0030] The Saccharomyces cerevisiae CGMCC No. 2266 strain was inoculated into the slant medium and cultured at 30°C for 6 days to prepare 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°C and 180 r / min for 24 h to obtain a seed solution. Inoculate 10 mL of seed liquid (the inoculum amount is 10% of the volume of the liquid medium) in a 250 mL Erlenmeyer flask containing 100 mL of liquid medium, and cultivate it at 30°C and 180 r / min for 24 h to obtain fermentation broth. Centrifuge the liquid to obtain enzyme-containing bacterial cells.
[0031] It was determined that the dry weight of the enzyme-containing bacterial cells per liter of fermentation broth in this example was 50 grams.
[0032] Add 200 mL of the fermentation broth obtained above into ten Erlenmeyer flasks containing 100 mL of pH 7.0 phosph...
Example Embodiment
[0036] Example 2
[0037] The Saccharomyces cerevisiae CGMCC No. 2266 strain was inoculated into the slant medium and cultured at 30°C for 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 it at 30 ℃ and 180 r / min for 24 h to obtain a seed solution. Inoculate 10 mL of seed solution (the inoculum amount is 10% of the volume of the liquid medium) in a 250 mL Erlenmeyer flask containing 100 mL of liquid medium, and cultivate it at 30 ℃ and 180 r / min for 24 h 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.
[0038] Add 200 mL of the fermentation broth obtained above into seven Erlenmeyer flasks containing 100 mL pH 7.0 phosphate buffer. The dry weigh...
Example Embodiment
[0042] Example 3
[0043] The Saccharomyces cerevisiae CGMCC No. 2266 strain was inoculated into the slant medium and cultured at 30°C for 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°C and 180 r / min for 24 h to obtain a seed solution. The seed solution was inoculated into a 250 mL Erlenmeyer flask containing 100 mL of liquid culture medium with 10% of the inoculum, and cultured at 30 ℃ and 180 r / min for 24 h to obtain fermentation broth. The fermentation broth was centrifuged to obtain enzyme-containing bacteria Somatic cells, the dry weight of enzyme-containing bacterial cells per liter of fermentation broth is 50 grams.
[0044] Add 200 mL of the fermentation broth obtained above into five Erlenmeyer flasks containing 100 mL of pH 7.0 phosphate buffer. The dry weight of the enzyme-containing bacteri...
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