A method for preparing (r)-1-(3-trifluoromethylphenyl)ethanol by biocatalysis
A trifluoromethyl phenyl, biocatalysis technology, applied in biochemical equipment and methods, microorganism-based methods, microorganisms, etc., can solve the problems of unsatisfactory catalytic efficiency and low concentration of catalytic substrates, and achieve product stereoselectivity. high performance, improved substrate concentration and yield, and excellent catalytic efficiency
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0022] Example 1: Using different types of microbial cells as biocatalysts, the asymmetric reduction reaction of m-trifluoromethylacetophenone was carried out.
[0023] 1. Catalyst
[0024] (1) Recombinant Escherichia coli BL21(DE3) / pET28a(+)-LXCAR wet cells
[0025]The construction process of recombinant Escherichia coli BL21(DE3) / pET28a(+)-LXCAR is detailed in Example 1 of the applicant's previous patent application (publication number: CN104212841A, publication date December 17, 2014).
[0026] The recombinant Escherichia coli BL21(DE3) / pET28a(+)-LXCAR obtained from the construction was inoculated in LB liquid medium containing 50 μg / mL kanamycin, cultured with shaking at 37°C and 200 rpm for 12 hours, and then injected with 1% volume concentration of The inoculum was inoculated into fresh LB liquid medium containing 50 μg / mL kanamycin, and cultured at 37°C and 200 rpm to the cell concentration OD 600 After the temperature is 0.6-0.9, isopropyl-β-D-thiogalactopyranoside (...
Embodiment 2
[0054] Embodiment 2: the influence of auxiliary substrate kind
[0055] Add 5mL of phosphate buffer (200mM, pH 7.0) to a 50mL Erlenmeyer flask, add recombinant Escherichia coli BL21(DE3) / pET28a(+)-LXCAR wet cells with a final concentration of 50g / L, different types of auxiliary substrates ( Table 2), the final concentration is 50mM substrate m-trifluoromethyl acetophenone to form a 5mL transformation system, and the transformation is carried out at 30°C and 200rpm for 12h. After the reaction, the method of Example 1 was used to measure the yield and e.e. value, and the results are shown in Table 2. The results show that the yield is the highest when using isopropanol as auxiliary substrate, reaching 86.2%, and the e.e. value of the product is greater than 99.9%.
[0056] Table 2 Effect of different auxiliary substrates on product yield and ee value
[0057]
[0058]
Embodiment 3
[0059] Embodiment 3: the impact of buffer pH value on conversion result
[0060] Add phosphate buffer (200mM) with a pH value of 6.0 to 8.0 in a 50mL Erlenmeyer flask, and add a final concentration of 50g / L recombinant Escherichia coli BL21(DE3) / pET28a(+)-LXCAR wet cells, 10% (v / v ) isopropanol as the auxiliary substrate, and the final concentration is 50 mM m-trifluoromethyl acetophenone substrate to form a 5 mL transformation system. The transformation was carried out at 30° C. and 200 rpm for 12 hours. After the reaction, the method of Example 1 was used to measure the yield and the e.e. value of the product, and the results are shown in Table 3. The results showed that the optimum buffer pH value was 7.5, and the yield was 89.6%.
[0061] The impact of table 3 buffer pH value on reaction yield and e.e. value
[0062]
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More