Biosynthesis method of borneol
A technology of biosynthesis and borneol, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of low concentration of borneol in the transformation liquid and no practical application value, and achieve improved selectivity and side effects The effect of less product
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Embodiment 1
[0023] Use 10g / L peptone, 10g / L glucose, 1.0g / L beef extract, 1.0g / L KH 2 PO 4 , 0.1g / L of MgSO 4 , 1.0g / L of NaCl and 0.5g / L of (NH 4 ) 2 SO 4 Prepare 40 mL of culture medium with pH 5.5, put it into a 250 mL Erlenmeyer flask, then insert the Klebsiella planticola (Klebsiella planticola) bacterial strain preserved on the slant, cultivate it in a shaker at 30°C and 150 rpm for 36 hours, and then place the The cultured bacterial solution was centrifuged at 4°C and 4500rpm for 10 minutes to collect bacterial cells, washed with pH 5.5 phosphate buffer solution twice, and stored in the refrigerator until use.
[0024] Add 20 mL of phosphate buffered saline with pH 6.5, 1 ml of ethanol (4% by volume), the aforementioned bacterial cells and 333 μL of bornyl acetate (85 mM) in sequence in a 250 mL Erlenmeyer flask, and place at 22°C , 2d in a 60rpm shaker. After the reaction, the biocatalyst was separated by filtration and reused, then extracted three times with 10 mL of petrol...
Embodiment 2
[0026] Use 10g / L peptone, 20g / L waste molasses, 5.0g / L beef extract, 1.0g / L KH 2 PO 4 , 0.5g / L MgSO 4 , 0.5g / L of NaCl and 1.0g / L of (NH 4 ) 2 SO 4 Prepare 100 mL of culture medium with pH 7.0, put it into a 1000 mL Erlenmeyer flask, insert Klebsiella planticola (Klebsiella planticola) strains, and culture at 40°C for 40 h in a shaker at 250 rpm. The bacterial solution was centrifuged at 4°C and 4500 rpm for 10 minutes to collect bacterial cells, and the bacterial cells were washed twice with pH 7.0 phosphate buffer solution, and stored in the refrigerator until use.
[0027] In a 500mL Erlenmeyer flask, add 50mL of disodium hydrogen phosphate-citric acid buffer solution at pH 7.0, 5mL of petroleum ether (that is, the volume content is 10%), the aforementioned bacterial cells and 1.5mL of bornyl ester (150mM) , and then placed in a 28°C, 150rpm shaker for 4d. After the reaction is over, the biocatalyst is filtered and separated for reuse, and then the borneol product is ...
Embodiment 3
[0029] With 15g / L corn flour, 15g / L glucose, 5.0g / L beef extract, 0.8g / L KH 2 PO 4 , 0.2g / L MgSO 4 , 1.5g / L of NaCl and 1.0g / L of (NH 4 ) 2 SO 4Prepare 100 mL of culture medium with pH 8.5, inoculate Klebsiella planticola (Klebsiella planticola) strains and culture in a shaker at 45°C and 350rpm for 70h, then place the cultured bacterial solution at 4°C and 4500rpm The bacterial cells were collected by centrifugation for 10 min, and the bacterial cells were washed twice with pH 8.5 phosphate buffer solution, and stored in the refrigerator until use.
[0030] In a 500mL Erlenmeyer flask, add 45mL, pH7.5 phosphate buffered saline, 5mL ethyl acetate (i.e. the volume content is 10%), cultured bacterial cells and 1.5mL bornyl ester (150mM), and then place 35 ° C, 300 rpm shaker reaction 6d. After the reaction is over, the biocatalyst is filtered and separated for repeated use, and then the borneol product is separated by recrystallization, and 150 mg of borneol solid is obtai...
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