Rhodococcussp.CCZU10-1 and method for synthesizing benzoyl formic acid through nitrile hydrolysis by using same
A technology of acetophenone acid and Rhodococcus, which is applied in the field of microorganisms and achieves the effects of no by-products, high conversion rate, simple process route and mild reaction conditions
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Embodiment 1
[0032] Rhodococcus Rhodococcus sp. CCZU10-1 grows cells and catalyzes the hydrolysis of benzoylcarbonitrile to produce acetophenone
[0033]Prepare seed medium (glucose 5 g, peptone 5 g, yeast extract 5 g, KH 2 PO 4 1 g, NaCl 0.5 g, MgSO 4 0.1 g, 1000 mL of water, pH 6.0), fill 200 mL of seed medium in a 1 L shake flask, and cultivate the seed liquid. Then prepare the fermentation medium (glucose 5 g, peptone 5 g, yeast extract 5 g, KH 2 PO 4 1 g, NaCl 0.5 g, MgSO 4 0.1 g, FeSO 4 . 7H 2 O 0.01 g, inducer caprolactam 0.01 g, water 1000 mL, pH 6.0). Put 2.5 L of fermentation medium in a 5 L fermenter, add 1% seed liquid after disinfection, ventilation rate 1:0.1 (v / v / m), stirring speed 200 rpm, temperature 20 °C, and carry out fermentation culture 15 h, add 100 mM benzoylformonitrile, sample HPLC analysis at different times, when the analysis yield of acetophenone acid is 96%, continue to add 100 mM benzoylformonitrile solution neutralized by sodium hydroxide, Wh...
Embodiment 2
[0035] Rhodococcus Rhodococcus Sp. CCZU10-1 quiescent cells catalyze the production of acetophenone from benzoylcarbonitrile
[0036] Prepare seed medium (glucose 15 g, peptone 20 g, yeast extract 20 g, KH 2 PO 4 5 g, NaCl 2.5 g, MgSO 4 2.5 g, water 1000 mL, pH 9.0), fill 200 mL seed medium in a 1 L shake flask, and cultivate the seed liquid. Then prepare the fermentation medium (glucose 15 g, peptone 20 g, yeast extract 20 g, KH 2 PO 4 5 g, NaCl 2.5 g, MgSO 4 2.5 g, FeSO 4 . 7H 2 O 0.1 g, inducer caprolactam 10 g, water 1000 mL, pH 9.0). Add an appropriate amount of fermentation medium accounting for 3.5L of the volume of the fermentation tank into the 5 L fermenter, and the inoculation amount accounts for 10% of the volume of the fermentation medium. After disinfection, inoculate into the 5 L fermenter, and the ventilation rate is 1:1:1 (v / v / m), the stirring speed was 450 rpm, the temperature was 40 °C, and the time was 120 h, centrifuged and washed to obtain...
Embodiment 3
[0038] fed-batch utilization of Rhodococcus Rhodococcus Sp. CCZU10-1 quiescent cells catalyze the production of acetophenone from benzoylcarbonitrile
[0039] Weigh 10 g of resting cells with a wet weight obtained from the culture in Example 2, suspend the cells in 1000 mL pH 8.0 potassium phosphate buffer solution-toluene (90%:10%, volume ratio), add benzoylcarbonitrile respectively, The final concentration was 100 mM, and the reaction was shaken on a constant temperature shaker at 40 °C and 160 rpm. After 12 h of reaction, 100 mM substrate benzoylcarbonitrile was fed in batches for the synthesis of acetophenone acid. figure 2 It showed that the substrate was fed 10 times (100 mM each time), and the cumulative concentration of acetophenone was 932 mM after 120 h of reaction. During the hydrolysis of benzoylcarbonitrile, no generation of benzoylformamide was found.
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