Prepn of isotopically labeled 15 N-L-aspartic acid and 15 N-L-alanine
A technology of isotope labeling and aspartic acid, applied in the direction of fermentation, etc., can solve problems that have not been seen yet, and achieve the effect of large-scale industrialization implementation prospects
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Embodiment 1
[0066] The strain Escherichia coli Asp-4 was cultured in a slant medium at 37°C for 24 to mature, then inserted into the aspartase enzyme liquid medium, filled with 50ml of medium in a 250ml shaker flask, and cultivated on a rotary shaker at 37°C at 220rpm for 24 hours. After the cultivation is completed, the culture solution is placed in a sterile centrifuge tube and centrifuged at 4000rpm for 15 minutes, and 0.5 g of the wet bacterial cells after centrifugation is added to a solution containing 5% fumaric acid, 8% 15 In 200 ml of N-ammonium sulfate and 0.0125% magnesium sulfate enzyme reaction solution, adjust the pH to 8.2-8.8 with NaOH, and react in a constant temperature incubator at 42° C. for 24 hours. After the reaction, the results are shown in Table 1:
[0067] Table 1
[0068] Item No.1 No.2 No.3 Average
[0069] 15 N-(NH 4 ) 2 SO 4 Amount added (g) 40 40 40
[0070] Amount of fumaric acid substrate added (g) 10 10 10
[...
Embodiment 2
[0078] Add 0.5 g of mature wet thalline cells containing 5% fumaric acid, 0.0125% magnesium sulfate and 3% 15 N-ammonium sulfate, 3% 15 In the mixed nitrogen source reaction liquid 200ml of N-ammonium chloride, use 15 Adjust the pH to 8.2-8.8 with N-ammonia water, and react in a constant temperature incubator at 42°C for 24 hours. Results The amount of aspartic acid produced was 52.0 g, the conversion rate of fumaric acid was 96.8%, and the molar conversion rate of nitrogen source was 62.4%.
Embodiment 3
[0080] The reaction solution completed in Example 1 was heated to 85-90° C., 0.5% activated carbon was added, stirred and kept for 45 minutes, vacuum filtered, and the filter cake was washed with distilled water. Combine the filtrates and concentrate the filtrate to 20% in vacuum at 65°C and 680mmHg, heat to 80°C, slowly add concentrated sulfuric acid while stirring, when the pH is 3.0, fine crystals are precipitated, stir and cool in a 100rpm constant speed stirrer to crystallize. When cooled to room temperature, refrigerate overnight at 5°C. The crystallization solution was vacuum filtered and crystal washed to obtain the finished wet crystal, which was dried at 60°C overnight. The results are shown in Table 2:
[0081] Table 2
[0082] Item No.1 No.2 No.3 Average
[0083] Reaction volume (ml) 625 630 618
[0084] Aspartic acid content (%) 7.93 8.14 8.25 8.11
[0085] Obtained product (g) 42.5 43.1 43.9
[0086] Total yield (%) 85.8 84....
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