Reaction method
A reaction and reaction tank technology, applied in the field of reaction, can solve the problems of fragmentation and wear, limited application range, large bed pressure drop, etc., and achieve the effects of avoiding fragmentation and wear, simple replacement steps, and improved reaction speed
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[0055] Example 1. Monolithic open-pore D-amino acid oxidase immobilized enzyme is directly used as a stirring blade for batch stirring reaction to prepare glutaryl-7-aminocephalosporanic acid (GL-7-ACA)
[0056] First, prepare the honeycomb-shaped immobilized D-amino acid oxidase block according to the following method.
[0057] D-amino acid oxidase converts cephalosporin C into glutaryl-7-aminocephalosporanic acid (GL-7-ACA). Prepare BL-HS-GHA [E.coli BL21(DE3)pLysS containing recombinant D-amino acid oxidase GHA] E. coli cells according to the following method.
[0058] The source of BL-HS-GHA:
[0059] According to the known Thermoanaerobacterium saccharolyticum glucoseisomerase DNA sequence (GenBank L09699), PCR primers were designed, specifically:
[0060] Upstream primer GI-NdeI:
[0061] 5’-AGCCTAGGTTAATTAACTTTAAGAAGGAGATATACATATGAATAAATATTTTGAGA
[0062] Downstream primer GI-EcoRI:
[0063] 5’-ATAAGCTCAGCGGCGCGCCTTATTCTGCAAACAAATAC
[0064] Using Thermoanaerobacterium saccharolytic...
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