Abstract
A cascade reaction combining the enzymatic hydrolysis of Penicillin G potassium salt (PGK) with the kinetically controlled enzymatic coupling of in situ formed 6-aminopenicillanic acid (6-APA) with p-hydroxyphenylglycine methyl ester (D-HPGM) to give amoxicillin as the final product by using a single enzyme has been demonstrated successfully. Ethylene glycol (EG) was employed as a component of reaction buffer to improve the synthesis yield. Reaction parameters, including different cosolvents, EG content, the loading of immobilized penicillin G acylase (IPA), and reaction temperature and time were studied to evaluate their effects on the reaction. The best result of 55.2% yield was obtained from the reaction which was carried out in the mixed media containing 40% sodium dihydrogen phosphate buffer (apparent pH 6.0) and 60% EG (v/v), with the initial concentration 150 mM and 450 mM of PGK and D-HPGM, respectively, catalyzed by 50 IU/mL IPA at 25 °C for 10 h. The IPA could be recycled for nine batches without obviously losing of catalytic activity. The important strategy will have potential application in the β-lactam antibiotics industry due to the advantages of saving the effort of isolating 6-APA, reducing usual enzymatic steps and the industrial cost of amoxicillin synthesis.
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Abbreviations
- PGK:
-
Penicillin G potassium salt
- 6-APA:
-
6-Aminopenicillanic acid
- D-HPGM:
-
p-Hydroxyphenylglycine methyl ester
- EG:
-
Ethylene glycol
- IPA:
-
Immobilized penicillin G acylase
- PGA:
-
Penicillin G acylase
- CLEAs:
-
Cross-linked penicillin acylase aggregates
- 7-ADCA:
-
7-Aminodeacetoxycephalosporanic Acid
- HPGA:
-
p-Hydroxyphenylglycine amide
- HPLC:
-
High-performance liquid chromatography
- UV:
-
Ultraviolet
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The financial supports from National Natural Science Foundation of China (No. 20704037) and Zhejiang Provincial Science and Technology Council (No. 2005C11023-03) are gratefully acknowledged.
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Wu, Q., Chen, CX., Du, LL. et al. Enzymatic Synthesis of Amoxicillin via a One-pot Enzymatic Hydrolysis and Condensation Cascade Process in the Presence of Organic Co-solvents. Appl Biochem Biotechnol 160, 2026–2035 (2010). https://doi.org/10.1007/s12010-009-8847-x
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DOI: https://doi.org/10.1007/s12010-009-8847-x