臺灣博碩士論文加值系統

Clavulanic acid是一種由Streptomyces clavuligerus發酵生產的β-lactamase抑制劑，醫療上經常合併使用clavulanic acid與amoxicillin。本論文利用批式與進料批式的操作，以S. clavuligerus進行clavulanic acid的發酵生產，探討大豆萃取液與clavulanic acid的前驅物甘油、ornithine、arginine對其生化合成的影響，並尋求最適的前驅物及其進料量，以及建立進料批式操作的方法。在搖瓶培養過程中，含有大豆萃取液(TKN=0.59 g/L)會有較佳的clavulanic acid產量，而進料批式操作的過程中，利用間歇進料的方式，將甘油持續進料到培養液內，其所生成clavulanic acid的最高濃度是未進行glycerol進料的2.3倍。同時進行甘油和ornithine或arginine的進料，所生成的clavulanic acid濃度分別是未進行glycerol進料的2.7、2.8倍。甘油的進料添加能夠提供clavulanic acid分子左側結構上β-lactam環的前驅物，而ornithine或arginine的添加則能提供clavulanic acid分子右側結構上C5-moiety的前驅物，這顯示甘油和ornithine或arginine能有效率地構成clavulanic acid。

Clavulanic acid is one of the fermentation products through secondary metabolism by Streptomyces clavuligerus. It is also a specific and irreversible inhibitor of a wide range of bacterial β-lactamases. Clavulanic acid has been used in human therapy for several years in combination with amoxicillin. The purpose of this study is to investigate the effects of different precursors and soy meal extract on the biosynthesis of clavulanic acid using S. clavuligerus in batch and fed-batch fermentations. A regularly intermittent feeding strategy was employed to obtain the maximum productivity of clavulanic acid when different feed medium was used to improve the production. The medium containing soy meal extract with the TKN concentration of 0.59 g/L was optimum. Intermittently feeding the medium containing glycerol to the shake flask cultures resulted in the maximum production of clavulanic acid being 2.3-fold higher than that with no additional feeding. Intermittent feeding simultaneously with glycerol and ornithine (or arginine) resulted in the maximum production of clavulanic acid being 2.7-fold (or 2.8-fold) higher than that with no feeding. The fed batch result indicates that glycerol is an important precursor of the β-lactam moiety(C-5,C-6, and C-7), whereas ornithine (or arginine) is an important precursor of C5-moiety. Both glycerol and ornithine (or arginine) exert a stimulation of the biosynthesis of clavulanic acid, and they are efficiently incorporated into clavulanic acid.

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