Verdartis Desenvolvimento Biotecnológico Ltda-ME, Rua dos Técnicos s/n sala 5, Incubadora Supera Campus USP, Riberão Preto, SP, Brazil.
Appl Biochem Biotechnol. 2012 Jan;166(1):208-21. doi: 10.1007/s12010-011-9417-6. Epub 2011 Nov 5.
A study was made for purification of cephamycin C from fermentation of Streptomyces clavuligerus. Initially, the culture broth was clarified by microfiltration and ultrafiltration, after which the resulting permeates were subjected to nonspecific adsorption and ion-exchange chromatography on resin columns. The antibiotic activity was measured by the biological method at each stage by assaying its activity against the Escherichia coli ESS, super sensitive to β-lactam antibiotic. The purification processes were assessed in relation to the variables affecting each step. The purification efficiency by nonspecific adsorption was monitored by UV spectrophotometry, while the ion-exchange adsorption fractions were assessed by NMR spectroscopy. Some of the fractions obtained during purification were also analyzed by mass spectrometry (LC/MS and LC/MS/MS) to identify the cephamycin C molecule. These preliminary results proved the process feasibility.
本研究旨在从链霉菌属棒状链霉菌的发酵液中提取头孢菌素 C。首先，通过微滤和超滤对发酵液进行澄清，然后将所得的透过液进行非特异性吸附和树脂柱离子交换层析。在每个阶段，通过测定其对超敏β-内酰胺抗生素大肠杆菌 ESS 的活性，用生物学方法测定抗生素活性。根据影响每个步骤的变量评估了纯化过程。通过紫外分光光度法监测非特异性吸附的纯化效率，而通过 NMR 光谱评估离子交换吸附级分。在纯化过程中获得的一些级分也通过质谱（LC/MS 和 LC/MS/MS）进行分析，以鉴定头孢菌素 C 分子。这些初步结果证明了该工艺的可行性。
