Patiño Luisa F, Caicedo-Montoya Carlos, Pinilla-Mendoza Laura, Cuartas Jaison H, Ríos-Estepa Rigoberto
Grupo de Bioprocesos, Departamento de Ingeniería Química, Universidad de Antioquia UdeA, Calle 70 No. 52-21, Medellín 050010, Colombia.
Independent Researcher, Medellín 050010, Colombia.
Metabolites. 2025 May 19;15(5):337. doi: 10.3390/metabo15050337.
Clavulanic acid (CA) is produced by cell suspension cultures of ATCC 27064, and is widely used as a beta-lactamase inhibitor to combat antibiotic resistance. CA titers are moderate due to bioprocess complexity, prompting ongoing efforts to overcome these limitations. In this study, we aimed to evaluate the effect of live and inactivated FZB42 cells on CA production in , and to explore the transcriptional response underlying this interaction using RNA-seq technology. The addition of dead and live cells of improved CA production by 1.4 and 2.0-fold, respectively. Furthermore, the transcriptome of obtained with live cells of FZB42 at the peak of maximum CA production, revealed that 410 genes were up-regulated and 594 were down-regulated under these conditions, with a < 0.05. Most of the genes from the cephamycin C and CA clusters were up-regulated, which correlates well with the increase in CA production. Likewise, ATCC 27064 enhanced the expression of genes encoding enzymes that scavenge endogenous HO, as well as other genes related to oxidative stress defense. Regarding downregulated genes, we found that decreased the expression of genes involved in the biosynthesis of terpenoids, polyketides, and lantibiotics, as well as the expression of the operon involved in the synthesis of the pyrroloquinoline quinone (PQQ) cofactor. These findings contribute to the understanding of metabolism and pave the way for future metabolic engineering efforts aimed at obtaining CA-overproducing strains.
棒酸（CA）由ATCC 27064的细胞悬浮培养物产生，被广泛用作β-内酰胺酶抑制剂以对抗抗生素耐药性。由于生物过程的复杂性，CA的滴度适中，这促使人们不断努力克服这些限制。在本研究中，我们旨在评估活的和灭活的FZB42细胞对ATCC 27064中CA产生的影响，并使用RNA测序技术探索这种相互作用背后的转录反应。添加死细胞和活细胞分别使CA产量提高了1.4倍和2.0倍。此外，在CA产量最高的峰值时用FZB42活细胞获得的ATCC 27064转录组显示，在这些条件下，410个基因上调，594个基因下调，P < 0.05。来自头霉素C和CA簇的大多数基因上调，这与CA产量的增加密切相关。同样，ATCC 27064增强了编码清除内源性HO的酶的基因以及其他与氧化应激防御相关的基因的表达。关于下调的基因，我们发现FZB42降低了参与萜类化合物、聚酮化合物和羊毛硫抗生素生物合成的基因的表达，以及参与合成吡咯并喹啉醌（PQQ）辅因子的操纵子的表达。这些发现有助于理解ATCC 27064的代谢，并为未来旨在获得CA高产菌株的代谢工程努力铺平道路。
