Key Laboratory of Systems Bioengineering, Ministry of Education and Department of Pharmaceutical Engineering, School of Chemical Engineering and Technology, Tianjin University, P.O. Box 6888, Tianjin 300072, People's Republic of China.
Appl Microbiol Biotechnol. 2011 Feb;89(4):1193-202. doi: 10.1007/s00253-010-2910-y. Epub 2010 Oct 13.
The disparity of secondary metabolites in Penicillium chrysogenum between two scales of penicillin G fermentation (50 L as pilot process and 150,000 L as industrial one) was investigated by ion-pair reversed-phase liquid chromatography tandemed with hybrid quadrupole time-of-flight mass spectrometry. In industrial process, the pools of intracellular L-α-aminoadipyl-L-cysteinyl-D-valine (LLD-ACV) and isopenicillin N (IPN) were remarkably less than that in the pilot one, which indicated that the productivity of penicillin G might be higher in the large scale of fermentation. This conclusion was supported by the higher intracellular penicillin G concentration as well as its higher yield per unit biomass in industrial cultivation. The different changing tendencies of IPN, 6-aminopenicillanic acid and 6-oxopiperide-2-carboxylic acid between two processes also suggested the same conclusion. The higher content of intracellular LLD-ACV in pilot process lead to a similarly higher concentration of bis-δ-(L-α-aminoadipyl)-L-cysteinyl-D-valine, which had an inhibitory effect on ACV synthetase and also subdued the activity of IPN synthetase. The interconversion of secondary metabolites and the influence they put on enzymes would intensify the discrepancy between two fermentations more largely. These findings provided new insight into the changes and regulation of secondary metabolites in P. chrysogenum under different fermentation sizes.
采用离子对反相液相色谱-串联混合四极杆飞行时间质谱法研究了青霉素 G 发酵(50 L 为中试规模和 15 万 L 为工业规模)中青霉素 G 发酵二级代谢产物的差异。在工业生产过程中,细胞内 L-α-氨酰基-L-半胱氨酸-D-缬氨酸(LLD-ACV)和青霉素 N(IPN)的池显著低于中试规模,这表明青霉素 G 的生产效率可能更高在大规模发酵中。这一结论得到了以下结果的支持:工业培养中细胞内青霉素 G 浓度更高,单位生物量的产率也更高。两个过程中 IPN、6-氨基青霉素酸和 6-氧代哌啶-2-羧酸的不同变化趋势也表明了同样的结论。中试过程中细胞内 LLD-ACV 含量较高,导致双-δ-(L-α-氨酰基)-L-半胱氨酸-D-缬氨酸的浓度也较高,这对 ACV 合成酶有抑制作用,也抑制了 IPN 合成酶的活性。次级代谢产物的相互转化及其对酶的影响会使两种发酵之间的差异进一步扩大。这些发现为不同发酵规模下青霉素 G 发酵中次级代谢产物的变化和调控提供了新的见解。