Azevedo Ana, Teixeira-Santos Rita, Gomes Luciana C, Duarte Sofia O D, Monteiro Gabriel A, Mergulhão Filipe J
LEPABE-Laboratory for Process Engineering, Environment, Biotechnology and Energy, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, 4200-465 Porto, Portugal.
ALICE-Associate Laboratory in Chemical Engineering, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, 4200-465 Porto, Portugal.
Molecules. 2025 May 2;30(9):2031. doi: 10.3390/molecules30092031.
Biofilms are emerging platforms for the production of valuable compounds. The present study is the first to assess the capacity of biofilms to produce curcumin through the expression of a biosynthetic pathway involving three genes: 4-coumarate-CoA ligase (), diketide-CoA synthase (), and curcumin synthase (). The effects of chemical induction with isopropyl β-d-1-thiogalactopyranoside (IPTG) and ferulic acid (FA), and the incubation temperature on biofilm formation and curcumin production were evaluated. Biofilms were formed in 12-well microtiter plates over three days and then induced with 1 mM IPTG and FA at 2 or 8 mM. After induction, the samples were incubated for two days at 26 or 30 °C. Total and culturable planktonic and biofilm cells, as well as biofilm thickness and volumetric and specific curcumin production, were assessed on days 3, 4, and 5. The results demonstrated that biofilms produced up to 10-fold higher curcumin levels (0.9-2.2 fg·cell) than their planktonic counterparts (0.1-0.3 fg·cell). The highest specific curcumin production (2.2 fg·cell) was achieved using 8 mM FA. However, no significant differences in curcumin production were observed between the induced samples incubated at the tested temperatures. These results validated the potential of biofilm systems for expressing a complete exogenous biosynthetic pathway using metabolic engineering, particularly for curcumin production.
生物膜正成为生产有价值化合物的新兴平台。本研究首次评估了生物膜通过表达由三个基因组成的生物合成途径来生产姜黄素的能力，这三个基因分别是：4-香豆酸-CoA连接酶、二酮-CoA合酶和姜黄素合酶。评估了用异丙基-β-D-1-硫代半乳糖苷（IPTG）和阿魏酸（FA）进行化学诱导以及培养温度对生物膜形成和姜黄素生产的影响。在12孔微量滴定板中培养三天形成生物膜，然后用1 mM IPTG和2或8 mM的FA进行诱导。诱导后，样品在26或30℃下孵育两天。在第3、4和5天评估总浮游细胞和可培养浮游细胞、生物膜细胞，以及生物膜厚度、姜黄素的体积产量和比产量。结果表明，生物膜产生的姜黄素水平（0.9-2.2 fg·细胞）比其浮游对应物（0.1-0.3 fg·细胞）高10倍。使用8 mM FA时获得了最高的姜黄素比产量（2.2 fg·细胞）。然而，在测试温度下孵育的诱导样品之间，未观察到姜黄素产量的显著差异。这些结果验证了生物膜系统利用代谢工程表达完整外源生物合成途径的潜力，特别是在姜黄素生产方面。
