Ana Azevedo, Rita Teixeira-Santos, Luciana C Gomes, Sofia O D Duarte, Gabriel A Monteiro, Filipe J Mergulhão
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引用次数: 0
Abstract
Biofilms are emerging platforms for the production of valuable compounds. The present study is the first to assess the capacity of Escherichia coli biofilms to produce curcumin through the expression of a biosynthetic pathway involving three genes: 4-coumarate-CoA ligase (4CL), diketide-CoA synthase (DCS), and curcumin synthase (CURS). 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-1) than their planktonic counterparts (0.1-0.3 fg·cell-1). The highest specific curcumin production (2.2 fg·cell-1) 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.
生物膜是生产有价值化合物的新兴平台。本研究首次评估了大肠杆菌生物膜通过表达涉及3个基因的生物合成途径产生姜黄素的能力:4-香豆酸-辅酶a连接酶(4CL)、二酮酸-辅酶a合成酶(DCS)和姜黄素合成酶(CURS)。考察了异丙基β-d-1-硫代半乳糖苷(IPTG)和阿魏酸(FA)的化学诱导以及培养温度对生物膜形成和姜黄素产量的影响。生物膜在12孔微滴板上形成3天,然后用1 mM IPTG和2或8 mM FA诱导。诱导后,样品在26或30°C下孵育2天。在第3、4和5天评估浮游生物和生物膜细胞总数和可培养细胞,以及生物膜厚度、体积和姜黄素产量。结果表明,生物膜产生的姜黄素水平(0.9-2.2 fg·cell-1)比浮游膜的姜黄素水平(0.1-0.3 fg·cell-1)高10倍。使用8 mM FA时,姜黄素比产量最高(2.2 fg·cell-1)。然而,在测试温度下培养的诱导样品之间没有观察到姜黄素产量的显着差异。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,特别是姜黄素的生产。
期刊介绍:
Molecules (ISSN 1420-3049, CODEN: MOLEFW) is an open access journal of synthetic organic chemistry and natural product chemistry. All articles are peer-reviewed and published continously upon acceptance. Molecules is published by MDPI, Basel, Switzerland. Our aim is to encourage chemists to publish as much as possible their experimental detail, particularly synthetic procedures and characterization information. There is no restriction on the length of the experimental section. In addition, availability of compound samples is published and considered as important information. Authors are encouraged to register or deposit their chemical samples through the non-profit international organization Molecular Diversity Preservation International (MDPI). Molecules has been launched in 1996 to preserve and exploit molecular diversity of both, chemical information and chemical substances.