Journal
LANGMUIR
Volume 37, Issue 25, Pages 7771-7779Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.langmuir.1c00934
Keywords
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Funding
- Sao Paulo Research Foundation (FAPESP) [2018/00291-9, 2019/01797-6, 2019/03239-0, 2018/22214-6]
- University of Miami (Miami, Florida)
- Federal University of Sao Paulo (Diadema, Sao Paulo)
- Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP) [19/03239-0, 18/22214-6] Funding Source: FAPESP
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This work explores the physicochemical properties of mixed stearic acid (HSt)/phenylalanine dehydrogenase enzyme (PheDH) Langmuir films and their immobilization onto solid supports as Langmuir-Blodgett films. The hydrophobic interactions between the enzyme and HSt were found to tune the secondary structure of PheDH. The stability of the mixed floating monolayers allowed their successful transfer to solid supports and formation of LB films, which is crucial for creating optical and electrochemical sensors for phenylalanine with controlled molecular architecture.
This work investigates the physicochemical properties of mixed stearic acid (HSt)/phenylalanine dehydrogenase enzyme (PheDH) Langmuir films and their immobilization onto solid supports as Langmuir-Blodgett (LB) films. PheDH from the aqueous subphase enters the surfactant matrix up to an exclusion surface pressure of 25.3 mN/m, leading to the formation of stable and highly condensed mixed Langmuir monolayers. Hydrophobic interactions between the enzyme and HSt nonpolar groups tuned the secondary structure of PheDH, evidenced by the presence of beta-sheet structures as demonstrated by infrared and circular dichroism spectra. The floating monolayers were successfully transferred to solid quartz supports, yielding Y-type LB films, and then characterized employing fluorescence, circular dichroism, and microscopic techniques, which indicated that PheDH was co-immobilized with HSt proportionally to the number of transferred layers. The enzyme fluidized the HSt monolayers, reducing their maximum dipoles when condensed to their maximum, and disorganized the alkyl chains of the fatty acid, as detected with infrared spectroscopy. The stability of the mixed floating monolayers enabled their transfer to solid supports as LB films, which is important for producing optical and electrochemical sensors for phenylalanine whose molecular architecture can be controlled with precision.
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