Lipase is an enzyme catalyzing reactions borne by triglycerides such as transesterification for biodiesel production and has been used in biosensors for detection of β-hydroxyacid esters [1] and triglycerides in blood serum [2]. Immobilization of the enzymes is essential for their industrial application, since it allows the development of continuous processes, easier separation of products, the reuse of the catalyst and, in some cases, it enhances enzyme properties such as pH and temperature stability and their catalytic activity in non-aqueous media [3]. MAPLE is a thin film deposition technique derived from Pulsed Laser Deposition (PLD) for deposition of delicate materials (biomolecules, polymers, etc.) in undamaged form. The main difference in comparison to classical PLD is the use of a frozen (usually by means of a liquid nitrogen flux) target obtained from a solution or a suspension of the guest material (to be deposited) in a matrix (a volatile solvent). In this way, the laser beam energy is mainly absorbed by the matrix while only the guest material reaches the substrate, since the solvent is pumped away by the vacuum system. By MAPLE technique it can be possible to “freeze” the conformation of the lipase as it is in solution, in such a way to tailor lipase properties in solution. In this way the lipase conformation, essential for its catalytic activity, would be independent on the support properties. Here we show that Matrix Assisted Pulsed Laser Evaporation (MAPLE) technique can be used to deposit lipase. 1. T. Kullick, R. Ulber, H.H. Meyer, T. Scheper, K. Schlügerl. Anal. Chim. Acta. 239 (1994) 271 2. Mohanasundaram Sulur Veeramani, Karuppiah Prakash Shyam, Noel Prashant Ratchagar, Anju Chadhabc and Enakshi Bhattacharya, Miniaturised silicon biosensors for the detection of triglyceride in blood serum, Anal. Methods, 2014, Advance Article; DOI: 10.1039/C3AY42274G. 3. P.M. Nielsen, J. Brask, L. Fjerbaek. Eur. J. Lipid Sci. Technol. 110 (2008) 692-700.
Matrix Assisted Pulsed Laser Evaporation of Biological Thin Films: Lipase / Aronne, Antonio; Bloisi, Francesco; Calabria, Raffaela; Califano, Valeria; Depero Laura, E; Fanelli, Esther; Federici, Stefania; Massoli, Patrizio; Vicari, LUCIANO ROSARIO MARIA. - (2014). (Intervento presentato al convegno EMRS Spring Meeting 2014 - Session J - Laser interaction with advanced materials: fundamentals and applications tenutosi a Lille, France nel May 26-30 2014).
Matrix Assisted Pulsed Laser Evaporation of Biological Thin Films: Lipase
ARONNE, ANTONIO;BLOISI, FRANCESCO;FANELLI, Esther;VICARI, LUCIANO ROSARIO MARIA
2014
Abstract
Lipase is an enzyme catalyzing reactions borne by triglycerides such as transesterification for biodiesel production and has been used in biosensors for detection of β-hydroxyacid esters [1] and triglycerides in blood serum [2]. Immobilization of the enzymes is essential for their industrial application, since it allows the development of continuous processes, easier separation of products, the reuse of the catalyst and, in some cases, it enhances enzyme properties such as pH and temperature stability and their catalytic activity in non-aqueous media [3]. MAPLE is a thin film deposition technique derived from Pulsed Laser Deposition (PLD) for deposition of delicate materials (biomolecules, polymers, etc.) in undamaged form. The main difference in comparison to classical PLD is the use of a frozen (usually by means of a liquid nitrogen flux) target obtained from a solution or a suspension of the guest material (to be deposited) in a matrix (a volatile solvent). In this way, the laser beam energy is mainly absorbed by the matrix while only the guest material reaches the substrate, since the solvent is pumped away by the vacuum system. By MAPLE technique it can be possible to “freeze” the conformation of the lipase as it is in solution, in such a way to tailor lipase properties in solution. In this way the lipase conformation, essential for its catalytic activity, would be independent on the support properties. Here we show that Matrix Assisted Pulsed Laser Evaporation (MAPLE) technique can be used to deposit lipase. 1. T. Kullick, R. Ulber, H.H. Meyer, T. Scheper, K. Schlügerl. Anal. Chim. Acta. 239 (1994) 271 2. Mohanasundaram Sulur Veeramani, Karuppiah Prakash Shyam, Noel Prashant Ratchagar, Anju Chadhabc and Enakshi Bhattacharya, Miniaturised silicon biosensors for the detection of triglyceride in blood serum, Anal. Methods, 2014, Advance Article; DOI: 10.1039/C3AY42274G. 3. P.M. Nielsen, J. Brask, L. Fjerbaek. Eur. J. Lipid Sci. Technol. 110 (2008) 692-700.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
