Surfactant enhanced lipase containing films characterized by confocal laser scanning microscopy

• Published in: Colloids and surfaces, B, Biointerfaces B, Biointerfaces, 2011-02, Vol.82 (2), p.291-296
• Main Authors: Jayawardena, Menuk B., Yee, Lachlan H., Rainbow, Ian J., Bergquist, Peter, Such, Christopher, Steinberg, Peter D., Kjelleberg, Staffan J.
• Format: Article
• Language: English
• Subjects: Acetates; additives; Ascomycota - enzymology; assays; Candida - enzymology; Candida antarctica; Catalysis; Catalytic coating; Coating; coatings; Confocal; Confocal laser scanning microscopy; Correlation; enzyme activity; Enzymes; films (materials); fluorescent dyes; fluorescent labeling; Humicola; Humicola lanuginosa; image analysis; Latex; Latex - chemistry; Lipase; Lipase - chemistry; Materials Testing; Microscopy, Confocal - methods; Pseudozyma antarctica; Scanning microscopy; Software; Surface enzyme activity; Surface Properties; Surface-Active Agents - chemistry; surfaces; Surfactant; Surfactants; Thermomyces lanuginosus; triacylglycerol lipase
• ISSN: 0927-7765; 1873-4367
• DOI: 10.1016/j.colsurfb.2010.08.042

Confocal laser scanning microscopy (CLSM) in combination with a fluorescently labeling enzyme dye, LavaPurple™, was demonstrated as a technique for the visualization of Thermomyces ( Humicola) lanuginosa lipase (LIP HLL) and Candida antarctica lipase A (LIP CA) within a transparent latex coating. Addition of Teric Surfactants (C 16 non-ionic Teric 475, 1.8% (w/w) or C 10 non-ionic Teric 460, 2.0% (w/w)) significantly increased the accumulation of both LIP HLL and LIP CA to the surface of a latex coating. An α-naphthyl acetate substrate assay was used to quantify the accumulated lipase. The results derived from the acetate assay correlated with the enzyme accumulation (at the surface) observed in the CLSM images of the latex coating. This correlation demonstrated that the increased enzyme accumulation within the top 2 μm of the latex film was responsible for the increase in surface enzymatic activity. The combination of CLSM imagery and quantifiable image analysis provided a valuable tool for the optimization of surfactant concentrations for maximizing the activity of an enzyme (and potentially other additives) within a latex coating.