Bioluminescent living organisms emit light through a specific biocatalyzed reaction involving a luciferin substrate and a luciferase enzyme. The present work investigated the possibility of creating optimal luminescence by immobilization of one or both the enzymes Luciferase (Luc) and FMN reductase (Red) involved in a bioluminescent bacterial system onto a plasma-activated microfibrous PET nonwoven. Parameters affecting the catalytic activity and efficiency of the bacterial system in aqueous medium were determined by luminescence intensity measurements using a luminometer. Two types of plasma, air atmospheric plasma (ATMP) and cold remote plasma (CRPNO) treatment, were used to activate the PET nonwoven. Further, one or both enzyme(s) were immobilized using a physical adsorption technique, without or with the use of natural biopolymers (gelatin and starch) and bovine serum albumin-BSA protein, to improve enzyme stability and activity. Coimmobilization of both Red and Luc enzymes on the CRPNO plasma-activated nonwoven in the presence of BSA led to the maximum luminescence. As high as 60,000 RLU equivalent to that of an LED light used for calibration was observed and showed stable intensity up to 6 min. Fiber surface analysis was tested using wettability tests (water contact angle and capillary uptake), while scanning electron microscopy, atomic force microscopy, and electron spectroscopy for chemical analysis showed changes in fiber surface morphology and chemical functional groups. A considerable increase in "N" atom content after coimmobilization of enzymes in the presence of BSA was detected. This study is the first successful attempt to use a biomimetic strategy for immobilization of enzymes involved in bacterial luminescence on a plasma-activated microfibrous nonwoven in an attempt to attain bioluminescent materials.
Keywords: Bioluminescence; bacterial luminescence; bioluminescent materials; enzyme; immobilization; microfibrous nonwoven PET; plasma.