Journal
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
Volume 103, Issue 7, Pages 3061-3071Publisher
SPRINGER
DOI: 10.1007/s00253-019-09678-2
Keywords
Immobilization; Chimera proteins; L-DOPA; Caffeic acid; Self-assembly
Categories
Funding
- University Federico II [000023_ALTRI_DR_409_2017_Ricerca Ateneo_GIARDINA]
- Ministry of Education, Universities and Research
- European Union [172]
Ask authors/readers for more resources
A simple and stable immobilization of a laccase from Pleurotus ostreatus was obtained through genetic fusion with a self-assembling and adhesive class I hydrophobin. The chimera protein was expressed in Pichia pastoris and secreted into the culture medium. The crude culture supernatant was directly used for coatings of polystyrene multi-well plates without additional treatments, a procedure that resulted in a less time-consuming and chemicals reduction. Furthermore, the gene fusion yielded a positive effect with respect to the wild-type recombinant enzyme in terms of both immobilization and stability. The multi-well plate with the immobilized chimera was used to develop an optical biosensor to monitor two phenolic compounds: L-DOPA ((S)-2-amino-3-(3,4-dihydroxyphenyl) propanoic acid) and caffeic acid (3-(3,4-dihydroxyphenyl)-2-propenoic acid); the estimation of which is a matter of interest in the pharmaceutics and food field. The method was based on the use of the analytes as competing inhibitors of the laccase-mediated ABTS oxidation. The main advantages of the developed biosensor are the ease of preparation, the use of small sample volumes, and the simultaneous analysis of multiple samples on a single platform.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available