TITLE:
Enzyme from Banana (Musa sp.) Extraction Procedures for Sensitive Adrenaline Biosensor Construction
AUTHORS:
Solange Carvalho da Silva, Célio Wisniewski, Pedro Orival Luccas, Cristiana Schmidt de Magalhães
KEYWORDS:
Enzymatic Extract; Epinephrine; Biosensor; Polyphenol Oxidase
JOURNAL NAME:
American Journal of Analytical Chemistry,
Vol.4 No.6,
June
20,
2013
ABSTRACT: For building a biosensor, at least two enzyme sources
can be employed. The pure enzyme has features with better selectivity
and low stability. Crude extract presents better stability and wrong
selectivity. Thus, one intermediate condition can be feasible joining both
benefits of crude extract and pure enzymes. For that result, several procedures
of extraction and semi purification of polyphenol oxidase (PPO) enzyme from
banana (Musa sp.) were studied. The
results showed that cleaned enzymatic extracts presented higher specific
activities than crude extracts (2.8 up to 5.3 fold), despite the total protein
concentration diminishing from 27% up to 72%, indicating that polyphenol
oxidase (PPO) enzyme was preserved in them. The biosensors with 125 AU mL-1 containing cleaned enzymatic
extracts performed better by grinding or grinding plus sonication for 30 s. They
were linear over the ranges of 5.9 × 10-6 mol·L-1 to 1.4 × 10-3 mol·L-1 and 7.9 × 10-8 mol·L-1 to 4.0 × 10-3 mol·L-1, respectively. The limit of
Detection (LOD) was 5.9 × 10-6 mol·L-1 and 7.9 × 10-8 mol·L-1, respectively. The LOD obtained is adequate to adrenaline
determination on blood and medicinal samples, and were
applied in medicinal samples with satisfactory results.