Abstract
The use of marine enzymes as catalysts for biotechnological applications is a topical subject. Marine enzymes usually display better operational properties than their animal, plant or bacterial counterparts, enlarging the range of possible biotechnological applications. Due to the fact that cytochrome P450 enzymes can degrade many different toxic environmental compounds, these enzymes have emerged as valuable tools in bioremediation processes. The present work describes the isolation, purification and biochemical characterization of a liver NADPH-dependent cytochrome P450 reductase (CPR) from the marine fish Liza klunzingeri (LkCPR). Experimental results revealed that LkCPR is a monomer of approximately 75 kDa that is active in a wide range of pH values (6-9) and temperatures (40-60 degrees C), showing the highest catalytic activity at pH 8 and 50 degrees C. The activation energy of the enzyme reaction was 16.3 kcal mol(-1) K-1. The K-M values for cytochrome C and NADPH were 8.83 mu M and 7.26 mu M, and the k(cat) values were 206.79 s(-1) and 202.93 s(-1), respectively. LkCPR displayed a specific activity versus cytochrome C of 402.07 mu mol min(-1) mg(1), the highest activity value described for a CPR up to date (3.2-4.7 times higher than the most active reported CPRs) and showed the highest thermostability described for a CPR. Taking into account all these remarkable catalytic features, LkCPR offers great potential to be used as a suitable biocatalyst.