Kcat of lipase B with p-Nitrophenol butyrate

Value 5.1 1/sec Range: ±0.2 1/sec
Organism Yeast Candida antarctica
Reference Qian Z, Lutz S. Improving the catalytic activity of Candida antarctica lipase B by circular permutation. J Am Chem Soc. 2005 Oct 5 127(39):13466-7. Table 1 Table - link PubMed ID16190688
Method Directed evolution: To examine the impact of circular permutation on catalysis, researchers selected eight functional lipase B from Candida antarctica (CALB) variants with termini in or near the active site for detailed kinetic characterization (Figure 2). Following overexpression in P. pastoris, the proteins were purified to homogeneity, and kinetic data for these variants were measured on two standard lipase substrates, p-nitrophenol butyrate (pNB) and 6.8-difluoro-4-methylumbelliferyl (DiFMU) octanoate (table link).
Comments 305±10 min^-1. Note-In 'The metabolic pathway engineering handbook' (CRC Press), ed. C.D. Smolke 2010, Chapter 2-23, Nair and Zhao point that researchers in the above experiment used ~5×10^5 mutants and increased catalytic efficiency of CALB for p-Nitrophenol butyrate over 12 fold (see table link).
Entered by Uri M
ID 105429