Cutinase activity

Acetate is known to be a good carbon source for fungi and would be expected to be the ultimate degradation product of cutin (14). The effect of acetate on the production of cutinase by the T-8 strain of F. solani was examined and compared with that of glucose. Since previous studies showed that hydrolysis of the artificial substrate p-nitrophenylbutyrate, PNB, was specifically hydrolyzed by cutinase in the T-8 strain, this activity was used to measure cutinase levels (8). Figure 1 illustrates that basal levels of cutinase activity were detected in the growth medium when T-8 was grown on... 

Next the parent and mutant were compared for their ability to induce cutinase by cutin and hydrolyzed cutin (consisting of small molecular weight inducers) after growth on glucose. As shown in Figure 5, cutinase activity increased over the three-day induction period for both strains in the presence of cutin (Panel A) or hydrolyzed cutin (Panel B) however, cutinase was induced less effectively in the mutant strain, as evidenced by an 80-90% reduction. Hydrolyzed cutin was a 10-fold less effective inducer than cutin in both strains. These data indicated that lack of induction in the mutant was not related to its inability to hydrolyze cutin to small molecular weight inducers. Consequently the defect observed was not related to the inability of the mutant to produce the inducer. 

The PNB-1 mutant is a leaky mutant that is partially defective in cutinase production, and apparently produced 80 to 90% less normal enzyme. The regulation of the residual enzyme in the mutant was unchanged. The enzyme was repressed by glucose, and was induced by cutin or hydrolyzed cutin after depletion of glucose. Cutinase activity increased over a 15-day time period when the mutant was grown on cutin as the sole carbon source. The mutant was also less virulent than the parental strain. The nature of the PNB-1 mutation remains to be elucidated and... 

Araujo R, Silva C, O Neill A et al (2007) Tailoring cutinase activity towards polyethylene terephthalate and polyamide 6,6 fibers. J Biotechnol 128 849-857... 

Korpecka J (2009) Cutinase activity of PET-hydrolases. In Proceedings of INTB 2009, Ghent, Belgium, September 2009... 

Araujo, R., Silva, C., O Neill, A., Micaelo, N., Guebitz, G., Soares, C.M., Casal, M., and Cavaco-Paulo, A. 2007. Tailoring cutinase activity towards polyethylene terephthalate and polyamide 6, 6 hbers. Journal of Biotechnology, 128 849-57. 

Fontes N, Almeida MC, Peres C, Garcia S, Grave J, Aires-Barros MR, Soares CM, Cabral IMS, Maycock CD, Barreiros S. Cutinase activity and enantioselectivity in supercritical fluids. Ind Eng Chem Res 1998 37 3189-3194. 

J., Cavaco-Paulo, A., Guebitz, G.M. (2009) Mechanistic insights in enzymatic functionalisation of synthetic polymers cutinase activity of PET hydrolases, Proceedings of 6th International Conference of Textile and Polymer Biotechnology (ISBN 9789081392419), pp. 230-235. 

Esterase and Cutinase Activity Released into the Extracellular Fluid During Spore Germination... 

PNB activity is expressed nmol min per 10 0 spores using -nitophenylbutyrate as substrate, Cutinase activity is expressed as the amount of ct min released h per 10 spores. 

Figure 8. Time course of appearance of cutinase as measured by immunochemical techniques (left) and cutinase activity as measured by -nitrophenyl butyrate hydrolysis (right) in the extracellular fluid of spore suspensions of F. solani f. sp. pisi induced with either cutin hydrolysate or purified Cie dihydroxy acid. (Reproduced with permission from Ref. 35. Copyright 1986 The National Academy of Sciences.)...

Goncalves, A.M., Serrob, A.P., Aires-Barros, M.R., Cabral, J.M. 2000. Effects of ionic surfactants used in reversed micelles on cutinase activity and stability. Biochim. Biophys. Acta 1480, 92-106. 

Gray GL, Power SD, Poulouse AJ (1995) Lipase from Pseudomonas Mendocina having cutinase activity. US Patent 5,389,536 Griffiths AD, Tawfik DS (2003) Directed evolution of an extremely fast phosphotriesterase by in vitro compartmentalization. EMBO J 22 24-35 Gusakov AV, Sinitsyn AP, Berlin AG, Markov AV, Ankudimova NV (2000) Surface hydrophobic amino acid residues in cellulase molecules as a structural factor responsible for their high denim-washing performance. Enzyme Microb Technol 27 664-671... 

Figure 7 Induction of cutinase activity in Pseudomonas putida cultures in nutrient broth-yeast extract with or without cutin hydroiysate (1 and2) and with cutin (3). Tracing A indicates release of iabeied monomers from H-labeled cutin.

Powdery mildew infection of wheat leaves was accompanied by increased production of cuticular waxes but did not result in consistent changes in the proportions of the component fractions. The amount of wax present per unit area of leaf surface of healthy plants was greater in resistant than in susceptible cultivars, while the partially resistant cultivar investigated was intermediate in this respect. Although exudates from Erysiphe graminis f sp. hordei have been shown to have esterase and cutinase activity which is likely to be of importance in the initiation of infection (Kunoh et a/., 1990 Nicholson et al, 1993), the present results support other evidence for the importance of the thickness of the cuticular wax layer in resistance to powdery mildew. 

---