Cutinase

Creveld, L., Amadei, A., Van Schaik, C., Pepermans, R., De Vlieg, J., Berendsen, H.J.C. Identification of functional and unfolding motions of cutinase as obtained from molecular dynamics computer simulations. Submitted (1998). 

Characterization of Substrate-specificity for Cutinase Produced by Novel Pseudozyma aphidis Isolated from Pea Leaves in South Korea... 

Enzymatic Degradation and Detoxification of Diethyl Phthalate by Fusarium oxysporum f. sp.pisi Cutinase... 

Cutinase is a hydrolytic enzyme that degrades cutin, the cuticular polymer of higher plants [4], Unlike the oflier lipolytic enzymes, such lipases and esterases, cutinase does not require interfacial activation for substrate binding and activity. Cutinases have been largely exploited for esterification and transesterification in chemical synthesis [5] and have also been applied in laundry or dishwashing detergent [6]. 

The purified fungal cutinase fiom F. oxysporum f. sp. pisi was kindly provided by Prof. C.M.J. Sagt in Utrecht University. The commercial product of esterase firom Candida cylindracea was purchased fi om Boehringer Mannheim (Germany). The enzymatic degradation of DEP... 

The estimation of the initial degradation constants, kcio and kEioo, for cutinase (10 mg f ) and esterase (100 mg l ), respectively, demonstrated that the initial DEP degradation by cutinase seemed to proceed almost 2.6 times faster. 

Fig. 2. Time-course variation in the amount of chemical compounds produced during DEP degradation by fungal cutinase (10 mg 1 ) (a) and yeast esterase (100 mg F) (b), analyzed through GC/MS chromatography.

Isolation of Fungal Cutinases and their Molecular Properties... 

When polyester-hydrolyzing activity was isolated using synthetic polyesters such as polycaprolactone, and the enzyme was examined in detail, it was found that it was a cutinase that was responsible for the hydrolysis [113]. Similarly, the polyester domains of suberin were found to be degraded by cutinase. Cutinase is a polyesterase, and similar enzymes may be widely distributed and can degrade a variety of natural and synthetic polyesters. Microbial polyhydroxy-alkanoic acids that are attracting increasing attention as biodegradable polyesters can be hydrolyzed by bacterial polyesterases that share some common features with cutinases [114] and this area is covered in another chapter [115]. 

Fungal cutinase catalyzes hydrolysis of model substrates and in particular p-nitrophenyl esters of short chain fatty acids, providing a convenient spectro-photometric assay for this enzyme activity [101,102,116]. Hydrolysis of model esters by this cutinase showed the high degree of preference of this enzyme for primary alcohol ester hydrolysis. Wax esters and methyl esters of fatty acids were hydrolyzed at low rates. Alkane-2-ol esters were hydrolyzed much more slowly than wax esters and esters of mid-chain secondary alcohols were not... 

Fungal cutinases show no free SH groups but have 4 Cys residues, indicating that they are in disulfide linkage [119]. The reaction of the native enzyme with DTE was extremely slow but in the presence of SDS at its CMC rapid reduction could be observed [102]. Reduction of the disulfide bridge resulted in irreversible inactivation of the enzyme and the protein tended to become insoluble. CD spectra of cutinase in the 205-230 nm region, before and after DTE reduc-... 

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