A1 Refereed original research article in a scientific journal
Screening of microbes for novel acid cutinases and cloning and expression of an acidic cutinase from Aspergillus niger CBS 513.88
Authors: Nyyssölä A, Pihlajaniemi V, Järvinen R, Mikander S, Kontkanen H, Kruus K, Kallio H Buchert J
Publication year: 2013
Journal: Enzyme and Microbial Technology
Journal acronym: Enz Microbiol Technol
Number in series: 4-5
Volume: 52
Issue: 4-5
First page : 272
Last page: 278
Number of pages: 7
ISSN: 0141-0229
DOI: https://doi.org/10.1016/j.enzmictec.2013.01.005
Abstract
Isolates from gardening waste compost and 38 culture collection microbes were grown on agar plates at pH 4.0 with the cutinase model substrate polycaprolactone as a carbon source. The strains showing polycaprolactone hydrolysis were cultivated in liq. at acidic pH and the cultivations were monitored by assaying the p-nitrophenyl butyrate esterase activities. Culture supernatants of four strains were analyzed for the hydrolysis of tritiated apple cutin at different pHs. Highest amts. of radioactive hydrolysis products were detected at pHs below 5. The hydrolysis of apple cutin by the culture supernatants at acidic pH was further confirmed by GC-MS anal. of the hydrolysis products. On the basis of screening, the acidic cutinase from Aspergillus niger CBS 513.88 was chosen for heterogeneous prodn. in Pichia pastoris and for anal. of the effects of pH on activity and stability. The recombinant enzyme showed activity over a broad range of pHs with maximal activity between pH 5.0 and 6.5. Activity could be detected still at pH 3.5.
Isolates from gardening waste compost and 38 culture collection microbes were grown on agar plates at pH 4.0 with the cutinase model substrate polycaprolactone as a carbon source. The strains showing polycaprolactone hydrolysis were cultivated in liq. at acidic pH and the cultivations were monitored by assaying the p-nitrophenyl butyrate esterase activities. Culture supernatants of four strains were analyzed for the hydrolysis of tritiated apple cutin at different pHs. Highest amts. of radioactive hydrolysis products were detected at pHs below 5. The hydrolysis of apple cutin by the culture supernatants at acidic pH was further confirmed by GC-MS anal. of the hydrolysis products. On the basis of screening, the acidic cutinase from Aspergillus niger CBS 513.88 was chosen for heterogeneous prodn. in Pichia pastoris and for anal. of the effects of pH on activity and stability. The recombinant enzyme showed activity over a broad range of pHs with maximal activity between pH 5.0 and 6.5. Activity could be detected still at pH 3.5.
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