Screening of microbes for novel acidic cutinases and cloning and expression of an acidic cutinase from Aspergillus niger CBS 513.88

Antti Nyyssölä (Corresponding Author), V. Pihlajaniemi, R. Järvinen, S. Mikander, Hanna Kontkanen, Kristiina Kruus, H. Kallio, J. Buchert

    Research output: Contribution to journalArticlepeer-review

    26 Citations (Scopus)

    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 liquid 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 amounts 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 analysis of the hydrolysis products. On the basis of screening, the acidic cutinase from Aspergillus niger CBS 513.88 was chosen for heterogeneous production in Pichia pastoris and for analysis 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.
    Original languageEnglish
    Pages (from-to)272-278
    Number of pages7
    JournalEnzyme and Microbial Technology
    Volume52
    Issue number4-5
    DOIs
    Publication statusPublished - 2013
    MoE publication typeA1 Journal article-refereed

    Keywords

    • cutin
    • cutinase
    • esterase
    • fungi
    • plant pathogen
    • polyester

    Fingerprint

    Dive into the research topics of 'Screening of microbes for novel acidic cutinases and cloning and expression of an acidic cutinase from <i>Aspergillus niger</i> CBS 513.88'. Together they form a unique fingerprint.

    Cite this