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

Paper ID Volume ID Publish Year Pages File Format Full-Text
17372 42663 2013 7 PDF Available
Title
Screening of microbes for novel acidic cutinases and cloning and expression of an acidic cutinase from Aspergillus niger CBS 513.88
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.

Graphical abstractFigure optionsDownload full-size imageDownload as PowerPoint slideHighlights► Microbes showing hydrolysis of the cutin analog polycaprolactone at acidic pH were discovered by screening on agar. ► The culture supernatants of the strains showed activity toward tritiated cutin at pHs below 4. ► Hydrolysis of natural cutin by the culture supernatants at acidic pH was confirmed by GC-MS. ► The novel acidic cutinase from Aspergillus niger was produced in Pichia pastoris in active form. ► The discovery of acidic cutinases enables new applications in biomass processing.

Keywords
Cutin; Polyester; Cutinase; Esterase; Plant pathogen; Fungi
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Screening of microbes for novel acidic cutinases and cloning and expression of an acidic cutinase from Aspergillus niger CBS 513.88
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 52, Issues 4–5, 10 April 2013, Pages 272–278
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us