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 journalArticleScientificpeer-review

24 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

Fingerprint

Aspergillus niger
Cloning
Aspergillus
Organism Cloning
Hydrolysis
Screening
Polycaprolactone
Malus
Gardening
Enzyme activity
Pichia
Agar
Carbon
Gas Chromatography-Mass Spectrometry
cutinase
Soil
Liquids
Substrates
Enzymes

Keywords

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

Cite this

Nyyssölä, Antti ; Pihlajaniemi, V. ; Järvinen, R. ; Mikander, S. ; Kontkanen, Hanna ; Kruus, Kristiina ; Kallio, H. ; Buchert, J. / Screening of microbes for novel acidic cutinases and cloning and expression of an acidic cutinase from Aspergillus niger CBS 513.88. In: Enzyme and Microbial Technology. 2013 ; Vol. 52, No. 4-5. pp. 272-278.
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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. / Nyyssölä, Antti (Corresponding Author); Pihlajaniemi, V.; Järvinen, R.; Mikander, S.; Kontkanen, Hanna; Kruus, Kristiina; Kallio, H.; Buchert, J.

In: Enzyme and Microbial Technology, Vol. 52, No. 4-5, 2013, p. 272-278.

Research output: Contribution to journalArticleScientificpeer-review

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AU - Nyyssölä, Antti

AU - Pihlajaniemi, V.

AU - Järvinen, R.

AU - Mikander, S.

AU - Kontkanen, Hanna

AU - Kruus, Kristiina

AU - Kallio, H.

AU - Buchert, J.

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KW - plant pathogen

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