Purification and properties of an alkaline proteinase of Fusarium culmorum

Anja Pekkarinen (Corresponding Author), Berne Jones, Marja-Leena Niku-Paavola

Research output: Contribution to journalArticleScientificpeer-review

44 Citations (Scopus)

Abstract

The disease Fusarium head blight (scab) causes severe problems for farmers and for the industries that use cereals. It is likely that the fungi that cause scab (Fusarium spp.) use various enzymes when they invade grains. We are studying enzymes that the fungi may use to hydrolyze grain proteins. To do this, Fusarium culmorum was grown in a gluten-containing medium from which an alkaline serine proteinase with a molecular mass of 28.7kDa was purified by size-exclusion and cation exchange chromatographies. The enzyme was maximally active at pH8.3-9.6 and 50°C, but was unstable under these conditions. It hydrolyzed the synthetic substrates N-succinyl-Ala-Ala-Pro-Phe p-nitroanilide and, to a lesser extent, N-succinyl-Ala-Ala-Pro-Leu p-nitroanilide. It was inhibited by phenylmethanesulfonyl fluoride and chymostatin, but not by soybean trypsin or Bowman-Birk inhibitors. Parts of the amino-acid sequence were up to 82% homologous with those of several fungal subtilisins. One of the active site amino acids was detected and it occupied the same relative position as in the other subtilisins. Therefore, on the basis of these characteristics, the proteinase is subtilisin-like. Purification of the enzyme was complicated by the fact that, when purified, it apparently underwent autolysis. The presence of extraneous protein stabilized the activity.
Original languageEnglish
Pages (from-to)798-807
JournalEuropean Journal of Biochemistry
Volume269
Issue number3
DOIs
Publication statusPublished - 2002
MoE publication typeA1 Journal article-refereed

Fingerprint

Fusarium
Purification
Subtilisins
Enzymes
Fungi
Bowman Birk Soybean Trypsin Inhibitor
Phenylmethylsulfonyl Fluoride
Amino Acids
Subtilisin
Autolysis
Glutens
Molecular mass
Serine Proteases
Chromatography
Trypsin
Cations
Amino Acid Sequence
Catalytic Domain
Industry
Proteins

Keywords

  • scab
  • head blight
  • Fusarium
  • fungi
  • cereals
  • proteinase

Cite this

Pekkarinen, Anja ; Jones, Berne ; Niku-Paavola, Marja-Leena. / Purification and properties of an alkaline proteinase of Fusarium culmorum. In: European Journal of Biochemistry. 2002 ; Vol. 269, No. 3. pp. 798-807.
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Purification and properties of an alkaline proteinase of Fusarium culmorum. / Pekkarinen, Anja (Corresponding Author); Jones, Berne; Niku-Paavola, Marja-Leena.

In: European Journal of Biochemistry, Vol. 269, No. 3, 2002, p. 798-807.

Research output: Contribution to journalArticleScientificpeer-review

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