Proteolytic resistance and its utilization in purification of hydrogenase from Thiocapsa roseopersicina

Kornel L. Kovacs, Csaba Bagyinka, Gabor Tigyi

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

The effect of various proteinases on the hydrogenase from Thiocapsa roseopersicina has been studied by activity measurements accompanied with electrophoretic separation of the protein components. The native enzyme cannot be digested by the following proteinases: thermolysin, subtilisin, trypsin, papain, pepsin, proteinase-K, pronase E, Bacillus subtilis proteinase, Staphylococcus aureus strain V8 proteinase under standard conditions for optimal proteolytic cleavage. Pretreatments which commonly make resistant proteins accessible to proteolysis have been found ineffective in destabilizing the hydrogenase. The native hydrogenase is readily cleavable and inactivated by CNBr. Also under conditions which irreversibly inactivate the hydrogenase (boiling for 10 min) the enzyme loses its active conformation and becomes susceptible to proteolysis. The protein, as isolated, apparently contains no protective lipids or carbohydrates. The results suggest that a stable hydrogenase apoprotein structure is responsible for the proteolytic resistance. The remarkable stability of this enzyme offers new ways for its purification. A rapid and effective procedure, suitable for scale-up, has been established.

Original languageEnglish (US)
Pages (from-to)166-172
Number of pages7
JournalBBA - Bioenergetics
Volume935
Issue number2
DOIs
StatePublished - Sep 14 1988

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Thiocapsa roseopersicina
Hydrogenase
Purification
Peptide Hydrolases
Proteolysis
Enzymes
Thermolysin
Subtilisin
Enzyme Stability
Endopeptidase K
Pronase
Proteins
Apoproteins
Papain
Pepsin A
Bacilli
Bacillus subtilis
Trypsin
Boiling liquids
Conformations

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Cell Biology

Cite this

Proteolytic resistance and its utilization in purification of hydrogenase from Thiocapsa roseopersicina. / Kovacs, Kornel L.; Bagyinka, Csaba; Tigyi, Gabor.

In: BBA - Bioenergetics, Vol. 935, No. 2, 14.09.1988, p. 166-172.

Research output: Contribution to journalArticle

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