Large scale production of the copper enzyme peptidylglycine monooxygenase using an automated bioreactor

Andrew T. Bauman, Martina Ralle, Ninian Blackburn

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Rat PHM (peptidylglycine α-hydroxylating monooxygenase; EC 1.14.17.3) expressed in CHO DG44 cells as a recombinant protein (rat PHMcc, residues 42-356 cloned in the pCIS vector, A.S. Kolhekar, H.T. Keutman, R. E. Mains, A.S.W. Quon, B.A. Eipper, Biochemistry 36 (1997) 10901-10909), was produced in two different bioreactors, a Cellmax 100 (B1) and an Accusyst-MiniMax (B2). B2 contains features not present in B1, which contribute to environmental control, and ease of operation, and was more successful at producing high quality PHM than B1 in both yield (B1: 5 mg/day, B2: 12-15 mg/day), activity (B1: 12-20 μmol O2/min/mg, B2: 24-36 μmol O2/min/mg), and viability (B1:

Original languageEnglish (US)
Pages (from-to)34-38
Number of pages5
JournalProtein Expression and Purification
Volume51
Issue number1
DOIs
StatePublished - Jan 2007

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Bioreactors
Rats
Copper
Biochemistry
CHO Cells
Enzymes
Mixed Function Oxygenases
Recombinant Proteins
peptidylglycine monooxygenase

Keywords

  • Bioreactor
  • Copper
  • Mass spectrometry
  • Monooxygenase
  • Peptidylglycine
  • PHM

ASJC Scopus subject areas

  • Biochemistry

Cite this

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title = "Large scale production of the copper enzyme peptidylglycine monooxygenase using an automated bioreactor",
abstract = "Rat PHM (peptidylglycine α-hydroxylating monooxygenase; EC 1.14.17.3) expressed in CHO DG44 cells as a recombinant protein (rat PHMcc, residues 42-356 cloned in the pCIS vector, A.S. Kolhekar, H.T. Keutman, R. E. Mains, A.S.W. Quon, B.A. Eipper, Biochemistry 36 (1997) 10901-10909), was produced in two different bioreactors, a Cellmax 100 (B1) and an Accusyst-MiniMax (B2). B2 contains features not present in B1, which contribute to environmental control, and ease of operation, and was more successful at producing high quality PHM than B1 in both yield (B1: 5 mg/day, B2: 12-15 mg/day), activity (B1: 12-20 μmol O2/min/mg, B2: 24-36 μmol O2/min/mg), and viability (B1:",
keywords = "Bioreactor, Copper, Mass spectrometry, Monooxygenase, Peptidylglycine, PHM",
author = "Bauman, {Andrew T.} and Martina Ralle and Ninian Blackburn",
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AU - Ralle, Martina

AU - Blackburn, Ninian

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KW - Bioreactor

KW - Copper

KW - Mass spectrometry

KW - Monooxygenase

KW - Peptidylglycine

KW - PHM

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