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Biotechnology and Applied Biochemistry (2003) 38, (123–130) (Printed in Great Britain)

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Maize (Zea mays)-derived bovine trypsin: characterization of the first large-scale, commercial protein product from transgenic plants

Susan L. Woodard*¹, Jocelyne M. Mayor*, Michele R. Bailey*, Donna K. Barker*, Robert T. Love*, Jeffrey R. Lane*, Donna E. Delaney*, Janet M. McComas-Wagner*, Hanuman D. Mallubhotla*, Elizabeth E. Hood*, Lawrence J. Dangott†, Shane E. Tichy‡ and John A. Howard*

*ProdiGene, Inc., 101 Gateway Blvd., Suite 100, College Station, TX 77845-4473, U.S.A., †The Protein Chemistry Laboratory, Texas A&M University, Department of Biochemistry, College Station, TX 77843, U.S.A., and ‡The Laboratory for Biological Mass Spectrometry, Department of Chemistry, Texas A&M University, College Station, TX 77843, U.S.A.

Key words: maize, processing enzyme, protease, recombinant enzyme, trypsinogen.

Abbreviations used: BAPNA, N-a-benzoyl-L-arginine 4-nitronilide; CTI, corn (maize) trypsin inhibitor; MALDI-TOF, matrix-assisted laser-desorption ionization–time-of-flight; STI, soya-bean trypsin inhibitor; TAME, p-tosyl-L-arginine methyl ester.

¹To whom correspondence should be addressed (e-mail swoodard@prodigene.com).

Bovine trypsin (EC 3.4.21.4) is an enzyme that is widely used for commercial purposes to digest or process other proteins, including some therapeutic proteins. The biopharmaceutical industry is trying to eliminate animal-derived proteins from manufacturing processes due to the possible contamination of these products by human pathogens. Recombinant trypsin has been produced in a number of systems, including cell culture, bacteria and yeast. To date, these expression systems have not produced trypsin on a scale sufficient to fulfill the need of biopharmaceutical manufacturers where kilogram quantities are often required. The present paper describes commercial-level production of trypsin in transgenic maize (Zea mays) and its physical and functional characterization. This protease, the first enzyme to be produced on a large-scale using transgenic plant technology, is functionally equivalent to native bovine pancreatic trypsin. The availability of this reagent should allow for the replacement of animal-derived trypsin in the processing of pharmaceutical proteins.

Received 3 February 2003/7 April 2003; accepted 15 May 2003

Published as Immediate Publication 15 May 2003, DOI 10.1042/BA20030026

© 2003 Portland Press Ltd