Lactococcus lactis as expression host for the biosynthetic incorporation of tryptophan analogues in recombinant proteins - Archive ouverte HAL Access content directly
Journal Articles Biochemical Journal Year : 2007

Lactococcus lactis as expression host for the biosynthetic incorporation of tryptophan analogues in recombinant proteins

Mohamed El Khattabi
  • Function : Author
Maarten L. Van Roosmalen
  • Function : Author
Dennis Jager
  • Function : Author
Heidi Metselaar
  • Function : Author
Hjalmar Permentier
  • Function : Author
Kees Leenhouts
  • Function : Correspondent author
  • PersonId : 870156

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Jaap Broos
  • Function : Author

Abstract

Incorporation of tryptophan (Trp) analogues into a protein may facilitate its structural analysis by spectroscopic techniques. Development of a biological system for the biosynthetic incorporation of such analogues into proteins is of considerable importance. The Gram-negative Escherichia coli is the only prokaryotic expression host regularly used for the incorporation of Trp analogues into recombinant proteins. Here we present the use of the versatile Gram-positive expression host Lactococcus lactis for the incorporation of Trp analogues. The availability of a tightly regulated expression system for this organism, the possibility to secrete modified proteins into the growth medium and the construction of the trp-synthetase deletion strain PA1002 of L. lactis rendered this organism potentially an efficient tool for the incorporation of Trp analogues in recombinant proteins. The Trp analogues 7-azatryptophan, 5-fluorotryptophan and 5-hydroxytryptophan were incorporated with efficiencies of >97%, >97% and 89%, respectively. Interestingly, 5-methyltryptophan could be incorporated with 92% efficiency. Successful biosynthetical incorporation of 5-methylTrp in recombinant proteins has not been reported before.

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hal-00478844 , version 1 (30-04-2010)

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Mohamed El Khattabi, Maarten L. Van Roosmalen, Dennis Jager, Heidi Metselaar, Hjalmar Permentier, et al.. Lactococcus lactis as expression host for the biosynthetic incorporation of tryptophan analogues in recombinant proteins. Biochemical Journal, 2007, 409 (1), pp.193-198. ⟨10.1042/BJ20070909⟩. ⟨hal-00478844⟩

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