The short form of the recombinant CAL-A-type lipase UM03410 from the smut fungus Ustilago maydis exhibits an inherent trans-fatty acid selectivity

Henrike Brundiek, Stefan Saß, Andrew Evitt, Robert Kourist, Uwe T Bornscheuer

Research output: Contribution to journalArticle

Abstract

The Ustilago maydis lipase UM03410 belongs to the mostly unexplored Candida antarctica lipase (CAL-A) subfamily. The two lipases with [corrected] the highest identity are a lipase from Sporisorium reilianum and the prototypic CAL-A. In contrast to the other CAL-A-type lipases, this hypothetical U. maydis lipase is annotated to possess a prolonged N-terminus of unknown function. Here, we show for the first time the recombinant expression of two versions of lipase UM03410: the full-length form (lipUMf) and an Nterminally truncated form (lipUMs). For comparison to the prototype, the expression of recombinant CAL-A in E. coli was investigated. Although both forms of lipase UM03410 could be expressed functionally in E. coli, the N-terminally truncated form (lipUMs) demonstrated significantly higher activities towards p-nitrophenyl esters. The functional expression of the N-terminally truncated lipase was further optimized by the appropriate choice of the E. coli strain, lowering the cultivation temperature to 20 °C and enrichment of the cultivation medium with glucose. Primary characteristics of the recombinant lipase are its pH optimum in the range of 6.5-7.0 and its temperature optimum at 55 °C. As is typical for lipases, lipUM03410 shows preference for long chain fatty acid esters with myristic acid ester (C14:0 ester) being the most preferred one.More importantly, lipUMs exhibits an inherent preference for C18:1Δ9 trans and C18:1Δ11 trans-fatty acid esters similar to CAL-A. Therefore, the short form of this U. maydis lipase is the only other currently known lipase with a distinct trans-fatty acid selectivity.

Original languageEnglish
Pages (from-to)141-50
Number of pages10
JournalApplied Microbiology and Biotechnology
Volume94
Issue number1
DOIs
Publication statusPublished - Apr 2012

Keywords

  • Enzyme Stability
  • Fungal Proteins
  • Hydrogen-Ion Concentration
  • Kinetics
  • Lipase
  • Plant Diseases
  • Protein Structure, Tertiary
  • Substrate Specificity
  • Trans Fatty Acids
  • Ustilago
  • Journal Article

Fields of Expertise

  • Human- & Biotechnology

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