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.

LanguageEnglish
Pages141-50
Number of pages10
JournalApplied Microbiology and Biotechnology
Volume94
Issue number1
DOIs
StatusPublished - Apr 2012

Fingerprint

Ustilago
Trans Fatty Acids
Lipase
Candida
Fungi
Esters
Escherichia coli
Temperature
Myristic Acid

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

Cite this

The short form of the recombinant CAL-A-type lipase UM03410 from the smut fungus Ustilago maydis exhibits an inherent trans-fatty acid selectivity. / Brundiek, Henrike; Saß, Stefan; Evitt, Andrew; Kourist, Robert; Bornscheuer, Uwe T.

In: Applied Microbiology and Biotechnology, Vol. 94, No. 1, 04.2012, p. 141-50.

Research output: Contribution to journalArticle

@article{5803a040993b458cb10fda30ab0371e5,
title = "The short form of the recombinant CAL-A-type lipase UM03410 from the smut fungus Ustilago maydis exhibits an inherent trans-fatty acid selectivity",
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.",
keywords = "Enzyme Stability, Fungal Proteins, Hydrogen-Ion Concentration, Kinetics, Lipase, Plant Diseases, Protein Structure, Tertiary, Substrate Specificity, Trans Fatty Acids, Ustilago, Journal Article",
author = "Henrike Brundiek and Stefan Sa{\ss} and Andrew Evitt and Robert Kourist and Bornscheuer, {Uwe T}",
note = "Brundiek, Henrike Sass, Stefan Evitt, Andrew Kourist, Robert Bornscheuer, Uwe T Germany Applied microbiology and biotechnology Appl Microbiol Biotechnol. 2012 Apr;94(1):141-50. Epub 2012 Feb 1.",
year = "2012",
month = "4",
doi = "10.1007/s00253-012-3903-9",
language = "English",
volume = "94",
pages = "141--50",
journal = "Applied Microbiology and Biotechnology",
issn = "0175-7598",
publisher = "Springer Verlag",
number = "1",

}

TY - JOUR

T1 - The short form of the recombinant CAL-A-type lipase UM03410 from the smut fungus Ustilago maydis exhibits an inherent trans-fatty acid selectivity

AU - Brundiek,Henrike

AU - Saß,Stefan

AU - Evitt,Andrew

AU - Kourist,Robert

AU - Bornscheuer,Uwe T

N1 - Brundiek, Henrike Sass, Stefan Evitt, Andrew Kourist, Robert Bornscheuer, Uwe T Germany Applied microbiology and biotechnology Appl Microbiol Biotechnol. 2012 Apr;94(1):141-50. Epub 2012 Feb 1.

PY - 2012/4

Y1 - 2012/4

N2 - 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.

AB - 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.

KW - Enzyme Stability

KW - Fungal Proteins

KW - Hydrogen-Ion Concentration

KW - Kinetics

KW - Lipase

KW - Plant Diseases

KW - Protein Structure, Tertiary

KW - Substrate Specificity

KW - Trans Fatty Acids

KW - Ustilago

KW - Journal Article

U2 - 10.1007/s00253-012-3903-9

DO - 10.1007/s00253-012-3903-9

M3 - Article

VL - 94

SP - 141

EP - 150

JO - Applied Microbiology and Biotechnology

T2 - Applied Microbiology and Biotechnology

JF - Applied Microbiology and Biotechnology

SN - 0175-7598

IS - 1

ER -