Cloning and characterization of a new delta-specific L-leucine dioxygenase from Anabaena variabilis

Raquel S. Correia Cordeiro, Junichi Enoki, Florian Busch, Carolin Mügge, Robert Kourist

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Optically pure hydroxy amino acids show several bioactivities and are valuable building blocks for the pharmaceutical industry. Fe(II)/α-ketoglutarate dependent dioxygenases catalyze the hydroxylation or sulfoxidation of L-amino acids with high regio- and stereoselectivity. While several β- and γ-specific enzymes have been described, only one δ-specific hydroxylase has been reported so far. Based on its similarity to the known L-leucine 5-hydroxylase from Nostoc punctiforme, an open reading frame from the cyanobacterium Anabaena variabilis was identified as putative L-leucine dioxygenase (AvLDO). Here we report the cloning and characterization of this dioxygenase. The enzyme showed a high preference for acidic conditions and moderate reaction temperatures. AvLDO catalyzed the regio- and stereoselective hydroxylation of several aliphatic amino acids in δ-position. In case of the sulfoxidation of L-methionine, AvLDO produced the opposite diastereomer than isoleucine dioxygenase. AvLDO is thus an interesting addition to the toolbox of Fe(II)/α-ketoglutarate dependent dioxygenases. On the genomic DNA of Anabaena variabilis ATCC 29413, the avldo gene is located on a gene cluster involved (2S,4S)-4-methylproline biosynthesis, which is contained in bioactive peptides often found from cyanobacteria. This fact suggests the metabolic functional role of this amino acid dioxygenase in cyanobacteria.

LanguageEnglish
Pages68-74
Number of pages7
JournalJournal of Biotechnology
Volume284
DOIs
StatusPublished - 20 Oct 2018

Fingerprint

Anabaena variabilis
Dioxygenases
Cloning
Leucine
Amino acids
Organism Cloning
Hydroxylation
Cyanobacteria
Enzymes
Genes
Amino Acids
Stereoselectivity
Regioselectivity
Mixed Function Oxygenases
Biosynthesis
Bioactivity
Drug products
Peptides
Nostoc
DNA

Keywords

  • Asymmetric oxidation
  • Biocatalysis
  • Cyanobacteria
  • Hydroxyamino acids
  • L-leucine 5-dioxygenase

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

Cloning and characterization of a new delta-specific L-leucine dioxygenase from Anabaena variabilis. / Correia Cordeiro, Raquel S.; Enoki, Junichi; Busch, Florian; Mügge, Carolin; Kourist, Robert.

In: Journal of Biotechnology, Vol. 284, 20.10.2018, p. 68-74.

Research output: Contribution to journalArticleResearchpeer-review

Correia Cordeiro, Raquel S. ; Enoki, Junichi ; Busch, Florian ; Mügge, Carolin ; Kourist, Robert. / Cloning and characterization of a new delta-specific L-leucine dioxygenase from Anabaena variabilis. In: Journal of Biotechnology. 2018 ; Vol. 284. pp. 68-74
@article{734f4fa658a34510b9c2d3edf3fac656,
title = "Cloning and characterization of a new delta-specific L-leucine dioxygenase from Anabaena variabilis",
abstract = "Optically pure hydroxy amino acids show several bioactivities and are valuable building blocks for the pharmaceutical industry. Fe(II)/α-ketoglutarate dependent dioxygenases catalyze the hydroxylation or sulfoxidation of L-amino acids with high regio- and stereoselectivity. While several β- and γ-specific enzymes have been described, only one δ-specific hydroxylase has been reported so far. Based on its similarity to the known L-leucine 5-hydroxylase from Nostoc punctiforme, an open reading frame from the cyanobacterium Anabaena variabilis was identified as putative L-leucine dioxygenase (AvLDO). Here we report the cloning and characterization of this dioxygenase. The enzyme showed a high preference for acidic conditions and moderate reaction temperatures. AvLDO catalyzed the regio- and stereoselective hydroxylation of several aliphatic amino acids in δ-position. In case of the sulfoxidation of L-methionine, AvLDO produced the opposite diastereomer than isoleucine dioxygenase. AvLDO is thus an interesting addition to the toolbox of Fe(II)/α-ketoglutarate dependent dioxygenases. On the genomic DNA of Anabaena variabilis ATCC 29413, the avldo gene is located on a gene cluster involved (2S,4S)-4-methylproline biosynthesis, which is contained in bioactive peptides often found from cyanobacteria. This fact suggests the metabolic functional role of this amino acid dioxygenase in cyanobacteria.",
keywords = "Asymmetric oxidation, Biocatalysis, Cyanobacteria, Hydroxyamino acids, L-leucine 5-dioxygenase",
author = "{Correia Cordeiro}, {Raquel S.} and Junichi Enoki and Florian Busch and Carolin M{\"u}gge and Robert Kourist",
year = "2018",
month = "10",
day = "20",
doi = "10.1016/j.jbiotec.2018.07.038",
language = "English",
volume = "284",
pages = "68--74",
journal = "Journal of Biotechnology",
issn = "0168-1656",
publisher = "Elsevier B.V.",

}

TY - JOUR

T1 - Cloning and characterization of a new delta-specific L-leucine dioxygenase from Anabaena variabilis

AU - Correia Cordeiro,Raquel S.

AU - Enoki,Junichi

AU - Busch,Florian

AU - Mügge,Carolin

AU - Kourist,Robert

PY - 2018/10/20

Y1 - 2018/10/20

N2 - Optically pure hydroxy amino acids show several bioactivities and are valuable building blocks for the pharmaceutical industry. Fe(II)/α-ketoglutarate dependent dioxygenases catalyze the hydroxylation or sulfoxidation of L-amino acids with high regio- and stereoselectivity. While several β- and γ-specific enzymes have been described, only one δ-specific hydroxylase has been reported so far. Based on its similarity to the known L-leucine 5-hydroxylase from Nostoc punctiforme, an open reading frame from the cyanobacterium Anabaena variabilis was identified as putative L-leucine dioxygenase (AvLDO). Here we report the cloning and characterization of this dioxygenase. The enzyme showed a high preference for acidic conditions and moderate reaction temperatures. AvLDO catalyzed the regio- and stereoselective hydroxylation of several aliphatic amino acids in δ-position. In case of the sulfoxidation of L-methionine, AvLDO produced the opposite diastereomer than isoleucine dioxygenase. AvLDO is thus an interesting addition to the toolbox of Fe(II)/α-ketoglutarate dependent dioxygenases. On the genomic DNA of Anabaena variabilis ATCC 29413, the avldo gene is located on a gene cluster involved (2S,4S)-4-methylproline biosynthesis, which is contained in bioactive peptides often found from cyanobacteria. This fact suggests the metabolic functional role of this amino acid dioxygenase in cyanobacteria.

AB - Optically pure hydroxy amino acids show several bioactivities and are valuable building blocks for the pharmaceutical industry. Fe(II)/α-ketoglutarate dependent dioxygenases catalyze the hydroxylation or sulfoxidation of L-amino acids with high regio- and stereoselectivity. While several β- and γ-specific enzymes have been described, only one δ-specific hydroxylase has been reported so far. Based on its similarity to the known L-leucine 5-hydroxylase from Nostoc punctiforme, an open reading frame from the cyanobacterium Anabaena variabilis was identified as putative L-leucine dioxygenase (AvLDO). Here we report the cloning and characterization of this dioxygenase. The enzyme showed a high preference for acidic conditions and moderate reaction temperatures. AvLDO catalyzed the regio- and stereoselective hydroxylation of several aliphatic amino acids in δ-position. In case of the sulfoxidation of L-methionine, AvLDO produced the opposite diastereomer than isoleucine dioxygenase. AvLDO is thus an interesting addition to the toolbox of Fe(II)/α-ketoglutarate dependent dioxygenases. On the genomic DNA of Anabaena variabilis ATCC 29413, the avldo gene is located on a gene cluster involved (2S,4S)-4-methylproline biosynthesis, which is contained in bioactive peptides often found from cyanobacteria. This fact suggests the metabolic functional role of this amino acid dioxygenase in cyanobacteria.

KW - Asymmetric oxidation

KW - Biocatalysis

KW - Cyanobacteria

KW - Hydroxyamino acids

KW - L-leucine 5-dioxygenase

UR - http://www.scopus.com/inward/record.url?scp=85052147882&partnerID=8YFLogxK

U2 - 10.1016/j.jbiotec.2018.07.038

DO - 10.1016/j.jbiotec.2018.07.038

M3 - Article

VL - 284

SP - 68

EP - 74

JO - Journal of Biotechnology

T2 - Journal of Biotechnology

JF - Journal of Biotechnology

SN - 0168-1656

ER -