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Four Amino Acid Residues Influence the Substrate Chain‐Length and Regioselectivity of Siganus canaliculatus Δ4 and Δ5/6 Desaturases
Author(s) -
Lim Ze Long,
Senger Toralf,
Vrinten Patricia
Publication year - 2014
Publication title -
lipids
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.601
H-Index - 120
eISSN - 1558-9307
pISSN - 0024-4201
DOI - 10.1007/s11745-014-3880-0
Subject(s) - biochemistry , amino acid , enzyme , heterologous expression , biology , substrate (aquarium) , fatty acid desaturase , fatty acid , linoleic acid , stereochemistry , polyunsaturated fatty acid , chemistry , gene , recombinant dna , ecology
Although ω3‐ and ω6‐ desaturases have been well studied in terms of substrate preference and regiospecificity, relatively little is known about the membrane‐bound, “front‐end” long chain fatty acid desaturases, such as ∆4, Δ5 or Δ6 desaturases. The first vertebrate ∆4 desaturase was recently identified in the marine teleost fish Siganus canaliculatus ( S. canaliculatus ), which also possesses a bifunctional Δ5/6 desaturase. These two long chain polyunsaturated fatty acid desaturases are very different in terms of regiospecificity and substrate chain‐length, but share an unusually high degree of amino acid identity (83 %). We took advantage of this similarity by constructing a series of chimeric enzymes, replacing regions of one enzyme with the corresponding sequence of the other. Heterologous expression of the chimeric series of enzymes in yeast indicated that the substitution of a four amino acid region was sufficient to convert a ∆4 desaturase to an enzyme with ∆6 desaturase activity, and convert a ∆5/6 desaturase to an enzyme with a low level of ∆4 desaturase activity. In addition, enzymes having both ∆4 and ∆6 desaturase activities were produced by single or double amino acid substitutions within this four‐amino acid region.