申请人:Johann Wolfgang Goethe-Universität
Frankfurt am Main
公开号:EP3075848A1
公开(公告)日:2016-10-05
The present invention relates to proteins involved in fatty acid synthesis, such as fatty acid synthases (FAS) variants, comprising one or more polypeptide chains, wherein said polypeptide chain(s) comprise one or more subunits comprising a malonyl/palmitoyl transferase domain (MPT domain), acetyl transferase domain (AT domain), and ketoacyl synthase domain (KS domain), and at least one amino acid substitution in the MPT domain at a position corresponding to R130, in the AT domain at a position corresponding to I306, and/or in the KS domain, preferably in the acyl binding channel, to modulate affinities of acyl intermediates, and optionally further amino acid substitution(s). The present invention relates to the respective polypeptide domains.
The present invention further relates to nucleic acid molecules encoding the proteins (or the polypeptide domains) and to host cells containing said nucleic acid molecules. The present invention further relates to a method for the production of short fatty acids, CoA esters of short fatty acids, ethyl esters of short fatty acids, esters of short fatty acids with other metabolites, and/or enzyme bound short fatty acids (C6 to C12), comprising the expression of said nucleic acid molecules, preferably in said host cells. The present invention further relates to a method for the production of biofuels, flavoring compounds and/or fine chemicals, comprising the expression of said nucleic acid molecules, preferably in said host cells. The present invention also relates to the use of the proteins, nucleic acids molecule or host cells for the bulk production of short fatty acids (C6 to C12), the specific production of C6 fatty acids and/or C8 fatty acids, the bulk production of CoA esters of short fatty acids (C6 to C12), the specific production of C6-CoA esters and/or C8-CoA esters, the bulk production of ethyl esters of short fatty acids (C6 to C12), the specific production of C6 fatty acid ethyl esters and/or C8 fatty acid ethyl esters, the bulk production of esters of short fatty acids (C6 to C12) with other metabolites, the specific production of C6 fatty acid esters with other metabolites and/or C8 fatty acid esters with other metabolites, the bulk production of enzyme bound short fatty acids (C6 to C12), the specific production of enzyme bound C6 fatty acids and/or enzyme bound C8 fatty acids, the production of biofuels, fine chemicals and/or flavoring substances.
本发明涉及参与脂肪酸合成的蛋白质,如脂肪酸合酶(FAS)变体,其包括一条或多条多肽链,其中所述多肽链包括一个或多个亚基,这些亚基包括丙二酰/棕榈酰转移酶结构域(MPT结构域)、乙酰转移酶结构域(AT结构域)和酮酰合成酶结构域(KS结构域)、在丙二酰/棕榈酰转移酶结构域(MPT 结构域)、乙酰转移酶结构域(AT 结构域)和酮酰合成酶结构域(KS 结构域)的一个或多个亚基,以及在丙二酰/棕榈酰转移酶结构域中对应于 R130 的位置、在乙酰转移酶结构域中对应于 I306 的位置和/或在 KS 结构域中,优选在酰基结合通道中的至少一个氨基酸取代,以调节酰基中间体的亲和性,以及可选的进一步氨基酸取代。本发明涉及各自的多肽结构域。
本发明还涉及编码蛋白质(或多肽结构域)的核酸分子以及含有所述核酸分子的宿主细胞。本发明进一步涉及一种生产短脂肪酸、短脂肪酸的 CoA 酯、短脂肪酸的乙酯、短脂肪酸与其它代谢物的酯和/或酶结合的短脂肪酸(C6 至 C12)的方法,包括优选在所述宿主细胞中表达所述核酸分子。本发明还涉及一种生产生物燃料、调味化合物和/或精细化学品的方法,包括所述核酸分子的表达,最好在所述宿主细胞中表达。本发明还涉及使用所述蛋白质、核酸分子或宿主细胞批量生产短脂肪酸(C6 至 C12)、特定生产 C6 脂肪酸和/或 C8 脂肪酸、大量生产短脂肪酸(C6 至 C12)的 CoA 酯,特定生产 C6-CoA 酯和/或 C8-CoA 酯,大量生产短脂肪酸(C6 至 C12)的乙酯,特定生产 C6 脂肪酸乙酯和/或 C8 脂肪酸乙酯、大量生产短脂肪酸(C6 至 C12)与其他代谢物的酯,特定生产 C6 脂肪酸酯与其他代谢物和/或 C8 脂肪酸酯与其他代谢物的酯,大量生产酶结合短脂肪酸(C6 至 C12),特定生产酶结合 C6 脂肪酸和/或酶结合 C8 脂肪酸,生产生物燃料、精细化学品和/或调味物质。