公开(公告)号：EP3112458A1

公开(公告)日：2017-01-04

申请号：EP15174342.4

申请日：2015-06-29

申请人： Johann Wolfgang Goethe-Universität Frankfurt am Main

发明人： Grininger, Martin ,  Gajewski, Jan ,  Boles, Eckard

IPC分类号： C12N9/10 ,  C07K14/34 ,  C07K14/345

CPC分类号： C12N9/10 ,  C12N9/1029 ,  C12P7/64 ,  C12P7/6454

摘要： 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/or ketoacyl synthase domain (KS domain), and at least one amino acid substitution in the MPT domain at a position corresponding to R90, in the AT domain at a position corresponding to I151, 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 (C 6 to C 12 ), 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 production of short fatty acids (C 6 to C 12 ), the production of CoA esters of short fatty acids (C 6 to C 12 ), the production of ethyl esters of short fatty acids (C 6 to C 12 ), the production of esters of short fatty acids (C 6 to C 12 ) with other metabolites , the production of enzyme bound short fatty acids (C 6 to C 12 ), the production of biofuels, fine chemicals and/or flavoring substances.

摘要翻译： 本发明涉及脂肪酸合成中涉及的蛋白质，例如包含一个或多个多肽链的脂肪酸合成酶（FAS）变体，其中所述多肽链包含一个或多个包含丙二酰/棕榈酰转移酶结构域（MPT）的亚单位 结构域），乙酰转移酶结构域（AT结构域）和/或酮酸酰基合成酶结构域（KS结构域），以及MPT结构域中对应于R90的位置的至少一个氨基酸取代，在AT结构域中对应于I151 ，和/或在KS结构域，优选在酰基结合通道中，调节酰基中间体的亲和力，以及任选地进一步的氨基酸取代。 本发明涉及相应的多肽结构域。 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。 本发明还涉及生产短脂肪酸的方法，短脂肪酸的辅酶A，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（C 6 至C 12），包括所述核酸分子的表达，优选在所述宿主细胞中。 本发明还涉及生物燃料，调味化合物和/或精细化学品的生产方法，其包括优选在所述宿主细胞中表达所述核酸分子。 本发明还涉及蛋白质，核酸分子或宿主细胞用于生产短脂肪酸（C 6至C 12），短脂肪酸（C 6至C 12）的CoA酯的生产， 生产短脂肪酸乙酯（C 6〜C 12），生产短脂肪酸（C 6〜C 12）与其他代谢产物，生成酶结合的短脂肪酸（C 6〜C 12 ），生产生物燃料，精细化学品和/或调味物质。

公开(公告)号：EP3075848A1

公开(公告)日：2016-10-05

申请号：EP15162192.7

申请日：2015-04-01

申请人： Johann Wolfgang Goethe-Universität Frankfurt am Main

发明人： Grininger, Martin ,  Gajewski, Jan ,  Boles, Eckard ,  Pavlovic, Renata

IPC分类号： C12N9/10 ,  C12N15/82

CPC分类号： C12N9/1029 ,  C12N15/82 ,  C12P7/04 ,  C12P7/6409 ,  C12P7/6436 ,  C12P7/649 ,  Y02E50/13

摘要： 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 (C 6 to C 12 ), 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 (C 6 to C 12 ), the specific production of C 6 fatty acids and/or C 8 fatty acids, the bulk production of CoA esters of short fatty acids (C 6 to C 12 ), the specific production of C 6 -CoA esters and/or C 8 -CoA esters, the bulk production of ethyl esters of short fatty acids (C 6 to C 12 ), the specific production of C 6 fatty acid ethyl esters and/or C 8 fatty acid ethyl esters, the bulk production of esters of short fatty acids (C 6 to C 12 ) with other metabolites, the specific production of C 6 fatty acid esters with other metabolites and/or C 8 fatty acid esters with other metabolites, the bulk production of enzyme bound short fatty acids (C 6 to C 12 ), the specific production of enzyme bound C 6 fatty acids and/or enzyme bound C 8 fatty acids, the production of biofuels, fine chemicals and/or flavoring substances.

摘要翻译： 本发明涉及脂肪酸合成中涉及的蛋白质，例如包含一个或多个多肽链的脂肪酸合成酶（FAS）变体，其中所述多肽链包含一个或多个包含丙二酰/棕榈酰转移酶结构域（MPT）的亚单位 结构域），乙酰转移酶结构域（AT结构域）和酮酰基合成酶结构域（KS结构域），以及在对应于I306的位置的AT结构域中，在对应于R130的位置的MPT结构域中至少一个氨基酸取代，以及 /或在KS结构域中，优选在酰基结合通道中，以调节酰基中间体的亲和力，以及任选地进一步的氨基酸取代。 本发明涉及相应的多肽结构域。 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。 本发明还涉及生产短脂肪酸的方法，短脂肪酸的辅酶A，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（C 6 至C 12），包括所述核酸分子的表达，优选在所述宿主细胞中。 本发明还涉及生物燃料，调味化合物和/或精细化学品的生产方法，其包括优选在所述宿主细胞中表达所述核酸分子。 本发明还涉及蛋白质，核酸分子或宿主细胞用于大量生产短脂肪酸（C 6至C 12），C 6脂肪酸和/或C 8脂肪酸的特异性产生， 大量生产短脂肪酸（C 6至C 12）的CoA酯，C 6 -CoA酯和/或C 8 -CoA酯的特异性生产，大量生产短脂肪酸的乙酯（C 6至 C 12），具体生产C 6脂肪酸乙酯和/或C 8脂肪酸乙酯，大量生产短脂肪酸酯（C 6〜C 12）与其他代谢产物，具体生产C 6 脂肪酸酯与其他代谢物和/或C 8脂肪酸酯与其他代谢物，大量生产酶结合的短脂肪酸（C 6至C 12），酶结合的C 6脂肪酸和/或酶结合的特异性产生 C 8脂肪酸，生物燃料，精细化学品和/或调味物质的生产。