Modified membrane permeability
View Patent ↗Provided are microorganisms genetically modified to overexpress porin polypeptides to enhance the production of lysine and lysine derivatives by the microorganism. Also provided are methods of generating such microorganism, and methods of producing lysine and lysine derivatives using the genetically modified microorganisms.
1. A genetically modified host cell comprising a heterologous nucleic acid encoding an OMP porin polypeptide, wherein the host cell overexpresses the OMP porin polypeptide and has increased production of an amino acid lysine and cadaverine, and wherein the OMP porin polypeptide is an OmpA, OmpC, OmpF, OmpX, OmpE, OmpG, or OmpW porin polypeptide set forth as SEQ ID NO: 4, 6, 8, 10, 12, 14, or 16 respectively; or wherein the OMP porin polypeptide is a conservatively modified porin polypeptide variant of SEQ ID NO: 4, 6, 8, 10, 12, 14, or 16 that encodes the mature OMP porin polypeptide.
2. The genetically modified host cell of claim 1 , wherein the heterologous nucleic acid encoding the OMP porin polypeptide is encoded by an expression vector introduced into the cell, wherein the expression vector comprises the heterologous nucleic acid operably linked to a promoter; and/or
wherein the OMP porin polypeptide is endogenous to the host cell; and/or
wherein the heterologous nucleic acid is integrated into the host chromosome; and/or
wherein the host cell overexpresses a lysine decarboxylase; and/or
wherein the host cell overexpresses one or more lysine biosynthesis polypeptides; and/or
wherein the host cell overexpresses a TetA polypeptide.
3. The genetically modified host cell of claim 1 , wherein the host cell is of the genus Escherichia, Hafnia , or Corynebacterium ; and/or wherein the host cell is Escherichia coli, Hafnia alvei , or Corynebacterium glutamicum ; and/or wherein the host cell is Escherichia coli or Hafnia alvei.
4. The genetically modified host cell of claim 3 , wherein the Omp porin polypeptide is an OmpA, OmpC, OmpF, or OmpW polypeptide.
5. The genetically modified host cell of claim 4 , wherein the host cell overexpresses a lysine decarboxylase polypeptide; and/or wherein the host cell overexpresses a LysC, DapA, LysA, Asd, DapB, AspC, or TetA polypeptide.
6. A method of producing an amino acid, the method comprising culturing a host cell of claim 1 under conditions in which the OMP porin polypeptide is overexpressed.