[Purification and production of the extracellular 5-aminolevulinate from recombiniant Escherichia coli expressing yeast ALAS].
Aminolevulinic acid (ALA) is biosynthesized by the enzyme ALA synthase (ALAS). The ALA production has been studied using the overproducing ALAS from several bacteria in Escherchia coil, respectively. However, ALAS from eucaryote expressed in E. coli for producing ALA in the culture is not known. The extracellular ALA production and cell growth were investageted respectively using the recombinant E. coli overproducing Saccharomyces cerevisiae ALAS in shake-flask fermentations. The ALAS activity from the cell extract was assayed. The extracellular ALA was purified by the national-made large-dimension resins and confirmed by the capillary electrophoresis measurements. At 12h after induction at 37 degrees C, the extracellular ALA production was up to 162mg per liter LB culture at initial pH 6.5 with exogenous levulinate, succinate and and glycine at the concentration of 20 mmol/L respectively. The purity of ALA after purification is up to 90%.
He
Xiao-Mei
XM
School of Life Sciences, Anhui Agricultural University, Hefei 230036, China.
Zhou
Jing
J
Cheng
Ying
Y
Fan
Jun
J
D000624:5-Aminolevulinate Synthetase / Q000235:genetics / Q000378:metabolism*; D000622:Aminolevulinic Acid / Q000302:isolation & purification / Q000378:metabolism*; D002455:Cell Division / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D019075:Electrophoresis, Capillary; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D005110:Extracellular Space / Q000187:drug effects / Q000378:metabolism; D005998:Glycine / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007982:Levulinic Acids / Q000494:pharmacology; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D019802:Succinic Acid / Q000494:pharmacology
D007982:Levulinic Acids; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D000622:Aminolevulinic Acid; D019802:Succinic Acid; D000624:5-Aminolevulinate Synthetase; C032246:levulinic acid; D005998:Glycine