RT Journal Article SR Electronic(1) A1 Romero, Mauricio A1 Guzmán-León, Simón A1 Aranda, Cristina A1 González-Halphen, Diego A1 Valenzuela, Lourdes A1 González, Alicia YR 2000 T1 Pathways for glutamate biosynthesis in the yeast Kluyveromyces lactis JF Microbiology, VO 146 IS 1 SP 239 OP 245 DO https://doi.org/10.1099/00221287-146-1-239 PB Microbiology Society, SN 1465-2080, AB Purified glutamate synthase (GOGAT) from Kluyveromyces lactis was characterized as a high-molecular-mass polypeptide, a distinction shared with previously described GOGATs from other eukaryotic micro-organisms. Using degenerate deoxyoligonucleotides, designed from conserved regions of the alfalfa, maize and Escherichia coli GOGAT genes, a 300 bp PCR fragment from the K. lactis GOGAT gene KlGLT1 was obtained. This fragment was used to construct null GOGAT mutants of K. lactis by gene replacement. These mutants showed no growth defect phenotype and were able to grow on ammonium as sole nitrogen source. Double mutants obtained from a cross between a previously described KlGDH1 mutant and the K. lactis null GOGAT strain were full glutamate auxotrophs. These results indicate that glutamate biosynthesis in K. lactis is afforded through the combined action of KlGDH1 and KlGLT1 products., UL https://www.microbiologyresearch.org/content/journal/micro/10.1099/00221287-146-1-239