Genetic engineering approaches for the fermentative production of phenylglycines

• Published in: Applied microbiology and biotechnology 2020-04, Vol.104 (8), p.3433-3444
• Main Authors: Moosmann, David, Mokeev, Vladislav, Kulik, Andreas, Osipenkov, Natalie, Kocadinc, Susann, Ort-Winklbauer, Regina, Handel, Franziska, Hennrich, Oliver, Youn, Jung-Won, Sprenger, Georg A., Mast, Yvonne
• Format: Article
• Language: English
• Subjects: Actinomycetes; Amides; Amino acids; Anti-Bacterial Agents - biosynthesis; Antibiotics; Applied Genetics and Molecular Biotechnology; Beta lactamases; Biomedical and Life Sciences; Biotechnology; Chemical industry; Chromosomes; Fermentation; Fine chemicals; Genes, Bacterial; Genetic engineering; Genetic Engineering - methods; Genetically modified organisms; Glycine - analogs & derivatives; Glycine - biosynthesis; Life Sciences; Mezlocillin; Microbial Genetics and Genomics; Microbiology; Operon; Optimization; Phenylglycine; Pivampicillin; Pristinamycin; Pseudomonas putida; Pseudomonas putida - enzymology; Pseudomonas putida - genetics; Stereoisomerism; Streptomyces - genetics; Streptomyces lividans - genetics; Synthetic Biology - methods; Virginiamycin; β-Lactam antibiotics
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-020-10447-9

L-phenylglycine (L-Phg) is a rare non-proteinogenic amino acid, which only occurs in some natural compounds, such as the streptogramin antibiotics pristinamycin I and virginiamycin S or the bicyclic peptide antibiotic dityromycin. Industrially, more interesting than L-Phg is the enantiomeric D-Phg as it plays an important role in the fine chemical industry, where it is used as a precursor for the production of semisynthetic β-lactam antibiotics. Based on the natural L-Phg operon from Streptomyces pristinaespiralis and the stereo-inverting aminotransferase gene hpgAT from Pseudomonas putida , an artificial D-Phg operon was constructed. The natural L-Phg operon, as well as the artificial D-Phg operon, was heterologously expressed in different actinomycetal host strains, which led to the successful production of Phg. By rational genetic engineering of the optimal producer strains S. pristinaespiralis and Streptomyces lividans , Phg production could be improved significantly. Here, we report on the development of a synthetic biology-derived D-Phg pathway and the optimization of fermentative Phg production in actinomycetes by genetic engineering approaches. Our data illustrate a promising alternative for the production of Phgs.