Directed evolution of the CpcA biosynthetic pathway and optimization of conditions for CpcA production and its properties

• Published in: Applied microbiology and biotechnology Vol. 98; no. 11; pp. 4995 - 5007
• Main Authors: Dong, Dalu, Pan, Hangtao, Yu, Ping
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01.06.2014; Springer; Springer Nature B.V
• Subjects: Amino acids; Analysis; Antioxidants; Bacterial proteins; beef extracts; Biomedical and Life Sciences; Biosynthesis; Biosynthetic Pathways; Biotechnologically Relevant Enzymes and Proteins; Biotechnology; carbon; Carbon - metabolism; Chemical engineering; Cloning; Culture Media - chemistry; Directed Molecular Evolution; E coli; Enzymes; Escherichia coli; evolution; Free radicals; Free Radicals - metabolism; Gene amplification; genes; Genetic aspects; Glycerol; Hydrogen-Ion Concentration; Industrial production; inhibitory concentration 50; Life Sciences; Metabolic Engineering; Microbial Genetics and Genomics; Microbiology; molecular weight; mutants; Mutation, Missense; Nickel; nitrogen; Nitrogen sources; Optimization; oxygen; Phycocyanin - biosynthesis; Phycocyanin - chemistry; Phycocyanin - genetics; Phycocyanin - isolation & purification; Point Mutation; polyacrylamide gel electrophoresis; polymerase chain reaction; Polypeptides; Properties; Protein Stability; response surface methodology; Sequence Alignment; sodium dodecyl sulfate; Spectrum Analysis; Spirulina - genetics; Spirulina - metabolism; Spirulina maxima; Studies; Sulfates; Temperature; Wavelengths; Yeast; yeast extract; China; Antioxidative activity; Directed evolution; Stability; CpcA; Error-prone PCR; Response surface methodology; Pathway engineering; Spectrum
• ISSN: 0175-7598; 1432-0614; 1432-0614
• DOI: 10.1007/s00253-014-5522-0

To improve the production of phycocyanin holo-α-subunit (CpcA) from Spirulina maxima , five genes and their spacer region sequences involved in its biosynthesis were subject to the directed evolution by error-prone PCR using the plasmid pETDuet-6 as the template. Mutants were screened by determining...