caa a22 4500 606212035 CHVBK 20210128101041.0 cr unu---uuuuu 210128e20150601xx s 000 0 eng 10.1007/s10295-015-1613-x doi (NATIONALLICENCE)springer-10.1007/s10295-015-1613-x Investigation into the misincorporation of norleucine into a recombinant protein vaccine candidate [Elektronische Daten] [Joyce Ni, Meg Gao, Andrew James, Jiansheng Yao, Tao Yuan, Bruce Carpick, Tony D'Amore, Patrick Farrell] A high level of norleucine misincorporation was detected in a recombinant methionine-rich protein vaccine candidate expressed in E. coli K12. An investigation was conducted to evaluate a simple remediation strategy to reduce norleucine misincorporation and to determine if the phenomenon was either (a) due to the depletion of methionine during fermentation, (b) a result of the cultivation environment, or (c) a strain-specific effect. While supplementation with exogenous methionine improved product quality, the undesirable biosynthesis of non-standard amino acids such as norleucine and norvaline persisted. In contrast, non-standard amino acid biosynthesis was quickly minimized upon selection of an appropriate fed-batch process control strategy, fermentation medium, and nutrient feed. By expressing the same protein in E. coli BL21(DE3), it was determined that the biosynthesis of norleucine and norvaline, and the misincorporation of norleucine into the protein were primarily attributed to the use of E. coli K12 as the host for protein expression. Society for Industrial Microbiology and Biotechnology, 2015 Norleucine (Nle) nationallicence Norvaline (Nva) nationallicence Misincorporation nationallicence Chemically defined medium (CDM) nationallicence Semi-defined medium (SDM) nationallicence Ni Joyce Bioprocess Research and Development NA, Sanofi Pasteur, 1755 Steeles Avenue West, M2R 3T4, Toronto, Canada aut Gao Meg Bioprocess Research and Development NA, Sanofi Pasteur, 1755 Steeles Avenue West, M2R 3T4, Toronto, Canada aut James Andrew Analytical Research and Development NA, Sanofi Pasteur, 1755 Steeles Avenue West, M2R 3T4, Toronto, Canada aut Yao Jiansheng Bioprocess Research and Development NA, Sanofi Pasteur, 1755 Steeles Avenue West, M2R 3T4, Toronto, Canada aut Yuan Tao Bioprocess Research and Development NA, Sanofi Pasteur, 1755 Steeles Avenue West, M2R 3T4, Toronto, Canada aut Carpick Bruce Analytical Research and Development NA, Sanofi Pasteur, 1755 Steeles Avenue West, M2R 3T4, Toronto, Canada aut D'Amore Tony Bioprocess Research and Development NA, Sanofi Pasteur, 1755 Steeles Avenue West, M2R 3T4, Toronto, Canada aut Farrell Patrick Bioprocess Research and Development NA, Sanofi Pasteur, 1755 Steeles Avenue West, M2R 3T4, Toronto, Canada aut Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/6(2015-06-01), 971-975 1367-5435 42:6<971 2015 42 10295 https://doi.org/10.1007/s10295-015-1613-x text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 brief-communication jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s10295-015-1613-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Ni Joyce Bioprocess Research and Development NA, Sanofi Pasteur, 1755 Steeles Avenue West, M2R 3T4, Toronto, Canada aut NATIONALLICENCE 700 1- Gao Meg Bioprocess Research and Development NA, Sanofi Pasteur, 1755 Steeles Avenue West, M2R 3T4, Toronto, Canada aut NATIONALLICENCE 700 1- James Andrew Analytical Research and Development NA, Sanofi Pasteur, 1755 Steeles Avenue West, M2R 3T4, Toronto, Canada aut NATIONALLICENCE 700 1- Yao Jiansheng Bioprocess Research and Development NA, Sanofi Pasteur, 1755 Steeles Avenue West, M2R 3T4, Toronto, Canada aut NATIONALLICENCE 700 1- Yuan Tao Bioprocess Research and Development NA, Sanofi Pasteur, 1755 Steeles Avenue West, M2R 3T4, Toronto, Canada aut NATIONALLICENCE 700 1- Carpick Bruce Analytical Research and Development NA, Sanofi Pasteur, 1755 Steeles Avenue West, M2R 3T4, Toronto, Canada aut NATIONALLICENCE 700 1- D'Amore Tony Bioprocess Research and Development NA, Sanofi Pasteur, 1755 Steeles Avenue West, M2R 3T4, Toronto, Canada aut NATIONALLICENCE 700 1- Farrell Patrick Bioprocess Research and Development NA, Sanofi Pasteur, 1755 Steeles Avenue West, M2R 3T4, Toronto, Canada aut NATIONALLICENCE 773 0- Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/6(2015-06-01), 971-975 1367-5435 42:6<971 2015 42 10295