caa a22 4500 606212663 CHVBK 20210128101043.0 cr unu---uuuuu 210128e20151001xx s 000 0 eng 10.1007/s10295-015-1663-0 doi (NATIONALLICENCE)springer-10.1007/s10295-015-1663-0 In silico model-driven cofactor engineering strategies for improving the overall NADP(H) turnover in microbial cell factories [Elektronische Daten] [Meiyappan Lakshmanan, Kai Yu, Lokanand Koduru, Dong-Yup Lee] Optimizing the overall NADPH turnover is one of the key challenges in various value-added biochemical syntheses. In this work, we first analyzed the NADPH regeneration potentials of common cell factories, including Escherichia coli, Saccharomyces cerevisiae, Bacillus subtilis, and Pichia pastoris across multiple environmental conditions and determined E. coli and glycerol as the best microbial chassis and most suitable carbon source, respectively. In addition, we identified optimal cofactor specificity engineering (CSE) enzyme targets, whose cofactors when switched from NAD(H) to NADP(H) improve the overall NADP(H) turnover. Among several enzyme targets, glyceraldehyde-3-phosphate dehydrogenase was recognized as a global candidate since its CSE improved the NADP(H) regeneration under most of the conditions examined. Finally, by analyzing the protein structures of all CSE enzyme targets via homology modeling, we established that the replacement of conserved glutamate or aspartate with serine in the loop region could change the cofactor dependence from NAD(H) to NADP(H). Society for Industrial Microbiology and Biotechnology, 2015 NADPH nationallicence Metabolic engineering nationallicence Cofactor specificity engineering (CSE) nationallicence Flux balance analysis (FBA) nationallicence Cofactor modification analysis (CMA) nationallicence Lakshmanan Meiyappan Bioprocessing Technology Institute, Agency for Science, Technology and Research (A*STAR), 20 Biopolis Way, #06-01, Centros, 138668, Singapore, Singapore aut Yu Kai Department of Chemical and Biomolecular Engineering, National University of Singapore, 4 Engineering Drive 4, 117585, Singapore, Singapore aut Koduru Lokanand Department of Chemical and Biomolecular Engineering, National University of Singapore, 4 Engineering Drive 4, 117585, Singapore, Singapore aut Lee Dong-Yup Bioprocessing Technology Institute, Agency for Science, Technology and Research (A*STAR), 20 Biopolis Way, #06-01, Centros, 138668, Singapore, Singapore aut Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/10(2015-10-01), 1401-1414 1367-5435 42:10<1401 2015 42 10295 https://doi.org/10.1007/s10295-015-1663-0 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s10295-015-1663-0 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Lakshmanan Meiyappan Bioprocessing Technology Institute, Agency for Science, Technology and Research (A*STAR), 20 Biopolis Way, #06-01, Centros, 138668, Singapore, Singapore aut NATIONALLICENCE 700 1- Yu Kai Department of Chemical and Biomolecular Engineering, National University of Singapore, 4 Engineering Drive 4, 117585, Singapore, Singapore aut NATIONALLICENCE 700 1- Koduru Lokanand Department of Chemical and Biomolecular Engineering, National University of Singapore, 4 Engineering Drive 4, 117585, Singapore, Singapore aut NATIONALLICENCE 700 1- Lee Dong-Yup Bioprocessing Technology Institute, Agency for Science, Technology and Research (A*STAR), 20 Biopolis Way, #06-01, Centros, 138668, Singapore, Singapore aut NATIONALLICENCE 773 0- Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/10(2015-10-01), 1401-1414 1367-5435 42:10<1401 2015 42 10295