From natural products discovery to commercialization: a success story

In order for a natural product to become a commercial reality, laboratory improvement of its production process is a necessity since titers produced by wild strains could never compete with the power of synthetic chemistry. Strain improvement by mutagenesis has been a major success. It has mainly been carried out by “brute force” screening or selection, but modern genetic technologies have entered the scene in recent years. For every new strain developed genetically, there is further opportunity to raise titers by medium modifications. Of major interest has been the nutritional control by induction, as well as inhibition and repression by sources of carbon, nitrogen, phosphate and end products. Both strain improvement and nutritional modification contribute to the new process, which is then scaled up by biochemical engineers into pilot scale and later into factory size fermentors.

[1]  V. Vinci,et al.  Improvement of microbial strains and fermentation processes , 2000, Applied Microbiology and Biotechnology.

[2]  M. Vrljic,et al.  A new type of transporter with a new type of cellular function: l‐lysine export from Corynebacterium glutamicum , 1996, Molecular microbiology.

[3]  Arnold L. Demain,et al.  The β-lactam antibiotics: past, present, and future , 2004, Antonie van Leeuwenhoek.

[4]  A. Demain.  Induction of microbial secondary metabolism. , 1998, International microbiology : the official journal of the Spanish Society for Microbiology.

[5]  A L Demain,et al.  Molecular genetics and industrial microbiology — 30 years of marriage , 2001, Journal of Industrial Microbiology and Biotechnology.

[6]  G Sermonti,et al.  Comutation in Streptomyces , 1973, Journal of bacteriology.

[7]  A L Demain,et al.  Small bugs, big business: the economic power of the microbe. , 2000, Biotechnology advances.

[8]  Sergio Sánchez,et al.  Metabolic regulation of fermentation processes , 2002 .

[9]  J. Barredo.  Microbial Processes and Products , 2005, Methods in Biotechnology.

[10]  M. Bibb,et al.  Engineering of Primary Carbon Metabolism for Improved Antibiotic Production in Streptomyces lividans , 2002, Applied and Environmental Microbiology.

[11]  E. Langley,et al.  Glucose repression of anthracycline formation in Streptomyces peucetius var. caesius , 1999, Applied Microbiology and Biotechnology.

[12]  Claes Gustafsson,et al.  Semi-synthetic DNA shuffling of aveC leads to improved industrial scale production of doramectin by Streptomyces avermitilis. , 2005, Metabolic engineering.

[13]  D J Newman,et al.  Natural products in drug discovery and development. , 1997, Journal of natural products.

[14]  K. VENKATASUBRAMANIAN,et al.  Genetic Engineering of Metabolic Pathways Applied to the Production of Phenylalanine , 1990, Annals of the New York Academy of Sciences.

[15]  J. Bérdy.  Bioactive Microbial Metabolites A Personal View , 2005 .

[16]  F. G. Jarvis,et al.  The role of the constituents of synthetic media for penicillin production. , 1947, Journal of the American Chemical Society.

[17]  H Umezawa,et al.  Low-molecular-weight enzyme inhibitors of microbial origin. , 1982, Annual review of microbiology.

[18]  K. Lewis,et al.  Isolating "Uncultivable" Microorganisms in Pure Culture in a Simulated Natural Environment , 2002, Science.

[19]  G. Turner,et al.  delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine synthetase is a rate limiting enzyme for penicillin production in Aspergillus nidulans. , 1996, Molecular & general genetics : MGG.

[20]  R. H. Baltz,et al.  Genes for the biosynthesis of spinosyns: applications for yield improvement in Saccharopolyspora spinosa , 2001, Journal of Industrial Microbiology and Biotechnology.

[21]  G. Turner,et al.  δ-(L-α-Aminoadipyl)-L-cysteinyl-D-valine synthetase is a rate limiting enzyme for penicillin production in Aspergillusnidulans , 1996, Molecular and General Genetics MGG.

[22]  Jens Nielsen,et al.  Metabolic engineering of beta-lactam production. , 2003, Metabolic engineering.

[23]  Albert A. de Graaf,et al.  Pathway Analysis and Metabolic Engineering in Corynebacterium glutamicum , 2000, Biological chemistry.

[24]  A. Demain.  Microbial biotechnology. , 2000, Trends in biotechnology.

[25]  A. Bull.  Microbial Diversity and Bioprospecting , 2003 .

[26]  M. J. Johnson,et al.  The effect of the carbohydrate nutrition on penicillin production by Penicillium chrysogenum Q-176. , 1953, Applied microbiology.

[27]  H. Ishizuka,et al.  Breeding of a mutant of Aureobasidium sp. with high erythritol production , 1989 .

[28]  K. Zengler,et al.  Cultivating the uncultured , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[29]  János Bérdy,et al.  Bioactive microbial metabolites. , 2005, The Journal of antibiotics.

[30]  A. Demain,et al.  Effects of Carboxymethylcellulose and Carboxypolymethylene on Morphology of Aspergillus fumigatus NRRL 2346 and Fumagillin Production , 2003, Current Microbiology.

[31]  Maria Papagianni,et al.  Fungal morphology and metabolite production in submerged mycelial processes. , 2004, Biotechnology advances.

[32]  J. Handelsman,et al.  Molecular biological access to the chemistry of unknown soil microbes: a new frontier for natural products. , 1998, Chemistry & biology.

[33]  A. Demain,et al.  Genetic improvement of processes yielding microbial products. , 2006, FEMS microbiology reviews.

[34]  A. Demain,et al.  Pharmaceutically active secondary metabolites of microorganisms , 1999, Applied Microbiology and Biotechnology.

[35]  T. Schmidt,et al.  New Strategies for Cultivation and Detection of Previously Uncultured Microbes , 2004, Applied and Environmental Microbiology.

[36]  C. Richard Hutchinson,et al.  The Streptomyces peucetius dpsY anddnrX Genes Govern Early and Late Steps of Daunorubicin and Doxorubicin Biosynthesis , 1998, Journal of bacteriology.

[37]  H. Kacser,et al.  A universal method for achieving increases in metabolite production. , 1993, European journal of biochemistry.

[38]  Jens Nielsen,et al.  Metabolic engineering of -lactam production , 2003 .

[39]  J. Gyimesi,et al.  Metabolites of gentamicin-producing Micromonospora species I. Isolation and identification of metabolites. , 1977, The Journal of antibiotics.

[40]  Arnold L. Demain,et al.  Manual of Industrial Microbiology and Biotechnology , 1986 .

[41]  A. Demain,et al.  Microbial Cells and Enzymes A Century of Progress , 2005 .

[42]  W. Stemmer,et al.  Genome shuffling leads to rapid phenotypic improvement in bacteria , 2002, Nature.

[43]  S. Parekh,et al.  Development of Improved Strains and Optimization of Fermentation Processes , 2005 .

[44]  J. Barredo.  Microbial Enzymes and Biotransformations , 2005, Methods in Biotechnology.

[45]  J. Nielsen,et al.  Effect of deletion of chitin synthase genes on mycelial morphology and culture viscosity in Aspergillus oryzae. , 2003, Biotechnology and bioengineering.

[46]  V. Vinci,et al.  Mutants of a lovastatin-hyperproducingAspergillus terreus deficient in the production of sulochrin , 1991, Journal of Industrial Microbiology.

[47]  A. Rodal,et al.  Expression of the Escherichia coli Catabolic Threonine Dehydratase in Corynebacterium glutamicum and Its Effect on Isoleucine Production , 1999, Applied and Environmental Microbiology.

[48]  H. Umezawa.  Enzyme inhibitors of microbial origin , 1972 .

[49]  H. Döhren,et al.  Products of secondary metabolism , 1997 .

[50]  S. Giovannoni,et al.  The uncultured microbial majority. , 2003, Annual review of microbiology.

[51]  Manor Askenazi,et al.  Integrating transcriptional and metabolite profiles to direct the engineering of lovastatin-producing fungal strains , 2003, Nature Biotechnology.

[52]  Mark R. Marten,et al.  Pulsed Feeding during Fed‐Batch Aspergillus oryzae Fermentation Leads to Improved Oxygen Mass Transfer , 2003, Biotechnology progress.

[53]  H. Sahm,et al.  Quantifying and directing metabolite flux: Application to amino acid overproduction , 1996 .