Application of l-glutamate to l-proline fermentation by Corynebacterium acetoacidophilum

Nakanishi, Toshihide; Hirao, Toshihiko; Azuma, Tomoki; Sakurai, Minoru; Hagino, Hiroshi

doi:10.1016/0385-6380(87)90157-9

Cited by 10 publications

(9 citation statements)

References 10 publications

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“…To demonstrate feasibility, we edited the γ-glutamyl kinase gene ( proB ) of C. glutamicum ATCC13032 to relieve feedback inhibition by L -proline (a useful chemical that is produced commercially by C. glutamicum 26), thereby facilitating L -proline production (Fig. 5a).…”

Section: Resultsmentioning

confidence: 99%

CRISPR-Cpf1 assisted genome editing of Corynebacterium glutamicum

et al. 2017

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Corynebacterium glutamicum is an important industrial metabolite producer that is difficult to genetically engineer. Although the Streptococcus pyogenes (Sp) CRISPR-Cas9 system has been adapted for genome editing of multiple bacteria, it cannot be introduced into C. glutamicum. Here we report a Francisella novicida (Fn) CRISPR-Cpf1-based genome-editing method for C. glutamicum. CRISPR-Cpf1, combined with single-stranded DNA (ssDNA) recombineering, precisely introduces small changes into the bacterial genome at efficiencies of 86–100%. Large gene deletions and insertions are also obtained using an all-in-one plasmid consisting of FnCpf1, CRISPR RNA, and homologous arms. The two CRISPR-Cpf1-assisted systems enable N iterative rounds of genome editing in 3N+4 or 3N+2 days. A proof-of-concept, codon saturation mutagenesis at G149 of γ-glutamyl kinase relieves L-proline inhibition using Cpf1-assisted ssDNA recombineering. Thus, CRISPR-Cpf1-based genome editing provides a highly efficient tool for genetic engineering of Corynebacterium and other bacteria that cannot utilize the Sp CRISPR-Cas9 system.

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Section: Resultsmentioning

confidence: 99%

CRISPR-Cpf1 assisted genome editing of Corynebacterium glutamicum

et al. 2017

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“…flavum produced 35 g l -1 proline . When a glutamate-producing strain of C. glutamicum was grown under modified conditions, it made 48 g l -1 (Nakanishi et al, 1973). A strain of Corynebacterium acetoacidophilum produced 108 g l -1 proline when grown in the presence of glutamate (Nakanishi et al, 1987).…”

Section: L-prolinementioning

confidence: 99%

Metabolic regulation and overproduction of primary metabolites

Sánchez

Demain

2008

Microbial Biotechnology

146

103

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SummaryOverproduction of microbial metabolites is related to developmental phases of microorganisms. Inducers, effectors, inhibitors and various signal molecules play a role in different types of overproduction. Biosynthesis of enzymes catalysing metabolic reactions in microbial cells is controlled by well‐known positive and negative mechanisms, e.g. induction, nutritional regulation (carbon or nitrogen source regulation), feedback regulation, etc. The microbial production of primary metabolites contributes significantly to the quality of life. Fermentative production of these compounds is still an important goal of modern biotechnology. Through fermentation, microorganisms growing on inexpensive carbon and nitrogen sources produce valuable products such as amino acids, nucleotides, organic acids and vitamins which can be added to food to enhance its flavour, or increase its nutritive values. The contribution of microorganisms goes well beyond the food and health industries with the renewed interest in solvent fermentations. Microorganisms have the potential to provide many petroleum‐derived products as well as the ethanol necessary for liquid fuel. Additional applications of primary metabolites lie in their impact as precursors of many pharmaceutical compounds. The roles of primary metabolites and the microbes which produce them will certainly increase in importance as time goes on. In the early years of fermentation processes, development of producing strains initially depended on classical strain breeding involving repeated random mutations, each followed by screening or selection. More recently, methods of molecular genetics have been used for the overproduction of primary metabolic products. The development of modern tools of molecular biology enabled more rational approaches for strain improvement. Techniques of transcriptome, proteome and metabolome analysis, as well as metabolic flux analysis. have recently been introduced in order to identify new and important target genes and to quantify metabolic activities necessary for further strain improvement.

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“…Additionally, further mutants are resistant to S-(2-aminoethyl)-L-cysteine or require L-alanine for cell growth. US20067008786 [40] discloses a Coryneform bacterium having resistance to the antibiotic monensin, whereas overproduction of L-lysine by resistant mutant strains of the genus Corynebacterium or Brevibacterium to purine and pyrimidine analogs or ethylene glycol is described in [41,42], respectively. L-Lysine producing strains of Corynebacterium or Brevibacterium sensitive to fluoropyruvic acid, in addition to homoserine auxotrophy and resistance to S-(2-aminoethyl)-L-cysteine (AEC), are described in [43] and to aspartic acid analogs and sulfa drugs in [2,39] describes coryneform Lglutamic acid and L-lysine producing mutants with a temperature-sensitive mutation (37 o C) with respect to a biotin action-suppressing agent.…”

Section: Classical Mutagenesismentioning

confidence: 99%

L-Lysine Fermentation

Anastassiadis

2007

BIOT

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Amino acids are the basic bioelements of proteins, which are the most important macromolecules for the functions of humans and animals. Out of the 20 L-amino acids, ecumenically found in most of living organisms, L-lysine is one of the 9 amino acids which are essential for human and animal nutrition. L-lysine is useful as medicament, chemical agent, food material (food industry) and feed additive (animal food). Its demand has been steadily increasing in recent years and several hundred thousands tones of L-lysine (about 800,000 tones/year) are annually produced worldwide almost by microbial fermentation. The stereospecificity of amino acids (the L isomer) makes the fermentation advantageous compared with synthetic processes. Mutant auxotrophic or resistant to certain chemicals strains of so-called gram positive coryneform bacteria are generally used, including the genera Brevibacterium and Corynebacterium, united to the genus. The significance of Research and Development increased rapidly since the discovery of fermentative amino acid production in the fifties (S. Kinoshita et al., Proceedings of the International Symposium on Enzyme Chemistry 2:464-468 (1957)), leading to innovative fermentation processes which replaced the classical manufacturing methods of L-lysine like acid hydrolysis. L-Lysine is separated and purified by suitable downstream processes involving classical separation or extraction methods (ultrafiltration or centrifugation, separation or ion exchange extraction, crystallization, drying) and is sold as a powder. Alternatively, spray dried pellets or liquid fermentation broth can be used as animal feed supplement. On behalf of today's strong competition in amino acid industry, Biotechnology companies are continuously aiming in innovative research developments and use complex management concepts and business strategies, towards gaining market leadership in the field of amino acid production.

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Application of l-glutamate to l-proline fermentation by Corynebacterium acetoacidophilum

Cited by 10 publications

References 10 publications

CRISPR-Cpf1 assisted genome editing of Corynebacterium glutamicum

CRISPR-Cpf1 assisted genome editing of Corynebacterium glutamicum

Metabolic regulation and overproduction of primary metabolites

L-Lysine Fermentation

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