This indicates that oxygen supply greatly affects l lysine productivity in continuous culture. From zero to herodesignbased systems metabolic engineering of corynebacterium glutamicum for l lysine production elsevier enhanced reader. Production of l lysine by genetically modified corynebacterium glutamicum strains ca 2455878 ca2455878a1 en 20010806. Heteroexpression of scrk increases llysine production in c. Bioprocess technology development and convalescing. A number of damino acids occur in nature, and there is growing interest in their function and metabolism, as well as in their production and use. Metabolic engineering of corynebacterium glutamicum for l. We constructed pep carboxylasenegative strains of the wildtype and of the llysine. Comparisons of potentials for llysine production among. In the present work, lysine production by corynebacterium glutamicum was improved by metabolic engineering of the tricarboxylic acid tca cycle. Engineering of a glycerol utilization pathway for amino. Native promoters of corynebacterium glutamicum and its. Thus, corynebacterium glutamicum, a highly efficient llysine producing microorganism, should be useful in the development of direct fermentative production of 5ava using llysine as a precursor for 5ava. Here we report the development of metabolically engineered corynebacterium glutamicum atcc 21831 for.
Shah 15 and ferreira and duarte 5, stated that the initial sugar concentration of 10% gave the highest production yield. Specifically, the coenzyme specificity of gapdh of corynebacterium glutamicum is systematically manipulated by rational protein design and the effect of the manipulation for cellular metabolism and lysine production is evaluated. The growth rate, glucose consumption rate, and production rate of an llysine producing corynebacterium glutamicum mutant were studied in batch and. In batch fermentation, higher llysine productivity 1. Transport and excretion of llysine in corynebacterium. Metabolic responses to pyruvate kinase deletion in lysine. Corynebacterium glutamicum cgmcc 3704, respectively. However, no mutation leading to increased l lysine production has been reported with respect to the pentose phosphate pathway. The producing strains are mostly derived from the wildtype of c. Rational modification of tricarboxylic acid cycle for. The present work describes the production of l lysine from corynebacterium glutamicum iibc9 through submerged fermentation process. The juice was found to have inadequate nutrients for growth of the auxotroph alone and was supplemented. All the kinetic parameters including yield coefficients and volumetric rates revealed the overproduction of lysine by the mutant using black strap molasses as a sole source of carbon. Approaches for improving the production of l lysine in corynebacterium glutam icum article pdf available in international journal of research in pharmaceutical sciences 24 january 2011 with.
Corynebacterium glutamicum is widely used for industrial production of amino acids such as glutamate and lysine. Us8048649b2 production of amino acids from sucrose in. Pdf native promoters of corynebacterium glutamicum and. Toward the elucidation of advanced mechanisms of llysine production by corynebacterium glutamicum, a highly developed industrial strain b6 was analyzed from the viewpoint of gene expression. The gram positive bacteriumcorynebacterium glutamicum is used for the production of llysine.
Lysine is nowadays produced aerobically by industrial strains of corynebacterium glutamicum, in an aerationintensive and sensitive process 1. Corynebacterium glutamicum is an industrially important microorganism used for amino acids production. Potential metabolic limitations in lysine production by. The powerful toolbox of genetic engineering allows the targeted modification of genes in c. Corynebacterium glutamicum was engineered for improvement of l. Glutamate is the precursor of the amino acid proline. Corynebacterium glutamicum lys9 was devoid of any detectable l. Corynebacterium glutamicum as a host for synthesis and export.
Engineering of a glycerol utilization pathway for amino acid. Variables for llysine production by corynebacterium glutamicum. Transcriptome analysis reveals global expression changes. Heterologous expression of lactose and galactoseutilizing pathways from lactic acid bacteria in corynebacterium glutamicum for production of lysine in whey. Attenuating l lysine production by deletion of ddh and lyse and their effect on l threonine and l isoleucine production in corynebacterium glutamicum. Pdf one of the most commonly used approaches for improving the. Metabolic engineering of corynebacterium glutamicum for. By flux analysis, this could be correlated to a flux shift from the tca cycle toward anaplerotic carboxylation. Llysine hcl does not give rise to any safety concern with regard to the genetic modification of the production strain. Heteroexpression of scrk increases l lysine production in c.
Us20090081740a1 production of amino acids from sucrose. Methods and compositions for increased production of amino acids from c. Here, we report the development of metabolically engineered c. Currently, lornithine production is focused on microorganism fermentation using escherichia coli or corynebacterium glutamicum. Evaluation of sweet sorghum juice for the production of. Bioprocess technology development and convalescing production. Production of llysine from starch by corynebacterium. Engineering of corynebacterium glutamicum for growth and. Hereby lysine is one of the major products, with worldwide production of about 400,000 tons per year and an annual market increase of about 10 to 15%.
Higher agitation or use of oxygenenriched air was critical for high productivity. Suboptimal flux partitioning at a metabolic branch point can result. In one aspect, increased production of llysine from c. Whereas pyruvate kinase deletion resulted in increased lysine production for different strains of the close relative brevibacterium flavum 6,11,12, and during the major production phase of a batch process with c. Phosphoenolpyruvate carboxylase in corynebacterium. Mutations in the transport system did not lead to overproduction of the amino acid. Corynebacterium glutamicum has been used since several decades for the largescale production of amino acids, esp. L lysine hcl does not give rise to any safety concern with regard to the genetic modification of the production strain. These promoters covered a strength range of 31fold with small increments and exhibited relatively stable activity during the whole growth phase using. Utilization of sweet sorghum juice as a feedstock for the production of lysine was investigated utilizing the auxotrophic mutant corynebacterium glutamicum atcc 215 in stirred tank bioreactors. A nonpathogenic species of coryneform bacteria, corynebacterium glutamicum, was originally isolated as an lglutamate producing bacterium and is now used for fermentative production of various amino acids. Llysine transport in corynebacterium glutamicum was investigated.
A defined l lysine producing mutant was generated by introduction of the lysc mutation t311i into each of six representative c. We constructed pep carboxylasenegative strains of the wildtype and of the llysine producer mh2022b by disruption of the respective gene. According to our previous results, total sugar concentration in beet molasses, supplied by cofco biochemical anhui co. Great effort has been undertaken in order to optimize producer strains. Ornithine cyclodeaminasebased proline production by. Metabolic pathways involved in the production of llysine by corynebacterium glutamicum and the mechanisms mediating its efflux and secretion are discussed. Pdf fermentative production of lysine by corynebacterium. Transport and excretion of llysine in corynebacterium glutamicum. L lysine fermentation recent patents on biotechnology 2007, vol. Comparison of growth and amino acid production, proteome and transcriptome differences between c. Prasad c a department of chemical engineering, national institute of technology srinagar190006, india b department of biotechnology, indian institute of technology roorkee247667, india c department of chemical. To identify useful native promoters of corynebacterium glutamicum for finetuning of gene expression in metabolic engineering. Sweet sorghum juice is a liquid sugar feedstock that can be produced in nontropical climates. Metabolic engineering of the tricarboxylic acid cycle for.
L scale applying the general approach of reaction engineering. Aug 21, 2003 the grampositive bacterium corynebacterium glutamicum is widely used for the industrial production of different amino acids. Barrett e, stanton c, zelder o, fitzgerald g, ross rp. Llysine monohydrochloride, technically pure, produced. Fedbatch production of llysine by corynebacterium glutamicum. At present, a various genes involved in llysine production were characterized at the molecular level, and subsequently, the llysine producers were achieved by genetic engineering of llysine biosynthetic pathway, central metabolic pathways as well as. The growth rate, glucose consumption rate, and production rate of an llysine producing corynebacterium glutamicum mutant were studied in batch and fedbatch cultures. Amino acid production by corynebacterium glutamicum. By heterologous expression of escherichia coli glycerol utilization genes, c. Corynebacterium glutamicum, lysine, fermentation, homoserine auxotrophs. Here we use the wellestablished laminoacidproducing bacterium corynebacterium glutamicum to study whether damino acid synthesis is possible and whether mechanisms for the export of these amino acids exist.
May 23, 2003 a comprehensive approach to c tracer studies, labeling measurements by gas chromatographymass spectrometry, metabolite balancing, and isotopomer modeling, was applied for comparative metabolic network analysis of lysine producing corynebacterium glutamicum on glucose or fructose. Corynebacterium glutamicum and related bacteria are industrially important microorganisms that are widely used for the production of various amino acids 1,2. Fermentative production of lysine by corynebacterium. Statistical optimization of process variables for llysine. Corynebacterium glutamicum variety producing larginine and method for fabricating the same de102007005072a1 en. Northern blot analysis showed that the lysc gene encoding aspartokinase, the key enzyme of llysine biosynthesis, was upregulated by several folds in strain b6, while no repression mechanism exists in l. Llysine monohydrochloride, technically pure, produced with. In one aspect, increased production of l lysine from c. Production of llysine was followed in two llysine accumulating mutants of c. The amino acidproducing organism corynebacterium glutamicum cannot utilize glycerol, a stoichiometric byproduct of biodiesel production. Its most prominent feature is probably its capacity to produce lglutamate, 1. Lysine secretion by wildtype corynebacterium glutamicum. While the classical mutagenic procedure has greatly.
L lysine transport in corynebacterium glutamicum was investigated. Nov 21, 2017 to identify useful native promoters of corynebacterium glutamicum for finetuning of gene expression in metabolic engineering. Analyses indicate that the lysine yield in corynebacterium glutamicum is limited by suboptimal flux partitioning at either the glucose 6. As the first complete compilation of major findings, handbook of. Optimization of an industrial llysine producing corynebacterium. The 70% decreased activity of isocitrate dehydrogenase, achieved by start codon exchange, resulted in a 40% improved lysine production. Transcriptome analysis reveals global expression changes in. Corynebacterium glutamicum is an industrially important organism that is most widely used for the production of various amino acids.
Corynebacterium glutamicum as a host for synthesis and. Aug 24, 2015 attenuating l lysine production by deletion of ddh and lyse and their effect on l threonine and l isoleucine production in corynebacterium glutamicum. This microorganism is a grampositive, nonsporulating, nonmotile, ellipsoidal bacteria and is aerobic. One of the most commonly used approaches for improving the production of llysine in corynebacterium glutamicum was classical mutagenesis which. Experimental optimization of medium composition components. Production of l lysine from starch by corynebacterium glutamicum displaying.
Comparative metabolic flux analysis of lysineproducing. Enhancement of anaerobic lysine production in corynebacterium. Frontiers efficient production of the dicarboxylic acid. In using ferricyanide as extracellular electron carrier, anaerobic growth was enabled and the feedback. Production of l lysine by genetically modified corynebacterium glutamicum strains ep20020760293 ep1414970a2 en 20010806. Nov 15, 2014 barrett e, stanton c, zelder o, fitzgerald g, ross rp. Metabolic flux analysis, which is considered to be a very powerful tool providing valuable information regarding bottlenecks in the production of desired metabolites, is also covered in. Metabolic engineering of carbohydrate metabolism systems in. Metabolic pathways involved in the production of l lysine by corynebacterium glutamicum and the mechanisms mediating its efflux and secretion are discussed. A novel methodology employing corynebacterium glutamicum. The approach followed by the feedap panel to assess the safety and the ef. Tateno, toshihiro, fukuda, hideki, kondo, akihiko source.
Due to its importance as an amino acid producer, c. Extensive investigation and optimization of lysineproducing strains of c. Indepth profiling of lysineproducing corynebacterium. The organism selected to carry out the production of l lysine is corynebacterium glutamicum. Alternatively, proline can be synthesised from ornithine, an intermediate of arginine biosynthesis. Organic acids were coproduced during the anaerobic fermentation as is also discussed in the next paragraph. Llysine, fermentation, corynebacterium glutamicum, process variables, optimization, response surface methodology ozet amac.
Understanding the high lvaline production in corynebacterium. Proline biosynthesis typically involves three enzymes and a spontaneous cyclisation reaction. Optimization of lysine production in an industrial relevant lysine producing corynebacterium glutamicum strain was done using a combination of mathematically modelling, metabolic engineering and metabolic flux estimation. Sucrose was assimilated and phosphorylated by sucrosepts. The bacterium was shown to possess a highly specific, energydependent system of active lysine transport. During the half century following its discovery, the lglutamateproducing microorganism corynebacterium glutamicum has played a leading role in the amino acid fermentation industry. Abstract llysine c6h14n2o2, one of the essential and commercially important amino acids, is found in naturallyoccurring proteins of all living organisms. Pdf approaches for improving the production of llysine in. Method for the production of llysine using an alkaliphilic bacterium ep2940039a1 en 20140430.
Analysis of the metabolic network of lysineproducing corynebacterium glutamicum showed that lysine yields are limited by the excess energy production in l. Native promoters of corynebacterium glutamicum and its application in l lysine production article pdf available in biotechnology letters 402. Llysine fermentation recent patents on biotechnology 2007, vol. Until now, research and information have been scattered among individual papers which are often difficult to locate in a timely manner. Microbial production of glutarate was achieved but not optimized in escherichia coli by using the llysine degradation pathway or 5aminovalerate 5ava pathway from pseudomonas putida kt2440 adkins et al. Fermentative production of lysine by corynebacterium glutamicum from 103 as the carbon sources in this study, and the production yield was compared in different time intervals 0120 h. Corynebacterium glutamicum is well known for its extraordinary lamino acid production properties. Larginine is an important amino acid for diverse industrial and health product applications. Lglutamate production by lysozymesensitive corynebacterium. Users may download and print one copy of any publication from the public.
Rational modification of tricarboxylic acid cycle for improving l. Transcriptome analysis reveals global expression changes in an industrial l lysine producer of corynebacterium glutamicum. Biology of llysine overproduction by corynebacterium glutamicum. Engineering a lysineon riboswitch for metabolic control of. The grampositive bacterium corynebacterium glutamicum is widely used for the industrial production of different amino acids. The system transferred lysine into the cells and exchanged intra and extracellular lysine. The strains of this bacteria typically form pale yellow colonies, however some creamwhite strands can also occur kjeldsen, 2008. Metabolic engineering of carbohydrate metabolism systems. Jun 28, 20 the soil bacterium corynebacterium glutamicum, best known for its glutamate producing ability, is suitable as a producer of a variety of bioproducts. Oct 16, 2001 a nonpathogenic species of coryneform bacteria, corynebacterium glutamicum, was originally isolated as an lglutamate producing bacterium and is now used for fermentative production of various amino acids. One of the most important organisms in biotechnology, corynebacterium glutamicum is currently used to produce 2 million tons of amino acids per year for a rapidly expanding market. Phosphoenolpyruvate carboxylase in corynebacterium glutamicum. Engineering a lysineon riboswitch for metabolic control. Phosphoenolpyruvate pep carboxylase is assumed to be of major importance as anaplerotic enzyme in the amino acid producing corynebacterium glutamicum.
659 949 1318 750 1250 1137 699 1023 270 470 898 619 331 70 848 823 1143 1268 849 1018 1177 1419 820 523 1355 611 895 544 9 210 4 1410 841 1220 579 1476 113 1098 198 1027