scholarly journals Fermentation Conditions and Media Optimization for Isocitric Acid Production from Ethanol by Yarrowia lipolytica

2018 ◽  
Vol 2018 ◽  
pp. 1-9 ◽  
Author(s):  
Svetlana V. Kamzolova ◽  
Roman V. Shamin ◽  
Nadezda N. Stepanova ◽  
Grigorii I. Morgunov ◽  
Julia N. Lunina ◽  
...  
Keyword(s):  
Yarrowia Lipolytica ◽  
Isocitrate Lyase ◽  
Krebs Cycle ◽  
Growth Period ◽  
Energy Yield ◽  
Good Growth ◽  
Acid Inhibitor ◽  
Isocitric Acid ◽  
Nutrition Medium ◽  
Key Enzyme

Isocitric acid exists in the form of four stereoisomers, of which only the threo-Ds-form (ICA) is a natural active compound, an intermediate of Krebs cycle, and suitable for nutritional and pharmaceutical use. In this paper, we propose a method for ICA production from ethanol by yeast Yarrowia lipolytica. The effects of temperature, pH of the medium, and aeration on the growth of the producer Y. lipolytica VKM Y-2373 and synthesis of ICA were studied. An optimal fermentation regime, which ensures a good growth of the producer and directed synthesis of the target product, was determined. The producer is advised to carry out cultivation at 29°C and various pH of the medium and the oxygen concentration (pH 5 and pO2 20–25% (of saturation) during the growth period and pH 6 and pO2 50–55% (of saturation) during the acid formation) on a nutrient medium containing an increased content of zinc (0.6 mg/L), iron (1.2 mg/L), and 30 mM itaconic acid (inhibitor of isocitrate lyase—the key enzyme of ICA metabolism) should also be introduced into the nutrition medium. Such fermentation production mode provides 90.5 g/L ICA with process selectivity of 80%, mass yield (YICA) of 0.77 g/g, and energy yield (ηICA) of 0.278 g/g.

scholarly journals Effects of Medium Components on Isocitric Acid Production by Yarrowia lipolytica Yeast

Fermentation ◽  
2020 ◽  
Vol 6 (4) ◽  
pp. 112
Author(s):  
Svetlana V. Kamzolova ◽  
Vladimir A. Samoilenko ◽  
Julia N. Lunina ◽  
Igor G. Morgunov
Keyword(s):  
Isocitrate Lyase ◽  
Nitrogen Deficiency ◽  
Aconitate Hydratase ◽  
Isocitric Acid ◽  
Medium Components ◽  
Regulatory Approach ◽  
Nutrition Medium ◽  
Key Enzyme ◽  
Nitrogen Phosphorus ◽  
Oxalic Acids

The microbiological production of isocitric acid (ICA) is more preferable for its application in medicine and food, because the resulting product contains only the natural isomer—threo-DS. The aim of the present work was to study ICA production by yeast using sunflower oil as carbon source. 30 taxonomically different yeast strains were assessed for their capability for ICA production, and Y. lipolytica VKM Y-2373 was selected as a promising producer. It was found that ICA production required: the limitation of Y. lipolytica growth by nitrogen, phosphorus, sulfur or magnesium, and an addition of iron, activating aconitate hydratase, a key enzyme of isocitrate synthesis. Another regulatory approach capable to shift acid formation to a predominant ICA synthesis is the use of inhibitors (itaconic and oxalic acids), which blocks the conversion of isocitrate at the level of isocitrate lyase. It is recommended to cultivate Y. lipolytica VKM Y-2373 under nitrogen deficiency conditions with addition of 1.5 mg/L iron and 30 mM itaconic acid. Such optimized nutrition medium provides 70.6 g/L ICA with a ratio between ICA and citric acid (CA) equal 4:1, a mass yield (YICA) of 1.25 g/g and volume productivity (QICA) of 1.19 g/L·h.

scholarly journals The Gluconeogenic Enzyme Fructose-1,6-Bisphosphatase Is Dispensable for Growth of the Yeast Yarrowia lipolytica in Gluconeogenic Substrates

2008 ◽  
Vol 7 (10) ◽  
pp. 1742-1749 ◽  
Author(s):  
Raquel Jardón ◽  
Carlos Gancedo ◽  
Carmen-Lisset Flores
Keyword(s):  
Yarrowia Lipolytica ◽  
Phosphoenolpyruvate Carboxykinase ◽  
Isocitrate Lyase ◽  
Carbon Sources ◽  
Subcellular Fractionation ◽  
Fructose 1,6 Bisphosphatase ◽  
Genes Encoding ◽  
Key Enzyme ◽  
Gluconeogenic Enzyme ◽  
Fructose 1,6 Bisphosphate

ABSTRACT The genes encoding gluconeogenic enzymes in the nonconventional yeast Yarrowia lipolytica were found to be differentially regulated. The expression of Y. lipolytica FBP1 (YlFBP1) encoding the key enzyme fructose-1,6-bisphosphatase was not repressed by glucose in contrast with the situation in other yeasts; however, this sugar markedly repressed the expression of YlPCK1, encoding phosphoenolpyruvate carboxykinase, and YlICL1, encoding isocitrate lyase. We constructed Y. lipolytica strains with two different disrupted versions of YlFBP1 and found that they grew much slower than the wild type in gluconeogenic carbon sources but that growth was not abolished as happens in most microorganisms. We attribute this growth to the existence of an alternative phosphatase with a high Km (2.3 mM) for fructose-1,6-bisphosphate. The gene YlFBP1 restored fructose-1,6-bisphosphatase activity and growth in gluconeogenic carbon sources to a Saccharomyces cerevisiae fbp1 mutant, but the introduction of the FBP1 gene from S. cerevisiae in the Ylfbp1 mutant did not produce fructose-1,6-bisphosphatase activity or growth complementation. Subcellular fractionation revealed the presence of fructose-1,6-bisphosphatase both in the cytoplasm and in the nucleus.

scholarly journals Effect of Metabolic Regulators and Aeration on Isocitric Acid Synthesis by Yarrowia lipolytica Grown on Ester-Aldehyde Fraction

Fermentation ◽  
2021 ◽  
Vol 7 (4) ◽  
pp. 283
Author(s):  
Svetlana V. Kamzolova ◽  
Igor G. Morgunov
Keyword(s):  
Yarrowia Lipolytica ◽  
Itaconic Acid ◽  
Isocitrate Lyase ◽  
Nitrogen Deficiency ◽  
Carbon Sources ◽  
Acid Synthesis ◽  
Aconitate Hydratase ◽  
Isocitric Acid ◽  
Metabolic Regulators ◽  
Putative Mechanism

Isocitric acid (ICA) has found wide application in medicine as a promising compound with powerful antioxidant activity to combat oxidative stress. In the known microbiological processes of ICA production by non-conventional yeast Yarrowia lipolytica, the pure carbon sources are commonly used. ICA can be also synthetized by Y. lipolytica from ester-aldehyde fraction (EAF)-waste of the ethanol production process. A highly effective method of ICA production from EAF based on regulation of key enzymes (aconitate hydratase and isocitrate lyase) by metabolic regulators (iron and itaconic acid) and aeration was developed. It is recommended to cultivate Y. lipolytica VKM Y-2373 under nitrogen deficiency conditions, a high aeration (60% of air saturation), an addition of 15 mM itaconic acid, and 2.4 mg/L iron. Under optimal conditions, Y. lipolytica VKM Y-2373 produced 83 g/L ICA with isocitrate to citrate ratio of 4.1:1 and mass yield of 1.1 g/g. The putative mechanism of ICA overproduction from EAF by Y. lipolytica was suggested.

Disruption of genes for the enhanced biosynthesis of α-ketoglutarate in Corynebacterium glutamicum

2012 ◽  
Vol 58 (3) ◽  
pp. 278-286 ◽  
Author(s):  
Jae-Hyung Jo ◽  
Hye-Young Seol ◽  
Yun-Bom Lee ◽  
Min-Hong Kim ◽  
Hyung-Hwan Hyun ◽  
...  
Keyword(s):  
Corynebacterium Glutamicum ◽  
Biosynthetic Pathway ◽  
Isocitrate Lyase ◽  
Glutamate Synthase ◽  
Flask Culture ◽  
Control Strain ◽  
Enhanced Production ◽  
Key Enzyme ◽  
Microbial Strains ◽  
Glyoxylate Pathway

The development of microbial strains for the enhanced production of α-ketoglutarate (α-KG) was investigated using a strain of Corynebacterium glutamicum that overproduces of l-glutamate, by disrupting three genes involved in the α-KG biosynthetic pathway. The pathways competing with the biosynthesis of α-KG were blocked by knocking out aceA (encoding isocitrate lyase, ICL), gdh (encoding glutamate dehydrogenase, l-gluDH), and gltB (encoding glutamate synthase or glutamate-2-oxoglutarate aminotransferase, GOGAT). The strain with aceA, gltB, and gdh disrupted showed reduced ICL activity and no GOGAT and l-gluDH activities, resulting in up to 16-fold more α-KG production than the control strain in flask culture. These results suggest that l-gluDH is the key enzyme in the conversion of α-KG to l-glutamate; therefore, prevention of this step could promote α-KG accumulation. The inactivation of ICL leads the carbon flow to α-KG by blocking the glyoxylate pathway. However, the disruption of gltB did not affect the biosynthesis of α-KG. Our results can be applied in the industrial production of α-KG by using C. glutamicum as producer.

Co-ordinate regulation of genes involved in storage lipid mobilization in Arabidopsis thaliana

2001 ◽  
Vol 29 (2) ◽  
pp. 283-286 ◽  
Author(s):  
E. L. Rylott ◽  
M. A. Hooks ◽  
I. A. Graham
Keyword(s):  
Arabidopsis Thaliana ◽  
Enzyme Activities ◽  
Phosphoenolpyruvate Carboxykinase ◽  
Isocitrate Lyase ◽  
Glyoxylate Cycle ◽  
Molecular Genetic ◽  
Regulation Of Gene Expression ◽  
Growth Period ◽  
Malate Synthase ◽  
The Glyoxylate Cycle

Molecular genetic approaches in the model plant Arabidopsis thaliana (ColO) are shedding new light on the role and control of the pathways associated with the mobilization of lipid reserves during oilseed germination and post-germinative growth. Numerous independent studies have reported on the expression of individual genes encoding enzymes from the three major pathways: β-oxidation, the glyoxylate cycle and gluconeogenesis. However, a single comprehensive study of representative genes and enzymes from the different pathways in a single plant species has not been done. Here we present results from Arabidopsis that demonstrate the co-ordinate regulation of gene expression and enzyme activities for the acyl-CoA oxidase- and 3-ketoacyl-CoA thiolasemediated steps of β-oxidation, the isocitrate lyase and malate synthase steps of the glyoxylate cycle and the phosphoenolpyruvate carboxykinase step of gluconeogenesis. The mRNA abundance and enzyme activities increase to a peak at stage 2, 48 h after the onset of seed germination, and decline thereafter either to undetectable levels (for malate synthase and isocitrate lyase) or low basal levels (for the genes of β-oxidation and gluconeogenesis). The co-ordinate induction of all these genes at the onset of germination raises the possibility that a global regulatory mechanism operates to induce the expression of genes associated with the mobilization of storage reserves during the heterotrophic growth period.

Comparison of glycolytic, pentose phosphate pathway, glyoxylate shunt, Krebs' cycle enzymes in Ganeo tigrinum parasitizing hibernating and non-hibernating Rana cyanophlyctis and R. tigrina

1983 ◽  
Vol 57 (1) ◽  
pp. 59-68 ◽  
Author(s):  
P. N. Sharma ◽  
Sushila Mandawat
Keyword(s):  
Isocitrate Lyase ◽  
Krebs Cycle ◽  
Pentose Phosphate ◽  
Glyoxylate Shunt ◽  
Strong Activity ◽  
Key Enzymes ◽  
Phosphogluconate Dehydrogenase ◽  
Weak Activity ◽  
Krebs Cycle Enzymes ◽  
Glucose 6 Phosphate Dehydrogenase

AbstractThe histochemical site and distribution of hexokinase, glycogen phosphorylase (GP Rylase), lactate dehydrogenase (LDH) (key enzymes of glycolysis), glucose-6-phosphate dehydrogenase (GPD) and 6-phosphogluconate dehydrogenase (6PGD) (pentose phosphate shunt enzymes), isocitrate dehydrogenase (IDH), succinate dehydrogenase (SDH), malate dehydrogenase (MDH), and α-ketoglutarate dehydrogenase (α-KDH) (key enzymes of Krebs' cycle), malate synthetase (MS) and isocitrate lyase (IL) (enzymes of glyoxylate shunt) in various tissues of Ganeo tigrinum from hibernating and non-hibernating Rana cyanophlyctis and R. tigrina were studied. Differences in their intensities were revealed. Weak activity of GP Rylase and strong activity of hexokinase in flukes from non-hibernating hosts indicates that they utilize glucose through glycolysis for energy turnover. Intense GP Rylase and weak hexokinase activity in worms from hibernating hosts indicates the utilization of glycogen. Strong activity of IDH, SDH, MDH, α-KGD, MS and IL was demonstrable in the tissues of flukes from non-hibernating hosts, suggesting that Krebs' cycle and glyoxylate shunt, respectively, were operating. Tissues of the fluke from hibernating hosts, on the other hand, displayed positive activity only for SDH and MDH; no activity for MS and IL, the enzymes of glyoxylate shunt, was observed, The activity of the above enzymes was found to be relatively low in worms from hibernating hosts.

scholarly journals Molasses as a new nutrition medium for Scenedsmus quadricauda growth and production of some bio compounds

Bionatura ◽  
2021 ◽  
Vol 6 (4) ◽  
pp. 2202-2208
Author(s):  
Yousef J.I. AL –Shahery ◽  
Israa N. AL-Asady
Keyword(s):  
Protein Content ◽  
Algal Growth ◽  
Carbohydrate Content ◽  
Good Growth ◽  
Human Beings ◽  
Green Pigment ◽  
Sugar Factory ◽  
L Protein ◽  
Growing Period ◽  
Nutrition Medium

Algae comprise a large group of Thallophyta, which may be used as direct nutrition of human beings. Molasses is the by-product of the sugar manufacturing facility. In this study, a locally isolated Scendsmus quadricauda from the environment of Mosul in the Shalalat region was obtained. Biomass of Scenedsmus was measurement by carried out and filtration then drying in an oven for 24 h and weighed, Estimation of chlorophyll and protein and carbohydrate content of Scenedsmus. The research has proved that the best growing period for Scendsmue quadricauda is 15 days when using sugar factory waste as a carbon source, the growth reached (1.42 nm) as optical density, biomass (1525 mg /L), chlorophyll (green), pigment (18 mg /l) protein content (396 mg /l ) and carbohydrates ( 501 mg / l ). The research showed that the use of sugar factory waste as a nutritional medium for algal growth in the dark (11.5%) achieved good growth of Scendesmues quadricauda ( 0.632 nm), biomass (820 mg / L), green pigment (Chlorophyll) (18 mg /L) protein content (235 mg / L ) and carbohydrates (401 mg/L). while using phosphor (0.018%) of K2HPO4 in dark medium achieved highest growth rate (0.91 nm) , biomass (1110 mg / L) chlorophyll ( 22 mg/L) protein (301mg/L) and carbohydrate (461 mg/L) . It is noted too , that using IAA (0.5 g/L) in dark medium support best growth (0.888 nm) , biomass (1010 mg/L) chlorophyll (25 mg/L) , protein (230mg/L) and carbohydrate (440 mg//L) . The study showed that thiamine (1 g/L) in dark medium achieved highest growth (0.750 nm) biomass (218 mg/L), chlorophyll (29mg/L), protein (220 mg/L), carbohydrate (340mg/L). Therefore, using Molasses can enhance the growth, biomass, chlorophyll, protein, and carbohydrate content in the S. quadricauda.

Screening of Anti-mycobacterial Phytochemical Compounds for Potential Inhibitors against Mycobacterium Tuberculosis Isocitrate Lyase

2019 ◽  
Vol 19 (8) ◽  
pp. 600-608 ◽  
Author(s):  
Ashish Tiwari ◽  
Akhil Kumar ◽  
Gaurava Srivastava ◽  
Ashok Sharma
Keyword(s):  
Binding Energy ◽  
Virtual Screening ◽  
Md Simulation ◽  
Isocitrate Lyase ◽  
Docking Study ◽  
High Morbidity ◽  
Average Binding Energy ◽  
Ic50 Value ◽  
Key Enzyme ◽  
Potential Inhibitors

Conclusion:Phytochemical based anti-mycobacterial compound can further developed into effective drugs against persistence tuberculosis with lesser toxicity and side effects.Results:Docking and MD simulation studies of top hit compounds have identified shinjudilactone (quassinoid), lecheronol A (pimarane) and caniojane (diterpene) as potential MtbICL inhibitors.Methods:Virtual screening, molecular docking and MD simulation study has been integrated for screening of phytochemical based anti-mycobacterial compounds. Docking study of reported MtbICL inhibitors has shown an average binding affinity score -7.30 Kcal/mol. In virtual screening, compounds exhibiting lower binding energy than calculated average binding energy were selected as top hit compounds followed by calculation of drug likeness property. Relationship between experimental IC50 value and calculated binding gibbs free energy of reported inhibitors was also calculated through regression analysis to predict IC50 value of potential inhibitors.Background and Introduction:Tuberculosis (TB) is a leading infectious disease caused by Mycobacterium tuberculosiswith high morbidity and mortality. Isocitrate lyase (MtbICL), a key enzyme of glyoxylate pathway has been shown to be involved in mycobacterial persistence, is attractive drug target against persistent tuberculosis.

scholarly journals Yarrowia lipolytica Mutants Devoid of Pyruvate Carboxylase Activity Show an Unusual Growth Phenotype

2005 ◽  
Vol 4 (2) ◽  
pp. 356-364 ◽  
Author(s):  
Carmen-Lisset Flores ◽  
Carlos Gancedo
Keyword(s):  
Yarrowia Lipolytica ◽  
Pyruvate Carboxylase ◽  
Glucose Repression ◽  
Isocitrate Lyase ◽  
Glyoxylate Cycle ◽  
Growth Defect ◽  
Carboxylase Activity ◽  
Spliceosomal Introns ◽  
Eukaryotic Microorganisms ◽  
Growth Phenotype

ABSTRACT We have cloned and characterized the gene PYC1, encoding the unique pyruvate carboxylase in the dimorphic yeast Yarrowia lipolytica. The protein putatively encoded by the cDNA has a length of 1,192 amino acids and shows around 70% identity with pyruvate carboxylases from other organisms. The corresponding genomic DNA possesses an intron of 269 bp located 133 bp downstream of the starting ATG. In the branch motif of the intron, the sequence CCCTAAC, not previously found at this place in spliceosomal introns of Y. lipolytica, was uncovered. Disruption of the PYC1 gene from Y. lipolytica did not abolish growth in glucose-ammonium medium, as is the case in other eukaryotic microorganisms. This unusual growth phenotype was due to an incomplete glucose repression of the function of the glyoxylate cycle, as shown by the lack of growth in that medium of double pyc1 icl1 mutants lacking both pyruvate carboxylase and isocitrate lyase activity. These mutants grew when glutamate, aspartate, or Casamino Acids were added to the glucose-ammonium medium. The cDNA from the Y. lipolytica PYC1 gene complemented the growth defect of a Saccharomyces cerevisiae pyc1 pyc2 mutant, but introduction of either the S. cerevisiae PYC1 or PYC2 gene into Y. lipolytica did not result in detectable pyruvate carboxylase activity or in growth on glucose-ammonium of a Y. lipolytica pyc1 icl1 double mutant.

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