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Isocitrate Lyase Plays Important Roles in Plant Salt Tolerance

Contents

Introduction

Background

Results

Discussion

References

Introduction to Development of Micro Bodies in Sunflower with Respect to Glyoxylate Cycle

Several plants and microorganisms work on acetate that produces, acetyl CoA. This can only be accomplished through a metabolic pathway that converts two carbon acetyl molecules to four carbon acetyl molecules. This byproduct of four carbon-acetyl units is called as succinate. The significance of this pathway is to synthesise energy. Therefore, the glyoxylate cycle is a different version of the TCA cycle i.e. the tricarboxylic acid cycle. This could be a replacement in the plants, bacterias and other organisms like the fungi and the protists. The Glyoxylate cycle is an anabolic pathway that produces energy and is necessary to take place for the synthesis of carbohydrates. Bacterias, fungi, protists and plants can carry out the conversion of fat into carbohydrates in successful amounts whereas the humans and the animals cannot produce carbohydrates from fat in considerable amounts. (Eprintsev, A. T., Fedorin, D. N., Sazonova, O. V., & Igamberdiev, A. U., 2018) In this experiment, sunflower germination was examined and the metabolic changes were considered for proving the certain laid points like the enzymatic activity and the sugar content change in context to the 14 days of seed germination.

Biology engineers have evolved the idea of genetic engineering or cloning and have come up with a new technique of increasing the yield of a few substances or materials to reduce the economic rates and also the time consumption. Different kinds of catabolic and anabolic cycles are modified and altered in the cells of a mammal, obstructing the regular cellular function which may result in positive changes but at times could change the metabolic significance. For instance, the cellular metabolism in a sheep has to be genetically engineered to yield a greater amount of wool. This yield is hampered due to less amount of glucose. In this assignment, we studied the engineered cycle, wherein the gene is extracted and further sequenced to incorporate new properties. The E.coli is considered for the experiment and then the AceA gene responsible for coding for isocitrate lyase enzyme followed by AceB gene responsible for encoding maltase enzyme is isolated and altered for introducing it in a mammalian cell like the mice or the liver cells. This engineering was a success but there was no impact seen for the complex animals.

The glyoxylate cycle begins with the formation of citrate by the condensation of oxaloacetate followed by acetyl CoA. This proceeds to the next step of the cycle accounting to the isomerisation and forming isocitrate. There are several enzymes involved in the process. Usually, the vertebrates use fatty acids as an energy source that is decomposed through beta-oxidation from the lipids forming acetate molecules.

Here comes up the coenzyme A, the acetate molecule formed combines with the thiol group of the coenzyme A producing acetyl CoA. This produced Acetyl CoA jumps into the TCA cycle and is further oxidized to carbon dioxide. To synthesise carbohydrates from fat, glyoxylate cycle is identical to the Citric acid or the TCA cycle in the initial steps and successfully converts fat molecules to carbohydrates in plants, bacteria, fungi, and protists like organisms. (Faraoni, P., Sereni, E., Gnerucci, A., Cialdai, F., Monici, M., & Ranaldi, F., 2019)

In plants, the glyoxylate pathway occurs in the cell organelle called glyoxysomes. With the help of gluconeogenesis and the TCA cycle together, succinate which is a by-product of the glyoxylate cycle can be changed into carbohydrates molecules. Therefore, the glyoxylate cycle provides an upper hand to the organism in metabolic variations. (Gonçalves, I. L., Mielniczki-Pereira, A. A., Borges, A. C. P., & Valduga, A. T., 2016)

In the vertebrates, the isocitric lyase and malate succinate enzymes are absent which makes them unable to undergo glyoxylate cycle, however, some studies show the presence of glyoxylate cycle in a few vertebrates like the hens.

2Acetyl CoA+ NAD+ 2H2O → succinate + 2CoA + NADH + 2H

Background of Development of Micro Bodies in Sunflower with Respect to Glyoxylate Cycle

Acetate is an inexpensive and predominantly found biochemical. In a lignocelluloses biomass pretreatment, it is obtained as a promising product. Apart from this, acetate is also generated from organic waste decomposition. During fermentation also, acetate is inevitably produced. But, due to the toxic composition of acetate to the microorganisms, it cannot be dealt with effectively. Several researchers have come up with the conversion of the acetate into beneficial chemicals that could prove to be successfully promising feedstock. This has to be achieved due to the financial and ethical problems by using the grain crop-based feedstock. So, the acetate feedstock could be a good source that is economic and does not hinder the ethics. (Wu, W. L., Hsiao, Y. Y., Lu, H. C., Liang, C. K., Fu, C. H., Huang, T. H., ... & Tsai, W. C., 2020)

Considering tyrosine, it is a non-essential amino acid that can efficiently be synthesized in the human body. In the pharmaceutical industry, cosmetic industries, and the food market, tyrosine is a boon. It can be of great use and therefore, multiple extraction techniques have been enquired and researched for tyrosine and using it economically. The plant and animal extract represented reduced amounts of tyrosine and therefore, an alternative source could be reliable. Extracting tyrosine from biomass-derived sugar has been tried. Apart from this, there have been successive experiments to produce tyrosine from genetically modified microorganisms. (Ma, Z., Marsolais, F., Bernards, M. A., Sumarah, M. W., Bykova, N. V., & Igamberdiev, A. U., 2016)

In this experimental research we, check the regulation of the enzyme isocitrate lyase in the germination of Helianthus Annus.

Results of Development of Micro Bodies in Sunflower with Respect to Glyoxylate Cycle

Oil is utterly crucial for many plants. When the plant completes germination, the stored oil amount is then transported to the plant’s required parts for its growth and development of the seed. Glyoxylate cycle provides an opportunity to the plant for producing carbohydrates from the lipids and the acetyl CoA successfully uses fatty acids for the further metabolic conversions. (Dwevedi, A., 2016) The enzyme isocitrate lyase makes the entire difference and hence, a mutant enzyme taking place of the isocitrate lyase enzyme has been introduced for testing in vivo glyoxylate cycle and its biochemical role.

The enzyme isocitrate lyase is the most significant enzyme while talking about the glyoxylate cycle. (Bhusal RP, Bashiri G, Kwai BX, Sperry J, Leung IK, July 2017). As stated previously, it plays a crucial role in germination followed by the growth of the plant. The enzymatic action drastically shoots at the time of seed germination but comes back to normal after a couple of days when the plants become photosynthetic competent. Since the isocitrate lyase has proven to be unruly to the experimental analysis by the biochemical trials. The major gene coding for the enzyme isocitrate lyase would come up with considerable data and details required for the protein by sequencing the DNA. A change can be observed with the levels of the isocitrate lyase and the germination time in sunflower. This can be demonstrated by the below graphical representation.

Discussion on Development of Micro Bodies in Sunflower with Respect to Glyoxylate Cycle

The above-mentioned pathways significantly demonstrate the differences amongst each other. The glyoxylate cycle allows plants and some microorganisms to grow on acetate because the cycle bypasses the decarboxylation steps of the citric acid cycle. The enzymes that permit the conversion of acetate into succinate-isocitrate lyase and malate synthase-are boxed in blue. (Fig.4)

The enzymes like glyoxisomes and peroxisomal enzymes are observed to display the varied role of action during germination as stated earlier, the action of isocitrate lyase enzyme heightens during a couple of days but then gets normalized till day 9 or 10. There is a significant reduction of 88 percent. When the sunflower plant is exposed to the sunlight the fall increase. (Roode, E. C. (2017). Considering the sucrose levels, the reducing sugar gradient improved in the initial stages of day 3 to 4. Further days, it stayed constant even when exposed to the light. There was no significant change after the fourth day. The complete action of the regular micro body marker, catalase, grown also to isocitrate lyase, yet diminished just 72% by day 9. The particular exercises of compounds (catalase, malate dehydrogenase, and aspartate aminotransferase) regular to both micro body frameworks were 10-to 1000-overlap more noteworthy than those of different chemicals. It is recommended that malate and aspartate might be engaged with hydrogen transport among micro bodies and other cell destinations. Glutamate-glyoxylate aminotransferase was dynamic in micro bodies from castor bean endosperm and sunflower cotyledons. The particular movement of this aminotransferase grew comparatively to glyoxysomal catalysts in obscurity however further expanded in the light, as did peroxisomal compounds. The microbody division of castor bean endosperm sprouted in obscurity for 5 days contained both glyoxysomal and peroxisomal compounds of comparative explicit movement.( Yuenyong, W., Sirikantaramas, S., Qu, L. J., & Buaboocha, T. 2019) Nearby the micro body portion on sucrose inclinations from sunflower cotyledons were etioplasts at marginally lower densities and protein bodies at comparative and higher densities. Their quality in the micro body parts brought about misleadingly low explicit exercises.

References for Development of Micro Bodies in Sunflower with Respect to Glyoxylate Cycle

Bhusal RP, Bashiri G, Kwai BX, Sperry J, Leung IK (July 2017). "Targeting isocitrate lyase for the treatment of latent tuberculosis". Drug Discovery Today. 22 (7): 1008–1016

Corpas, F. J. (2019). Peroxisomes in higher plants: an example of metabolic adaptability. Botany Letters, 166(3), 298-308.

Dwevedi, A. (2016). Enzyme immobilization: advances in industry, agriculture, medicine, and the environment. Springer.

Eprintsev, A. T., Fedorin, D. N., Sazonova, O. V., & Igamberdiev, A. U. (2018). Expression and properties of the mitochondrial and cytosolic forms of fumarase in sunflower cotyledons. Plant Physiology and Biochemistry, 129, 305-309.

Faraoni, P., Sereni, E., Gnerucci, A., Cialdai, F., Monici, M., & Ranaldi, F. (2019). Glyoxylate cycle activity in Pinus pinea seeds during germination in altered gravity conditions. Plant Physiology and Biochemistry, 139, 389-394.

Gonçalves, I. L., Mielniczki-Pereira, A. A., Borges, A. C. P., & Valduga, A. T. (2016). Metabolic modeling and comparative biochemistry in glyoxylate cycle. Acta Scientiarum. Biological Sciences, 38(1), 1-6.

Ma, Z., Marsolais, F., Bernards, M. A., Sumarah, M. W., Bykova, N. V., & Igamberdiev, A. U. (2016). Glyoxylate cycle and metabolism of organic acids in the scutellum of barley seeds during germination. Plant Science, 248, 37-44.

Roode, E. C. (2017). The effect of exogenous DIM on Brassica napus and its role in response to heavy metal stress.

Wu, W. L., Hsiao, Y. Y., Lu, H. C., Liang, C. K., Fu, C. H., Huang, T. H., ... & Tsai, W. C. (2020). Expression regulation of MALATE SYNTHASE involved in glyoxylate cycle during protocorm development in Phalaenopsis aphrodite (Orchidaceae). Scientific reports, 10(1), 1-16.

Yuenyong, W., Sirikantaramas, S., Qu, L. J., & Buaboocha, T. (2019). Isocitrate lyase plays important roles in plant salt tolerance. BMC plant biology, 19(1), 472.

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