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Biokim 11 Nitrate Assimilation
Biokim 11 Nitrate Assimilation
What are the net reactions of nitrogen fixation and assimilation by plants and bacteria?
Nitrogen fixation is mediated by the nitrogenase enzyme complex:
1.Fiksasi nitrogen dari udara; 2.Assimilasi nitrate dari ammonia yang terkandung dalam air atau tanah.
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Nitrate assimilation in the roots and leaves of a plant. Nitrat harus diubah menjadi NH4+ di dalam tumbuhan sebelum nitrogen masuk ke asam amino dan senyawa nitrogen lainnya Nitrate diambil dari tanah oleh akar. Nitrat dapat disimpan sementara dalam vacuola dari sel akar atau direduksi dalam sel epidermis dan cortex dari akar Kelebihan nitrat dibawa via pembuluh xylem ke sel mesophyll, dimana nitrate dapat disimpan sementara dalam vacuole.
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Nitrate direduksi menjadi nitrite dalam cytosol dan kemudian nitrite direduksi lebih lanjut dalam chloroplast menjadi NH4+, dari mana asam amino terbentuk NH4+ ini digunakan untuk mensintesis glutamine dan asparagine (yang umumnya dinamakan amida)
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Dua asam amino (glutamine dan asparagine) dapat dipindahkan ke daun melalui pembuluh xylem. Saat kapasitas asimilasi nitrat dalam akar berlebihan nitrat dikeluarkan dari akar ke dalam pembuluh xylem dan terbawa ke daun akibat transpirasi. Sejumlah besar nitrat dapat disimpan dalam daun pada vakuola. Terkadang penyimpanan vakuola dapat habis karena asimilasi nitrat di siang hari dan terisi lagi pada saat malam hari Sebagai contoh, daun bayam memiliki kandungan nitrat tertinggi ditemukan pada KRT-2011 pagi hari.
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Nitrate dalam mesophyll cells direduksi menjadi nitrite oleh nitrate reductase yang ada dalam cytosol dan selanjutnya menjadi NH4+oleh nitrite reductase dalam chloroplasts
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KRT-2011
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Reduksi nitrite menjadi ammonia membutuhkan enam elektron Reaksi ini dikatalisis oleh satu enzim, yaitu: the nitrite reductase , yang banyak terdapat dalam plastids. Enzim ini memanfaatkan reduced ferredoxin sebagai electron donor, yang disediakan oleh photosystem I sebagai hasil photosynthetic electron transport
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ATP yang dibutuhkan untuk glutamine synthesis dapat dihasilkan oleh mitochondria dan dipindahkan ke dalam leucoplasts oleh suatu plastid ATP translocator. Glutamate synthase dari leucoplasts juga digunakan mereduksi ferredoxin sebagai redox partner, meskipun beberapa leucoplasts juga mengandung glutamate synthase Reduksi nitrate di dalam akar memberikan organic nitrogen compounds terutama dalam bentuk glutamine dan asparagine pada tunas melalui aliran transpirasi dalam pembuluh xylem. This is also the case bila NH4+ merupakan sumber nitrogen dalam tanah.
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Nitrate assimilation
Nitrate (NO3-) + NADPH + H+ + 2e(uptake in roots) NO2- + H2O + NADP+ Cytoplasm
Nitrate Reductase
Nitrite Reductase
Nitrate assimilation
Nitrate (NO3-) + NADPH + H+ +2e(uptake in roots) NO2- + H2O + NADP+ Cytoplasm
Nitrate Reductase
Nitrate assimilation
Nitrite (NO2-) + 8H+ + 6Fdred + 6e(toxic) NH4+ + 6Fdox + 2H2O Chloroplasts
Nitrite Reductase
Iron-sulfur
Photosynthesis ETC
serine dan glycine, yang terbentuk sebagai intermediate products dalam photorespiratory cycle, juga menunjukkan bagian penting dari total asam amino yang ada dalam mesophyll cells. Sejumlah besar alanine sering terbentuk dalam C4 plants
KRT-2011
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CO2 assimilation provides the carbon skeletons to synthesize the end products of nitrate assimilation CO2 assimilation menyediakan carbon skeletons yang dibutuhkan untuk synthesis bermacam-macam amino acids. Gambar disamping menunjukkan ringkasan dari awal carbon skeletons dari individual amino acids.
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3-Phosphoglycerate merupakan carbon precursor yang sangat penting untuk synthesis of amino acids. Ini dihasilkan dalam Calvin cycle dan diekspor dari chloroplasts ke cytosol melalui triose phosphate-phosphate translocator dalam perubahan phosphate. 3-Phosphoglycerate diubah dalam cytosol oleh phosphoglycerate mutase dan enolase menjadi phosphoenolpyruvate (PEP) From PEP two pathways branch off, the reaction via pyruvate kinase leading to pyruvate, and via PEP-carboxylase to oxaloacetate.
KRT-2011
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Yang berwarna merah adalah: Carbon skeletons untuk synthesis amino acids yang Diperoleh melalui asimilasi CO2 Merupakan Prekursor penting untuk sintesis amino acid
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carbamoyl-phosphate synthetase I
Carbamoyl-phosphate synthetase I catalyzes one of the steps in the urea cycle. Two ATP are consumed : (1) in the activation of HCO3- for reaction with ammonium, (2) in the phosphorylation of the carbamate
glutamate dehydrogenase
Glutamate dehydrogenase (GDH) catalyzes the reductive amination of a-ketoglutarate to yield glutamate.
glutamine synthetase
Glutamine synthetase (GS) catalyzes the ATPdependent amidation of the gamma-carboxyl group of glutamate to form glutamine. The reaction proceeds via a gamma-glutamylphosphate intermediate, and GS activity depends on the presence of divalent cations such as Mg2+ (Figure 26.10).
Glutamine
Glutamine major N donor in the biosynthesis of many organic N compounds such as purines, pyrimidines, and other amino acids. The amide-N of glutamine provides the nitrogen atom in these biosyntheses. GDH and GS responsible for most of the ammonium assimilated into organic compounds.
Ammonium Assimilation
Two principal pathways Principal route: GDH+GS in organisms rich in N Secondary route: GS+GOGAT in organisms confronting N limitation GOGAT is glutamate synthase glutamate:oxo-glutarate amino transferase
Ammonium Assimilation
(2 Principles Pathway)
In organisms that enjoy environments rich in nitrogen, GDH and GS acting in sequence furnish the principal route of NH4+ incorporation GDH generates glutamate & GS use glutamate to form glutamine. Green plants which grow under conditions where little NH4+ is available GDH is not effective need another option to generate glutamate which GS can use to form glutamine.
Ammonium Assimilation
(2 Principles Pathway) Limited availibility of NH4+ creates the need for an alternative mode of glutamate synthesis to replenish the glutamate consumed by the GS reaction. This need is filled by glutamate synthase (also known as GOGAT, the acronym for the other name of this enzyme - glutamate:oxo-glutarate aminotransferase). Glutamate synthase catalyzes the reductive amination of a-keto-glutarate using the amide-N of glutamine as the N donor to generate glutamate
Two glutamates are formed from amination of aketoglutarate and the other from deamidation of Gln These glutamates can now serve as ammonium acceptors for glutamine synthesis by GS. Organisms variously use NADH, NADPH, or reduced ferredoxin as reductant.
Together, GS and GOGAT constitute a second pathway of ammonium assimilation, in which GS is the only NH4+-fixing step; the role of GOGAT is to regenerate glutamate (Figure 26.13).
Note that most often, this reaction releases NH4+ from glutamate in other contexts.
In this mechanism, the enzyme glutamine synthetase (GS) uses ATP in a coupled reaction to form glutamine from glutamate using NH4+.
Importantly, the newly acquired nitrogen in glutamate and glutamine is used to synthesize a variety of other amino acids through aminotransferase enzymes such as aspartate aminotransferase.
Biosynthesis of proline and arginine from glutamate in bacteria. All five carbon atoms of proline arise from glutamate. In many organisms, glutamate dehydrogenase is unusual in that it uses either NADH or NADPH as a cofactor.
The -semialdehyde in the proline pathway undergoes a rapid, reversible cyclization to 1pyrroline-5carboxylate (P5C), with the equilibrium favoring P5C formation. Cyclization is averted in the ornithine/arginine pathway by acetylation of the amino group of glutamate in the first step and removal of the acetyl group after the transamination.
Biosynthesis of serine from 3phosphoglycerate and of glycine from serine in all organisms. Glycine is also made from CO2 and NH4+ by the action of glycine synthase, with N5,N10-methylenetetrahydrofolate as methyl group donor
Biosynthesis of cysteine from serine in bacteria and plants. The origin of reduced sulfur is shown in the pathway on the right.
Biosynthesis of cysteine from homocysteine and serine in mammals. The homocysteine is formed from methionine.