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9/7/2009

Amino acid biosynthesis in plants

Aromatic amino acids and herbicides


Plants, algae, bacteria, fungi Function? Origin of carbon sceleton? Why important to know about their synthesis? Knowledge from bacteria is not enough. Why? Amino acid biosynthesis as a target for herbicides

Glycolysis

Calvin cycle Oxidative pentose Phosphate pathway

Citric acid cycle

GS/GOGAT cycle
Mito/ chloro

Aminotransferases

Chloroplast

Km 10-80 mM

Km 3-5M

GS1: cytosol, low in leaf high in roots, phloem GS2: chloroplast, mesophyll cells, leaves non-photosynthetic

Subcellular localization: Isoenzymes in cytosol, mitochondria, Chloroplasts and peroxisomes Km values and substrate specificity varies

Transaminases have pyridoxal phosphate as cofoactor

Biosynthesis of aromatic amino acids


Plants are autotrophic: Able to synthesize all required organic compounds from carbon dioxide and inorganic nutrients

Pyridoxal phosphate Plus transaminase

Human and animals are heterotrophic: Vitamins, essential amino acids

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Biosynthesis of aromatic amino acids


Phenylalanine, tyrosine, tryptophan Shikimic acid pathway Phenylalanine and tryptophan are essential amino acids like histidin, lysine, arginine, threonin, valine, leucine, isoleucine and methionine

Biosynthesis of aromatic amino acids


Shikimic acid pathway: Plants, fungi and bacteria
What are aromatic amino acids used for in plants and microorganisms?

Biosynthesis of aromatic amino acids


Microorganisms use about 90% of their energy for protein synthesis In plants, about 35% of plant dry matter is based on aromatic amino acids derived from the shikimate pathway 20% of carbon fixed by photosynthesis passes through the shikimate pathway 7 x 1012 ton C per year

Biosynthesis of aromatic amino acids


Primary metabolism:
protein synthesis vitamin E and K precursors folic acid precursors ubiquinone and plastoquinone precursors

Secondary metabolism:
flavonoids alkaloids phenylpropanoids cyanogenic glucosides lignin

Biosynthesis of aromatic amino acids


Marques et al 2007 Inhibition of 5-enolpuruvylshikimate-3-phosphate synthase as a model for novel antimicrobials Tuberculosis: chemotherapy with front line drugs Expensive Multi drug resistant strains Malaria

Biosynthesis of aromatic amino acids


Apicomplexan Parasites: Eucaryotic, unicellular spore forming parasites of animals Protists possessing a unique organelle called the apicoplast Apicoplast is a chloroplast related organelle Genome sequencing revealed the presence of genes related to aromatic amino acid synthesis Target as selective inhibitors

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Shikimate pathway

Tree of life

glycolysis

Calvin cycle

branchpoint

Tetrahydrofolate Vitamin A

1.6 billion years

IAA

Chorismate biosynthesis
4 1 5 2 6 3 7 Bacteria: Separate enzymes catalysing each of the seven steps Fungi: Step 2 to 7 catalyzed by a single polypeptide Plants: Step 3 and 4 Catalyzed by the same enzyme

DAHP reaction mechanism

Aldole condensation

Chorismate biosynthesis
4 1 5 2
Cofactors: NAD, Co2+

Bacteria: Separate enzymes catalysing each of the seven steps Fungi: Step 2 to 7 catalyzed by a single polypeptide

6 x

3 x 7

Plants: Step 3 and 4 Catalyzed by the same enzyme

Arogenate pathway in plants

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Chorismate mutase reaction mechanism

Arogenate pathway in plants

Prephenate to phe and tyr

Transfer of hydrogen to the pyridin ring as a hydride ion

NAD or NADP

NADH or NADPH

Shikimate pathway

Chorismate mutase is a committing enzyme

branchpoint

Tetrahydrofolate Vitamin A

Two isoforms CMI feed back inhibited by phenylalanine and tyrosine, activated by tryptophan CMII not regulated by aromatic amino acids

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Chorismate mutase isoforms have different subcellular localization


CMI targeted to Chloroplast CMII Cytoplasm Possible function???

Arogenate dehydratase and dehydrogenase is inhibited by phe and tyr

Arogenate pathway

Round-Up reaction mechanisms

Amino acid synthesis proceeds in the chloroplasts Why has this been so difficult to resolve?

Shikimate pathway

EPSP synthase

Marques, p. 447

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Model of EPSP synthase

C N

Two similar domains each with three domains of folds Only other enzyme with similar architechture is UDP-N-acetylglucosamine enolpyruvyl transferase

Glyphosate inhibition

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Round-Up degradation pathways