The Metabolism of Labelled Lysine and Pipecolic Acid by Acacia Phyllodes

C¹⁴-lysine and C¹⁴- and H³-pipecolic acids have been used tostudy the metabolism of the lysine family of amino-acids inAcacia, which contains ₄-hydroxypipecolic acid as a characteristiccomponent of the soluble-nitrogen fraction. Degradation of C¹⁴-lysinewas rapid and was far more extensive than that observed earlierin higher plants. Pipecolic acid was the major radioactive productin short-term experiments. After longer metabolic periods, radioactivitywas distributed over a wide range of amino-acids, organic acids,and sugars. A tentative metabolic scheme is produced to explainthese observations concerning lysine degradation. The distributionof radioactivity in the amino-acids of the protein present inthe phyllodes was determined 24 hours after supplying C¹⁴-lysine.Specific activities of free and bound amino-acids are comparedat this time. Hydroxyproline forms a notable component of thephyllode protein. Pipecolic acid degradation has been demonstrated for the firsttime in a biological system. The breakdown pathway was studiedin Acacia phyllodes using H²-pipecolic acid. Substances tentativelyidentified as '-piperdine-2-carboxylic acid and -amino--hydroxycaproicacid were amongst the early degradation products. Ultimately,lysine and -aminoadipic acid became labelled. In contrast tothe experiments with C organic acids and sugars did not becomeradioactive. The explanation of this finding is probably tobe found in the ease with which H³ atoms present in certainchemical groupings may undergo exchange with normal hydrogenatoms of water molecules. The biosynthesis of 4-hydroxypipecolic acid probably involvesa direct hydroxylation of the parent imino-acid.     

The Incorporation of C14-labelled Substrates into the Amino-acids of Groundnut Plants (Arachis hypogaea)

The amino-acid metabolism of groundnut plants has been studiedwith special reference to -methyleneglutamic acid (-MGA) andy-methyleneglutamine (-MG), constituents not found in the greatmajority of plant species. The sequence in which C¹⁴ from radioactive C¹⁴-carbon dioxideenters the amino-acids of leaves was determined. The patternof labelling was very similar to that found for leaves of otherspecies. The metabolic relationships existing between photosynthesisand amino-acid synthesis therefore do not seem to be affectedby the large quantities of -MGA and -MG present in the leaves.-MGA and -MG only gained traces of radioactivity. Experiments designed to study the incorporation of C¹⁴ fromuniformly labelled C¹⁴-alanine into the amino-acids of roots,immature leaves and cotyledons of seedlings and young plantsindicated that the main site of synthesis of -MGA was the cotyledons. Various C^I4-labelled substrates were fed to germinating seedsand, after a period of growth, the specific activities of theamino-acids of seedlings receiving different treatments weredetermined. Comparison of the specific activities enabled certaindeductions to be made concerning the probable biosynthetic pathwaysleading to -MGA and -MG. The results were consistent with theintact incorporation of pyruvate molecules, or another related3-carbon-atom containing compound, into -MGA.     

Tramsamination and the Oxidation of Some Amino-acids by Mung Bean Mitochondria

The transaminases for the reaction of -ketoglutarate with theamino-acids L-alanine, L-aspartic acid, glycine, L-leucine,L-ornithine, and L-valine have been shown to be associated withmung bean mitochondria. Aspartate-glutamate and alanine-glutamatetransaminases are two different enzymes both of which are reversibleand stimulated by pyridoxal-5'-phosphate. Aspartate is deaminated, with associated oxygen uptake, by mitochondriafrom 1-day 8eedlings, but there is no oxygen uptake with 2-daymitochondria and aspartate, nor with 1-or 2-day mitochondriaand L-alanine, glycine, L-leucine, or L-valine although theseamino-acids are all deaminated. The change in aspartate metabolismwith increased age of the seedlings is probably due to the reorganizationof the enzyme systems which resulted in DPNH being largely oxidizedby malic dehydrogenase and oxaloacetate, instead of mainly bya DPNH oxidase.     

Protein Composition in Relation to Age of Daffodil Leaves

Daffodil foliage leaves were divided into sections along theirlength; the basal sections then contained the youngest, growingregions of the leaves, and the other sections represented progressivelyolder tissue as the leaf apex was approached. Representativeprotein fractions were isolated from some of these sections,and after hydrolysis their amino-acid compositions were compared.Protein from bulb scale leaves was also analysed. Within thefoliage leaf, age did not markedly affect the composition ofthe proteins. Larger differences of composition were found whenthe proteins of the bulb scale, a typical storage tissue, werecompared with those of the foliage leaves. The free amino-acid complements of the different sections ofthe foliage leaves were also compared. Variation of compositionwith leaf age did occur, but no generalizations can be madethat are applicable to all amino-acids.     

{gamma}-Amiobutyric Acid Metabolism in Plants: Part 1. Metabolism in Yeasts

The metabolism of -aminobutyric acid (AB) by two yeasts, Saccharomycescerevisiae and Torulopsis utilis, was investigated. Both yeastsgrew well upon AB as a sole source of nitrogen (N), and thelag phase for Torulopsis was shorter than when provided the N-source. The metabolism of AB by Torulopsis, whichwas associated with an increased O₂ uptake, was adaptive incharacter. The enzyme whose formation was induced by the supplyof AB was a transaminase, which was apparently specific forAB as the amino donor. Small amounts of transaminase were presentin unadapted, -grown cells. The optimum pH, equilibrium constant, Michaelis' constant, and coenzyme requirementwere investigated for the transamination reaction involving-ketoglutaric acid (KG) as amino group acceptor. Succinic semi-aldehyde(SSA) was a product of this transamination reaction.The possibility;that some AB was converted into SSA by a direct oxidative deaminationremained unconfirmed. The further conversion of SSA into succinic acid was establishedusing intact. cells for both yeasts. This oxidation processwas shown to be linked to the reduction of pyridine nucleotidesvising extracts of Saccharomyces as a source of SSA dehydrogenase.Dehydrogenase activity could be ascribed to two separate enzymes,one linked to DPN, and the other utilizing TPN and requiringMg⁺⁺ as an activator. The properties of the former enzyme, whichwas more important quantitatively, were investigated and comparedwith those described in the literature for an aldehyde dehydrogenaseof baker's yeast and for SSA dehydro-genases of Pseudomonas.Torulopsis extracts could catalyse the reduction of SSA to -hydroxybutyricacid (OHB); the OHB dehydrogenase involved required TPNH asa coenzyme. Certain other properties of this enzyme are recorded. The possibility is discussed that AB and SSA act as intermediatesin a metabolic pathway that may form a by-pass of the KG-succinatestage of the tricarboxylic acid cycle.     

The Deamidase of Groundnut Plants (Arachis hypogaea)

The deamidase enzyme system present in extracts of groundnutplant tissues has been studied. The distribution of the enzymewithin the different organs of the plant were determined, andsome of the properties of the enzyme present in leaf extractsare recorded. From its substrate specificity the enzyme is probablybest regarded as a -methyleneglutaininase, although it catalysesthe hydrolysis of glutamine at about one-fifth of the rate of-methyleneglutamine. The enzyme may play an important role inthe over-all nitrogen metabolism of the plant, as well as controllingthe relative concentrations of -methyleneglutamine and -methyleneglutamicacid in the different organs. The properties of the enzyme havebeen compared with those recorded in the literature of glutaminasesand asparaginases, and certain common features are apparent.The new deamidase was, however, more stable to certain denaturingtreatments than were the other types of deamidase.