Key Enzymes Of Carbon Metabolism Research Articles

Effect of Ca2+ signal on the activity of key enzymes of carbon metabolism and related gene expression in yeast under high sugar fermentation.

Saccharomyces cerevisiae is a fungus widely used in the food industry and biofuel industry, whereas it is usually exposed to high sugar stress during the fermentation process. Ca2+ is a key second messenger of the cell, it can regulate cell metabolism. The present study investigated the effect of the Ca2+ signal on the activity of key enzymes of carbon metabolism and related gene expression in yeast under high sugar fermentation. The expression of genes encoding hexokinase was up-regulated in the high sugar environment, the activity of hexokinase was increased, glucose transmembrane transport capacity was enhanced, the ability of glucose to enter into glycolytic metabolism was increased, and the expression of genes related to pentose phosphate metabolism, glycerol metabolism and trehalose metabolism was up-regulated in the high glucose with Ca2+ group. Ca2+ signal regulates the cellular metabolism of glycerol and trehalose and optimizes the allocation of carbon flow by regulating the key enzymes and related gene expression to enhance the resistance of yeast to high sugar stress. © 2024 Society of Chemical Industry.

Effects of elevated nitrogen fertilizer on the multi-level structure and thermal properties of rice starch granules and their relationship with chalkiness traits.

Chalkiness in rice reduces its market value and affects consumer acceptance. Research on the mechanism of chalkiness formation has focused primarily on the activity of key enzymes of carbon metabolism and starch accumulation. The relationship between the formation of chalkiness induced by N fertilizer and rice starch's multi-level structure and thermal properties still needs to be fully elucidated. In this study, the rates of chalky grains and degree of chalkiness decreased with the increase in N fertilizer dosage. This was attributed to an increased proportion of short chains, ordered structure carbon chains, small starch granules, and branched starches, and a higher degree of crystallinity and ΔHg in protein, and a decreased proportion of amylose, large starch granules, and weighted average diameter of starch granule surface area and volume. Application of N fertilizer promoted an increased proportion of short-branched chain amylopectin to develop a more ordered carbohydrate structure and crystalline lamella. These effects enhanced the normal development and compactness of starch granules in grains, and improved their arrangement morphology, thereby reducing the chalkiness in rice. These changes in starch multi-level structure and protein improve the physicochemical characteristics of starch and enhance the fullness, crystallinity and compactness of starch granules, while synergistically increasing the regularity and homogeneity of starch granules and thus optimizing the stacking pattern of starch granules, leading to a reduction in rice chalkiness under nitrogen fertilization and thus improving the appearance of rice. © 2023 Society of Chemical Industry.

Irrigation regimes modulate non-structural carbohydrate remobilization and improve grain filling in rice (Oryza sativa L.) by regulating starch metabolism

Recently developed ‘super’ rice cultivars with greater yield potentials often suffer from the problem of poor grain filling, especially in inferior spikelets. Here, we studied the activities of enzymes related to starch metabolism in rice stems and grains, and the microstructures related to carbohydrate accumulation and transportation to investigate the effects of different water regimes on grain filling. Two ‘super’ rice cultivars were grown under two irrigation regimes of well-watered (WW) and alternate wetting and moderate soil drying (AWMD). Compared with the WW treatment, the activities of ADP glucose pyrophosphorylase (AGPase), starch synthase (StSase) and starch branching enzyme (SBE), and the accumulation of non-structural carbohydrates (NSCs) in the stems before heading were significantly improved, and more starch granules were stored in the stems in the AWMD treatment. After heading, the activities of α-amylase, β-amylase, sucrose phosphate synthase (SPS) and sucrose synthase in the synthetic direction (SSs) were increased in the stems to promote the remobilization of NSCs for grain filling under AWMD. During grain filling, the enzymatic activities of sucrose synthase in the cleavage direction (SSc), AGPase, StSase and SBE in the inferior spikelets were increased, which promoted grain filling, especially for the inferior spikelets under AWMD. However, there were no significant differences in vascular microstructures. The grain yield and grain weight could be improved by 13.1 and 7.5%, respectively, by optimizing of the irrigation regime. We concluded that the low activities of key enzymes in carbon metabolism is the key limitation for the poor grain filling, as opposed to the vascular microstructures, and AWMD can increase the amount of NSC accumulation in the stems before heading, improve the utilization rate of NSCs after heading, and increase the grain filling, especially in the inferior spikelets, by altering the activities of key enzymes in carbon metabolism.

Metabolomic analysis reveals the impact of ketoprofen on carbon and nitrogen metabolism in rice (Oryza sativa L.) seedling leaves.

Pharmacologically active compounds (PACs) are becoming common pollutants in the natural environment, posing potential risks to crop quality; however, the toxic effects and metabolic changes that they cause in agricultural plants remain unclear. Here, we investigated the effects of ketoprofen on respiration rate, ATP synthesis, carbon and nitrogen metabolism, and metabolomics in rice seedling leaves. The results showed that ketoprofen treatment adversely affected the respiration rate, ATP content, H+-ATPase activity and induced changes in the contents of carbon assimilation products (soluble sugar, reducing sugar, sucrose, and starch) and the activities of key enzymes in carbon metabolism (sucrose synthase (SS), sucrose phosphate synthase (SPS), and sucrose invertase (InV)). The contents of nitrate, ammonium, and free amino acids, and the activities of key enzymes involved in nitrogen metabolism (nitrate reductase (NR), nitrite reductase (NiR), glutamine synthetase (GS), glutamate synthase (GOGAT), and glutamate dehydrogenase (GDH)) were also affected in a concentration-dependent manner. Metabolomics analysis showed that ketoprofen disturbed the type and content of metabolites (amino acids, carbohydrates, and secondary metabolites) to varying degrees and perturbed key metabolic pathways (substance synthesis and energy metabolism), ultimately resulting in the reduction of rice seedling biomass. This study provides important information and a useful reference for the accurate assessment of the environmental risks of PACs.

Quantitative Proteomics Reveals Metabolic Reprogramming in Host Cells Induced by Trophozoites and Intermediate Subunit of Gal/GalNAc Lectins from Entamoeba histolytica.

ABSTRACTEntamoeba histolytica is an intestinal protozoan parasite with remarkable ability to kill and phagocytose host cells, causing amoebic colitis and extraintestinal abscesses. The intermediate subunit (Igl) of galactose (Gal)- and N-acetyl-d-galactosamine (GalNAc)-specific lectins is considered an important surface antigen involved in the pathogenesis of E. histolytica. Here, we applied mass spectrometry-based quantitative proteomics technology to analyze the protein expression profile changes occurring in host Caco2 cells incubated with E. histolytica trophozoites or stimulated by purified native Igl protein. The expression levels of 1,490 and 489 proteins were significantly altered in the E. histolytica-treated and Igl-treated groups, respectively, among 6,875 proteins totally identified. Intriguingly, central carbon metabolism of host cells was suppressed in both E. histolytica-treated and Igl-treated groups, with evidence of decreased expression levels of several key enzymes, including pyruvate kinase muscle type 2, presenting a Warburg-like effect in host cells. Besides, Igl had potential physical interactions with central carbon metabolism enzymes and the proteolytic degradation family members proteasome subunit alpha and beta, which may be responsible for the degradation of key enzymes in carbon metabolism. These results provided a novel perspective on the pathogenic mechanism of E. histolytica and compelling evidence supporting the important role of Igl in the virulence of E. histolytica.IMPORTANCE Metabolic reprogramming is considered a hallmark of some infectious diseases. However, in amoebiasis, a neglected tropical disease caused by protozoan parasite E. histolytica, metabolic changes in host cells have yet to be proven. In this study, advanced data-independent acquisition mass spectrometry-based quantitative proteomics was applied to investigate the overall host cellular metabolic changes as high-throughput proteomics could measure molecular changes in a cell or tissue with high efficiency. Enrichment analysis of differentially expressed proteins showed biological processes and cellular pathways related to amoeba infection and Igl cytotoxicity. Specifically, central carbon metabolism of host cells was dramatically suppressed in both E. histolytica-treated and Igl-treated groups, indicating the occurrence of a Warburg-like effect induced by trophozoites or Igl from E. histolytica. Distinct differences in ubiquitin-mediated proteolysis, rapamycin (mTOR) signaling pathway, autophagy, endocytosis, and tight junctions provided novel perspectives on the pathogenic mechanism of E. histolytica.

Effects of Drought Stress on Key Enzymes of Carbon Metabolism, Photosynthetic Characteristics and Agronomic Traits of Soybean at the Flowering Stage under Different Soil Substrates

Soybean is an important legume food crop, and its seeds are rich in nutrients, providing humans and animals with edible oil and protein feed. However, soybean is sensitive to water requirements, and drought is an important factor limiting soybean yield and quality. This study used Heinong 84 (drought resistant variety) and Hefeng 46 (intermediate variety) as tested varieties planted in chernozem, albic, and black soils. The effects of drought stress on the activities of key enzymes in carbon metabolism and photosynthetic characteristics of soybean were studied during the flowering stage, most sensitive to water. (1) The activities of SS-1, 6PGDH, and G6PDH enzymes in soybean leaves first increased and then decreased under drought stress. The enzyme activity was the highest under moderate drought stress and weakest in the blank group. (2) Drought stress increased Phi2, PhiNO, and Fm in soybean leaves and reached the highest value under severe drought; with the increase in drought stress, PhiNPQ and Fv/Fm of soybean leaves gradually decreased, reaching the lowest under severe drought. (3) With the increase in drought stress, F0 and Fs of soybean leaves showed a single peak curve, and the maximum was at moderate drought. (4) Correlation analysis showed that F0 was greatly affected by varieties and soil types; Fs, F0, and Fm soil varieties had a great influence, and chlorophyll fluorescence parameters were affected differently under drought stress with different drought degrees. (5) Drought stress changed the agronomic traits and yield of soybean. With the increase of drought degree, plant height, node number of main stem, effective pod number, 100-seed weight and total yield decreased continuously. (6) Drought stress affected the dry matter accumulation of soybean. With the increase of drought degree, the dry matter accumulation gradually decreased. Among them, the leaf was most seriously affected by drought, and SD decreased by about 55% compared with CK. Under the condition of black soil, the dry matter accumulation of soybean was least affected by drought.

Comparative Proteomic Analysis of Cucumber Fruits Under Nitrogen Deficiency At the Fruiting Stage

Nitrogen deficiency is a major factor that affects the yield and quality of horticultural crops. Differentially expressed proteins were identified in cucumber fruit under nitrogen deficiency treatment, and are expected to be highly useful for dissecting carbon, nitrogen and ascorbate metabolism in cucumber and for improving nitrogen fertilizer utilization. Cucumber plants grown hydroponically in Hoagland solution were transferred to nitrogen-free conditions for 3 or 5 d after the blossom of female flowers. Fruit proteome profiles were assessed using two-dimensional electrophoresis. The relative expression of the genes encoding 50 selected differentially expressed proteins and 4 key enzymes in carbon metabolism was analyzed by qRT-PCR. Resulted showed that most of the identified differentially expressed proteins were mainly associated with carbon metabolism, amino acid biosynthesis, ascorbate metabolism, and proteasomes. Glucose phosphorylation was enhanced, while the pentose phosphate pathway was inhibited. Carbon metabolism and the synthesis of most amino acids was affected significantly, and ascorbate might be accumulated under nitrogen deficiency in cucumber fruit.

Pleiotropic Effect of GNP1 Underlying Grain Number per Panicle on Sink, Source and Flow in Rice.

Rice yield potential is largely determined by the balance among source capacity, sink strength, and flow fluency. Our previous study indicated that the gene GNP1 encoding gibberellin biosynthesis gene GA20ox1 affects grain number per panicle (GNP) in rice, thus resulting in increase of grain yield. To clarify GNP1 effect on sink, source and flow in regulating rice grain yield, we compared Lemont, a japonica (geng) cultivar, with its near-isogenic line (NIL-GNP1TQ) in Lemont background with introgression of the allele at GNP1 from Teqing, a high-yielding indica (xian) cultivar. NIL-GNP1TQ exhibited averagely 32.8% more GNP than Lemont with the compensation by reduced seed setting rate, panicle number and single-grain weight. However, NIL-GNP1TQ still produced averagely 7.2% higher grain yield than Lemont in two years, mainly attributed to significantly more filled grain number per panicle, and greater vascular system contributing to photoassimilates transport to spikelets. The significantly decreased grain weight of superior spikelets (SS) in NIL-GNP1TQ was ascribed to a significant decrease of grain size while the significantly decreased grain weight of inferior spikelets (IS) ascribed to both grain size and poor grain-filling as compared with Lemont. The low activities of key enzymes of carbon metabolism might account for the poor grain-filling in IS, which resulted in more unfilled grains or small grain bulk density in NIL-GNP1TQ. In addition, low seed setting rate and grain weight of IS in NIL-GNP1TQ might be partially resulted from significantly lower carbohydrate accumulation in culms and leaf sheath before heading compared with Lemont. Our results indicated that significantly increased GNP from introgression of GNP1TQ into Lemont did not highly significantly improve grain yield of NIL-GNP1TQ as expected, due primarily to significant low sink activities in IS and possible insufficient source supply which didn’t fully meet the increased sink capacity. The results provided useful information for improving rice yield potential through reasonably introgressing or pyramiding the favorable alleles underlying source-related or panicle number traits by marker-assisted selection.

Corosolic acid isolated from Eriobotrya japonica leaves reduces glucose level in human hepatocellular carcinoma cells, zebrafish and rats

Type 2 diabetes (T2D) with high morbidity and mortality is characterized by abnormal glucose and lipid metabolism due in part to insulin resistance in liver, which lead to elevated hyperglycemia and hyperlipidemia. This study sough to explore the effects of corosolic acid (CA) in different T2D models and explored the underlying mechanism. Separated from Eriobotrya japonica leaves, CA purity was above 95% measured by a HPLC method. Compared with cAMP and DEX induced T2D HepG2 model, CA significantly stimulated glucose consumption and improved glycogen accumulation by inhibiting PEPCK mRNA expression. And in cAMP and DEX induced T2D zebrafish model, CA reduced glycogen degradation and increased glucose consumption by regulating some key enzymes in carbon metabolism including GLUT1, GLUT2, GLUT3, LDHA, LDHB, GP, G6Pase, GYS1, and PFKFB3. In addition, insulin receptor signals were also involved in CA-regulated hypoglycemic action. Furthermore, in STZ-induced T2D rat model, compared with diabetic control groups, CA remarkably downregulated the levels of serum lipid, blood glucose, ICAM-1, malonaldehyde and insulin resistance index, while upregulated SOD activity and impaired glucose tolerance. In a conclusion, CA can regulate glucose and lipid metabolic adaptation in T2D like HepG2, zebrafish and rat models partly through reducing inflammation and oxidative stress and suppressing PEPCK.

Genome-wide analysis of the invertase gene family from maize.

The recent release of the maize genome (AGPv4) contains annotation errors of invertase genes and therefore the enzymes are bestly curated manually at the protein level in a comprehensible fashion The synthesis, transport and degradation of sucrose are determining factors for biomass allocation and yield of crop plants. Invertase (INV) is a key enzyme of carbon metabolism in both source and sink tissues. Current releases of the maize genome correctly annotates only two vacuolar invertases (ivr1 and ivr2) and four cell wall invertases (incw1, incw2 (mn1), incw3, and incw4). Our comprehensive survey identified 21 INV isogenes for which we propose a standard nomenclature grouped phylogenetically by amino acid similarity: three vacuolar (INVVR), eight cell wall (INVCW), and ten alkaline/neutral (INVAN) isogenes which form separate dendogram branches due to distinct molecular features. The acidic enzymes were curated for the presence of the DPN tripeptide which is coded by one of the smallest exons reported in plants. Particular attention was placed on the molecular role of INV in vascular tissues such as the nodes, internodes, leaf sheath, husk leaves and roots. We report the expression profile of most members of the maize INV family in nine tissues in two developmental stages, R1 and R3. INVCW7, INVVR2, INVAN8, INVAN9, INVAN10, and INVAN3 displayed the highest absolute expressions in most tissues. INVVR3, INVCW5, INVCW8, and INVAN1 showed low mRNA levels. Expressions of most INVs were repressed from stage R1 to R3, except for INVCW7 which increased significantly in all tissues after flowering. The mRNA levels of INVCW7 in the vegetative stem correlated with a higher transport rate of assimilates from leaves to the cob which led to starch accumulation and growth of the female reproductive organs.

Exogenous 6‐benzyladenine improves antioxidative system and carbon metabolism of summer maize waterlogged in the field

AbstractA field experiment was performed to study the effects of spraying exogenous 6‐benzyladenine (6‐BA) after waterlogging for 6 days at the third leaf stage and the sixth leaf stage on the activities of key enzymes in carbon metabolism, leaf gas exchange parameters, fluorescent characteristic, antioxidant enzymes activities and MDA content of summer maize hybrids Denghai605 (DH605) and Zhengdan958 (ZD958), in order to illuminate the regulations of exogenous spraying 6‐BA to antioxidative system and carbon metabolism of waterlogged summer maize. Results showed that exogenous application of 6‐BA effectively alleviated the adverse effects of waterlogging on carbon metabolism by improving photosynthetic characteristics and the activities of RuBPCase (in the range of 48%–81%) and PEPCase (in the range of 14%–46%). Treatments with spraying exogenous 6‐BA resulted in significant and substantial increases in net photosynthetic rate (around 49%) and ΦPSII (around 16%) compared to those of waterlogging treatments. In addition, spraying 6‐BA effectively alleviated waterlogging damages on antioxidative enzyme activity and decreased MDA content, delaying leaf senescence, resulted in significant and substantial (around 19%) increases in grain yield. Visibly, spraying exogenous 6‐BA effectively alleviated waterlogging damages on antioxidative system and carbon metabolism of summer maize, resulting in yield increase in waterlogged maize.

Effect of Ethanol Accumulation on Porcine Interferon-α Production by Pichia pastoris and Activities of Key Enzymes in Carbon Metabolism.

In production of porcine interferon α (pIFN-α) by Pichia pastoris, improper glycerol feeding strategy leads to ethanol accumulation in the last stage of growth phase. In the present study, taking two runs with low ethanol accumulation under 2 g/L as control group, effects of long-term (>4 h) and instantaneous high ethanol concentration (>10 g/L) on pIFN-α production, and activities of key enzymes in carbon metabolism were discussed. As a result, compared with control group, pIFN-α expression level was decreased about 4~12 folds under long-term high ethanol concentration, from the level above 3 g/L to the level under 1 g/L; pIFN-α expression level was decreased about 8 folds under instantaneous high ethanol concentration, reaching to the low level of 0.42 g/L. The low production of pIFN-α was caused by the severe inhibitory effect of ethanol on these enzymes.

Betaine homocysteine methyltransferase (BHMT)-dependent remethylation pathway in human healthy and tumoral liver

Carcinogenesis is a multi-step and multifactorial process. It includes genetic, epigenetic, nutritional and environmental factors, which are closely interconnected. Human hepatocellular carcinoma (HCC) is among the most frequent and lethal cancers. Imbalance in the S-adenosylmethionine (SAM) concentration, the main methyl group donor, strongly influences the development of HCC. Key enzymes of carbon metabolism are greatly reduced in patients with cirrhosis and HCC. These alterations play a role in genetic instability and epigenetic modifications (DNA methylation, and histone modifications), however, the molecular underlying mechanisms are still poorly understood. We aimed to investigate betaine homocysteine methyltransferase (BHMT) expression in HepG2 cells and human hepatocarcinoma tissues. Tumor and surrounding healthy tissue were compared. HepG2 cells and tumor samples showed a strong decrease in BHMT transcripts resulting from the transcription of a splicing variant that contained a frameshift mutation generating a premature termination codon and gene loss of function. This splicing variant, not detected in normal adult and fetal liver, cannot be explained by any mechanism involving the known splicing consensus sequences. BHMT activity was abolished in HepG2 cells and protein expression was detected neither in HepG2 cells nor in five of the six tumor samples investigated. Further investigation is needed to elucidate whether this abnormal BHMT transcription is part of cause or consequence of liver carcinogenesis.

Regulation of AtSUS2 and AtSUS3 by glucose and the transcription factor LEC2 in different tissues and at different stages of Arabidopsis seed development

Sucrose synthase (SUS) is a key enzyme of carbon metabolism in heterotrophic tissues of plants. The Arabidopsis genome contains six SUS genes. Two members of this family, namely AtSUS2 (At5g49190) and AtSUS3 (At4g02280) are strongly and differentially expressed in Arabidopsis seed. Expression analysis was carried out using SUS:promoter-GUS fusion lines in a wild-type genetic background or in a mutant carrying a lesion in the transcription factor LEAFY COTYLEDON 2 (LEC2; At1g28300). The accumulation patterns of mRNA, protein, and SUS activity were altered in the lec2 mutant during seed development 9-18 days after flowering. This indicates that LEC2 acts epistatically on the expression of AtSUS2 and AtSUS3. It appears that LEC2 is required for cotyledon-specific expression of both SUS genes but it is not responsible for expression in the radicle tip during embryo development. The AtSUS2 promoter was induced in planta by feeding of glucose but less so by sucrose and trehalose. Non-phosphorylable glucose analogs such as 3-O-methyl-glucose and 2-deoxyglucose also caused an induction, suggesting that sugar signaling proceeds by a hexokinase-independent pathway, possibly involving hexose sensing. Analysis of transgenic lines carrying of truncated versions of the AtSUS2:promoter fused to Beta-glucuronidase activity revealed an internal 421 bp region that was responsible for expression in seeds. Bioinformatic sequence analysis revealed regulatory cis-elements putatively responsible for the spatio-temporal pattern of AtSUS2 expression such as the SEF3 (aaccca) and W-box (ttgact) motifs. These findings are discussed in relation to the roles played by AtSUS2, AtSUS3 and LEC2 in the biosynthesis of seed storage products in Arabidopsis.

Grain-filling problem in ‘super’ rice

Modern rice (Oryza sativa L.) cultivars, especially the newly bred 'super' rice, have numerous spikelets on a panicle with a large yield capacity. However, these cultivars often fail to achieve their high yield potential due to poor grain-filling of later-flowering inferior spikelets (in contrast to the earlier-flowering superior spikelets). Conventional thinking to explain the poor grain-filling is the consequence of carbon limitation. Recent studies, however, have shown that carbohydrate supply should not be the major problem because they have adequate sucrose at their initial grain-filling stage. The low activities of key enzymes in carbon metabolism may contribute to the poor grain-filling. Proper field practices, such as moderate soil drying during mid- and late grain-filling stages, could solve some problems in poor grain-filling. Further studies are needed by molecular approaches to investigate the signal transport, the hormonal action, the gene expressions, and the biochemical processes in inferior spikelets.

A protector effect of cytokinin preparations on the photosynthetic apparatus of wheat plants under water deficiency conditions

The protective effects of the cytokinin 6-benzylaminopurine and the compounds thidiazuron and kartolin, displaying a cytokinin activity, on the photosynthetic apparatus of young seedlings and leaves of adult plants of two wheat (Triticum aestivum L.) cultivars, Mironovskaya 808 (more drought tolerant) and Lutescens 758 (less tolerant to water stress), were compared on the background of an increasing water deficiency. At the stages of drought and subsequent rehydration, cartolin preparations were the most efficient protectors, enhancing a less pronounced decrease in the intensity of photosynthesis, carboxylating activity of the key enzyme of carbon metabolism--ribulose bisphosphate carboxylase/oxygenase (EC 4.1.1.39)-and the activity of NADP-glyceraldehyde phosphate dehydrogenase--the enzyme complex comprising phosphoglycerate kinase (EC 2.7.2.3) and glyceraldehyde phosphate dehydrogenase (EC 1.2.1.13). This effect also included an increase in the leaf specific density and plant productivity. The negative influence of water stress on the photosynthetic apparatus was more pronounced in a less tolerant cultivar Lutescens 758 and in the seedlings as compared with the adult plants.

Purification and Characterization of Cytosolic Glyceraldehyde-3-phosphate Dehydrogenase from the Dromedary Camel

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (EC 1.2.1.12), a key enzyme of carbon metabolism, was purified and characterized to homogeneity from skeletal muscle of Camelus dromedarius. The protein was purified approximately 26.8 folds by conventional ammonium sulphate fractionation followed by Blue Sepharose CL-6B chromatography, and its physical and kinetic properties were investigated. The native protein is a homotetramer with an apparent molecular weight of approximately 146 kDa. Isoelectric focusing analysis showed the presence of only one GAPDH isoform with an isoelectric point of 7.2. The optimum pH of the purified enzyme was 7.8. Studies on the effect of temperature on enzyme activity revealed an optimal value of approximately 28-32 degrees with activation energy of 4.9 kcal/mol. The apparent K(m) values for NAD(+) and DL-glyceraldehyde-3-phophate were estimated to be 0.025+/-0.040 mM and 0.21+/-0.08 mM, respectively. The V(max) of the purified protein was estimated to be 52.7+/-5.9 U/mg. These kinetic parameter values were different from those described previously, reflecting protein differences between species.

Effects of Cytokinin Preparations on the Stability of the Photosynthetic Apparatus of Two Wheat Cultivars Experiencing Water Deficiency

Carboxylase activities of the key enzyme of carbon metabolism, ribulose-bisphosphate carboxylase/oxygenase (RuBisCO; EC 4.1.1.39), and phosphoenolpyruvate carboxylase (PEPC; EC 4.1.1.31), as well as intensities of carbon dioxide photosynthetic assimilation in young seedlings and adult leaves of the wheat Triticum aestivum L. cultivars Mironovskaya 808 (a more tolerant) and Lyutestsens 758 (a less tolerant), were compared under conditions of progressive water deficiency. The water stress had more pronounced negative effects on all the studied characteristics of photosynthetic apparatus of cultivar Lyutestsens 758 photosynthetic machinery of the cultivar Lyutestsens 758. Its seedlings were more sensitive to water stress. Compounds with a cytokinin activity (6-benzylaminopurine, thidiazuron, cartolin 2, and cartolin 4) played a protective role, increasing the stability of the photosynthetic machinery under conditions of water deficiency. Preparations of cartolins displayed the maximum protective effect.

Effect of cytokinin preparations on the pools of pigments and proteins of wheat cultivars differing in their tolerance to water stress

Contents of chlorophylls, carotenoids, soluble leaf proteins, and the key enzyme of carbon metabolism--ribulose bisphosphate carboxylase/oxygenase (RuBisCO; EC 4.1.1.39)--in young seedlings and adult leaves of the wheat Triticum aestivum L. cultivars Mironovskaya 808 and Lyutestsens 758, contrasting in their water stress tolerances, were compared under conditions of normal available water supply, water deficiency, and subsequent rehydration. It was discovered that compounds displaying a cytokinin activity (6-benzylaminopurine, thidiazuron, cartolin-2, and cartolin-4) reduced the decreases in contents of chlorophylls, carotenoids, soluble leaf proteins, and RuBisCO progressing with development of water stress. These compounds with cytokinin activity also accelerated restoration of the compounds studied to their initial concentrations during rehydration. The cartolin preparations caused a maximal protective effect. Water stress had a more pronounced negative effect on cultivar Lyutestsens 758. Dehydration resulted in a more extensive destruction of seedlings compared to leaves of adult plants.

Effects of the loss of triose phosphate isomerase activity on carbon metabolism in Kluyveromyces lactis

The effect of the loss of triose phosphate isomerase activity on carbon metabolism in Kluyveromyces lactis was studied in batch and in continuous cultures. The Kltpi1 mutant was able to grow on media containing glucose as the sole carbon source both in batch and in continuous culture, unlike the corresponding S. cerevisiae mutant. In K. lactis tpi1 mutant no glycerol production was detected in chemostat cultivations. DHAP accumulation triggers glycerol production only when glucose is the sole carbon source in excess. The analysis of the activities of some key enzymes of carbon metabolism shows that in chemostat cultivations on mixed-substrates the activities of enzymes involved in ethanol assimilation are higher both in K. lactis wild type and mutant strains than in S. cerevisiae.