Increase In Alcohol Dehydrogenase Activity Research Articles

Growth Responses and Adventitious Root Formation of Cucumber Hybrid Lines in a Waterlogged Condition

Cucumber (Cucumis sativus L.) F1 hybrids are grown commercially in open-field or greenhouse conditions. Hybrids are well adapted to these settings due to directed breeding. In protected cultivation systems, a small rhizosphere volume and intensive, continuous fertigation predispose the roots to waterlogging (WL) conditions and potentially to hypoxia. However, high productivity is expected and achieved under these conditions. The aim of this study was to identify traits that play a role in this surprising behavior. Initial observations revealed the presence of a significantly higher number of adventitious roots (ARs) in three greenhouse (7–14) vs. three open-field cultivars (less than two) grown under normal conditions. Further on, two contrasting representative hybrids typically grown in open-field and in greenhouse conditions were subjected to WL stress. Declining oxygen levels in the media and increased alcohol dehydrogenase activity (ADH) in the roots were experienced during the WL treatment in both hybrids, with anaerobic metabolism triggered less intensively (~4-fold less ADH activity) in the greenhouse-type ‘Oitol’. The induction characteristics of cysteine oxidase (CysOx) genes, key components of the hypoxia sensing pathway, were used to confirm the hypoxic stress experienced by the roots. The lower extent of upregulation in CysOx genes expression agreed with the milder level of hypoxic stress in the roots of ‘Oitol’ than in ‘Joker’. The more efficient induction of AR formation with a ~50% increase upon waterlogging stress was found to be a prominent trait in ‘Oitol’, apparently helping root internal aeration and mitigating hypoxia. The shoot growth of neither hybrid was set back by hypoxic root conditions. ‘Joker’ plants maintained the same growth rate as that of the control, while the growth of ‘Oitol’ accelerated when its root system was flooded with nutrient solution. Acclimation processes to hypoxia were proposed to explain the lack of growth retardation in both varieties. This corresponded well with a general abundance of AR development in greenhouse-type (slicing) cucumbers that are typically cultivated in soilless systems.

Preparation of Dihydromyricetin-Loaded Self-Emulsifying Drug Delivery System and Its Anti-Alcoholism Effect.

Intraperitoneal injection of dihydromyricetin (DMY) has shown promising potential in the treatment of alcoholism. However, its therapeutic effect is limited due to its low solubility, poor stability, and high gut-liver first-pass metabolism, resulting in very low oral bioavailability. In this study, we developed a DMY-loaded self-emulsifying drug delivery system (DMY-SEDDS) to enhance the oral bioavailability and anti-alcoholism effect of DMY. DMY-SEDDS improved the oral absorption of DMY by facilitating lymphatic transport. The area under the concentration-time curve (AUC) of DMY in the DMY-SEDDS group was 4.13-fold higher than in the DMY suspension group. Furthermore, treatment with DMY-SEDDS significantly enhanced the activities of alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) in the liver of mice (p < 0.05). Interestingly, DMY-SEDDS also increased ADH activity in the stomach of mice with alcoholism (p < 0.01), thereby enhancing ethanol metabolism in the gastrointestinal tract and reducing ethanol absorption into the bloodstream. As a result, the blood alcohol concentration of mice with alcoholism was significantly decreased after DMY-SEDDS treatment (p < 0.01). In the acute alcoholism mice model, compared to saline treatment, DMY-SEDDS prolonged the onset of LORR (loss of righting reflex) (p < 0.05) and significantly shortened the duration of LORR (p < 0.01). Additionally, DMY-SEDDS treatment significantly reduced gastric injury in acute alcoholism mice. Collectively, these findings demonstrate the potential of DMY-SEDDS as a treatment in the treatment of alcoholism.

Plant stem cells under low oxygen: metabolic rewiring by phytoglobin underlies stem cell functionality.

Root growth in maize (Zea mays L.) is regulated by the activity of the quiescent center (QC) stem cells located within the root apical meristem. Here, we show that despite being highly hypoxic under normal oxygen tension, QC stem cells are vulnerable to hypoxic stress, which causes their degradation with subsequent inhibition of root growth. Under low oxygen, QC stem cells became depleted of starch and soluble sugars and exhibited reliance on glycolytic fermentation with the impairment of the TCA cycle through the depressed activity of several enzymes, including pyruvate dehydrogenase (PDH). This finding suggests that carbohydrate delivery from the shoot might be insufficient to meet the metabolic demand of QC stem cells during stress. Some metabolic changes characteristic of the hypoxic response in mature root cells were not observed in the QC. Hypoxia-responsive genes, such as PYRUVATE DECARBOXYLASE (PDC) and ALCOHOL DEHYDROGENASE (ADH), were not activated in response to hypoxia, despite an increase in ADH activity. Increases in phosphoenolpyruvate (PEP) with little change in steady-state levels of succinate were also atypical responses to low-oxygen tensions. Overexpression of PHYTOGLOBIN 1 (ZmPgb1.1) preserved the functionality of the QC stem cells during stress. The QC stem cell preservation was underpinned by extensive metabolic rewiring centered around activation of the TCA cycle and retention of carbohydrate storage products, denoting a more efficient energy production and diminished demand for carbohydrates under conditions where nutrient transport may be limiting. Overall, this study provides an overview of metabolic responses occurring in plant stem cells during oxygen deficiency.

Cross‐disciplinary insights into the mechanisms of plant cold hardiness: From molecules to ecosystems

Cross‐disciplinary insights into the mechanisms of plant cold hardiness: From molecules to ecosystems

Hypoxia-Induced Aquaporins and Regulation of Redox Homeostasis by a Trans-Plasma Membrane Electron Transport System in Maize Roots

In plants, flooding-induced oxygen deficiency causes severe stress, leading to growth reduction and yield loss. It is therefore important to understand the molecular mechanisms for adaptation to hypoxia. Aquaporins at the plasma membrane play a crucial role in water uptake. However, their role during hypoxia and membrane redox changes is still not fully understood. The influence of 24 h hypoxia induction on hydroponically grown maize (Zea mays L.) was investigated using an oil-based setup. Analyses of physiological parameters revealed typical flooding symptoms such as increased ethylene and H2O2 levels, an increased alcohol dehydrogenase activity, and an increased redox activity at the plasma membrane along with decreased oxygen of the medium. Transcriptomic analysis and shotgun proteomics of plasma membranes and soluble fractions were performed to determine alterations in maize roots. RNA-sequencing data confirmed the upregulation of genes involved in anaerobic metabolism, biosynthesis of the phytohormone ethylene, and its receptors. Transcripts of several antioxidative systems and other oxidoreductases were regulated. Mass spectrometry analysis of the plasma membrane proteome revealed alterations in redox systems and an increased abundance of aquaporins. Here, we discuss the importance of plasma membrane aquaporins and redox systems in hypoxia stress response, including the regulation of plant growth and redox homeostasis.

Phenotypic plasticity facilitates initial colonization of a novel environment.

Phenotypic plasticity can allow organisms to respond to environmental changes by producing better matching phenotypes without any genetic change. Because of this, plasticity is predicted to be a major mechanism by which a population can survive the initial stage of colonizing a novel environment. We tested this prediction by challenging wild Drosophila melanogaster with increasingly extreme larval environments and then examining expression of alcohol dehydrogenase (ADH) and its relationship to larval survival in the first generation of encountering a novel environment. We found that most families responded in the adaptive direction of increased ADH activity in higher alcohol environments and families with higher plasticity were also more likely to survive in the highest alcohol environment. Thus, plasticity of ADH activity was positively selected in the most extreme environment and was a key trait influencing fitness. Furthermore, there was significant heritability of ADH plasticity that can allow plasticity to evolve in subsequent generations after initial colonization. The adaptive value of plasticity, however, was only evident in the most extreme environment and had little impact on fitness in less extreme environments. The results provide one of the first direct tests of the adaptive role of phenotypic plasticity in colonizing a novel environment.

Metabolic and biochemical responses of Potamogeton anguillanus Koidz. (Potamogetonaceae) to low oxygen conditions

Oxygen availability in water is considered one of the most important factors for growth and productivity in aquatic submerged macrophytes. In the present study, the growth, stress responses, and metabolic changes in Potamogeton anguillanus Koidz. (Potamogetonaceae) were assessed after a 21-day exposure to low (hypoxia; dissolved oxygen, DO < 1 mg/L) or null (anoxia) oxygen concentrations in water. High growth rates and an increased indole acetic acid (IAA) content in P. anguillanus were observed under the hypoxic conditions (4-fold to control) compared to the anoxic conditions. In addition, the activation of glycolysis and fermentation processes was further recorded, given the increase in alcohol dehydrogenase activity and pyruvate concentration on the studied plants that were exposed to low oxygen concentrations. Moreover, the positive correlations of antioxidative enzyme activities, catalase (CAT) and guaiacol peroxidase (POD) with hydrogen peroxide (H2O2) confirmed the species ability to scavenge excess H2O2 under low oxygen stress. The capillary electrophoresis–mass spectrometry (CE-MS) analysis of the metabolome identified metabolite accumulations (e.g., glutamate, glutamine, aspartate, asparagine, valine, malate, lactate, citrate, isocitrate, proline and γ-amino butyric acid) in response to the anoxia.

Relationship between Polyamines and Anaerobic Respiration of Wheat Seedling Root under Water-Logging Stress

To elucidate the relationship between polyamines and anaerobic respiration of wheat (Triticum aestivum L.) seedling root under water-logging stress, the contents of polyamines (PAs), lactate and alcohol, and the activities of anaerobic respiration enzymes were investigated in seedling roots of two wheat cultivars, Yumai no. 18 and Yangmai no. 9. On the 5th day after water-logging treatment, spermidine (Spd) and spermine (Spm) contents increased significantly, pyruvate decarboxylase (PDC) activity increased and there was no difference between two cultivars. Alcohol dehydrogenase (ADH) activity and alcohol content in Yangmai no. 9 increased more markedly than Yumai no. 18, while lactate dehydrogenase (LDH) activity and the lactate content in the Yumai no. 18 increased more markedly than Yangmai no. 9. Treatments with exogenous Spd and Spm resulted in enhancing the increases in ADH activity, alcohol content, and the levels of Spd and Spm. This concomitantly inhibited the increases in LDH activity and lactate content in Yumai no. 18 under water-logging stress, alleviating stress-induced injury to the seedlings. Treatment with exogenous inhibitor methylglyoxyl-bis-guanylhydrazone (MGBG), resulted in reducing the increases in ADH activity, alcohol content, and Spd and Spm levels, promoting the increases in LDH activity and lactate content in Yangmai no. 9 under water-logging stress, and aggravating the stress-induced injury to the seedlings. The results suggested that under water-logging stress, increased Spd and Spm could facilitate the tolerance of wheat seedling to the stress by enhancing the increases in ADH activity and alcohol content, and inhibiting the increases in LDH activity and lactate content.

Deletion of the hfsB gene increases ethanol production in Thermoanaerobacterium saccharolyticum and several other thermophilic anaerobic bacteria

BackgroundWith the discovery of interspecies hydrogen transfer in the late 1960s (Bryant et al. in Arch Microbiol 59:20–31, 1967), it was shown that reducing the partial pressure of hydrogen could cause mixed acid fermenting organisms to produce acetate at the expense of ethanol. Hydrogen and ethanol are both more reduced than glucose. Thus there is a tradeoff between production of these compounds imposed by electron balancing requirements; however, the mechanism is not fully known.ResultsDeletion of the hfsA or B subunits resulted in a roughly 1.8-fold increase in ethanol yield. The increase in ethanol production appears to be associated with an increase in alcohol dehydrogenase activity, which appears to be due, at least in part, to increased expression of the adhE gene, and may suggest a regulatory linkage between hfsB and adhE. We studied this system most intensively in the organism Thermoanaerobacterium saccharolyticum; however, deletion of hfsB also increases ethanol production in other thermophilic bacteria suggesting that this could be used as a general technique for engineering thermophilic bacteria for improved ethanol production in organisms with hfs-type hydrogenases.ConclusionSince its discovery by Shaw et al. (JAMA 191:6457–64, 2009), the hfs hydrogenase has been suspected to act as a regulator due to the presence of a PAS domain. We provide additional support for the presence of a regulatory phenomenon. In addition, we find a practical application for this scientific insight, namely increasing ethanol yield in strains that are of interest for ethanol production from cellulose or hemicellulose. In two of these organisms (T. xylanolyticum and T. thermosaccharolyticum), the ethanol yields are the highest reported to date.

An Alcohol Dehydrogenase Gene from Synechocystis sp. Confers Salt Tolerance in Transgenic Tobacco.

Synechocystis salt-responsive gene 1 (sysr1) was engineered for expression in higher plants, and gene construction was stably incorporated into tobacco plants. We investigated the role of Sysr1 [a member of the alcohol dehydrogenase (ADH) superfamily] by examining the salt tolerance of sysr1-overexpressing (sysr1-OX) tobacco plants using quantitative real-time polymerase chain reactions, gas chromatography-mass spectrometry, and bioassays. The sysr1-OX plants exhibited considerably increased ADH activity and tolerance to salt stress conditions. Additionally, the expression levels of several stress-responsive genes were upregulated. Moreover, airborne signals from salt-stressed sysr1-OX plants triggered salinity tolerance in neighboring wild-type (WT) plants. Therefore, Sysr1 enhanced the interconversion of aldehydes to alcohols, and this occurrence might affect the quality of green leaf volatiles (GLVs) in sysr1-OX plants. Actually, the Z-3-hexenol level was approximately twofold higher in sysr1-OX plants than in WT plants within 1–2 h of wounding. Furthermore, analyses of WT plants treated with vaporized GLVs indicated that Z-3-hexenol was a stronger inducer of stress-related gene expression and salt tolerance than E-2-hexenal. The results of the study suggested that increased C6 alcohol (Z-3-hexenol) induced the expression of resistance genes, thereby enhancing salt tolerance of transgenic plants. Our results revealed a role for ADH in salinity stress responses, and the results provided a genetic engineering strategy that could improve the salt tolerance of crops.

Enhanced production of 3-methylthiopropionic ethyl ester in native Iranian Cucumis melo L. Group dudaim under regulated deficit irrigation

Sulfur esters such as 3-methylthiopropionic ethyl ester (MTPE) significantly influence taste and fragrance of melons. Nutraceutical properties of MTPE have been demonstrated. GC–Mass analysis confirmed MTPE presence in two Cucumis melo L. Group dudaim cultivars of Iran, Birjand and Kerman. Considering potential effect of abiotic stresses on biosynthesis of esters, both cultivars were subjected to regulated deficit irrigation (RDI) at three levels of matric potentials [control (−50kPa), moderate (−65kPa), high (−75kPa)]. MTPE production rose from 1.55 and 2.8μg/kg of fresh pulp under control RDI to 7.65 and 9.1 under moderate and to 19.3 and 27.8μg/kg under high RDI in Birjand and Kerman samples, respectively. MTPE higher production was accompanied by higher ethylene production and increased alcohol dehydrogenase activity. Ten-fold increase in MTPE production was assumingly owing to higher activities of the esterification enzymes which were supported by sulfur substrates coming from the accelerated methionine catabolism.

Alpha-ketoglutarate reduces ethanol toxicity in Drosophila melanogaster by enhancing alcohol dehydrogenase activity and antioxidant capacity.

Ethanol at low concentrations (<4%) can serve as a food source for fruit fly Drosophila melanogaster, whereas at higher concentrations it may be toxic. In this work, protective effects of dietary alpha-ketoglutarate (AKG) against ethanol toxicity were studied. Food supplementation with 10-mM AKG alleviated toxic effects of 8% ethanol added to food, and improved fly development. Two-day-old adult flies, reared on diet containing both AKG and ethanol, possessed higher alcohol dehydrogenase (ADH) activity as compared with those reared on control diet or diet with ethanol only. Native gel electrophoresis data suggested that this combination diet might promote post-translational modifications of ADH protein with the formation of a highly active ADH form. The ethanol-containing diet led to significantly higher levels of triacylglycerides stored in adult flies, and this parameter was not altered by AKG supplement. The influence of diet on antioxidant defenses was also assessed. In ethanol-fed flies, catalase activity was higher in males and the levels of low molecular mass thiols were unchanged in both sexes compared to control values. Feeding on a mixture of AKG and ethanol did not affect catalase activity but caused a higher level of low molecular mass thiols compared to ethanol-fed flies. It can be concluded that both a stimulation of some components of antioxidant defense and the increase in ADH activity may be responsible for the protective effects of AKG diet supplementation in combination with ethanol. The results suggest that AKG might be useful as a treatment option to neutralize toxic effects of excessive ethanol intake and to improve the physiological state of D.melanogaster and other animals, potentially including humans.

발효 전후 구기자열매 추출물의 ADH 및 ALDH 활성인자 분리 및 특성

Lycii fructus has been traditionally used as a preventive and therapeutic medicine to treat enervation and diverse chronic diseases. In this study, we investigated whether fermentation of Lycii fructus extract (LE) increases the enzymatic activity of the alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH). The fermentation of LE by Bacillus subtilis subsp. subtilis and Saccharomyces cerevisiae IFO 2376 was shown to increase the enzymatic activity of ADH and ALDH. TLC analysis of LE and fermented LE (FLE) showed that ADH and ALDH activities increased in different spots. Fraction No. 66 of LE and fraction No. 68 of FLE by Silica gel chromatography showed increased ADH activity of 129.1% and 148.9%, respectively. Fractions No. 128 of LE and FLE by Silica gel chromatography showed increased ALDH activity of 134.1% and 148.1%, respectively. The fraction No. 68 of FLE obtained by HPLC showed new peaks at Rt 11.938min, Rt 22.072min and Rt 28.842min, indicating that ADH activity was increased. The LE and FLE fractions with the greatest increases in ADH activity peaked at the same time (Rt 13min),whereas the LE and FLE fractions with the greatest increases in ALDH activity peaked at different times (Rt 16.307min and Rt 36.640min, respectively).

Acclimation of Hydrilla verticillata to sediment anoxia in vegetation restoration in eutrophic waters.

Sediment anoxia generally results from intense organic enrichment and is a limiting factor in the restoration of vegetation in eutrophic waters. To investigate the effect of sediment anoxia on a typical pollution-tolerant submerged macrophyte species, Hydrilla verticillata, and acclimation mechanisms in the plant, a gradient of sediment anoxia was simulated with additions of sucrose to the sediment, which can stimulate increased concentrations of total nitrogen, NH4(+) and Fe in pore water. H. verticillata growth was significantly affected by highly anoxic conditions, as indicated by reduced total biomass in the 0.5 and 1% sucrose treatments. However, slight anoxia (0.1% sucrose addition) promoted growth, and the shoot biomass was 22.64% higher than in the control. In addition to morphologic alterations, H. verticillata showed physiological acclimations to anoxia, including increased anaerobic respiration and changes in carbon and nitrogen metabolism in roots. The soluble protein and soluble carbohydrate contents in roots of the 1% treatment were both significantly higher compared with those in the control. The increase in alcohol dehydrogenase activity and pyruvate content in the roots suggested that H. verticillata has a well-developed capacity for anaerobic fermentation. This study suggests that highly anoxic sediments inhibit the growth of H. verticillata and the species has a degree of tolerance to anoxic conditions. Further in situ investigations should be conducted on the interactions between sediment conditions and macrophytes to comprehensively evaluate the roles of sediment in the restoration of vegetation in eutrophic waters.

Effect of waterlogging on photosynthetic and biochemical parameters in pigeonpea

We studied the effect of waterlogging stress on photosynthetic and biochemical parameters in pigeonpea [Cajanus cajan (L.) Mill sp.] genotypes viz. ICPL-84023 (waterlogging resistant) and MAL-18 (waterlogging susceptible). Plants at early growth stage (20 days) were subjected to stress by keeping the pots in water filled containers. Stress was imposed for six days. Waterlogging reduced carbon exchange rate, stomatal conductance, intercellular CO2 concentration and transpiration rate. Photosynthesis was limited by stomatal as well as nonstomatal components under waterlogging. Reduction in chlorophyll, starch and increase in alcohol dehydrogenase activity along with the membrane injury were observed under waterlogging. High carboxylation efficiency, more chlorophyll content, starch availability, alcohol dehydrogenase activity, and membrane stability were associated with better survival of ICPL-84023 under waterlogging.

The Adh1 gene of the fungus Metarhizium anisopliae is expressed during insect colonization and required for full virulence

Zymography of alcohol dehydrogenase (ADH) activity in the entomopathogenic fungus Metarhizium anisopliae grown under various conditions revealed that micro-aerobic growth was associated with increased ADH activity. The major ADH protein, AdhIp, was purified to homogeneity by affinity chromatography and has an estimated molecular weight of 41kDa and an isoelectric point (pI) of 6.4. Peptide mass fingerprint analysis allowed the identification and cloning of the gene that encodes this protein, Adh1, as annotated in the M. anisopliae genome database. AdhIp is related to the medium-chain dehydrogenase/reductase (MDR)/zinc-dependent alcohol dehydrogenase-like family and contains conserved ADH sequence motifs, such as the zinc-containing ADH signature, the FAD/NAD binding domain and amino acid residues that are conserved in most microbial ADHs. Semi-quantitative RT-PCR analysis revealed that Adh1 gene expression occurs at low levels during early Plutella xylostella infection and that the Adh1 gene was primarily expressed at larval death and as mycelia emerge from the insect cuticle before conidiation. Antisense-RNA experiments indicated that NAD+-dependent ADH activity was diminished by 20–75% in the transformants, and the transformants that had lower ADH activity showed allyl alcohol resistance, which indicates that reduction in ADH activity also occurs in vivo. Bioassays performed using antisense adh1 transformants, which have lower ADH activity, showed that LC50 values were two to five times higher than the wild-type, indicating that AdhIp is required for full capability of the fungus to penetrate and/or colonize the insect.

Acute and chronic ethanol exposure differentially alters alcohol dehydrogenase and aldehyde dehydrogenase activity in the zebrafish liver.

Chronic ethanol exposure paradigms have been successfully used in the past to induce behavioral and central nervous system related changes in zebrafish. However, it is currently unknown whether chronic ethanol exposure alters ethanol metabolism in adult zebrafish. In the current study we examine the effect of acute ethanol exposure on adult zebrafish behavioral responses, as well as alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) activity in the liver. We then examine how two different chronic ethanol exposure paradigms (continuous and repeated ethanol exposure) alter behavioral responses and liver enzyme activity during a subsequent acute ethanol challenge. Acute ethanol exposure increased locomotor activity in a dose-dependent manner. ADH activity was shown to exhibit an inverted U-shaped curve and ALDH activity was decreased by ethanol exposure at all doses. During the acute ethanol challenge, animals that were continuously housed in ethanol exhibited a significantly reduced locomotor response and increased ADH activity, however, ALDH activity did not change. Zebrafish that were repeatedly exposed to ethanol demonstrated a small but significant attenuation of the locomotor response during the acute ethanol challenge but ADH and ALDH activity was similar to controls. Overall, we identified two different chronic ethanol exposure paradigms that differentially alter behavioral and physiological responses in zebrafish. We speculate that these two paradigms may allow dissociation of central nervous system-related and liver enzyme-dependent ethanol induced changes in zebrafish.

Encapsulation of Alcohol Dehydrogenase in Mannitol by Spray Drying

The retention of the enzyme activity of alcohol dehydrogenase (ADH) has been studied in various drying processes such as spray drying. The aim of this study is to encapsulate ADH in mannitol, either with or without additive in order to limit the thermal denaturation of the enzyme during the drying process. The retention of ADH activity was investigated at different drying temperatures. When mannitol was used, the encapsulated ADH was found inactive in all the dried powders. This is presumably due to the quick crystallization of mannitol during spray drying that resulted in the impairment of enzyme protection ability in comparison to its amorphous form. Maltodextin (dextrose equivalent = 11) was used to reduce the crystallization of mannitol. The addition of maltodextrin increased ADH activity and drastically changed the powder X-ray diffractogram of the spray-dried powders.

Waterlogging tolerance on a New Zealand saltmarsh

The importance of waterlogging in saltmarshes, and especially its role in small-scale patterning, are little known. A physiological/chorological approach was taken to investigate this in a New Zealand salt meadow. The effect of waterlogging on the activities of enzymes associated with anaerobic metabolism, especially alcohol dehydrogenase (ADH), was investigated in a greenhouse experiment with five representative salt meadow species: Puccinellia stricta, Isolepis cernua, Samolus repens, Selliera radicans and Triglochin striata. All species showed a significant increase in ADH activity in either shoots or roots in response to waterlogging. In P. stricta, sucrose synthase and four out of the five enzymes involved in fermentation increased under waterlogging. However, the photosynthetic efficiency (Fv/Fm) of the species did not change under this regime. The species composition gradient on the saltmarsh was related primarily to elevation, a proxy for water inundation, and the distribution of species along that gradient was related to their tolerance of waterlogging as measured by stimulation of ADH activity. However, the species-composition gradient was also related to water-ponding depressions, demonstrating the role of microtopography, with depression mimicking higher elevations, probably due to rainwater ponding.

Effect of Different Variables on the Efficiency of the Baker's Yeast Cell Disruption Process to Obtain Alcohol Dehydrogenase Activity

Cell disruption process of dry baker's yeast was studied in this work to obtain maximum activity of alcohol dehydrogenase (ADH). Disruption by ultrasonication, glass beads, and combination of these two methods was compared. A 1.8-fold increase of ADH activity can be achieved by combining glass beads with ultrasonication in comparison to ultrasonication. To achieve maximum volume activity of ADH, the effect of different variables on the cell disruption process was investigated (time, glass bead diameter, mass of glass beads, and ultrasound amplitude). Using the Design-Expert© software, 2⁴ factorial experimental design was performed. Two ultrasound probes were tested: MS 73 and KE 76. Optimal conditions (process variables) for cell disruption process were obtained. Optimal ADH activities after cell disruption with MS 73 and KE 76 probes were 1,890.9 and 1,531.7 U cm⁻³, respectively. Necessary ultrasonication time and ultrasound amplitude should be at the maximum values in the investigated variable range (30 min and 62 %). Bead size should be at maximum (4 mm) when using MS 73 probe and at minimum (0.3 mm) when using KE 76 probe. Partial purification of the enzyme was carried out and it was kinetically characterized using several oxidation and reduction systems.