Specific Activity Of Alcohol Dehydrogenase Research Articles

The redox-sensing transcriptional repressor Rex is important for regulating the products distribution in Thermoanaerobacterium aotearoense SCUT27.

The redox-sensing transcriptional repressor Rex (Rex) displayed diverse functions in different microbial species. Nowadays, only part function of rex has been verified in vitro and alcohol dehydrogenase gene (adhE) as the target of Rex has been widely reported. In this study, rex was knocked out in Thermoanaerobacterium aotearoense SCUT27 (GDMCC 60765) and the carbon metabolic distribution analysis was performed. Results showed that the ethanol yield (mol product/mol carbon) of SCUT27(Δrex) had increased by 75.00-90.91%, cell growth improved by 27.27-36.36%, and acetic acid and lactic acid decreased by 58.33-61.54% accompanied with the yield of hydrogen decreased by 46.15-58.35% within different carbon sources. The ability of sugar consumption of SCUT27(Δrex) had improved about 74.19-130.55% with the improvement of total ATP concentration and the cofactors NADH and NAD+ concentrations. In addition, the specific activities of alcohol dehydrogenase of SCUT27(Δrex) with NADH and NADPH as cofactors were improved by 119.26-140.28% and 35.66-47.69%, respectively. After ldh was further knocked out in SCUT27(Δrex), SCUT27(ΔldhΔrex) showed higher ethanol production and yield when various carbon resources were used as substrates (including glucose, xylose, glucose/xylose mixture and three kinds of lignocellulosic hydrolysates). This study confirms that Rex is an important regulator for determining products distribution in SCUT27 and deletion of rex and ldh is a promising strategy for enhanced ethanol production.

Elucidation of the Regulation of Ethanol Catabolic Genes and ptsG Using a glxR and Adenylate Cyclase Gene (cyaB) Deletion Mutants of Corynebacterium glutamicum ATCC 13032

The cyclic AMP receptor protein (CRP) homolog, GlxR, controls the expression of several genes involved in the regulation of diverse physiological processes in Corynebacterium glutamicum. In silico analysis has revealed the presence of glxR binding sites upstream of genes adhA, ald, and ptsG, encoding glucose-specific phosphotransferase system protein, alcohol dehydrogenase (ADH), and acetaldehyde dehydrogenase (ALDH), respectively. However, the involvement of the GlxR-cAMP complex on the expression of these genes has been explored only in vitro. In this study, the expressions of ptsG, adhA, and ald were analyzed in detail using an adenylate cyclase gene (cyaB) deletion mutant and glxR deletion mutant. The specific activities of ADH and ALDH were increased in both the mutants in glucose and glucose plus ethanol media, in contrast to the wild type. In accordance, the promoter activities of adhA and ald were derepressed in the cyaB mutant, indicating that glxR acts as a repressor of adhA. Similarly, both the mutants exhibited derepression of ptsG regardless of the carbon source. These results confirm the involvement of GlxR on the expression of important carbon metabolic genes; adhA, ald, and ptsG.

Screening of allyl alcohol resistant mutant of Rhizopus oryzae and its fermentation characterization

Ethanol is a main by-product in the fermentation broth of Rhizopus oryzae during the production of high-optical purity L-lactic acid. By screening the lower activity of alcohol dehydrogenase (ADH) mutant, thus decreasing the flux of pyruvic acid to ethanol may be a virtual method for increasing the conversion rate of glucose to L-lactic acid. Mutagenesis of R. oryzae As3.3461 was conducted in this study, and a high-producing lactic acid mutant HBF-12 was screened out with selective medium of Yeast-Peptone-Dextrose (YPD) containing allyl alcohol. This mutant showed higher conversion rate of 88.4 g/l to lactate in a 7 L bioreactor, which was an increase of 30.1% compared to original strain of 67.95 g/l, and the conversion rate to ethanol decreased 73.6%. The zymological analysis indicated HBF-12 had a lower ADH specific activity, but an appreciably higher lactate dehydrogenase (LDH) specific activity. The HBF-12 mutant also had a higher consumption rate of glucose and higher biomass collection.

Influence of postharvest water stress on lipoxygenase and alcohol dehydrogenase activities, and on the composition of some volatile compounds of Gewürztraminer grapes dehydrated under controlled and uncontrolled thermohygrometric conditions

Gewurztraminer grapes with a sugar content of around 212 g/L (21.7oBrix) were dried at 17oC, 40% relative humidity and 1.5 m/sec air flow in a 300 L thermo-conditioned tunnel. Control grapes were dried traditionally in a window ventilated room, under uncontrolled environmental conditions varying with outside climate. Tunnel-dried grapes reached the desired sugar concentration (305 g/L, 29.5oBrix) in 17 days, loosing 36% of their weight. Control grapes lost only 22% of their weight and grey mould developed in several bunches at the last sampling. Titratable acidity decreased for tunnel-dried and control grapes from 6.5 g/L to 4 g/L and 5 g/L, respectively. Lipoxygenase (LOX) activity declined in both samples from 120 to 90 U/mg protein dw, with a subsequent significant increase after 20% weight loss in tunnel-treated grapes while the control grapes showed a small peak (150 U/mg protein dw) at 13% weight loss. Six carbon compound evolution showed a loose correlation with LOX activity. Alcohol dehydrogenase specific activity and the concentrations of ethanol and of acetaldehyde plus ethyl acetate showed fluctuating patterns of change, with the evolution of these three variables showing similarity, particularly evident in the tunnel-dried grapes. Carotenoids declined significantly, to increase slightly at the end of the experiment in both samples, with the decline more rapid in the control grapes. Traditional, uncontrolled conditions, did not permit constant dehydration, and provoked a rapid stress to the berries (10% of weight loss). Controlled conditions permitted uniform dehydration, postponed water stress, giving a higher quality product without loss of berries.

Ethanol Catabolism in Corynebacterium glutamicum

Corynebacterium glutamicum grows on a variety of carbohydrates and organic acids as single or combined sources of carbon and energy. Here we show the ability of C. glutamicum to grow on ethanol with growth rates up to 0.24 h^–1 and biomass yields up to 0.47 g dry weight (g ethanol)^–1. Mutants of C. glutamicum deficient in phosphotransacetylase (PTA), isocitrate lyase (ICL) and malate synthase (MS) were unable to grow on ethanol, indicating that acetate activation and the glyoxylate cycle are essential for utilization of this substrate. In accordance, the expression profile of ethanol-grown C. glutamicum cells compared to that of glucose-grown cells revealed an increased expression of genes encoding acetate kinase (AK), PTA, ICL and MS. Furthermore, the specific activities of these four enzymes as well as those of alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) were found to be high in ethanol-grown and low in glucose-grown cells. Growth of C. glutamicum on a mixture of glucose and ethanol led to a biphasic growth behavior, which was due to the sequential utilization of glucose before ethanol. Accordingly, the specific activities of ADH, ALDH, AK, PTA, ICL and MS in cells grown in medium containing both substrates were as low as in glucose-grown cells in the first growth phase, but increased 5- to 100-fold during the second growth phase. The results indicate that ethanol catabolism in C. glutamicum is subject to carbon source-dependent regulation, i.e., to a carbon catabolite control.

Enantioselective reduction of carbonyl compounds by whole-cell biotransformation, combining a formate dehydrogenase and a (R)-specific alcohol dehydrogenase

A whole-cell biotransformation system for the reduction of prochiral carbonyl compounds, such as methyl acetoacetate, to chiral hydroxy acid derivatives [methyl (R)-3-hydroxy butanoate] was developed in Escherichia coli by construction of a recombinant oxidation/reduction cycle. Alcohol dehydrogenase from Lactobacillus brevis catalyzes a highly regioselective and enantioselective reduction of several ketones or keto acid derivatives to chiral alcohols or hydroxy acid esters. The adh gene encoding for the alcohol dehydrogenase of L. brevis was expressed in E. coli. As expected, whole cells of the recombinant strain produced only low quantities of methyl (R)-3-hydroxy butanoate from the substrate methyl acetoacetate. Therefore, the fdh gene from Mycobacterium vaccae N10, encoding NAD+-dependent formate dehydrogenase, was functionally coexpressed. The resulting two-fold recombinant strain exhibited an in vitro catalytic alcohol dehydrogenase activity of 6.5 units mg-1 protein in reducing methyl acetoacetate to methyl (R)-3-hydroxy butanoate with NADPH as the cofactor and 0.7 units mg-1 protein with NADH. The in vitro formate dehydrogenase activity was 1.3 units mg-1 protein. Whole resting cells of this strain catalyzed the formation of 40 mM methyl (R)-3-hydroxy butanoate from methyl acetoacetate. The product yield was 100 mol% at a productivity of 200 micromol g-1 (cell dry weight) min-1. In the presence of formate, the intracellular [NADH]/[NAD+] ratio of the cells increased seven-fold. Thus, the functional overexpression of alcohol dehydrogenase in the presence of formate dehydrogenase was sufficient to enable and sustain the desired reduction reaction via the relatively low specific activity of alcohol dehydrogenase with NADH, instead of NADPH, as a cofactor.

Gluconic acid consumption in wines by Schizosaccharomyces pombe and its effect on the concentrations of major volatile compounds and polyols.

Schizosaccharomyces pombe 1379 (ATCC 26760) yeast strain in wine substantially increases acetaldehyde and 1,1-diethoxyethane concentrations and to decreases levo-2,3-butanediol, glycerol, acetoin, and gluconic acid concentrations. In this study, S. pombe has been used for the first time to reduce gluconic acid in wine under aerobic conditions. Only acetaldehyde and acetoin exhibited significantly higher levels in the wines containing gluconic acid. The high in vitro specific activity of alcohol dehydrogenase observed may be directly related to the high production of acetaldehyde by the studied fission yeast.

In VitroSpecific Activities of Alcohol and Aldehyde Dehydrogenases from Two Flor Yeasts during Controlled Wine Aging

Two flor yeast strains of Saccharomyces cerevisiae (S. cerevisiae strains capensis and bayanus) which form velum on the surface of sherry wine during biological aging have been used. Aldehyde and alcohol (isoenzymes I and II) dehydrogenases were detected in vitro during the entire wine-aging process in the flor yeast strains. All enzymatic activities decreased during the first 155 days of wine aging, and after this period, an increase was observed. Ethanol consumption in the wine and the specific activity of alcohol dehydrogenase I were independent of the S. cerevisiae strain. The greater activity of alcohol dehydrogenase II is directly related to the higher acetaldehyde production by S. cerevisiae race bayanus in the wine. This strain has a slower and prolonged growth in the flor film, which permits a continued accumulation of acetaldehyde in the wine. The higher activity of aldehyde dehydrogenase in capensis strain during the flor formation may be related to the production and consumption of large amoun...

RESPONSE DIFFERENCES BETWEEN AVOCADO AND MANGO FRUIT EXPOSED TO INSECTICIDAL MODIFIED ATMOSPHERE

Atmospheres with very low levels of O2 (≤0.5%) and/or very high levels of CO2 (≥50%) were reported to be insecticidal, and thus have a potential for postharvest insect control in horticultural crops. Our previous studies have shown that avocado fruit (Persea americana Mill cv. Hass) is very sensitive, while mango fruit (Mangifera indica L. cv Keitt) is very tolerant to these extreme atmospheres. The objective of this study was to investigate the differences in responses of the two fruits to insecticidal atmospheres. Fruits were evaluated after storage in a static modified atmosphere (MA) of < 0.4% O2 and up to 80% CO2 for 0, 1, 2, and 3 days at 20°C. Avocado produced intensive fermentative odor in the second day of storage while mango did not present any throughout the 3 days. Pyruvate concentration was very low in both fruits. Lactate concentration was higher in avocado and has accumulated more in MA than in air stored fruits. Avocado had higher activity of alcohol dehydrogenase (ADH) and lactate dehydrogenase (LDH), but less activity of pyruvate decarboxylase (PDC) than mango. There were no major differences in the specific activity of ADH, PDC, and LDH between air and MA stored fruits. Avocado and mango fruit differ in their biochemical responses to insecticidal (extreme) atmosphere. The basis of these differences in relation to fruit sensitivity/tolerance will be discussed.

Effect of age and gender on in vivo ethanol elimination, hepatic alcohol dehydrogenase activity, and NAD+ availability in F344 rats.

It has been reported that aging strikingly decreases in vivo ethanol metabolism in F344 rats without major effects on hepatic alcohol dehydrogenase (ADH) activity. Because hepatic ADH activity is not always rate limiting in the oxidation of ethanol, we measured in vivo ethanol elimination rate (EER), hepatic ADH activity, and the hepatic-cytoplasmic and mitochondrial redox states after acute ethanol application in 2- and 12-month-old F344 rats of both sexes. In male, but not in female, animals EER decreased with age significantly, by 28% (P less than 0.01). The body-to-liver weight ratio was significantly increased in male (39.4 +/- 1.5 vs 46.5 +/- 2.0; P less than 0.05), but not in female, animals with age. Specific activity of ADH was not significantly changed by age, while the activity was significantly reduced with age in male, but not female, rats when related to body weight (5.1 +/- 0.4 vs 3.9 +/- 0.3 mumoles/100 g b.wt./min; P less than 0.05). The cytoplasmic, but not the mitochondrial (NAD+) to (NADH), ratio was significantly decreased with age in male livers (317 +/- 48 vs 793 +/- 128, P less than 0.05), while this was not the case in female livers. In summary, the data show a sex dependence of the effect of age on ethanol metabolism. The observed reduction in in vivo EER with age in male animals is due at least in part to an increased body-to-liver weight ratio, decreased hepatic ADH activity, and reduced availability of NAD+, the cofactor of the ADH reaction. The cause of this may be decreased transport of reducing equivalents through the mitochondrial membrane due to a lack of shuttle systems or a change in the physicochemical properties of the mitochondrial membrane, or decreased reutilization of NADH as NADPH resulting from a reduction of microsomal ethanol oxidation with age.

The Effect of Aluminum on Enzyme Activities in Wheat Roots

The effect of aluminum (Al) on the extractable activity of enzymes of carbon metabolism from wheat ( Triticum aestivum L. cv. Vulcan) roots has been studied. The addition of 75 µM Al to nutrient solutions supporting growth of 4-day-old seedlings inhibited root extension after 24 hand 48 h by 64% and 73%, respectively, compared with untreated seedlings. The most notable change observed with this concentration of Al was an increase in the amount of alcohol dehydrogenase activity in the wheat roots. The specific activity of alcohol dehydrogenase increased by approximately 50% after 24 h, and more than doubled after 48 h. The specific activities of sucrose synthase and lactate dehydrogenase were also increased in extracts from roots of Al-treated seedlings. The results indicate that an early effect of Al on wheat roots may be a shift from aerobic to anaerobic metabolism.

Developmental profile and tissue distribution of Drosophila alcohol dehydrogenase: an immunochemical analysis with monoclonal antibodies.

To analyze Drosophila alcohol dehydrogenase gene (Adh) expression and tissue distribution at various developmental stages, we devised several immunochemical techniques making use of monoclonal antibodies against Drosophila alcohol dehydrogenase (ADH), which had been obtained previously. We here report their application to analyze the expression of Adh in a wild-type strain of D. melanogaster. s-ELISA tests were performed to evaluate fluctuations in ADH content and specific activity during development in individual organs as well as in whole individuals. In all cases, ADH specific activity appeared to be quite constant, which implies that variations in enzyme activity reflect differences in protein content. Immunoblottings of crude homogenates revealed immunoreactive low relative molecular mass peptides in addition to the 27 KD monomeric band, showing a conserved banding pattern in different organs and developmental stages. Immunohistochemical assays on whole organs were used to analyze the general pattern of ADH distribution. Immunoperoxidase staining of cryosections proved to be of crucial relevance, as it yielded full details of the tissue localization of ADH within the ADH-positive organs. We have shown not only that ADH displays a specific distribution in some organs but also that the enzyme is restricted to certain cell types.

Synthesis of superoxide dismutase, catalase and other enzymes and oxygen and superoxide toxicity during changes in oxygen concentration in cultures of brewing yeast

AbstractThe physiological effects on brewing yeast, growing in semi‐defined wort medium, of a sudden transition from aerobiosis to anaerobiosis were studied. Two yeast strains were examined, used for ale and lager fermentations respectively. The reverse transition (from anaerobiosis to aerobiosis) was also examined. Transitions were applied by changing the sparging gas during growth or in stationary phase, and the effects on the specific activities of certain key enzymes and on the viability of the cultures were examined.Neither type of transition led to significant changes in growth rate, the rate of ethanol production or the specific activities of alcohol dehydrogenase and pyruvate decarboxylase. The most significant change was in the specific activity of CuZn‐superoxide dismutase, which showed a rapid increase in activity on transition from anaerobiosis to aerobiosis, and a decrease in activity on the reverse transition. Catalase activity in the ale yeast generally followed that of CuZn‐superoxide dismutase, whereas in the lager yeast it remained unchanged by the transitions. The transition from anaerobiosis to aerobiosis caused increases in citrate synthase and Mn‐superoxide dismutase, though only after a significant lag period. Aerobic to anaerobic transitions caused a decrease in Mn‐superoxide dismutase activity, while citrate synthase remained unchanged.Anaerobically grown cells showed a rapid loss in viability on exposure to oxygen (5–7% in the first hour), while aerobically grown cells were unaffected. When anaerobically grown cells were exposed to 0·25 mM‐potassium superoxide, there was an 8% loss of viability within 10 min, whereas aerobic cells were not affected.It is concluded that the toxic effect of oxygen is due to superoxide (or a species derived from it) and that the CuZn‐superoxide dismutase (but not the Mn‐isoenzyme) plays a role in protecting the cells. The de novo synthesis of the CuZn‐enzyme is not always rapid enough to confer full protection.

DEVELOPMENTAL REGULATION OF THE EXPRESSION OF ALCOHOL DEHYDROGENASE IN RIPENING TOMATO FRUITS

In each of 5 tomato (Lycopersicon esculentum Mill) cultivars examined, alcohol dehydrogenase (E.C.1.1.1.1) specific activity decreased during the early stages of ripening and then increased in the postclimacteric period. Alcohol dehydrogenase specific activity also increased when immature, mature or pink fruit were placed in an atmosphere of 3% (V/V) oxygen. Measurements of oxygen in the internal tissues and the respiratory quotient throughout ripening established that fruit ripening in air does not suffer an oxygen stress. Increases in alcohol dehydrogenase activity were also observed in pink fruit exposed to 10% (V/V) carbon dioxide. Such atmospheres are known to result in a lowering of the cytoplasmic pH in plants and it is suggested that alcohol dehydrogenase activity is induced during ripening in response to cytoplasmic acid stress. The increased ADH activity during ripening may contribute to flavor development.

Inter-specific analysis of Drosophila alcohol dehydrogenase by an immunoenzymatic assay using monoclonal antibodies

Three Drosophila alcohol dehydrogenase monoclonal antibodies have been prepared and characterized. These antibodies cross-react with alcohol dehydrogenase from different species as revealed by immunoblotting assay. An enzyme-linked immunosorbent assay has been devised to quantify alcohol dehydrogenase in several species, different strains and individual larval organs. The assay detects alcohol dehydrogenase via a double-antibody sandwich assay technique giving strictly proportional values for antigen concentration and optical densities in the range of 3–30 ng of antigen per 100 μl of sample. When alcohol dehydrogenase specific activity is compared in different larval organs a remarkable similarity is observed, whereas protein distribution varies substantially. Larval fat body and larval alimentary canal contribute 63% and 26% respectively to recovered alcohol dehydrogenase.

Changes in activities of several enzymes involved in carbohydrate metabolism during the cell cycle of Saccharomyces cerevisiae.

Activity changes of a number of enzymes involved in carbohydrate metabolism were determined in cell extracts of fractionated exponential-phase populations of Saccharomyces cerevisiae grown under excess glucose. Cell-size fractionation was achieved by an improved centrifugal elutriation procedure. Evidence that the yeast populations had been fractionated according to age in the cell cycle was obtained by examining the various cell fractions for their volume distribution and their microscopic appearance and by flow cytometric analysis of the distribution patterns of cellular DNA and protein contents. Trehalase, hexokinase, pyruvate kinase, phosphofructokinase 1, and fructose-1,6-diphosphatase showed changes in specific activities throughout the cell cycle, whereas the specific activities of alcohol dehydrogenase and glucose-6-phosphate dehydrogenase remained constant. The basal trehalase activity increased substantially (about 20-fold) with bud emergence and decreased again in binucleated cells. However, when the enzyme was activated by pretreatment of the cell extracts with cyclic AMP-dependent protein kinase, no significant fluctuations in activity were seen. These observations strongly favor posttranslational modification through phosphorylation-dephosphorylation as the mechanism underlying the periodic changes in trehalase activity during the cell cycle. As observed for trehalase, the specific activities of hexokinase and phosphofructokinase 1 rose from the beginning of bud formation onward, finally leading to more than eightfold higher values at the end of the S phase. Subsequently, the enzyme activities dropped markedly at later stages of the cycle. Pyruvate kinase activity was relatively low during the G1 phase and the S phase, but increased dramatically (more than 50-fold) during G2. In contrast to the three glycolytic enzymes investigated, the highest specific activity of the gluconeogenic enzyme fructose-1, 6-diphosphatase 1 was found in fractions enriched in either unbudded cells with a single nucleus or binucleated cells. The observed changes in enzyme activities most likely underlie pronounced alterations in carbohydrate metabolism during the cell cycle.

The role of alcohol dehydrogenase in the fermentation of D-xylose byCandida shehatae ATCC 22984

When the oxygen supply to an aerobic chemostat culture ofCandida shehatae growing on D-xylose was reduced to oxygen-limited and anoxic conditions, accumulation of ethanol occurred, the specific activity of alcohol dehydrogenase (ADH) increased up to four-fold and the number of ADH isozymes detected increased from one to three. ADH in crude extracts prepared from anoxically-grown cells had a lower affinity for but was more tolerant to ethanol than in extracts prepared from aerobically grown cells. ADH activity appeared to be sufficient for ethanol production byC. shehatae under anoxic conditions.

Allyl Alcohol Selection for Lower Alcohol Dehydrogenase Activity in Nicotiana plumbaginifolia Cultured Cells

One cell strain with stable tolerance to allyl alcohol (AA(r)) was selected from 6 x 10(8) suspension cultured Nicotiana plumbaginifolia Viviani cells. The selected strain contained one-half the alcohol dehydrogenase (ADH) activity of the wild type (NP) due to the loss of two of three bands of ADH activity seen on starch gels following electrophoresis of wild-type cell extracts. Anaerobic conditions, simulated by not shaking the suspension cultures, increased the ADH specific activity to more than 3-fold the initial level in both strains but did not change the number of activity bands or the relative levels of activity. The cell strain with decreased ADH activity lost viability more rapidly than the wild type under the anaerobic conditions. The AA(r) cells were 10 times more tolerant to ethanol than the NP cells and were also somewhat more tolerant to acetaldehyde and antimycin A. The substrate specificities of the ADH enzymes from both strains were very similar. Further selection of AA(r) cells with allyl alcohol produced strains with even lower ADH activity and selection under anaerobic conditions produced strains with increased ADH activity. Genetic studies indicate that the N. plumbaginifolia ADH activity bands arise from subunits produced by two nonallelic genes. This is the first example of the use of allyl alcohol to select for decreased ADH using cultured plant cells.

Testicular atrophy induced by di(2-ethylhexyl)phthalate: changes in histology, cell specific enzyme activities and zinc concentrations in rat testis.

Daily administration of 2g/kg/day di(2-ethylhexyl)phthalate (DEHP) to immature rats was found to cause testicular atrophy and reduce zinc concentration. Specific activities of testicular enzymes associated with postmeiotic spermatogenic cells, such as lactate dehydrogenase isozyme-X, hyaluronidase and sorbitol dehydrogenase, were lower than those of control by day 10, coincident with degeneration of spermatogenic cells. The specific activities of enzymes associated with premeiotic spermatogenic cells, Sertoli cells or interstitial cells (beta-glucuronidase, gamma-glutamyl transpeptidase and malate dehydrogenase) were higher than those of control by day 10. The specific activities of alcohol dehydrogenase and aldolase, zinc containing enzymes, increased after DEHP treatment in spite of the decrease in zinc concentration in the testis. In conclusion, changes in several testicular cell-specific enzymes appear to be useful biochemical markers of testicular injury induced by testicular toxicants such as DEHP. However, these changes occurred after or simultaneous with massive histological or morphological changes rather than prior to such changes.

Characterization of alcohol dehydrogenase in young soybean seedlings

Molecular properties of alcohol dehydrogenase (ADH) were examined in young soybean seedlings. Soybean radicle tissue is ADH-rich. Enzyme specific activity decreases slowly with the development of roots and becomes almost undetectable when the first true leaves appear. Soybean ADH was not found to be inducible by flooding. 2,4-Dichlorophenoxyacetic acid (2,4-D) treatment increased ADH specific activity as much as 14-fold. Only one ADH isozyme was detected by isoelectric focusing. By DNA-DNA hydridization, soybean ADH genomic sequences were shown to be partly homologous to maize ADH1 cDNA. The presence of more than one Adh gene in soybean is discussed.