Economical one-pot synthesis of isoquercetin and D-allulose from quercetin and sucrose using whole-cell biocatalyst.

Isoquercetin and D-allulose have diverse applications and significant value in antioxidant, antibacterial, antiviral, and lipid metabolism. Isoquercetin can be synthesized from quercetin, while D-allulose is converted from D-fructose. However, their production scale and overall quality are relatively low, leading to high production costs. In this study, we have devised a cost-effective one-pot method for biosynthesizing isoquercetin and D-allulose using a whole-cell biocatalyst derived from quercetin and sucrose. To achieve this, the optimized isoquercetin synthase and D-allulose-3-epimerase were initially identified through isofunctional gene screening. In order to reduce the cost of uridine diphosphate glucose (UDPG) during isoquercetin synthesis and ensure a continuous supply of UDPG, sucrose synthase is introduced to enable the self-circulation of UDPG. At the same time, the inclusion of sucrose permease was utilized to successfully facilitate the catalytic production of D-allulose in whole cells. Finally, the recombinant strain BL21/UGT-SUS+DAE-SUP, which overexpresses MiF3GTMUT, GmSUS, EcSUP, and DAEase, was obtained. This strain co-produced 41±2.4 mg/L of isoquercetin and 5.7±0.8 g/L of D-allulose using 120 mg/L of quercetin and 20 g/L of sucrose as substrates for 5 h after optimization. This is the first green synthesis method that can simultaneously produce flavonoid compounds and rare sugars. These findings provide valuable insights and potential for future industrial production, as well as practical applications in factories.

Valid Assessment of Carbohydrate Intolerance and the Need for a Distinction to Carbohydrate Malabsorption. Comment on "Roles of Lactose and Fructose Malabsorption and Dietary Outcomes in Children Presenting with Chronic Abdominal Pain.", Nutrients, 2019, 11, 3063.

We have read with interest the recent paper by Posovszky and Roesler et al [1] that reports, based on the doctoral thesis by Dr. Roesler [2] [...].

A Robust and Cost-Effective Luminescent-Based High-Throughput Assay for Fructose-1,6-Bisphosphate Aldolase A.

Hypoxic solid tumors induce the stabilization of hypoxia-inducible factor 1 alpha (HIF1α), which stimulates the expression of many glycolytic enzymes and hypoxia-responsive genes. A high rate of glycolysis supports the energetic and material needs for tumors to grow. Fructose-1,6-bisphosphate aldolase A (ALDOA) is an enzyme in the glycolytic pathway that promotes the expression of HIF1α. Therefore, inhibition of ALDOA activity represents a potential therapeutic approach for a range of cancers by blocking two critical cancer survival mechanisms. Here, we present a luminescence-based strategy to determine ALDOA activity. The assay platform was developed by integrating a previously established ALDOA activity assay with a commercial NAD/NADH detection kit, resulting in a significant (>12-fold) improvement in signal/background (S/B) compared with previous assay platforms. A screening campaign using a mixture-based compound library exhibited excellent statistical parameters of Z' (>0.8) and S/B (~20), confirming its robustness and readiness for high-throughput screening (HTS) application. This assay platform provides a cost-effective method for identifying ALDOA inhibitors using a large-scale HTS campaign.

Use of Graph Based Descriptors for Determination of Structural Features Causing Modulation of Fructose-1,6-Bisphosphatase.

Fructose-1,6-bisphosphatase performs a significant function in regulating the blood glucose level in type 2 diabetes by controlling the process gluconeogenesis. In this research work optimal descriptor (graph) based quantitative structural activity relationship studies of a set of 203 fructose-1,6-bisphosphatase has been performed with the help of Monte Carlo optimization. Distribution of compounds into different sets such as training set, invisible training set, calibration set and validation sets resulted in formation of splits. Statistical parameters obtained from quantitative structural activity relationship modeling were good for various designed splits. The statistical parameters such as R2 and Q2 for calibration and validation sets of best split developed were found to be 0.8338, 0.7908 & 0.7920 and 0.7036 respectively. Based on the results obtained for correlation weights, different structural attributes were described as promoters and demoters of the endpoint. Further these structural attributes were used in designing of new fructose-1,6-bisphosphatase inhibitors and molecular docking study was accomplished for the determination of interactions of designed molecules with the enzyme.

Functionalized polyhydroxyalkanoate nano-beads as a stable biocatalyst for cost-effective production of the rare sugar d-allulose.

d-Allulose is a promising low-calorie sweetener especially for diabetes and obesity patients. The functionalized polyhydroxyalkanoate (PHA) nano-beads decorated with d-tagatose 3-epimerase (DTE) was produced in recombinant endotoxin-free ClearColi, whereby the expression, purification, and immobilization of the active DTE were efficiently combined into one step. The immobilized DTE exhibited remarkable enzyme activity of 649.3 U/g beads and extremely high stability at a harsh working condition (pH 7.0-8.0, 65 °C). When DTE-PHA beads were subjected to enzymatic synthesis of d-allulose, a maximum conversion rate of 33% can be achieved at pH 7.0 and 65 °C for 3 h, and DTE-PHA beads retained about 80% of its initial activity after 8 continuous cycles. Moreover, the d-allulose/d-fructose binary mixture can be simply separated by a single cation exchange resin-equipped chromatography. Taken together, DTE-PHA beads are promising and robust nano-biocatalysts that will remarkably simplify the production procedures of d-allulose, contributing to its cost-effective production.

Assessment the viability properties of Lactobacillus casei strain using labneh as a carrier.

BACKGROUND: Our study was conducted in two stages; the first stage was to examine the fructose fermentation profile by Lactobacillus (Lb.) casei FEGY9973. The second stage was to investigate the viability properties of Lb. casei either during cold storage of labneh or under simulated gastrointestinal tract (GIT) conditions. METHODS: Labneh as a carrier medium was classified into four treatments; the first one con- tained 2% free cells of Lb. casei as a control. The second, third and fourth treatments used 2% of encapsulated cells of Lb. casei with different capsule materials, including alginate-milk, sodium alginate and κ-carrageenan served as T1, T2 and T3 respectively. The physiochemical, microbiological and sensory properties of labneh during 15 days of cold storage were shown. Moreover, the viability of free and encapsulated Lb. casei sub- jected to some manufacturing and simulated GIT conditions was tested. RESULTS: It was revealed that lactate was the major metabolite in the medium for colonic fermentation, where- as no amounts of ethanol could be detected. Moreover, labneh samples including free cells of Lb. casei had lower pH values than treatments containing microcapsules of Lb. casei. The levels of moisture, acetaldehyde and diacetyle in treatments with different encapsulated materials were increased during the cold storage period. Accordingly, labneh samples with encapsulated Lb. casei had higher sensory scores than the control. In addition, labneh samples with Lb. casei in milk-alginate microcapsules showed a high viability during cold storage and under simulated GIT conditions. A significant decrease in the viability of free or encapsulated Lb. casei was observed at 15 days of cold storage. CONCLUSIONS: Encapsulated Lb. casei by alginate-milk was more resistant during the cold storage period and under simulated gastric conditions than the other two treatments.

Statistical optimization of process parameters for inulinase production from Tithonia weed by Arthrobacter mysorens strain no.1.

Tithonia rotundifolia is an easily available and abundant inulin rich weed reported to be competitive and allelopathic. This weed inulin is hydrolyzed by inulinase into fructose. Response surface methodology was employed to optimize culture conditions for the inulinase production from Arthrobacter mysorens strain no.1 isolated from rhizospheric area of Tithonia weed. Initially, Plackett- Burman design was used for screening 11 nutritional parameters for inulinase production including inulin containing weeds as cost effective substrate. The experiment shows that amongst the 11 parameters studied, K2HPO4, Inulin, Agave sisalana extract and Tithonia rotundifolia were the most significant variables for inulinase production. Quantitative effects of these 4 factors were further investigated using Box Behnken design. The medium having 0.27% K2HPO4, 2.54% Inulin, 6.57% Agave sisalana extract and 7.27% Tithonia rotundifolia extract were found to be optimum for maximum inulinase production. The optimization strategies used showed 2.12 fold increase in inulinase yield (1669.45 EU/ml) compared to non-optimized medium (787 EU/ml). Fructose produced by the action of inulinase was further confirmed by spectrophotometer, osazone, HPTLC and FTIR methods. Thus Tithonia rotundifolia can be used as an eco-friendly, economically feasible and promising alternative substrate for commercial inulinase production yielding fructose from Arthrobacter mysorens strain no.1.

Enhanced Inulin Saccharification by Self-Produced Inulinase from a Newly Isolated Penicillium sp. and its Application in D-Lactic Acid Production.

In order to find an alternative for commercial inulinase, a strain XL01 identified as Penicillium sp. was screened for inulinase production. The broth after cultivated was centrifuged, filtered, and used as crude enzyme for the following saccharification. At pH 5.0 and 50 °C, the crude enzyme released 84.9 g/L fructose and 20.7 g/L glucose from 120 g/L inulin in 72 h. In addition, simultaneous saccharification and fermentation of chicory flour for D-lactic acid production was carried out using the self-produced crude inulinase and Lactobacillus bulgaricus CGMCC 1.6970. A high D-lactic acid titer and productivity of 122.0 g/L and 1.69 g/(L h) was achieved from 120 g/L chicory flour in 72 h. The simplicity for inulinase production and the high efficiency for D-lactic acid fermentation provide a perspective and profitable industrial biotechnology for utilization of the inulin-rich biomass.

Risk assessment of silica nanoparticles on liver injury in metabolic syndrome mice induced by fructose.

This study aims to assess the effects and the mechanisms of silica nanoparticles (SiNPs) on hepatotoxicity in both normal and metabolic syndrome mouse models induced by fructose. Here, we found that SiNPs exposure lead to improved insulin resistance in metabolic syndrome mice, but markedly worsened hepatic ballooning, inflammation infiltration, and fibrosis. Moreover, SiNPs exposure aggravated liver injury in metabolic syndrome mice by causing serious DNA damage. Following SiNPs exposure, liver superoxide dismutase and catalase activities in metabolic syndrome mice were stimulated, which is accompanied by significantly increased malondialdehyde and 8-hydroxy-2-deoxyguanosine levels as compared to normal mice. Scanning electron microscope (SEM) revealed that SiNPs were more readily deposited in the liver mitochondria of metabolic syndrome mice, resulting in more severe mitochondrial injury as compared to normal mice. We speculated that SiNPs-induced mitochondrial injury might be the cause of hepatic oxidative stress, which further lead to a series of liver lesions as observed in mice following SiNPs exposure. Based on these results, it is likely that SiNPs will increase the risk and severity of liver disease in individuals with metabolic syndrome. Therefore, SiNPs should be used cautiously in food additives and clinical settings.

Recent advances in microbial production of mannitol: utilization of low-cost substrates, strain development and regulation strategies.

Mannitol has been widely used in fine chemicals, pharmaceutical industries, as well as functional foods due to its excellent characteristics, such as antioxidant protecting, regulation of osmotic pressure and non-metabolizable feature. Mannitol can be naturally produced by microorganisms. Compared with chemical manufacturing, microbial production of mannitol provides high yield and convenience in products separation; however the fermentative process has not been widely adopted yet. A major obstacle to microbial production of mannitol under industrial-scale lies in the low economical efficiency, owing to the high cost of fermentation medium, leakage of fructose, low mannitol productivity. In this review, recent advances in improving the economical efficiency of microbial production of mannitol were reviewed, including utilization of low-cost substrates, strain development for high mannitol yield and process regulation strategies for high productivity.

Assessment of Sugar Components and Genes Involved in the Regulation of Sucrose Accumulation in Peach Fruit.

Soluble sugar contents in mature fruits of 45 peach accessions were quantified using gas chromatography analysis. Sucrose is the predominant sugar in mature fruit, followed by glucose and fructose, which have similar concentrations. Overall, sucrose metabolism and accumulation are crucial determinants of sugar content in peach fruit, and there is a wide range of sucrose concentrations among peach genotypes. To understand the mechanisms regulating sucrose accumulation in peach fruit, expression profiles of genes involved in sucrose metabolism and transport were compared among four genotypes. Two sucrose-cleaving enzyme genes (SUS4 and NINV8), one gene involved in sucrose resynthesis (SPS3), and three sugar transporter genes (SUT2, SUT4, and TMT2) were prevalently expressed in peach fruit, and their expression levels are significantly correlated with sucrose accumulation. In contrast, the VAINV genes responsible for sucrose cleavage in the vacuole were weakly expressed in mature fruit, suggesting that the sucrose-cleaving reaction is not active in the vacuole of sink cells of mature peach fruit. This study suggests that sucrose accumulation in peach fruit involves the coordinated interaction of genes related to sucrose cleavage, resynthesis, and transport, which could be helpful for future peach breeding.

Impact of zinc supplementation on the improved fructose/xylose utilization and butanol production during acetone-butanol-ethanol fermentation.

Lignocellulosic biomass and dedicated energy crops such as Jerusalem artichoke are promising alternatives for biobutanol production by solventogenic clostridia. However, fermentable sugars such as fructose or xylose released from the hydrolysis of these feedstocks were subjected to the incomplete utilization by the strains, leading to relatively low butanol production and productivity. When 0.001 g/L ZnSO4·7H2O was supplemented into the medium containing fructose as sole carbon source, 12.8 g/L of butanol was achieved with butanol productivity of 0.089 g/L/h compared to only 4.5 g/L of butanol produced with butanol productivity of 0.028 g/L/h in the control without zinc supplementation. Micronutrient zinc also led to the improved butanol production up to 8.3 g/L derived from 45.2 g/L xylose as sole carbon source with increasing butanol productivity by 31.7%. Moreover, the decreased acids production was observed under the zinc supplementation condition, resulting in the increased butanol yields of 0.202 g/g-fructose and 0.184 g/g-xylose, respectively. Similar improvements were also observed with increasing butanol production by 130.2 % and 8.5 %, butanol productivity by 203.4% and 18.4%, respectively, in acetone-butanol-ethanol fermentations from sugar mixtures of fructose/glucose (4:1) and xylose/glucose (1:2) simulating the hydrolysates of Jerusalem artichoke tubers and corn stover. The results obtained from transcriptional analysis revealed that zinc may have regulatory mechanisms for the sugar transport and metabolism of Clostridium acetobutylicum L7. Therefore, micronutrient zinc supplementation could be an effective way for economic development of butanol production derived from these low-cost agricultural feedstocks.

Multi-objective optimization for the economic production of d-psicose using simulated moving bed chromatography.

The biocatalytic production of rare carbohydrates from available sugar sources rapidly gains interest as a route to acquire industrial amounts of rare sugars for food and fine chemical applications. Here we present a multi-objective optimization procedure for a simulated moving bed (SMB) process for the production of the rare sugar d-psicose from enzymatically produced mixtures with its epimer d-fructose. First, model parameters were determined using the inverse method and experimentally validated on a 2-2-2-2 lab-scale SMB plant. The obtained experimental purities (PUs) were in excellent agreement with the simulated data derived from a transport-dispersive true-moving bed model demonstrating the feasibility of the proposed design. In the second part the performance of the separation was investigated in a multi-objective optimization study addressing the cost-contributing performance parameters productivity (PR) and desorbent requirement (DR) as a function of temperature. While rare sugar SMB operation under conditions of low desorbent consumption was found to be widely unaffected by temperature, SMB operation focusing on increased PR significantly benefited from high temperatures, with possible productivities increasing from 3.4kg(Lday)(-1) at 20°C to 5kg(Lday)(-1) at 70°C, indicating that decreased selectivity at higher temperatures could be fully compensated for by the higher mass transfer rates, as they translate into reduced switch times and hence higher PR. A DR/PR Pareto optimization suggested a similar but even more pronounced trend also under relaxed PU requirements, with the PR increasing from 4.3kg(Lday)(-1) to a maximum of 7.8kg(Lday)(-1) for SMB operation at 50°C when the PU of the non-product stream was reduced from 99.5% to 90%. Based on the in silico optimization results experimental SMB runs were performed yielding considerable PRs of 1.9 (30°C), 2.4 (50°C) and 2.6kg(Lday)(-1) (70°C) with rather low DR (27L per kg of rare sugar produced) on a lab-scale SMB installation.

Jerusalem artichoke as low-cost fructose-rich feedstock for fossil fuels desulphurization by a fructophilic bacterium.

AIMS: Through biodesulphurization (BDS) is possible to remove the sulphur present in fossil fuels to carry out the very strict legislation. However, this biological process is limited by the cost of the culture medium, and thus, it is important to explore cheaper alternative carbon sources, such as Jerusalem artichoke (JA). These carbon sources usually contain sulphates which interfere with the BDS process. The goal of this work was to remove the sulphates from Jerusalem artichoke juice (JAJ) through BaCl2 precipitation viewing the optimization of dibenzothiophene (DBT) desulphurization by Gordonia alkanivorans strain 1B. METHODS AND RESULTS: Using a statistical design (Doehlert distribution), the effect of BaCl2 concentration (0.125-0.625%) and pH (5-9) was studied on sulphate concentration in hydrolysed JAJ. A validated surface response derived from data indicated that zero sulphates can be achieved with 0.5-0.55% (w/v) BaCl2 at pH 7; however, parallel BDS assays showed that the highest desulphurization was obtained with the juice treated with 0.5% (w/v) BaCl2 at pH 8.73. Further assays demonstrated that enhanced DBT desulphurization was achieved using hydrolysed JAJ treated in these optimal conditions. A total conversion of 400 µmol l(-1) DBT into 2-hydroxybiphenyl (2-HBP) in <90 h was observed, attaining a 2-HBP maximum production rate of 28.2 µmol l(-1) h(-1) and a specific production rate of 5.06 µmol(-1) g(-1) (DCW) h(-1) . CONCLUSIONS: These results highlight the efficacy of the treatment applied to JAJ in making this agromaterial a promising low-cost renewable feedstock for improved BDS by the fructophilic strain 1B. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is a fundamental step viewing BDS application at the industrial level as it accounts a cost-effective production of the biocatalysts, one of the main drawbacks for BDS scale-up.

A GC-MS-based metabolomics study on the tubers of commercial potato cultivars upon storage.

Using gas chromatography-mass spectrometry (GC-MS) as a system for the detection of amino acids, organic acids, sugars, sugar alcohols, and fatty acids, we characterised six commercial potato cultivars (Hópehely, Katica, Lorett, Somogyi kifli, Vénusz Gold, and White Lady) with different pedigrees, starch contents, cooking types, and dormancy periods, in five developmental stages from harvest to sprouting. The tubers were stored at 20-22°C in the dark. The metabolite data were subjected to principal component analysis. No correlation between metabolite contents of freshly harvested tubers and starch content or cooking type of the cultivars was detected. The storage decreased the fructose and sucrose and increased the proline concentrations of tubers. Irrespective of the length of dormancy a substantial difference in metabolite composition at each time point upon storage was detected in each cultivar except Somogyi kifli, the only cultivar amongst those tested with a pure Solanum tuberosum origin and A cooking type.

Obesity and energy balance: is the tail wagging the dog?

The scientific study of obesity has been dominated throughout the twentieth century by the concept of energy balance. This conceptual approach, based on fundamental thermodynamic principles, states that energy cannot be destroyed, and can only be gained, lost or stored by an organism. Its application in obesity research has emphasised excessive appetite (gluttony), or insufficient physical activity (sloth), as the primary determinants of excess weight gain, reflected in current guidelines for obesity prevention and treatment. This model cannot explain why weight accumulates persistently rather than reaching a plateau, and underplays the effect of variability in dietary constituents on energy and intermediary metabolism. An alternative model emphasises the capacity of fructose and fructose-derived sweeteners (sucrose, high-fructose corn syrup) to perturb cellular metabolism via modification of the adenosine monophosphate (AMP)/adenosine triphosphate (ATP) ratio, activation of AMP kinase and compensatory mechanisms, which favour adipose tissue accretion and increased appetite while depressing physical activity. This conceptual model implicates chronic hyperinsulinaemia in the presence of a paradoxical state of 'cellular starvation' as a key driver of the metabolic modifications inducing chronic weight gain. We combine evidence from in vitro and in vivo experiments to formulate a perspective on obesity aetiology that emphasises metabolic flexibility and dietary composition rather than energy balance. Using this model, we question the direction of causation of reported associations between obesity and sleep duration or childhood growth. Our perspective generates new hypotheses, which can be tested to improve our understanding of the current obesity epidemic, and to identify novel strategies for prevention or treatment.

A novel methodology independent of fermentation rate for assessment of the fructophilic character of wine yeast strains.

The yeast Saccharomyces cerevisiae has a fundamental role in fermenting grape juice to wine. During alcoholic fermentation its catabolic activity converts sugars (which in grape juice are a near equal ratio of glucose and fructose) and other grape compounds into ethanol, carbon dioxide and sensorily important metabolites. However, S. cerevisiae typically utilises glucose and fructose with different efficiency: glucose is preferred and is consumed at a higher rate than fructose. This results in an increasing difference between the concentrations of glucose and fructose during fermentation. In this study 20 commercially available strains were investigated to determine their relative abilities to utilise glucose and fructose. Parameters measured included fermentation duration and the kinetics of utilisation of fructose when supplied as sole carbon source or in an equimolar mix with glucose. The data were then analysed using mathematical calculations in an effort to identify fermentation attributes which were indicative of overall fructose utilisation and fermentation performance. Fermentation durations ranged from 74.6 to over 150 h, with clear differences in the degree to which glucose utilisation was preferential. Given this variability we sought to gain a more holistic indication of strain performance that was independent of fermentation rate and therefore utilized the area under the curve (AUC) of fermentation of individual or combined sugars. In this way it was possible to rank the 20 strains for their ability to consume fructose relative to glucose. Moreover, it was shown that fermentations performed in media containing fructose as sole carbon source did not predict the fructophilicity of strains in wine-like conditions (equimolar mixture of glucose and fructose). This work provides important information for programs which seek to generate strains that are faster or more reliable fermenters.

Haemolymph sugar levels in a nectar-feeding ant: dependence on metabolic expenditure and carbohydrate deprivation.

In nectar-feeding insects, sugars are an important source of fuel and energy storage. Here, we analyzed the haemolymph sugar levels in foragers of the ant Camponotus rufipes trained to collect nectar from an artificial feeder, and their dependence on the metabolic rate during feeding. The main sugar found was trehalose, followed by glucose and traces of fructose and sucrose. In foragers, trehalose level was independent of their activity and metabolic rate while feeding. Carbohydrate deprivation of the colony had a strong effect on the haemolymph sugar levels of workers, with a significant decrease in trehalose and glucose with increasing starvation. We also found a correlation between haemolymph sugar levels and behavioral states, with immobile workers having higher trehalose and fructose levels than active ones. It is suggested that under food deprivation, inside-nest workers initially stay completely immobile as a strategy to save energy, and only become active and start to search for food when the trehalose levels decrease even more. Based on a conservative estimation, well-fed ants could travel up to 500 m, or spend more than 20 h inactive at 25 degrees C, using only the energy provided by the haemolymph trehalose, before reaching the levels found in starved nest-mates.

Assessment of insulin resistance in fructose-fed rats with 125I-6-deoxy-6-iodo-D-glucose, a new tracer of glucose transport.

PURPOSE: Insulin resistance, characterised by an insulin-stimulated glucose transport defect, is an important feature of the pre-diabetic state that has been observed in numerous pathological disorders. The purpose of this study was to assess variations in glucose transport in rats using (125)I-6-deoxy-6-iodo-D-glucose (6DIG), a new tracer of glucose transport proposed as an imaging tool to assess insulin resistance in vivo. METHODS: Two protocols were performed, a hyperinsulinaemic-euglycaemic clamp and a normoinsulinaemic-normoglycaemic protocol, in awake control and insulin-resistant fructose-fed rats. The tracer was injected at steady state, and activity in 11 tissues and the blood was assessed ex vivo at several time points. A multicompartmental mathematical model was developed to obtain fractional transfer coefficients of 6DIG from the blood to the organs. RESULTS: Insulin sensitivity of fructose-fed rats, estimated by the glucose infusion rate, was reduced by 40% compared with control rats. At steady state, 6DIG uptake was significantly stimulated by insulin in insulin-sensitive tissues of control rats (basal versus insulin: diaphragm, p < 0.01; muscle, p<0.05; heart, p<0.001), whereas insulin did not stimulate 6DIG uptake in insulin-resistant fructose-fed rats. Moreover, in these tissues, the fractional transfer coefficients of entrance were significantly increased with insulin in control rats (basal vs insulin: diaphragm, p<0.001; muscle, p<0.001; heart, p<0.01) whereas no significant changes were observed in fructose-fed rats. CONCLUSION: This study sets the stage for the future use of 6DIG as a non-invasive means for the evaluation of insulin resistance by nuclear imaging.

Performance assessment of malolactic fermenting bacteria Oenococcus oeni and Lactobacillus brevis in continuous culture.

The growth performance of malolactic fermenting bacteria Oenococcus oeni NCIMB 11648 and Lactobacillus brevis X(2) was assessed in continuous culture. O. oeni grew at a dilution rate range of 0.007 to 0.052 h(-1) in a mixture of 5:6 (g l(-1)) of glucose/fructose at an optimal pH of 4.5, and L. brevis X(2) grew at 0.010 to 0.089 h(-1) in 10 g l(-1) glucose at an optimal pH of 5.5 in a simple and safe medium. The cell dry weight, substrate uptake and product formation were monitored, as well as growth kinetics, yield parameters and fermentation balances were also evaluated under pH control conditions. A comparison of growth characteristics of two strains was made, and this showed significantly different performance. O. oeni has lower maximum specific growth rate (mu(max)=0.073 h(-1)), lower maximum cell productivity (Q (x) (max)=17.6 mg cell l(-1) h(-1)), lower maximum biomass yield (Y (x/s) (max)=7.93 g cell mol(-1) sugar) and higher maintenance coefficient (m (s)=0.45 mmol(-1) sugar g(-1) cell h(-1)) as compared with L. brevis X(2) (mu(max)=0.110 h(-1); Q (x) (max)=93.2 g(-1) cell mol(-1) glucose; Y (x/s) (max)=22.3 g cell mol(-1) glucose; m (s)=0.21 mmol(-1) glucose g(-1) cell h(-1)). These data suggest a possible more productive strategy for their combined use in maturation of cider and wine.