Cost-Efficient Production of the Sphingan WL Gum by Sphingomonas sp. WG Using Molasses and Sucrose as the Carbon Sources.

The polysaccharide WL gum is produced by the marine microorganism Sphingomonas sp. WG and presents great commercial utility potential in many industries especially in oil industries. However, the high fermentation cost limits its wide application. Therefore, an efficient production system at a lower cost was established using beet molasses to partially replace the commonly used carbon sources. Four different molasses were screened and their composition was investigated. One-factor design and RSM statistical analysis were employed to optimize the WL gum fermentation medium. The effects of molasses on the rheological properties and gene expression of WL gum were also investigated. The results showed that the pretreated beet molasses generated both high broth viscosity and WL gum production (12.94 Pa·s and 11.16 g/L). Heavy metal ions and ash were found to be the key factors in unpretreated and pretreated molasses affecting WL production. The cost-efficient production medium contained (g/L): sucrose 61.79, molasses 9.95, yeast extract 1.23, K2HPO4 1, MgSO4 0.1, ZnSO4 0.1 and the WL gum production reached 40.25 ± 1.15 g/L. The WL gum product WL-molasses showed the higher apparent viscosity, and viscous modulus and elastic modulus than WL-sucrose and WL-mix, which might be related to its highest molecular mass. The higher expressional level of genes such as pgm, ugp, ugd, rmlA, welS, and welG in WL gum synthesis in the mixed carbon source medium caused the high production and broth viscosity. This work provided a cost-efficient method for WL gum production.

Zinc oxide nanoparticles (ZnONPs) influenced seed development, grain quality, and remobilization by affecting the transcription of microRNA 171 (miR171), miR156, NAM, and SUT genes in wheat (Triticum aestivum): a biological advantage and risk assessment study.

Limited studies have been conducted on the role of microRNAs (miRs) and transcription factors in regulating plant cell responses to nanoparticles. This study attempted to address whether the foliar application of zinc oxide nanoparticles (ZnONPs; 0, 10, 25, and 50 mgL-1) can affect miRs, gene expression, and wheat grain quality. The seedlings were sprayed with ZnONPs (0, 10, 25, and 50 mgL-1) or bulk counterpart (BZnO) five times at 72 h intervals. The application of ZnONPs at 10 mgL-1 increased the number of spikelets and seed weight, while the nano-supplement at 50 mgL-1 was accompanied by severe restriction on developing spikes and grains. ZnONPs, in a dose-dependent manner, transcriptionally influenced miR156 and miR171. The expression of miR171 showed a similar trend to that of miR156. The ZnONPs at optimum concentration upregulated the NAM transcription factor and sucrose transporter (SUT) at transcriptional levels. However, the transcription of both NAM and SUT genes displayed a downward trend in response to the toxic dose of ZnONPs (50 mgL-1). Utilization of ZnONPs increased proline and total soluble phenolic content. Monitoring the accumulation of carbohydrates, including fructan, glucose, fructose, and sucrose, revealed that ZnONPs at 10 mgL-1 modified the source/sink communication and nutrient remobilization. The molecular and physiological data revealed that the expression of miR156 and miR171 is tightly linked to seed grain development, remobilization of carbohydrates, and genes involved in nutrient transportation. This study establishes a novel strategy for obtaining higher yields in crops. This biological risk assessment investigation also displays the potential hazard of applying ZnONPs at the flowering developmental phase.

Biotransformation of sucrose rich Maple syrups into fructooligosaccharides, oligolevans and levans using levansucrase biocatalyst: Bioprocess optimization and prebiotic activity assessment.

Maple syrup was investigated as a source to produce FOSs and ß-(2-6)-linked-oligolevans/levans. The modulation of this biotransformation was achieved through the control of Maple syrup °Bx and reaction conditions. Reaction time was identified as the most influential factor for the oligolevans/FOSs production in Maple syrup 30°Bx reaction system as well as for the oligolevans/levans synthesis in the 66°Bx one. In the predictive model of oligolevans/levans production in Maple syrup 60°Bx, the interactive effect between levansucrase unit and reaction time was significant (p-value of 0.0008). The optimal conditions for oligolevans/FOSs production (109.20 g/L) in Maple syrup 30°Bx were 3.73 U/mL, pH 6.60 and 23.12 h; while 5 U/mL, pH 6.04 and 29.92 h were identified as the optimal conditions for oligolevans/levans production (147.09 g/L) in Maple syrup 66°Bx. As compared to inulin-type commercial FOSs, the fermentation of oligolevans/FOSs from Maple syrup led to a higher count of Lactobacillus acidophilus and Bifidobacterium lactis and resulted in a higher production of lactic acid. This study lays the foundation for the biotransformation of Maple syrups into functional prebiotic ingredients.

Sucrose Utilization for Improved Crop Yields: A Review Article.

Photosynthetic carbon converted to sucrose is vital for plant growth. Sucrose acts as a signaling molecule and a primary energy source that coordinates the source and sink development. Alteration in source-sink balance halts the physiological and developmental processes of plants, since plant growth is mostly triggered when the primary assimilates in the source leaf balance with the metabolic needs of the heterotrophic sinks. To measure up with the sink organ's metabolic needs, the improvement of photosynthetic carbon to synthesis sucrose, its remobilization, and utilization at the sink level becomes imperative. However, environmental cues that influence sucrose balance within these plant organs, limiting positive yield prospects, have also been a rising issue over the past few decades. Thus, this review discusses strategies to improve photosynthetic carbon assimilation, the pathways actively involved in the transport of sucrose from source to sink organs, and their utilization at the sink organ. We further emphasize the impact of various environmental cues on sucrose transport and utilization, and the strategic yield improvement approaches under such conditions.

Plant cell cultures as food-aspects of sustainability and safety.

KEY MESSAGE: Sustainability and safety aspects of plant cell cultures as food are presented. Applicability of dairy side streams as carbon source and use of natural growth enhancers in cultivation are shown. Biotechnologically produced cellular products are currently emerging to replace and add into the portfolio of agriculturally derived commodities. Plant cell cultures used for food could supplement current food production. However, still many aspects need to be resolved before this new food concept can enter the market. Issues related to sustainability and safety for human consumption are relevant for both consumers and regulators. In this study, two plant cell cultures, deriving from arctic bramble (Rubus arcticus) and birch (Betula pendula), were cultivated using lactose-rich dairy side streams as alternative carbon sources to replace sucrose. Biomasses were comparable to those of original plant cell culture media when up to 83% and 75% of the original sucrose was replaced by these side streams for arctic bramble and birch cell cultures, respectively. Furthermore, nutritional composition or sensory properties were not compromised. Synthetic plant growth regulators were replaced by natural components, such as coconut water and IAA for several subculture cycles. Finally, it was shown that only trace amounts of free growth regulators are present in the cells at the harvesting point and assessment by freshwater crustaceans assay indicated that toxicity of the cells was not exceeding that of traditionally consumed bilberry fruit.

Cytosolic/Plastid Glyceraldehyde-3-Phosphate Dehydrogenase Is a Negative Regulator of Strawberry Fruit Ripening.

Cytosolic glyceraldehyde-3-phosphate dehydrogenase (GAPC) and plastid glyceraldehyde-3-phosphate dehydrogenase (GAPCp) are key enzymes in glycolysis. Besides their catalytic function, GAPC/GAPCp participates in the regulation of plant stress response and growth and development. However, the involvement of GAPC/GAPCp in the regulation of fruit ripening is unclear. In this study, FaGAPC2 and FaGAPCp1 in strawberries were isolated and analyzed. FaGAPC2 and FaGAPCp1 transcripts showed high transcript levels in the fruit. Transient overexpression of FaGAPC2 and FaGAPCp1 delayed fruit ripening, whereas RNA interference promoted fruit ripening and affected fruit anthocyanins and sucrose levels. Change in the expression patterns of FaGAPC2 and FaGAPCp1 also influenced the expression of several glycolysis-related and ripening-related genes such as CEL1, CEL2, SS, ANS, MYB5, NCED1, ABI1, ALDO, PK, and G6PDH, and H2O2 level and reduced glutathione (GSH)/glutathione disulfide (GSSG) redox potential. Meanwhile, metabolomics experiments showed that transient overexpression of FaGAPCp1 resulted in a decrease in anthocyanins, flavonoids, organic acid, amino acids, and their derivatives. In addition, abscisic acid (ABA) and sucrose treatment induced the production of large amounts of H2O2 and inhibited the expression of FaGAPC2/FaGAPCp1 in strawberry fruit. These results revealed that FaGAPC2/FaGAPCp1 is a negative regulator of ABA and sucrose mediated fruit ripening which can be regulated by oxidative stress.

Assessment of sugar and sugar accumulation-related gene expression profiles reveal new insight into the formation of low sugar accumulation trait in a sweet orange (Citrus sinensis) bud mutant.

The accumulation of soluble sugars in fleshy fruits largely determines their sweetness or taste. A spontaneous sweet orange mutant 'Hong Anliu' (HAL, Citrus sinensis) accumulates low soluble sugar content in fruit juice sacs than its wild type, 'Anliu' (AL) orange; however, the cause of reduced sugar content in 'HAL' fruit remains unclear. In this study, sugar content and expression profiles of genes involved in sugar metabolism and transport were compared between 'HAL' and 'AL' fruit juice sacs. In both cultivars, fructose and glucose displayed the increasing trends with significantly lower contents in 'HAL' than 'AL' after 160 DAF; moreover, sucrose had a declining trend in 'HAL' and increasing trend in 'AL' with fruit development. On the other hand, transcript levels of VINV, CWINV1, CWINV2, SUS4, SUS5, SPS1, SPS2, VPP-1, VPP-2, and some sugar transporter genes were significantly decreased in 'HAL' compared with 'AL' after 100 DAF or 160 DAF. Interestingly, the transcript levels of SPS2 and SUT2 exhibited a similar trend as it was found for sucrose content in both cultivars. These results suggested that the low sugar accumulation in 'HAL' fruit JS is accompanied by the reduced sink strength, sucrose-synthesis ability, and vacuolar storage ability compared with 'AL'; reduction of CWINVs, VINV, SPS2, SUT2, VPP-1, and VPP-2 transcript levels possibly plays a key role in the low storage of soluble sugars in the vacuoles of mutant juice sacs.

Assessment of malathion toxicity on cytophysiological activity, DNA damage and antioxidant enzymes in root of Allium cepa model.

The current study was emphasized to assess the effect of malathion on root system (cell division and kinetics of the root elongation) and stress related parameters in Allium cepa L. The roots were exposed to different concentrations (0.05, 0.13, 0.26, 0.39 and 0.52 g/L) of malathion for different treatment periods (4, 8 and 18 h). The results revealed that malathion application affected the growth rate and cell division in root tips. The root elongation kinetics were impaired at 0.13 to 0.52 g/L concentrations. Reduction in tissue water content (TWC) indicated the limited osmotic adjustment due to membrane damage. Further, a decrease in sucrose content was observed in contrast to the accumulation of proline (upto 0.39 g/L). Moreover, malathion exposure elevated the levels of lipid peroxidation followed by changes in antioxidant enzymes status. The activities of ascorbate peroxidase (APX) and glutathione reductase (GR) were down-regulated whereas the activities of catalase (CAT), glutathione-S-transferase (GST) and superoxide dismutase (SOD) were up-regulated except in 0.52 g/L malathion. The molecular docking study of malathion with CAT, GST, SOD, APX and GR also supported of above results for their activity. All these physiological responses varied with increasing malathion concentration and duration of treatment. The single cell gel electrophoresis results showed that all concentrations of malathion induced DNA damage in root cells. The findings depicted that malathion application induces cytotoxic and phytotoxic effects mediated through oxidative stress and subsequent injuries.

Levan production from sucrose using chicken feather peptone as a low cost supplemental nutrient source.

Chicken feather peptone (CFP) derived from poultry waste is a rich source of essential minerals and amino acids. This, along with suitable carbon source, can be used as a low cost complex supplemental nutrient source for microbial fermentation. In the present work, CFP blended with sucrose was evaluated for the production of levan using Bacillus subtilis MTCC 441. Amount of CFP added to the medium significantly influenced levan production and it was found that at a concentration 2 g/L, maximum levan yield of 0.26 ±â€¯0.04 g/g sucrose was obtained. The levan yield obtained with CFP as a low cost supplemental nutrient source was comparable with that obtained from commercial medium (0.31 ±â€¯0.02 g/g sucrose). Levan produced using CFP was tested on primary cell lines at various concentrations (100-1000 µM) and found to be non-toxic and bio-compatible in nature. This indicates that CFP could be used as low cost nutrient source for levan production.

Energy Expenditure, Carbohydrate Oxidation and Appetitive Responses to Sucrose or Sucralose in Humans: A Pilot Study.

BACKGROUND: In light of obesity, replacing sugar with non-nutritive sweeteners is commonly used to reduce sugar content of food products. This study aimed to compare human energy expenditure (EE), carbohydrate oxidation and food intake after the ingestion of test foods sweetened with sucrose or a non-nutritive sweetener. METHODS: This was an acute crossover feeding study that entailed consumption of three test foods: jelly sweetened with 50 g sucrose (SUCROSE), with 120 mg of sucralose only (NNS), or 120 mg sucralose but matched in carbohydrate with 50 g maltodextrin (MALT). On test days, participants arrived at the research facility after an overnight fast. Resting energy expenditure (indirect calorimeter) was measured for 30 min followed by jelly consumption. Participants' EE and substrate oxidation were measured for 90 min subsequently. After EE assessment, participants completed a meal challenge before leaving the research facility, and recorded food intake for the remaining day. Subjective appetite ratings were assessed before and after test foods and meal challenge. RESULTS: Eleven participants completed the study. EE was higher in SUCROSE and MALT than NNS, but not statistically significant. Carbohydrate oxidation was SUCROSE > MALT > NNS (p < 0.001). Earlier and bigger rise in carbohydrate oxidation was observed in SUCROSE than MALT, although both were carbohydrate-matched. NNS did not promote energy expenditure, carbohydrate oxidation or stimulate appetite. CONCLUSIONS: Foods sweetened with sucrose or non-nutritive sweeteners but matched in carbohydrate content have different effects on human EE and carbohydrate oxidation. Sucralose alone did not affect EE, but lower energy in the test food from sugar replacement was eventually fully compensated. Findings from this pilot study should be verified with bigger clinical studies in the future to establish clinical relevance.

Commercially available non-nutritive sweeteners modulate the antioxidant status of type 2 diabetic rats.

Non-nutritive sweeteners (NNS) are increasingly being used by diabetics, but little is known about their effects on antioxidant status. We investigated the effects of ad libitum consumption of commercially available NNS (aspartame, saccharin, sucralose, and cyclamate-based sweeteners) on antioxidative markers in a rat model of type 2 diabetes (T2D). NNS consumption reduced (p < 0.05) T2D-induced lipid peroxidation and boosted serum, hepatic, renal, cardiac, and pancreatic glutathione (GSH) levels. Catalase, glutathione reductase, superoxide dismutase, and glutathione peroxidase activity was increased in the serum and most organs upon diabetes induction, perhaps due to adaptative antioxidant response to the diabetes-induced lipid peroxidation. NNS showed varying effects on serum and tissue antioxidant enzymes of animals. An antioxidant capacity scores sheet of NNS, suggest that aspartame-based NNS may not exert antioxidant effects in diabetics, while saccharin-based NNS may be a potent antioxidative sweetener as seen in the animal model of T2D. PRACTICAL APPLICATIONS: The use of NNS is becoming more popular, especially for diabetic individuals. While there are several commercial NNS available in the market, little is known about how they affect the antioxidant status of consumers. We therefore investigated how some commercially available NNS affect the antioxidant status of diabetic rats. Observed data revealed varying effects of NNS on serum and different organs, which suggest that some NNS may be better than others for diabetic oxidative stress and thus may be recommended for consumers. However, this finding is subject to additional corroborative clinical studies.

Cost-effective fibrinolytic enzyme production by Bacillus subtilis WR350 using medium supplemented with corn steep powder and sucrose.

The goal of this study was to develop a cheap and simple medium and to optimize fermentation parameters for fibrinolytic enzyme production by Bacillus subtilis WR350. A low-cost medium containing 35 g/L sucrose, 20 g/L corn steep powder and 2 g/L MgSO4·7H2O was developed via single-factor and orthogonal experiments. A cheap nitrogen source, corn steep powder, was used to replace the soy peptone present in the initial medium. The highest fibrinolytic activity of 5865 U/mL was achieved using the optimized medium in a 100-L fermenter with an aeration rate of 1.0 vvm and an agitation speed of 200 rpm. The resulting enzyme yield was among the highest described in the literature with respect to fibrinolytic activity, as determined by the fibrin plate method. Techno-economic evaluation indicated that the cost of the optimized medium was only 8.5% of the cost of the initial medium, and the total fermentation cost of fibrinolytic enzyme production using the optimized medium was 23.35% of the cost of using the initial medium.

Reinforcing effectiveness of midazolam, ethanol, and sucrose: behavioral economic comparison of a mixture relative to its component solutions.

Benzodiazepines (BZs) are relatively safe when administered alone. However, these drugs can produce severe side effects when coadministered with ethanol. Despite these adverse consequences, rates of concurrent BZ and ethanol misuse are increasing, and it is unclear whether this behavior is maintained by an enhanced reinforcing effect of the mixture. To address this issue, the current study compared the reinforcing effectiveness of sucrose solutions mixed with midazolam, ethanol, or both. Eight male rats were trained to orally self-administer solutions of either sucrose (S), sucrose+midazolam (SM), sucrose+ethanol (SE), or sucrose+midazolam+ethanol (SME). The response requirement was increased between sessions until the number of reinforcers earned was zero and the relationship between response requirement and reinforcers earned was analyzed using the exponential model of demand. Although baseline intake was similar across drug conditions, consumption of SM was least affected by increases in cost, indicating that it possessed the highest reinforcing effectiveness (i.e. least elastic). The reinforcing effectiveness of S, SE, and SME did not differ significantly. The finding that the reinforcing effectiveness of the SME was less than that of SM does not support the supposition that BZ and ethanol coadministration is maintained by a higher reinforcing effectiveness of the mixture.

Production of poly(malic acid) from sugarcane juice in fermentation by Aureobasidium pullulans: Kinetics and process economics.

Poly(ß-l-malic acid) (PMA) is a biodegradable polymer with many potential biomedical applications. PMA can be readily hydrolyzed to malic acid (MA), which is widely used as an acidulant in foods and pharmaceuticals. PMA production from sucrose and sugarcane juice by Aureobasidium pullulans ZX-10 was studied in shake-flasks and bioreactors, confirming that sugarcane juice can be used as an economical substrate without any pretreatment or nutrients supplementation. A high PMA titer of 116.3g/L and yield of 0.41g/g were achieved in fed-batch fermentation. A high productivity of 0.66g/L·h was achieved in repeated-batch fermentation with cell recycle. These results compared favorably with those obtained from glucose and other biomass feedstocks. A process economic analysis showed that PMA could be produced from sugarcane juice at a cost of $1.33/kg, offering a cost-competitive bio-based PMA for industrial applications.

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.

Development of a commercial scale process for production of 1,4-butanediol from sugar.

A sustainable bioprocess for the production of 1,4-butanediol (BDO) from carbohydrate feedstocks was developed. BDO is a chemical intermediate that goes into a variety of products including automotive parts, electronics, and apparel, and is currently manufactured commercially through energy-intensive petrochemical processes using fossil raw materials. This review highlights the development of an Escherichia coli strain and an overall process that successfully performed at commercial scale for direct production of bio-BDO from dextrose. Achieving such high level performance required an integrated technology platform enabling detailed engineering of enzyme, pathway, metabolic network, and organism, as well as development of effective fermentation and downstream recovery processes.

A Novel Production Method for High-Fructose Glucose Syrup from Sucrose-Containing Biomass by a Newly Isolated Strain of Osmotolerant Meyerozyma guilliermondii.

One osmotolerant strain from among 44 yeast isolates was selected based on its growth abilities in media containing high concentrations of sucrose. This selected strain, named SKENNY, was identified as Meyerozyma guilliermondii by sequencing the internal transcribed spacer regions and partial D1/D2 large-subunit domains of the 26S ribosomal RNA. SK-ENNY was utilized to produce high-fructose glucose syrup (HFGS) from sucrose-containing biomass. Conversion rates to HFGS from 310-610 g/l of pure sucrose and from 75-310 g/l of sugar beet molasses were 73.5-94.1% and 76.2-91.1%, respectively. In the syrups produced, fructose yields were 89.4-100% and 96.5-100% and glucose yields were 57.6-82.5% and 55.3-79.5% of the theoretical values for pure sucrose and molasses sugars, respectively. This is the first report of employing M. guilliermondii for production of HFGS from sucrose-containing biomass.

Economic analysis and environmental impact assessment of three different fermentation processes for fructooligosaccharides production.

Three different fermentation processes for the production of fructooligosaccharides (FOS) were evaluated and compared in terms of economic aspects and environmental impact. The processes included: submerged fermentation of sucrose solution by Aspergillus japonicus using free cells or using the cells immobilized in corn cobs, and solid-state fermentation (SSF) using coffee silverskin as support material and nutrient source. The scale-up was designed using data obtained at laboratory scale and considering an annual productivity goal of 200 t. SSF was the most attractive process in both economic and environmental aspects since it is able to generate FOS with higher annual productivity (232.6 t) and purity (98.6%) than the other processes; reaches the highest annual profit (6.55 M€); presents the lowest payback time (2.27 years); and is more favourable environmentally causing a lower carbon footprint (0.728 kg/kg, expressed in mass of CO2 equivalent per mass of FOS) and the lowest wastewater generation.

A re-assessment of sucrose signaling involved in cluster-root formation and function in phosphate-deficient white lupin (Lupinus albus).

Apart from substrate functions, a signaling role of sucrose in root growth regulation is well established. This raised the question whether sucrose signals might also be involved in formation of cluster-roots (CRs) under phosphate (Pi) limitation, mediating exudation of phosphorus (P)-mobilizing root exudates, e.g. in Lupinus albus and members of the Proteaceae. Earlier studies demonstrated that CR formation in L. albus was mimicked to some extent by external application of high sucrose concentrations (25 mM) in the presence of extremely high P supply (1-10 mM), usually suppressing CR formation. In this study, we re-addressed this question using an axenic hydroponic culture system with normal P supply (0.1 mM) and a range of sucrose applications (0.25-25 mM). The 2.5 mM sucrose concentration was comparable with internal sucrose levels in the zone of CR initiation in first-order laterals of P-deficient plants (3.4 mM) and induced the same CR morphology. Similar to earlier studies, high sucrose concentrations (25 mM) resulted in root thickening and inhibition of root elongation, associated with a 10-fold increase of the internal sucrose level. The sucrose analog palatinose and a combination of glucose/fructose failed to stimulate CR formation under P-sufficient conditions, demonstrating a signal function of sucrose and excluding osmotic or carbon source effects. In contrast to earlier findings, sucrose was able to induce CR formation but had no effect on CR functioning with respect to citrate exudation, in vitro activity and expression of genes encoding phosphoenolpyruvate carboxylase, secretory acid phosphatase and MATE transporters, mediating P-mobilizing functions of CRs.