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Sugar malabsorption in functional bowel disease: clinical implications.

OBJECTIVE: To investigate the relationship of sugar malabsorption to the development of clinical symptoms in functional bowel disease. METHODS: Twenty-five consecutive outpatients [five men, 20 women; mean age 38.7 +/- 2.6 (SEM) yr] with functional bowel disease and symptoms suggestive of carbohydrate malabsorption were studied. Twelve healthy subjects [six men, six women; mean age 35.7 +/- 3.7 (SEM) yr] acted as the control group. Sugar malabsorption was assessed by breath-hydrogen test after an oral load of various solutions containing lactose (50 g), fructose (25 g), sorbitol (5 g), fructose plus sorbitol (25 + 5 g), and sucrose (50 g). The severity of symptoms developing after sugar challenge was studied. In addition, the effect on clinical symptoms of a diet free of the offending sugars, compared to a low-fat diet, was assessed. RESULTS: Frequency of sugar malabsorption was high in both patients and controls, with malabsorption of at least one sugar in more than 90% of the subjects. Median symptom scores after both lactose [median 6; interquartile (IQ) range 3-7] and fructose plus sorbitol (median 2; IQ range 0-4) malabsorption were significantly higher than after sucrose load (median 1; IQ range 0-1.5) in functional bowel disease patients (p = 0.001 and p = 0.007, respectively). However, there were no differences in healthy controls. In addition, symptoms score after both lactose and fructose plus sorbitol malabsorption was significantly higher in patients than in control subjects (p = 0.02 and p = 0.008, respectively). On the other hand, H2 production capacity, as measured following lactulose load, was significantly higher in patients than in controls. The clinical symptoms improved in 40% of the evaluated patients after restriction of the offending sugars. CONCLUSIONS: These results suggest that sugar malabsorption may be implicated in the development of abdominal distress in at least a subset of patients with functional bowel disease.

Adult↗

Influence of age and salivary secretion rate on oral sugar clearance.

The influence of age and salivary flow on oral sugar clearance was studied. Saliva was sampled with small circular paper discs, absorbing a certain amount of saliva. Glucose concentration was analysed enzymatically. Salivary glucose clearance was estimated in: (i) children of various ages and adults, (ii) hospitalised elderly patients, elderly people living at home and middle-aged adults, and (iii) elderly individuals with various types of prosthodontic reconstructions. Furthermore, an experimental model with pharmacologically reduced salivary flow was used to evaluate the relationship between salivary glucose clearance and salivary flow, also taking into consideration pH changes in dental plaque and the sugar intake concentration. In addition, an attempt to speed up prolonged oral sugar clearance in a group of hospitalised elderly patients was made with an individual programme. The main findings were that 3-year-olds exhibited higher salivary glucose concentration levels and greater variations in glucose clearance between different food products than older children and adults. Elderly people, especially hospitalised patients, cleared sugar from the oral cavity less effectively than young and middle-aged adults. Removable dentures, particularly complete dentures, contributed to higher salivary sugar concentration in elderly people. Higher glucose levels in saliva and more pronounced pH drops in dental plaque were obtained at low than at normal salivary flow. There was a bi-linear relationship between salivary glucose clearance sublingually and salivary secretion rate. Small changes in salivary flow below the border line values--which correspond to the intersection between the two phases--resulted in relatively large differences in clearance. It was possible to speed up prolonged oral sugar clearance in hospitalised elderly patients by using an individual training programme. To conclude, oral sugar clearance was comparatively slow in young children and in hospitalised elderly patients. A low salivary secretion rate was found to affect oral sugar clearance negatively.

Adolescent↗

Provision of sugar-free medicines for young children: the views of a sample of parents in Northern Ireland.

Sugar in liquid medicines is a factor in dental caries for many children. There has been an increase in the availability of sugar-free medicines in recent years, but they need to be prescribed specifically, because most generic liquid medicines still contain sugar. The object of the present study was to examine parental and child factors which may be linked to the provision of sugar-free alternatives by their medical practitioner. A random sample of 291 parents of children aged four years was obtained. A questionnaire was used to ascertain the extent of their knowledge concerning liquid medicines and their effect on dental health and the frequency with which such medicines are requested by them was assessed. The levels of provision of sugar-free medication and the frequency of used common types of medication were recorded and long term users identified. There was a total of 212 children in the study representing an inclusion rate of 73 per cent of the original sample. Just more than 17 per cent were always provided with a sugar-free liquid medicine, 58.3 per cent sometimes received this form of medication and 24.6 per cent never received it. The strongest individual predictor of a high frequency of provision of the medication in sugar-free form was the frequency of parents requesting this type of medication from their medical doctor and the next strongest predictor was the frequency of use of antibiotics. Parental awareness or knowledge of the importance of the issue for dental health did not affect the outcome in the present study. It is concluded that there is a need for specific campaigns to promote the use of sugar-free medications and further increases in their availability should be sought from manufacturers.

Anti-Bacterial Agents↗

Sugar signalling and gene expression in relation to carbohydrate metabolism under abiotic stresses in plants.

Sucrose is required for plant growth and development. The sugar status of plant cells is sensed by sensor proteins. The signal generated by signal transduction cascades, which could involve mitogen-activated protein kinases, protein phosphatases, Ca 2+ and calmodulins, results in appropriate gene expression. A variety of genes are either induced or repressed depending upon the status of soluble sugars. Abiotic stresses to plants result in major alterations in sugar status and hence affect the expression of various genes by down- and up-regulating their expression. Hexokinase-dependent and hexokinase-independent pathways are involved in sugar sensing. Sucrose also acts as a signal molecule as it affects the activity of a proton-sucrose symporter. The sucrose trans-porter acts as a sucrose sensor and is involved in phloem loading. Fructokinase may represent an additional sensor that bypasses hexokinase phosphorylation especially when sucrose synthase is dominant. Mutants isolated on the basis of response of germination and seedling growth to sugars and reporter-based screening protocols are being used to study the response of altered sugar status on gene expression. Common cis-acting elements in sugar signalling pathways have been identified. Transgenic plants with elevated levels of sugars/sugar alcohols like fructans, raffinose series oligosaccharides, trehalose and mannitol are tolerant to different stresses but have usually impaired growth. Efforts need to be made to have transgenic plants in which abiotic stress responsive genes are expressed only at the time of adverse environmental conditions instead of being constitutively synthesized.

Carbohydrate Metabolism↗

Versatile sugar and valerate metabolic pathways in Paraburkholderia xenovorans LB400 enable tailored poly(3-hydroxybutyrate-co-3-hydroxyvalerate) production.

Poly(3-hydroxybutyrate) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) polymers are accumulated by diverse prokaryotes. Their distinct monomer compositions enable their use as tailored bioplastics. The aims were to characterize the poly(3-hydroxybutyrate) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) synthesis by Paraburkholderia xenovorans LB400 using different sugars and valerate, and to gain genome-oriented insights into polyhydroxyalkanoate production. d-Glucose, d-mannitol, d-gluconate, and d-xylose were evaluated as sole carbon sources or supplemented with valerate. Polyhydroxyalkanoates synthesized by strain LB400 were characterized through GC-MS, GC-FID, FTIR, and 1H and 13C-NMR. P. xenovorans LB400 reached 1.00-1.39 g L-1 of dry cell weight (DCW) with a P(3HB) content of 21-43% w w-1 when grown on different sugars. The addition of valerate to the sugar-grown LB400 cultures yielded a DCW of 1.79 to 2.29 g L-1 and a P(3HB-co-3HV) content of 50.0‒51.2% w w-1, with varying 3HV compositions (28‒43 mol%). The highest 3HV incorporation was observed with d-xylose and valerate. Genomic analyses of strain LB400 revealed key elements of sugar metabolism influencing growth, polymer accumulation, and monomer composition. LB400 genome encodes the PhaJ-like R-specific hydratase and FadJ epimerase, which are potentially useful for modulating copolymer composition. PHA production under bioreactor conditions was evaluated. In a bioreactor fed with d-glucose, LB400 achieved a P(3HB) concentration of 2.2 g L-1. These findings highlight the metabolic versatility of P. xenovorans LB400 in utilizing diverse sugars to produce either P(3HB) or tailor-made P(3HB-co-3HV), supporting the development of bioplastics for specific applications. KEY POINTS: • Strain LB400 produced P(3HB-co-3HV) from various sugars and valerate. • Sugar type drives LB400 PHA copolymer synthesis and composition. • Strain LB400 PHA production was scaled up to a bioreactor.

Polyesters↗

Use of catabolite repression mutants for fermentation of sugar mixtures to ethanol.

Use of agricultural biomass, other than corn-starch, to produce fuel ethanol requires a microorganism that can ferment the mixture of sugars derived from hemicellulose. Escherichia coli metabolizes a wide range of substrates and has been engineered to produce ethanol in high yield from sugar mixtures. E. coli metabolizes glucose in preference to other sugars and, as a result, utilization of the pentoses in hemicellulose-derived sugar mixtures is delayed and may be incomplete. Residual sugar lowers the ethanol yield and is problematic for downstream processing of fermentation products. Therefore, a catabolite repression mutant that simultaneously utilizes glucose and pentoses would be useful for fermentation of complex substrate mixtures. We constructed ethanologenic E. coli strains with a glucose phosphotransferase (ptsG) mutation and used the mutants to ferment glucose, arabinose, and xylose, singly and in mixtures, to ethanol. Yields were 87-94% of theoretical for both the wild type and mutants, but the mutants had an altered pattern of mixed sugar utilization. Phosphotransferase mutants metabolized the pentoses simultaneously with glucose, rather than sequentially. Based upon fermentations of sugar mixtures, a catabolite-repression mutant of ethanologenic E. coli is expected to provide more efficient fermentation of hemicellulose hydrolysates by allowing direct utilization of pentoses.

Biomass↗

Control of sugar utilization in the oral bacteria Streptococcus salivarius and Streptococcus sanguis by the phosphoenolpyruvate: glucose phosphotransferase system.

Three different Strep. salivarius (G2, G5 and G29) and two Strep. sanguis (GS3 and GS12) mutants affected in the phosphoenolpyruvate: glucose phosphotransferase system were selected on agar plates containing lactose and 2-deoxyglucose. All 5 were defective in a membrane-bound component of the transport system and grew less rapidly than the parent strain in 5 mM glucose-containing medium. Mutants G2 and G29 grew poorly in the presence of 5 mM mannose. Growth on mixed substrates revealed that the mutants and wild-type parents behaved differently. Wild-type strains in medium containing glucose plus another sugar (lactose, galactose, melibiose, raffinose or trehalose for Strep. salivarius and lactose, galactose or trehalose for Strep. sanguis) always exhausted most of the glucose before utilizing the other sugar. The mutants used the second sugar concurrently or preferentially to glucose. In medium containing glucose plus fructose or mannose, the wild types consumed both sugars concurrently whereas the mutants utilized the second sugar before glucose. Mutants G2 and G5 were insensitive to repression by fructose and released glucose into the medium when grown in the presence of 0.4 per cent lactose. Mutant G5 also released galactose. Sugar release was not detected with the wild types. The Strep. salivarius mutants contained normal levels of glucokinase and beta-galactosidase but G5 was almost totally devoid of galactokinase activity after growth on lactose. On galactose, the activity was restored. It seems that the phosphoenolpyruvate: glucose phosphotransferase system is involved in the regulation of sugar utilization in these two streptococci.

Glucose↗

Mechanism of O2- (-) and H2O2-induced stimulation of sugar transport in mouse fibroblast BALB/3T3 cells.

Xanthine/xanthine oxidase and H2O2 stimulated sugar transport. Application of superoxide dismutase and catalase to the cells showed an inhibitory effect on these agent-stimulated sugar transports. Addition of amiloride and 4-acetamide-4'-isothiocyanostilbene-2,2'-disulfonic acid (SITS), which abolish the cytoplasmic alkalinization, inhibited the stimulation of sugar transport by xanthine/xanthine oxidase in the presence of catalase. The calmodulin antagonists, N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide (W-7) and trifluoperazine inhibited H2O2-stimulated sugar transport. These results suggest that O2- stimulates sugar transport in an intracellular pH-dependent manner and that H2O2 stimulates sugar transport in a calcium-calmodulin-dependent manner. These mechanisms may be involved in sugar-transport stimulation in mouse fibroblast BALB/3T3 cells by the tumor-promoting phorbol ester phorbol-12,13-dibutyrate and insulin, since the stimulatory effects of these agents were inhibited by scavengers of oxygen radicals.

4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfo↗

Introduction of a new scale into reversed-phase high-performance liquid chromatography of pyridylamino sugar chains for structural assignment.

Addition of a monosaccharide residue to a pyridylaminated (PA)-N-linked sugar chain results in an increment or decrement in the elution time on reversed-phase HPLC, the difference being defined as the partial elution time of the residue. Based on this principle, an empirical rule was deduced, which states that the elution time is roughly equal to the sum of the partial elution times of the component sugar residues [Anal. Biochem., 167 (1987) 321-326]. In practice, however, some partial elution times obtained from different pairs of mother PA-sugar chains are found to deviate, and consequently the closeness of the elution times of PA-sugar chains calculated therefrom to the observed times is reduced in such cases. To improve the reliability of the additivity rule and to generalize elution times so that they are less dependent on minor alterations in the elution conditions, we have devised a new scale for elution time, which we have named a reversed-phase scale. The elution times on the reversed-phase scale (the R values) are read from a conversion curve constructed using the elution times of eight selected standard PA-sugar chains. The partial elution times on the reversed-phase scale of 22 monosaccharide residues were calculated from the R values of 93 PA-sugar chains. The R values obtained by summing the partial elution times of all the component monosaccharide residues became much closer to the R values obtained from the reversed-phase scale, compared to the results obtained using the previous method. In addition, the R values were less influenced by minor change in the elution conditions. These features of the new scale allow more accurate structural assignment of sugar chains.

Amino Sugars↗

Hexokinase activity alters sugar-nucleotide formation in maize root homogenates.

Two pools of hexokinase activities differing in sensitivity to ADP inhibition were characterised in maize roots. In order to evaluate how glucose utilisation could be affected by these hexokinases, glucose-6-P and NDP-5'-sugar levels were measured after a D-[U-14C]glucose pulse in root extracts in the presence of 0 or 1 mM ADP. Analysis of radio-labelled activated sugars by paper chromatography revealed that: (1) without ADP, nearly 20% of the 14C appeared in NDP-5'-sugars; (2) 0.1 mM ADP inhibited 14C-NDP-5'-sugar formation by 85%; and (3) with 1 mM ADP, 14C-NDP-5'-sugars were undetectable, but substantial (14%) 14C accumulated as glucose-6-P. Mannoheptulose, a hexokinase inhibitor, blocked the NDP-5'-sugar formation, but did not modify the amount of 14C-glucose-6-P in root extracts either with or without ADP. The analysis of the hexokinase activities with 0.8 mM glucose in maize root extracts showed that: (1) mitochondrial hexokinase activity was totally inhibited by 30 mM mannoheptulose; and (2) the cytosolic hexokinase was inhibited by only 30%. These data suggest that NDP-5'-sugar synthesis is sensitive to ADP fluctuations and that mannoheptulose affects preferentially the mitochondrial-bound hexokinase, but the cytosolic form is less sensitive. We propose that the mitochondrial hexokinase is the main energy charge sensor in this pathway in maize.

Adenosine Diphosphate↗

Analysis of transport activity of Arabidopsis sugar alcohol permease homolog AtPLT5.

The transporters responsible for sugar uptake into non-photosynthetic sink tissues in plants, such as roots and flowers, have not been fully identified and analyzed. Plants encode around 100 putative sugar transporters within the major facilitator superfamily, yet only a few have been studied. Here we report the analysis of a sugar alcohol permease homolog (AtPLT5, At3g18830) from Arabidopsis. A wide range of sugars including hexoses, pentoses, tetroses, a sugar acid, and sugar alcohols but not disaccharides induced inward currents in oocytes expressing AtPLT5. AtPLT5 expression also resulted in 14C-labeled substrate uptake in oocytes, indicating that AtPLT5 encodes an ion-coupled uptake transporter. K(0.5) values for glucose and sorbitol were highly dependent on external pH. Expression of AtPLT5 was found primarily in sink tissues: in the elongation zone of roots, in the inflorescence stem, and several floral structures, especially in the floral abscission zone. Expression was induced by mechanical wounding and insect feeding. Analysis of transport properties and expression in Arabidopsis indicate that AtPLT5 functions to transport a wide range of sugars into specific sink tissues in the plant.

Animals↗

The validity of assessing changes in intestinal absorption mechanisms for dietary sugars with non-metabolizable analogues (glucalogues).

1. Transfer potentials were obtained from everted jejunal sacs prepared from fed euthyroid, fasted euthyroid, fed hypothyroid and fasted hypothyroid rats by addition of serial concentrations of the dietary sugars glucose and galactose and the glucalogues 3-0-methyl glucose, alpha-methyl glucoside and 6-deoxy-D-glucose to the incubation fluids. The kinetic parameters of 'apparent Michaelis constant' (apparent Km) and maximum transfer potential difference (pdmax) obtained from the results were used to characterize the changes in the electrogenic transfer mechanisms for these substrates. 2. Analysis of the significant differential changes in values for 'apparent Km' and pdmax for the two dietary sugars and the three glucalogues indicated heterogeneity in the mechanisms for sugar transfer across the intestine and suggested a minimum of four possible carriers. 3. The validity of using glucalogues to characterize changes in the transfer mechanisms for the dietary sugars in different dietary and hormonal states was assessed. None of the kinetic parameters for electrogenic glucalogue transfer matched those for the dietary sugars in all the experimental conditions. The employment of glucalogues to assess changes in electrogenic transfer mechanisms for dietary sugars can thus lead to invalid conclusions. 4. Fasting decreased the 'apparent Km' of dietary sugars and the glucalogues. However, the pdmax values for glucose, galactose, and alpha-methyl glucoside decreased whereas those for 3-0-methyl glucose and 6-deoxy-D-glucose did not. 5. Hypothyroidism showed different effects in fed and fasted intestine. Because hypothyroidism induced a reduction in food intake, separation of the direct effects of the condition on electrogenic transfer from reduced food intake effect was not possible.

Animals↗

Transcriptional landscape and dynamics involved in sugar and acid accumulation during apple fruit development.

In fleshy fruit, sugars and acids are central components of fruit flavor and quality. To date, the mechanisms underlying transcriptional regulation of sugar and acid during fruit development remain largely unknown. Here, we combined ATAC-seq with RNA-seq to investigate the genome-wide chromatin accessibility and to identify putative transcription factors related to sugar and acid accumulation during apple (Malus domestica) fruit development. By integrating the differentially accessible regions and differentially expressed genes, we generated a global data set of promoter-accessibility and expression-increased genes. Using this strategy, we constructed a transcriptional regulatory network enabling screening for key transcription factors and target genes involved in sugar and acid accumulation. Among these transcription factors, 5 fruit-specific DNA binding with one finger genes were selected to confirm their regulatory effects, and our results showed that they could affect sugar or acid concentration by regulating the expression of sugar or acid metabolism-related genes in apple fruits. Our transcriptional regulatory network provides a suitable platform to identify candidate genes that control sugar and acid accumulation. Meanwhile, our data set will aid in analyzing other characteristics of apple fruit that have not been illuminated previously. Overall, these findings support a better understanding of the regulatory dynamics during apple fruit development and lay a foundation for quality improvement of apple.

Malus↗

Regulation of exopolysaccharide production by Lactococcus lactis subsp. cremoris By the sugar source.

Lactococcus lactis produced more exopolysaccharide (EPS) on glucose than on fructose as the sugar substrate, although the transcription level of the eps gene cluster was independent of the sugar source. A major difference between cells grown on the two substrates was the capacity to produce sugar nucleotides, the EPS precursors. However, the activities of the enzymes required for the synthesis of nucleotide sugars were not changed upon growth on different sugars. The activity of fructosebisphosphatase (FBPase) was by far the lowest of the enzymes involved in precursor formation under all conditions. FBPase catalyzes the conversion of fructose-1, 6-diphosphate into fructose-6-phosphate, which is an essential step in the biosynthesis of sugar nucleotides from fructose but not from glucose. By overexpression of the fbp gene, which resulted in increased EPS synthesis on fructose, it was proven that the low activity of FBPase is indeed limiting not only for EPS production but also for growth on fructose as a sugar source.

Cell Wall↗

Engineering of carbon distribution between glycolysis and sugar nucleotide biosynthesis in Lactococcus lactis.

We describe the effects of modulating the activities of glucokinase, phosphofructokinase, and phosphoglucomutase on the branching point between sugar degradation and the biosynthesis of sugar nucleotides involved in the production of exopolysaccharide biosynthesis by Lactococcus lactis. This was realized by using a described isogenic L. lactis mutant with reduced enzyme activities or by controlled expression of the well-characterized genes for phosphoglucomutase or glucokinase from Escherichia coli or Bacillus subtilis, respectively. The role of decreased metabolic flux was studied in L. lactis strains with decreased phosphofructokinase activities. The concomitant reduction of the activities of phosphofructokinase and other enzymes encoded by the las operon (lactate dehydrogenase and pyruvate kinase) resulted in significant changes in the concentrations of sugar-phosphates. In contrast, a >25-fold overproduction of glucokinase resulted in 7-fold-increased fructose-6-phosphate levels and 2-fold-reduced glucose-1-phosphate and glucose-6-phosphate levels. However, these increased sugar-phosphate concentrations did not affect the levels of sugar nucleotides. Finally, an approximately 100-fold overproduction of phosphoglucomutase resulted in 5-fold-increased levels of both UDP-glucose and UDP-galactose. While the increased concentrations of sugar-phosphates or sugar nucleotides did not significantly affect the production of exopolysaccharides, they demonstrate the metabolic flexibility of L. lactis.

Carbon↗

Enzymes II of the phosphotransferase system do not catalyze sugar transport in the absence of phosphorylation.

In Salmonella typhimurium, glucose, mannose, and fructose are normally transported and phosphorylated by the phosphoenolpyruvate:sugar phosphotransferase system. We have investigated the transport of these sugars and their non-metabolizable analogs in mutant strains lacking the phospho-carrier proteins of the phosphoenolpyruvate:sugar phosphotransferase system, the enzymes I and HPr, to determine whether the sugar-specific, membrane-bound components of the phosphonenolpyruvate: sugar phosphotransferase system, the enzymes II, can catalyze the uptake of these sugars in the absence of phosphorylation. This process does not occur. We have also isolated mutant strains which lack enzyme I and HPr, but have regained the ability to grow on mannose or fructose. These mutants contained elevated levels of mannokinase (fructokinase). In addition, growth on mannose required constitutive synthesis of the galactose permease. When strains were constructed which lacked the galactose permease, they were unable to grow even on high concentrations of mannose, although elevated levels of mannokinase (fructokinase) were present. These results substantiate the conclusion that the enzymes II of the phosphoenolpyruvate:sugar phosphotransferase system are unable to carry out facilitated diffusion.

Biological Transport, Active↗

An esterase gene from Lactobacillus casei cotranscribed with genes encoding a phosphoenolpyruvate:sugar phosphotransferase system and regulated by a LevR-like activator and sigma54 factor.

A new esterase-encoding gene was found in the draft genome sequence of Lactobacillus casei BL23 (CECT5275). It is located in an operon together with genes encoding the EIIA, EIIB, EIIC, and EIID proteins of a mannose class phosphoenolpyruvate:sugar phosphotransferase system. After overproduction in Escherichia coli and purification, the esterase could hydrolyze acetyl sugars, hence the operon was named esu for esterase-sugar uptake genes. Upstream of the genes encoding the EII components (esuABCD) and the esterase (esuE), two genes transcribed in the opposite sense were found which encode a Bacillus subtilis LevR-like transcriptional activator (esuR) and a sigma54-like transcriptional factor (rpoN). As compared with the wild-type strain, elevated fructose phosphorylation was detected in L. casei mutants constitutively expressing the esu operon. However, none of the many sugars tested could induce the esu operon. The fact that EsuE exhibits esterase activity on acetyl sugars suggests that this operon could be involved in the uptake and metabolism of esterified sugars. Expression of the esu operon is similar to that of the B. subtilis lev operon: it contains a -12,-24 consensus promoter typical of sigma54-regulated genes, and EsuR and RpoN are essential for its transcription which is negatively regulated by EIIB(Esu). The esuABCDE transcription unit represents the first sigma54-regulated operon in lactobacilli. Furthermore, replacement of His852 in the phosphoenolpyruvate:sugar phosphotransferase system regulation domain II of EsuR with Ala indicated that the transcription activator function of EsuR is inhibited by EIIB(Esu)-mediated phosphorylation at His852.

DNA-Binding Proteins↗

Over expression of modified Isomaltulose Synthase Gene II (ImSyGII) under single and double promoters drive unprecedented sugar contents in sugarcane.

Sugarcane has been grown all around the world to meet sugar demands for industrial sector. The current sugar recovery percentage in sugarcane cultivars is dismally low which demands scientific efforts for improvements. Multiple approaches were adopted to enhance sugar contents in commercial sugarcane plants in contrast to conventional plant breeding methods. The exploitation of biotechnological methods and exploration of isomaltulose synthetic genes presented a promising solution to increase the existing low level of sugar recovery percentage in Saccharum officinarum L. Isomaltulose synthase gene II was employed and integrated into plant expression vector driven under the leaf and stem specific promoters terminated by nopaline synthase gene in a cloning strategy shown in the present study. Three gene constructs were developed in various combinations driven under promoters Zea mays ubiquitin and Cestrum Yellow Leaf Curl virus in the single and double combined stacked system. The transformation was executed in multiple formats with single transformed events, double promoter transformation events and triple construct stacked promoters in sugarcane induced calli via the particle gene gun. The transformation of ImSyGII in sugarcane genotype HSF-240 was confirmed by molecular gene analysis while expression quantification was determined through Real Time PCR. Furthermore, HPLC was also done to harvest the increased amounts of Isomaltulose in transgenic sugarcane juice. The present work upheld the enhanced ImSyGII expression in leaves owing to the exploitation of ubiquitin, while the Cestrum Yellow Leaf Curl virus promoter enhanced gene expression in sugarcane stems. The employment of three gene constructs collectively produced elite sugar lines producing more than 78% enhancements in whole sugar recovery percentage. The mature internode proved highly efficient and receptive regarding the production of isomaltulose. Quantifications and sugar contents evaluations upheld an increased Brix ratio of transgenic sugarcane lines than control lines.

Saccharum↗