Production of mucic acid from pectin-derived D-galacturonic acid: a review / 生物工程学报

Mucic acid is a hexaric acid that can be biosynthesized by oxidation of D-galacturonic acid, which is the main constituent of pectin. The structure and properties of mucic acid are similar to that of glucaric acid, and can be widely applied in the preparation of important platform compounds, polymers and macromolecular materials. Pectin is a cheap and abundant renewable biomass resource, thus developing a process enabling production of mucic acid from pectin would be of important economic value and environmental significance. This review summarized the structure and hydrolysis of pectin, the catabolism and regulation of D-galacturonic acid in microorganisms, and the strategy for mucic acid production based on engineering of corresponding pathways. The future application of mucic acid are prospected, and future directions for the preparation of mucic acid by biological method are also proposed.

Pectin lyase overproduction by Penicillium griseoroseum mutants resistant to catabolite repression

Abstract Expression of pectinolytic genes is regulated by catabolic repression limiting the production of pectin lyase (PL) if the natural inducer, pectin, is missing from the growth medium. Here, we report the isolation of Penicillium griseoroseum mutants resistant to 2-deoxy-d-glucose (DG) that show resistance to catabolite repression and overproduce PL. Three spontaneous and nine UV-induced mutants were obtained. Some mutants produced sectors (segments morphologically different) that were also studied. The mutants were analyzed for pectinases production on pectinase-agar plates and five mutants and two sectors showing larger clearing zones than the wild type were selected for quantitative assay. Although PL production higher than the wild type has been found, phenotype instability was observed for most of the mutants and, after transfers to nonselective medium, the DG resistance was no longer present. Only mutants M03 and M04 were stable maintaining the DG-resistance phenotype. When growing for 120 h in liquid medium containing glucose with or without pectin, both mutants showed higher PL production. In the presence of glucose as sole carbon source, the mutant M03 produced 7.8-fold more PL than the wild type. Due its phenotypic stability and PL overproduction, the mutant M03 presents potential for industrial applications.

Draft genome sequence of pectic polysaccharide-degrading moderate thermophilic bacterium Geobacillus thermodenitrificans DSM 101594

Abstract Geobacillus thermodenitrificans DSM 101594 was isolated as a producer of extracellular thermostable pectic polysaccharide degrading enzymes. The completely sequenced genome was 3.6 Mb in length with GC content of 48.86%. A number of genes encoding enzymatic active against the high molecular weight polysaccharides of potential biotechnological importance were identified in the genome.

Characterization of pectinase activity for enology from yeasts occurring in Argentine Bonarda grape

Pectinolytic enzymes are greatly important in winemaking due to their ability to degrade pectic polymers from grape, contributing to enhance process efficiency and wine quality. This study aimed to analyze the occurrence of pectinolytic yeasts during spontaneous fermentation of Argentine Bonarda grape, to select yeasts that produce extracellular pectinases and to characterize their pectinolytic activity under wine-like conditions. Isolated yeasts were grouped using PCR-DGGE and identified by partial sequencing of 26S rRNA gene. Isolates comprised 7 genera, with Aureobasidium pullulans as the most predominant pectinolytic species, followed by Rhodotorula dairenensis and Cryptococcus saitoi. No pectinolytic activity was detected among ascomycetous yeasts isolated on grapes and during fermentation, suggesting a low occurrence of pectinolytic yeast species in wine fermentation ecosystem. This is the first study reporting R. dairenensis and Cr. saitoi species with pectinolytic activity. R. dairenensis GM-15 produced pectinases that proved to be highly active at grape pH, at 12 °C, and under ethanol and SO₂ concentrations usually found in vinifications (pectinase activity around 1.1 U/mL). This strain also produced cellulase activity at 12 °C and pH 3.5, but did not produce β-glucosidase activity under these conditions. The strain showed encouraging enological properties for its potential use in low-temperature winemaking.

Impact of pectolytic enzyme on the fermentation time and free-run juice rate of Sharad red wine.

We evaluated the effect of addition of commercially available pectolytic enzyme preparations in the must from Sharad variety, cultivated in India and its influence on some oenological parameters of red wine. The small scale fermentations demonstrated marked difference of organoleptic (colour) and rheologic characteristics (filterability, settling rates) and aroma between enzyme treated samples and control samples. We observed 29% enhancement in free-run juice yield and a remarkable 43% reduction in the fermentation time compared to the control. This biotechnological approach has demonstrated the economic feasibility and the benefits of adding 0.05 g. Kg-1 grapes pectolytic enzyme in terms of yield, aroma, colour, clarity and fermentation period.

Kinetic study on inducibility of polygalacturonases from Aspergillus flavipes FP-500

The aim of this work was to describe growth dynamics, substrate depletion and polygalacturonases production by Aspergillus flavipes FP-500 in batch cultures by means of unstructured models. The microorganism was cultivated on several mono- di- and poly- saccharides, and then the culture development modeled with Monod and Leudeking-Piret equations. The kinetic parameters related to the models (µmax, ãx/s, alpha and beta) were obtained by minimizing the quadratic residuals function with a simplex algorithm. An accurate description of experimental data was attained with the proposed models. Besides, modeling provided significant kinetic information on microbial degradation of complex substrates, such as the correlation between specific growth rate µmax and production yield á, suggesting that A. flavipes FP-500 polygalacturonases are actually constitutive, but also that there is a certain degree of induciblility in these enzymatic activities.

Avaliação da pectina fosfatada aplicada na formação de filmes isolados: material candidato a novos sistemas para liberação modificada de fármacos / Phosphated pectin application in the development of films for drug delivery systems: evaluation of permeability properties and swelling

Pectina fosfatada (Pect-TMFT) juntamente com o α-Gluco-oligossacarídeo (Bioecolians®) foram incorporados à dispersão aquosa de polimetacrilato (Eudragit® RS 30 D) para obtenção de filmes isolados pelo processo "casting" (50 ºC) em placa de Teflon®. A Pect-TMFT e o Bioecolians® foram adicionados à dispersão de Eudragit® RS 30 D nas concentrações: 90: 05: 05, 80: 10: 10 e 70: 20: 10 (4 por cento p/v). Citrato de trietila (TEC) foi utilizado como plastificante (20 por cento da massa do polimetacrilato). As dispersões propostas apresentaram habilidades filmogênicas. Os filmes isolados foram caracterizados pela determinação da permeabilidade ao vapor d'água (TVA), pelo índice de intumescimento (Ii por cento) em fluidos de simulação gástrica (FSG) e intestinal (FSI) e por microscopia eletrônica de varredura (MEV). O aumento do polissacarídeo modificado e do Bioecolians® nos filmes favoreceram a permeabilidade ao vapor d'água e o grau de hidratação quando em FSI (pH= 6,8). Dessa forma, o filme 70: 20: 10, poderá impedir a liberação prematura do fármaco em regiões superiores do TGI quando aplicado como revestimento de sistemas sólidos orais. Além disso, a presença da Pect-TMFT e do Bioecolians®, poderá favorecer a degradação específica da película por enzimas produzidas pela microflora colônica, possibilitando uma cinética de liberação modificada de fármacos apesar das variações inter-individuais de pH que possam existir.

Phosphated pectin (Pect-TMFT) together with α-gluco-oligossacaride (Bioecolians®) were incorporated into aqueous dispersion of polymethacrylate (Eudragit® RS 30 D) to obtain free films by the casting process (50ºC) in Teflon® plate. Pect-TMFT and Bioecolians® were added into dispersions of Eudragit ® RS 30 D at different rates: 90:05:05, 80:10:10 and 70:20:10 (4 percent p/v). Triethyl citrate (TEC) was used as plasticizer (20 percent of mass of the polymethacrylate). The proposed dispersions showed film formation ability. The free films were characterized by the determination of water vapour transmission (WVT), by the swelling index (Ii percent) in fluids of gastric simulation (FGS) and intestinal (FIS) and by scanning electron microscopy (SEM). The increase of modified polysaccharide and Bioecolians® in the films favored their permeability to the water vapour and their hydration degree when in FIS (pH = 6.8). In that way, the obtained film in the concentration 70:20:10, can prevent the premature release of the drug in the up GIT when applied to develop oral solid systems coating. Besides, the presence of Pect-TMFT and Bioecolians® can contribute to the specific membrane degradation by colonic microflora enzymes, making possible a modified release kinetics of drugs even with the existence of inter-individual variations of pH.

Role of pectin from cucumber (Cucumis sativus) in modulation of protein kinase C activity and regulation of glycogen metabolism in rats.

The regulatory role of protein kinase C (PKC) in glycogen metabolism in pectin fed rats was investigated. Administration of pectin (5 g/kg body wt/day) from cucumber (Cucumis sativius L.) led to inhibitory effects on PKC activity in the liver of rats. In the brain and pancreas, PKC activity was significantly higher in pectin-treated rats as compared to the control group. Level of blood glucose was significantly lowered and the level of glycogen in the liver was significantly increased in pectin-administered rats. Glycogen synthase activity was enhanced, while glycogen phosphorylase enzyme showed inhibition in pectin-treated rats. Results indicated that pectin administration might have caused an increase in the secretion of the insulin, which, in turn, had a stimulatory effect on the PKC activity in the pancreas. The decreased PKC activity in the liver and increased PKC activity in the brain and pancreas on pectin administration indicated enhanced glycogenesis and reduced glycogenolysis.

Distribution of pectins in the pollen apertures of Oenothera hookeri: velans ster/+ster

Cell wall pectins are some of the most complex biopolymers known, and yet their functions remain largely mysterious. The aim of this paper was to deepen the study of the spatial pattern of pectin distribution in the aperture of Oenothera hookeri.velans ster/+ster fertile pollen. We used "in situ" immunocytochemical techniques at electron microscopy, involving monoclonal antibodies JIM5 and JIM7 directed against pectin epitopes in fertile pollen grains of Oenothera hookeri.velans ster/+ster. The same region was also analyzed by classical cytochemistry for polysaccharide detection. Immunogold labelling at the JIM7 epitope showed only in mature pollen labelling mainly located at the intine endo-aperture region. Cytoplasmic structures near the plasma membrane of the vegetative cell showed no labelling gold grains. In the same pollen stge the labelling at the JIM5 epitope was mostly confined to a layer located in the limit between the endexine and the ektexine at the level of the border of the oncus. Some tubuli at the base of the ektexine showed also an accumulation of gold particles. No JIM5 label was demonstrated in the aperture chamber and either in any cytoplasmic structure of the pollen grains. The immunocytochemical technique, when compared with the traditional methods for non-cellulose polysaccharide cytochemistry is fare more sensitive and allows the univocal determination of temporal and spatial location of pectins recognized by the JIM7 and JIM5 MAbs.

Actividad enzimática de algunas saprolegniales, oomycetes / Enzymatic activity of some saprolegniales, oomycetes

Se ensayó la actividad enzimática de algunas Saprolegniales (Achlya americana, A. apiculata, A. oligacantha, A. polyandra, A. prolifera, Dictyuchus monosporus, Isoachlyatouroloides, Saprolegnia diclina y S. parasitica), en distintos medios sólidos. Estas especies fueron aisladas de materia orgánica flotante, en Río Santiago (Pdo. Ensenada, Provincia de Buenos Aires, Argentina), y se demostró que poseen actividad enzimática, in vitro, para la descomposición de pectina, celulosa, hemicelulosa, lignina y almidón

Modulation of postprandial glycaemia and insulinaemia by dietary fat.

The present study was designed to examine the effect of corn oil (Co) on postprandial glycaemia and insulinaemia when ingested with glucose (G), casein (Cs), cellulose (Cl) and pectin (P) in various combinations. The study was conducted on six healthy male volunteers, on each of whom six meal tolerance tests were performed. The meals were isocaloric and consisted of G; G and Co; G, Co and Cs; G, Co and P; G, Co, Cs and P; and G, Co, Cs and Cl. The meals were administered after an overnight fast. In addition to a fasting blood sample, blood was collected 0.5, 1.0, 1.5 and 2.0 h after ingestion for measurement of serum glucose and insulin levels. The glycaemic response to GCo was comparable to that to G, but the insulinaemic response was significantly lower. The glycaemic response to GCoCs was significantly lower than that to G but the insulinaemic response to both was comparable. The cellulose containing meal GCoCsCl showed a further reduction in the glycaemic response but not in the insulinaemic response. The pectin containing meals GCoP and GCoCsP gave the lowest glycaemic and insulinaemic responses, the responses to the latter being lower. Corn oil by itself has only a modest effect on the postprandial metabolic response to glucose. Addition of protein and fibre, specially pectin, leads to significant attenuation of glycaemic and insulinaemic responses.