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851.
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853.
André M. Almeida Enrique Villalobos Susana S. Araújo Luis A. Cardoso Dulce M. Santos Maria A. Santos Pedro S. Fevereiro José M. Torné 《Acta Physiologiae Plantarum》2007,29(3):273-281
Trehalose is a non-reducing disaccharide of glucose that occurs in a large number of organisms, playing an important role
in desiccation and heat stress protection. Trehalose accumulation has proven to be an effective way of increasing drought
tolerance in both model plants such as tobacco and important crops such as potato or rice. In this work we aim to genetically
engineer maize with the Arabidopsis thaliana trehalose phosphate synthase gene (AtTPS1), involved in trehalose biosynthesis via electroporation. A cassette harboring the
AtTPS1 gene under the control of the CaMV35S promoter and the Bialaphos resistance gene Bar as a selective agent was inserted
in the plasmid vector pGreen0229 and used to transform maize inbred line Pa91 via electroporation. Fifteen putative transgenic
plants (T0 generation) were obtained. Transgene integration in T0 plants was analyzed by Southern-blot analysis. T0 plants
had normal phenotypes, although smaller than wild type plants. Contrary to wild type plants, when sexual organs emerged, tassels
appeared at least 15 days earlier than ears in the same plant, rendering impossible the self-pollination of the T0 plant.
These plants were then crossed with wild type plants and in some cases T1 seeds were obtained. T1 seeds presented deformities,
especially the lack of endosperm, but it was still possible to germinate some of these seeds. The so obtained plants were
tested by Northern blot but no AtTPS1 gene expression was detected, a fact possibly due to the incomplete insertion of the
AtTPS1 gene or an extremely low gene expression level. 相似文献
854.
Luis J. Lpez Giraldo Mickaël Laguerre Jrme Lecomte Maria-Cruz Figueroa-Espinoza Nathalie Barouh Bruno Bara Pierre Villeneuve 《Enzyme and microbial technology》2007,41(6-7):721-726
Chlorogenic acid (5-caffeoylquinic acid or 5-CQA) is an hydrophilic phenolic compound with antioxidant properties. Because of its high polarity, its antioxidant properties may be altered when formulated in oil based food or cosmetic preparations. Therefore, there is an interest in trying to enhance its hydrophobicity by grafting of an aliphatic chain. Such lipophilization reactions can be generally achieved through enzymatic catalysis. Our study consisted in synthesizing fatty cholorogenate esters in a two steps reaction. Firstly, 5-CQA was chemically esterified by methanol using an Amberlite IR120 H resin to obtain methyl chlorogenate that is more soluble in the fatty alcohols than 5-CQA. Secondly, this chlorogenate intermediate was transesterified with fatty alcohols of various chain lengths (C4, C8, C12, or C16) in the presence of Candida antarctica B lipase. Under optimal reaction conditions (aw = 0.05; 5% (w/w) of biocatalyst), the transesterification rates were until two-fold higher than in the direct lipase-catalyzed esterification of chlorogenic acid by the same alcohols. The two-step reaction overall yield was between 61 and 93% depending on the alcohol chain length, whereas it was 40–60% for the direct esterification with the same alcohols. 相似文献
855.
Redondo-Nieto M Pulido L Reguera M Bonilla I Bolaños L 《Plant, cell & environment》2007,30(11):1436-1443
The peribacteroid membrane (PBM) of symbiosomes from pea root nodules developed in the presence of boron (+B) was labelled by anti-rhamnogalacturonan II (RGII) (anti-rhamnogalacturonan II pectin polysaccharide) antiserum. However, in nodules from plants grown at low boron (-B), anti-RGII pectin polysaccharide did not stain PBMs. Given that RGII pectin binds to borate, and that symbiosomes differentiate aberrantly in -B nodules because of abnormal vesicle traffic, anti-RGII pectin polysaccharide antigens were further analysed. Following electrophoresis and electroblotting, anti-RGII pectin polysaccharide immunostained three bands in +B but not in -B nodule-derived PBMs. A similar banding pattern was observed after the immunostaining of membrane fractions from uninfected roots, indicating that anti-RGII pectin polysaccharide antigens are common to both peribacteroid and plasma membranes. Protease treatment of samples led to disappearance of anti-RGII pectin polysaccharide labelling, indicating that the three immunostained bands correspond to proteins or glycoproteins. The immunochemical study of RGII antigen distribution during nodule development showed that it is strongly present on the PBM of dividing (undifferentiated) symbiosomes but progressively disappeared during symbiosome maturation. In B-deficient nodules, PBMs were never decorated with RGII antigens, and there was an abnormal targeting of vesicles containing pectic polysaccharide (homogalacturanan) to cell membranes. Overall, these results indicate that RGII, boron and certain membrane (glyco)-proteins may interact closely and function cooperatively in membrane processes associated with symbiosome division and general cell growth. 相似文献
856.
Ferrer EG López Tévez LL Baeza N Correa MJ Okulik N Lezama L Rojo T Castellano EE Piro OE Williams PA 《Journal of inorganic biochemistry》2007,101(5):741-749
Two Cu(II) complexes with cyanoguanidine (cnge) and o-phenanthroline, [Cu(o-phen)(2)(cnge)](NO(3))(2).2H(2)O (1) and [Cu(o-phen)(cnge)(H(2)O)(NO(3))(2)] (2), have been synthesized using different experimental techniques and characterized by elemental analyses, FTIR, diffuse and UV-vis spectra and EPR and magnetic moment measurements techniques. The crystal structures of both complexes were solved by X-ray diffraction methods. Complex (1) crystallizes in the monoclinic space group C2/c with a=12.621(5), b=31.968(3), c=15.39(1)A, beta=111.68(4) degrees, and Z=8 and complex (2) in the monoclinic space group P2(1)/n with a=10.245(1), b=13.923(2), c=12.391(2)A, beta=98.07(1) degrees, and Z=4. The environments of the copper(II) center are trigonal bipyramidal (TBP) for [Cu(o-phen)(2)(cnge)](2+) and an elongated octahedron for [Cu(o-phen)(cnge)(H(2)O)(NO(3))(2)]. Solution studies have been performed to determine the species distribution. The superoxide dismutase (SOD) activities of both complexes have also been tested in order to determine if these compounds mimic the enzymatic action of the enzyme SOD that protects cells against peroxide radicals. 相似文献
857.
Effect of selective growth media on the differentiation of Salmonella enterica serovars by Fourier-Transform Mid-Infrared Spectroscopy 总被引:1,自引:0,他引:1
Baldauf NA Rodriguez-Romo LA Männig A Yousef AE Rodriguez-Saona LE 《Journal of microbiological methods》2007,68(1):106-114
Salmonella enterica serovars are prevalent foodborne pathogens responsible for high numbers of salmonellosis each year. Complex Fourier-transform infrared (FTIR) spectra offer unique biochemical fingerprints of bacteria with bands due to major cellular components. Growth media effects on discrimination of Salmonella serovars by FTIR spectroscopy were investigated and a novel sample preparation technique was developed. S. enterica strains from six serovars were grown on xylose lysine desoxycholate (XLD), Miller-Mallinson (MM), and plate count (PCA) agar as a control (37 degrees C, 24 h). Isolated colonies were suspended in 50% acetonitrile and centrifuged; the remaining pellet was placed on an AMTIR (attenuated total reflectance) crystal and dried under vacuum. Classification models (Soft Independent Modeling of Class Analogy, SIMCA), generated from derivatized infrared spectra (1300-900 cm-1 or 1200-900 cm-1), successfully discriminated among Salmonella strains with major discrimination from 1000-970 cm-1 associated to stretching modes of O-specific polysaccharide chains of lipopolysaccharides. Sample treatment with acetonitrile enhanced safe handling of the bacteria, removed interfering signals and improved the discriminating ability of SIMCA. All media were able to discriminate the S. enterica strains studied, varying in discriminating peaks and class distances in SIMCA classification. This methodology, with the production of large libraries of pathogenic bacteria, could be applied for the rapid monitoring of bacterial contamination in food with minimal sample manipulation. 相似文献
858.
Production of a Rhizopus oryzae lipase from Pichia pastoris using alternative operational strategies
Surribas A Stahn R Montesinos JL Enfors SO Valero F Jahic M 《Journal of biotechnology》2007,130(3):291-299
Different cultivation strategies have been compared for the production of Rhizopus oryzae lipase (ROL) from Pichia pastoris. Several drawbacks have been found using a methanol non-limited fed-batch. On the one hand, oxygen limitation appeared at early cell dry weights and, on the other hand, high cell death was observed. A temperature limited fed-batch has been proposed to solve both problems. However, in our case study a methanol non-limited fed-batch results in better productivities. Finally, a lower salt medium were used to overcome cell death problems and a temperature limited fed-batch was applied thereafter to solve oxygen transfer limitations. This combined strategy has resulted in lower productivities when compared to a methanol non-limited fed-batch. However the culture could be longer prolonged and a 1.3-fold purer final product was obtained mainly due to cell death reduction. 相似文献
859.
Encarnação JM Rosa L Rodrigues R Pedro L da Silva FA Gonçalves J Ferreira GN 《Journal of biotechnology》2007,132(2):142-148
In this work three piezoelectric sensors modified with anti-HIV-1 Vif (virion infectivity factor) single fragment antibodies (4BL scFV), single domains (VH) and camelized single domains (VHD) were constructed and used to detect HIV1 Vif in liquid samples. Dithio-bis-succinimidyl-undecanoate (DSU) and 11-hydroxy-1-undecanethiol (HUT) mixed self assembled monolayers (SAM) were generated at the sensors surface onto which the antibodies were immobilized. All sensors detected specifically the target HIV1-Vif antigen in solution and no unspecific binding was monitored. Impedance analysis was performed to quantify electroacoustic and viscoelastic interferences during antibody immobilization and antigen recognition. The elimination of such interferences enabled the quantitative use of the piezoelectric immunosensors to estimate the antibody surface density as well as antigen binding and equilibrium constants. In spite of the possible limitation regarding mass transport and other related molecular phenomena, which were not considered in the binding model used, this work demonstrates the usefulness of piezoelectric biosensors in biorecognition analysis and evidences the advantages on using simultaneous impedance analysis to bring analytical significance to measured data, and thus to improve piezoelectric sensors sensitivity and applicability. 相似文献
860.