Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: 'ZP domain' of human zona pellucida glycoprotein-1 binds to human spermatozoa and induces acrosomal exocytosis

doi: 10.1186/1477-7827-8-110

Figure Lengend Snippet: Profile of lectins binding to baculovirus-expressed human ZP1 273-551aa in ELISA . Microtitration plates were coated with the baculovirus-expressed recombinant ZP1 273-551aa (open bar; 500 ng/well) and processed for evaluation of binding to 21 different biotinylated lectins in an ELISA as described in Methods . As an internal control, microtitration plates coated with same amount of E. coli -expressed recombinant human ZP1 273-551aa (grey bar; unpublished observations) were also used. Values are expressed as absorbance obtained with various lectins binding to the respective recombinant protein, after deducting the non-specific binding of the lectins to the uncoated wells. Each bar represents a mean of duplicate experiments and standard deviation of the absorbance values. The lectins tested were GSL-I: Griffon simplicifolia lectin I, SBA: Soybean agglutinin, DSL: Datura stramonium lectin, PSA: Pisum sativum agglutinin, ConA: Concanavalin A, ECL: Erythrina cristagalli lectin, LCA: Lens culinaris agglutinin, DBA: Dolichos biflorus agglutinin, LEL: Lycopersicon esculentum lectin, PHA-L: Phaseolus vulgaris leucoagglutinin, UEA-I: Ulex europaeus agglutinin I, STL: Solanum tuberosum lectin, PHA-E: Phaseolus vulgaris erythroagglutinin, RCA: Ricinus communis agglutinin, VVA: Vicia villosa agglutinin, SJA: Sophora japonica agglutinin, PNA: Peanut agglutinin, S- WGA: Succinylated Wheat germ agglutinin, Jacalin, WGA: Wheat Germ agglutinin and GSL II: Griffonia simplicifolia lectin II.

Article Snippet: The plate was blocked with 0.1% Tween-20 in PBS (PBST, 200 μl/well) for 90 min at 37°C followed by incubation with 21 biotinylated lectins (1 μg/ml; 100 μl/well) at 37°C for 1 h. The biotinylated lectins available in the Lectin kit-I, II and III (Vector Laboratories, Burlingame, CA, USA) were used in the lectin binding assay.

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Recombinant, Standard Deviation

Journal: Biotechnology for Biofuels

Article Title: Metabolic engineering of Corynebacterium glutamicum for efficient production of succinate from lignocellulosic hydrolysate

doi: 10.1186/s13068-018-1094-z

Figure Lengend Snippet: Succinate biosynthesis pathway of C. glutamicum . The bold black arrows indicate metabolic fluxes increased by overexpression or introduction of the corresponding genes. The gray arrows indicate the reactions leading to a byproduct or presumably irrelevant reactions. Deleted genes are indicated with crosses. Metabolites: G6P glucose-6-phosphate, 6PGL 6-phosphoglucono-1,5-lactone, 6PG 6-phosphogluconate, Ru5P ribulose-5-phosphate, Xu5P xylulose-5-phosphate, R5P ribose-5-phosphate, G3P glyceraldehyde-3-phosphate, S7P sedoheptulose-7-phosphate, F6P fructose-6-phosphate, FBP fructose-1,6-bisphosphate, E4P erythrose-4-phosphate, DHAP dihydroxyacetone, DPG glycerate-1,3-diphosphate, 3PG glycerate-3-phosphate, 2PG glycerate-2-phosphate, PEP phosphoenolpyruvate, PYR pyruvate, AcP acetyl phosphate, AcCoA acetyl-CoA. Genes and their encoded enzymes: iolT encoding myo-inositol permease, glk encoding glucokinase, ptsG encoding glucose-EII of phosphoenolpyruvate phosphotransferase system (PTS), pgi encoding glucose-6-phosphate isomerase, araE encoding a H + symporter protein, xylA encoding xylose isomerase, xylB encoding xylulokinase, zwf and opcA encoding glucose-6-phosphate dehydrogenase, devB encoding 6-phosphogluconolactonase, gnd encoding 6-phosphogluconate dehydrogenase, tkt encoding transketolase, tal encoding transaldolase, pqo encoding pyruvate: quinone oxidoreductase, pta encoding phosphotransacetylase, ackA encoding acetate kinase, cat encoding acetyl-CoA:CoA transferase, aceE encoding pyruvate complex dehydrogenase E1 component, ppc encoding phosphoenolpyruvate carboxylase, pyc encoding pyruvate carboxylase, mdh encoding malate dehydrogenase, gltA encoding citrate synthase, sucE encoding succinate exporter

Article Snippet: ATCC 13032 , C. glutamicum wild-type biotin auxotrophic , ATCC.

Techniques: Over Expression

Journal: Biotechnology for Biofuels

Article Title: Metabolic engineering of Corynebacterium glutamicum for efficient production of succinate from lignocellulosic hydrolysate

doi: 10.1186/s13068-018-1094-z

Figure Lengend Snippet: Strains and plasmids used in this study

Article Snippet: ATCC 13032 , C. glutamicum wild-type biotin auxotrophic , ATCC.

Techniques: Plasmid Preparation, Control, Derivative Assay, Over Expression

Journal: Biotechnology for Biofuels

Article Title: Metabolic engineering of Corynebacterium glutamicum for efficient production of succinate from lignocellulosic hydrolysate

doi: 10.1186/s13068-018-1094-z

Figure Lengend Snippet: Batch cultivation of C. glutamicum I-pXMJ19 (squares), I-eco (diamonds), I-ppm (circles), I-sco (upward triangles), and I-xcb (downward triangles). a Profiles of cell growth (filled symbols) and xylose consumption (open symbols) under aerobic conditions. The strains were cultured in CGXIIA medium containing 20 g L −1 xylose in 500-mL flasks at 30 °C and 220 rpm with an initial OD 600 of 0.8, and were induced with 0.5 mM IPTG. b SDS–PAGE analysis of intracellular proteins extracted from I-eco, I-ppm, I-sco and I-xcb from cultures with an OD 600 of 5. Proteins were separated on a 12% SDS-PAGE gel. c Profiles of xylose consumption (open symbols) under anaerobic conditions with an initial OD 600 of 30. 30 g L −1 4MgCO 3 ·Mg(OH) 2 ·5H 2 O, 200 mM sodium bicarbonate and 1 mM IPTG were added into the CGXIIB medium

Article Snippet: ATCC 13032 , C. glutamicum wild-type biotin auxotrophic , ATCC.

Techniques: Cell Culture, SDS Page

Journal: Biotechnology for Biofuels

Article Title: Metabolic engineering of Corynebacterium glutamicum for efficient production of succinate from lignocellulosic hydrolysate

doi: 10.1186/s13068-018-1094-z

Figure Lengend Snippet: Batch fermentation of C. glutamicum CGS1 (squares), CGS3 (circles) and CGS5 (upward triangles). a Profiles of cell growth (filled symbols) and xylose consumption (open symbols) in CGXIIA medium with 30 g L −1 xylose. b Profiles of succinate production (filled symbols) and xylose consumption (open symbols) under anaerobic conditions with an initial OD 600 of 30. 30 g L −1 4MgCO 3 ·Mg(OH) 2 ·5H 2 O, 200 mM sodium bicarbonate, and 1 mM IPTG were added into the CGXIIB medium. c Profiles of succinate yield (black bars), xylose consumption rate (white bars, calculated at 8 h), and succinate productivity (grey bars, calculated at 8 h) under anaerobic conditions

Article Snippet: ATCC 13032 , C. glutamicum wild-type biotin auxotrophic , ATCC.

Techniques:

Journal: Biotechnology for Biofuels

Article Title: Metabolic engineering of Corynebacterium glutamicum for efficient production of succinate from lignocellulosic hydrolysate

doi: 10.1186/s13068-018-1094-z

Figure Lengend Snippet: Production of succinate from non-food lignocellulosic biomass

Article Snippet: ATCC 13032 , C. glutamicum wild-type biotin auxotrophic , ATCC.

Techniques: