Amino acid sequences of trypsin inhibitors from oriental pickling melon (Cucumis melo L. var. Conomon Makino) seeds.
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Three inhibitors (CMCTI-I, II, and III) were isolated from oriental pickling melon (Cucumis melo L. var. Conomon Makino) seeds by acetone precipitation, gel filtration, and reversed phase chromatography. The amino acid sequences of these inhibitors were: [table; see text] The reactive sites (P1 and P1' sites) of these inhibitors are presumed to be the Lys-Ile indicated by an arrow, comparing them with other squash family inhibitors. All three inhibitors can inhibit lysyl endopeptidase and trypsin at the enzyme-inhibitor ratio of 1:1. (+info)
Epitope mapping on fragments of beet necrotic yellow vein virus coat protein.
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The location of five SDS-stable epitopes on the coat protein (CP) of beet necrotic yellow vein virus was determined by reacting Escherichia coli-expressed free CP, as well as fusion proteins (FP) containing fragments of the CP, with polyclonal and monoclonal antibodies on Western blots. Epitope 1, which has previously been found to be exposed on only one extremity of the virus particle, was located in the region between amino acids (aa) 1 and 7, i.e. on the N terminus of the CP. It was blocked when the N terminus of the CP was linked to a portion of the beta-galactosidase sequence in an FP. Epitope 3, which has previously been found to be exposed on the opposite extremity of the particle, was located in the region between aa 37 and 59. Epitope 4, which is exposed along the entire length of the particle, occurs on the C terminus of CP (aa 183 to 188). Two previously unknown epitopes were identified in the regions between aa 115 and 125 and 125 and 140, respectively. The former was located on the same extremity of the particle as epitope 3, the latter became accessible only after denaturation of the particle. Nothing is known about the probably non-adjacent aa sequences that participate in the formation of the two SDS-labile epitopes (epitopes 2 and 5) which are found on one extremity and along the entire length of the particle, respectively. (+info)
Growth of Streptococcus faecalis var. liquefaciens on plants.
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The proliferation of Streptococcus faecalis var. liquefaciens on two varieties of beans, and on corn, rye, and cabbage was investigated. Comparisons were made with growth patterns on these same plants exhibited by S. lactis and Lactobacillus plantarum. The ability of each of the bacteria to multiply and to spread to new plant parts as they developed from seed was studied under several environmental conditions. Plants were grown aseptically in glass culture and in sterilized and non-sterilized soil in the greenhouse. Quantitative estimations of increase in bacterial numbers were made. S. faecalis established commensal growth on each of five plants, although selectivity was noted for some plant parts. The organism increased in numbers on the plants equally as well as did the control bacteria, both alone, and in competition with the control bacteria and the microflora of the soil. (+info)
Protein synthesis by isolated pea nucleoli.
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A new method is described for the preparation of active, nucleus-free nucleoli and chromatin in relatively high purity and in sufficient quantities to permit biochemical and electron microscopic investigation. This method consists of disintegrating previously isolated nuclei by grinding with glass beads in an isotonic medium thus liberating structurally intact nucleoli and chromatin threads. Nucleoli and chromatin are then purified by differential centrifugation in Ficoll solutions. A study of the chemical composition, submicroscopic structure, and biological activity of the nucleolar preparation has been made. An equivalent study of the chromatin material has also been carried out in order to assess the significance of chromosomal contamination in nucleolar protein synthesis. The isolated nucleoli rapidly incorporate leucine-C(14) into acid and base stable compounds in vitro. Such incorporation lasts for 20 minutes at 37 degrees C and is enhanced by the addition of an energy-regenerating system and a complete amino acid mixture. It is independent of the nuclear Ph 5 enzymes. The bulk of the incorporated label is recovered in the residual, ribosome-like nucleolar protein fraction and a small percentage is found in the acid-extractable basic proteins. The rate of protein synthesis by isolated nucleoli is more rapid than that occurring in the chromatin fraction. This is taken as an additional proof that the nucleolus is the principal site of protein synthesis in the interphase pea nucleus. (+info)
QUICK SEROLOGICAL METHOD OF CLASSIFYING STRAINS OF RHIZOBIUM JAPONICUM IN NODULES.
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Means, Ura M. (U.S. Department of Agriculture, Beltsville, Md.), Herbert W. Johnson, and R. A. Date. Quick serological method of classifying strains of Rhizobium japonicum in nodules. J. Bacteriol. 87:547-553. 1964.-A new method of classifying strains of Rhizobium japonicum serologically by utilizing the contents of nodules as antigens is described. The method differs from standard procedures only in the preparation of antigens, but it is less time-consuming. Agglutination reactions of culture and nodule antigens were identical for 15 of the 17 strains investigated. Nodule antigens of ineffective strain 24 failed to agglutinate with any of the 17 antisera employed, and nodule antigens of strain 46 reacted with two antisera in addition to the two with which the culture antigens reacted. Host genotypes affected the reactions of nodule antigens of strain 46, with the reactions ranging from those typical for culture antigens for some soybean (Glycine max) varieties to reactions with heterologous antisera only for other soybean varieties and six cowpea (Vigna sinensis) varieties. Although nodule antigens of most strains of R. japonicum reacted the same as culture antigens, the inconsistent reactions of nodule antigens of strain 46 suggest precautions in the general use of nodule antigens. Indicated precautions are discussed. (+info)
COLIFORMS ASSOCIATED WITH SUGARCANE PLANTS AND JUICES.
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A study of the coliforms found on sugarcane plants throughout the growing season indicated that members of the genus Aerobacter comprise a part of the epiphytic microflora. When quantitative determinations were made of the coliforms by means of the most probable number technique, no completed tests were obtained on young sugarcane plants during the test period of February through May; however, Paracolobactrum aerogenoides was isolated frequently. Nine different IMViC types of coliforms were identified among the 198 isolates chosen for study. The predominating coliform found at the top, middle, and bottom portions of the sugarcane plants was A. cloacae, but the type predominating in leaf-sheath water was A. aerogenes. Only once was Escherichia coli found associated with the plants at the high dilutions necessary for determining most probable numbers. The numbers of different coliforms occurring in sugarcane juice were generally divided equally among A. aerogenes, A. cloacae, and E. coli. More variety among the IMViC types seemed to occur on the cane plants than in the leaf-sheath water or the cane juice in the mill. (+info)
Vitamin B-6 depletion followed by repletion with animal- or plant-source diets and calcium and magnesium metabolism in young women.
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An 84-98-d study was conducted in young women to determine the effect of vitamin B-6-deficient diets on calcium and magnesium metabolism. A vitamin B-6-deficient formula diet fed initially was followed by either animal- or plant-source protein food diets containing four increasing amounts of vitamin B-6. Calcium balance was negative during vitamin B-6 depletion. Serum calcium was higher and calcium balance negative with the plant protein diets. Magnesium balance was negative during vitamin B-6 depletion due to increased urinary magnesium excretion. Urinary calcium decreased during vitamin B-6 depletion and increased as dietary vitamin B-6 increased. Urinary oxalate was significantly higher at the end than at the beginning of vitamin B-6 depletion and was higher with plant than animal protein diets. The results suggest that vitamin B-6 depletion may alter calcium and magnesium metabolism and that dietary components associated with the protein source may influence calcium retention. (+info)
EFFECT OF WATER EXTRACTS OF CAROB PODS, TANNIC ACID, AND THEIR DERIVATIVES ON THE MORPHOLOGY AND GROWTH OF MICROORGANISMS.
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The effect of aqueous extracts of carob (Ceratonia siliqua) pods, gallotannic acid, gallic acid, and catechol on several microorganisms was studied. Carob pod extract and tannic acid showed a strong antimicrobial activity toward some cellulolytic bacteria. On the basis of tannin content, to which antimicrobial effect was related, carob pod extracts inhibited Cellvibrio fulvus and Clostridium cellulosolvens at 15 mug/ml, Sporocytophaga myxococcoides at 45 mug/ml, and Bacillus subtilis at 75 mug/ml. The inhibiting concentrations for tannic acid were found to be 12, 10, 45, and 30 mug/ml, respectively. Gallic acid and catechol were much less effective. Tannic acid and the tannin fraction of carob extract exerted both bacteriostatic and bactericidal effects on C. fulvus. Respiration of C. fulvus in the presence of bactericidal concentrations of tannic acid or tannin fraction of carob extract was inhibited less than 30%. A partial formation of "protoplasts" by C. fulvus was obtained after 2 hr of incubation in a growth medium to which 20% sucrose, 0.15% MgSO(4).7H(2)O, and 10 to 50 mug/ml of tannic acid or 500mug/ml of penicillin, or both, had been added. Tannic acid and the tannin fraction of carob extract protected C. fulvus from metabolic lysis in sucrose solution. Although the growth of other microorganisms tested was only slightly affected, the morphology of some of them was drastically changed in the presence of subinhibitory concentrations of carob pod extracts of tannic acid. It is suggested that the site of action of tannins on sensitive microorganisms is primarily the cell envelope. (+info)