Study of the lipolytic enzymes of cotton seeds IV. Triacetinase

Summary A triacetinase with mol. wt. 36,000 has been isolated from cotton seeds by ion-exchange chromatography on DEAE-cellulose and gel filtration on Sephadex G-100.The homogeneity of the protein has been confirmed by the results of rechromatography and ultracentrifugation.Institute of the Chemistry of Plant Substances, Academy of Sciences of the Uzbek SSR. Translated from Khimiya Prirodnykh Soedinenii, No. 5, pp. 628–632, September–October, 1974.     

Primary structure of triacetinase — An esterase from cotton seeds III. Peptides from tryptic hydrolysis

Summary 1. The peptides from tryptic hydrolysis of triacetinase have been isolated and characterized and their amino-acid sequences have been determined.2. On the basis of the results of the determination of the amino-acid sequences of the peptides of the tryptic hydrolyzate the sequence of peptides of the cyanogen bromide hydrolysis of triacetinase has been established or confirmed.Institute of the Chemistry of Plant Substances, Academy of Sciences of the Uzbek SSR, Tashkent. Translated from Khimiya Prirodnykh Soedinenii, No. 2, pp. 241–248, March–April, 1978.     

Lipolytic enzymes of cotton seeds V. Triacetinase

Chemistry of Natural Compounds - A triacetinase has been isolated from cotton seeds by the methods of gel filtration and ion-exchange chromatography. The homogeneity of the enzyme was shown by...     

Competitive bindings of histones with DNA in formation of plant nucleohistones

An acid form of ricin — ricin T — with a molecular weight of 58 kDa, an isoelectric point, pI, of 7.0–7.1, and a sedimentation coefficient of 4.60 S has been isolated from the seeds of the Central Asian castor-oil plantRicinus communis by gel filtration on Sephadex G-75 followed by ion exchange chromatography on DEAE- and CM-celluloses. The amino acid composition and the N-terminal amino acid residues of ricin T have been determined.Institute of the Chemistry of Plant Substances, Uzbek Academy of Sciences, Tashkent. Translated from Khimiya Prirodnykh Soedinenii, No. 6, pp. 883–887, November–December, 1987.     

Isolation,purification, and study of some properties of the extracellular lignin peroxidases of the wood-destroying fungusPleurotus ostreatus

Abstact Two bands with lignin peroxidase activity have been detected by isoelectric focusing in a total enzyme preparation obtained from a 15-day filtrate of the culture liquid of the fungusPleurotus ostreatus by fractionation with ammonium sulfate. Two homogeneous forms of the enzymes — LGP-I and LGP-II — have been obtained by gel filtration on Sephadex G-100, ion-exchange chromatography on DEAF-Toyopearl 650 M gel, and rechromatography on Sephadex G-75, and also by electrophoresis in PAAG. The specific activities of the purified lignin peroxidases LGP-I and LGP-H amounted to 36.5 and 54.3 units/mg, their degrees of purification being 8.7 and 12.9, respectively. The molecular masses of LGP-I and LGP-II, determined by electrophoresis in PAAG in the presence of Na-DS and by gel filtration on TSK HW–65 gel were 42–44 and 61–63 kDa. The isoelectric points of LGP-I and LGP-II were 3.4 and 4.1, their pH optima 2.7 and 3.4, and the, temperatures of their optimum enzymatic action 28 and 34°C, respectively. The isoenzymes differed from one another substantially with respect to pH stability and resistance to heat. The values of K_M determined from the rates of hydrolysis of the substrate by the enzymes in the presence of H₂O₂ at pH 3.7 were 0.09 mM for LGP-I and 0.07 mM for LGP-II. The values of K_M with respect to veratryl alcohol, determined by the Lineweaver—Burk method, were 0.117 mM for LGP-II and 0.132 mM for LGP-II.Institute of Microbiology, Academy of Sciences of the Republic of Uzbekistan, Tashkent, fax (3712) 41 71 29. Translated from Khimiya Prirodnykh Soedinenii, No. 3, pp. 394–402, May–June, 1996. Original article submitted November 11, 1994.     

Isolation and study of an inhibitor of the intrinsic proteolytic enzyme of cotton seeds

A scheme has been developed for the isolation of an inhibitor of cottonseed protease A using affinity chromatography on protease-A—Sepharose 4B followed by gel filtration on Sephadex G-150. The molecular mass of the inhibitor is 20 kDa. The protein molecule consists of two subunits with different molecular masses.Institute of the Chemistry of Plant Substances, Academy of Sciences of the Republic of Uzbekistan, Tashkent, Fax (3712) 62 73 48. Translated from Khimiya Prirodnykh Soedinenii, No. 2, pp. 274–277, March–April, 1994.     

Purification of the intracellular triacylglycerol lipase ofOospora lactis

A method for the isolation of homogeneous triacylglycerol hydrolase from the mycelium of the fungusOospora lactis is described. The homogeneity of the enzyme has been shown by gel filtration through Sephadex G-100, by ultracentrifugation, and by disc electrophoresis in polyacrylamide gel. Some properties of the enzyme have been studied: molecular weight, 43,000; optimum action temperature, 35–37°C; pH optimum, 7.5.     

Determination of size and shape of micelles by gel filtration chromatography

Summary The size and shape of a series of nonionic surfactants (polyoxyethylene nonyl phenols with number of ethyleneoxide groups 10, 13 and 18) were determined by gel filtration chromatography. TheStokes radius (R) was determined from the retention time. The hydration of micelles was estimated from the frictional ratios. The number of hydrating water molecules per oxyethylene unit was found to be 3–5, which agrees with the results reported by other investigators using other methods. It is thus shown that gel filtration chromatography can be applied to the micellar systems.

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Zusammenfassung Größe und Gestalt einer Serie von nichtionischen oberfldchenaktiven Substanzen (Polyoxydthylennonylphenolen mit 10, 13 und 18 Äthylenoxydgrappen) wurde mit Gelfiltrationschromatographie bestimmt. DerStokes-Radius wurde mit Hilfe der Rückhaltezeit bestimmt. Die Hydratation der Mizellen aus den Reibungszahlen geschdtzt, Zahl der Hydratwassermoleküle pro Oxydthylen ist 3–5, übereinstimmend mit Angaben anderer Autoren nach anderen Methoden. Damit ist die Anwendbarkeit der Gelfiltrationschromatographie auf mizellare Systeme erwiesen.

     

Isolation,purification, and some physicochemical properties of glucose isomerase from Streptomyces atratus

A highly purified glucose isomerase with a specific activity 23.5 times greater than that of a homogenate of the mycelium has been obtained fromStreptomyces atratus by methods of homogenization from the freeze-dried biomass, heat treatment (70°C, 10 min), ion-exchange chromatography, and gel filtration. The yield of enzyme on the initial biomass was 51.8%. The molecular mass of the enzyme has been determined by various methods as 160 kDa but in the presence of sodium dodecyl sulfate in thin-layer PAG it amounts to 40 kDa.Institute of Microbiology, Academy of Sciences of the Uzbek SSR, Tashkent. Translated from Khimiya Prirodnykh Soedinenii, No. 4, pp. 524–528, July–August, 1990.     

Use of disk electrophoresis for the separation of large peptide fragments of proteins

Summary The disk electrophoresis of peptide fragments from porcine pepsin obtained by cleaving the pepsin with cyanogen bromide has been performed. A method has been developed for extracting the colored peptide fractions after electrophoresis in polyacrylamide gel. An amino-acid analysis has been performed of peptide fragments B1, B2, and B4 isolated by disk electrophoresis. The results obtained have been compared with the amino-acid composition calculated from the structures of these peptides. The possibility has been shown of using the method described during structural investigations of both proteins and their low-molecular-weight fragments.The peptide preparations were supplied to us by I. Surova, V. Ostoslavskaya, L. Revina, I. Puracheva, É. Vakhitova, and G. Muratova. The amino-acid analysis was performed by E. Timokhina, A. Balduev, G. Fedyukina, O. Khodova, and N. Ivashechkina.All-Union Scientific-Research Institute of the Genetics and Breeding of Industrial Microorganisms. Translated from Khimiya Prirodnykh Soedinenii, No. 2, pp. 219–222, March–April, 1975.     

Detection and separation of two forms of cotton-plant pyrophosphatase

Two pyrophosphatase isoenzymes with identical molecular weights but differing in the pH optima of their action and also in their senstivity to some bivalent metal ions have been isolated from extracts of the cotyledons of three-day cotton-plant shoots by gel filtration and ion-exchange chromatography.V. I. Lenin Tashkent State University. Translated from Khimiya Prirodnykh Soedinenii, No. 3, pp. 375–378, May–June, 1985.     

Investigation of the toxins of the venom of the spiderLinothele sp.

Two low-molecular-mass toxins — Lsl and Ls2 — have ben isolated from the venom of a spiderLinothele sp. (fam. Dipluridae) with the aid of gel filtration and reversed phase high-performance liquid chromatography (HPLC). It hs been shown that they are highly toxic on cerebroventricular injection into white mice: the LD-50 values of toxins Lsl and Ls2 are 24 and 19 g/kg, respectively, which are 25 and 31 time greater than the toxicity, of the whole venon, the LD-50 of which is 0.6 mg/kg. According to mass spectrometry the molecular masses of toxins Ls1 and Ls2 are 337 and 505 Da, while their yields were, respectively, 0.21 and 0.43 % of the weight of the whole venom.Translated from Khimiya Prirodnykh Soedinenii, No. 2, pp. 229–233, March–April, 1996. Original article submitted October 6, 1995.     

Isolation of protease B from cotton seeds

Protease B has been isolated from dormant cotton seeds by fractionation with ammonium sulfate, ion-exchange chromatography on CM-cellulose, and gel filtration through Acrilex P-10 and Sephadex G-75, with 128-fold purification. The enzyme exists in dimeric and monomeric forms. According to the results of gel filtration, their molecular weights are 72,000 and 36,000, respectively. The enzyme consists of a single polypeptide chain including sugars. The N-terminal amino acid of protease B is alanine. The enzyme possesses proteolytic activity in the pH range from 4 to 6.Institute of the Chemistry of Plant Substances, Academy of Sciences of the Uzbek SSR, Tashkent. Translated from Khimiya Prirodnykh Soedinenii, No. 4, pp. 506–510, July–August, 1984.     

Phospholipases a of the venom of the Central Asian cobra

Conclusions Two phospholipases A with molecular weights of 11,000±1000 and 18,000±1000, consisting of individual compounds according to rechromatography and disk electrophoresis, have been isolated by filtration through Sephadex G-75 gel and by ion-exchange chromatography on CM-cellulose from the venom of the Central Asian cobra.V. I. Lenin Tashkent Order of the Red Banner of Labor State University. Institute of Biochemistry of the Academy of Sciences of the Uzbek SSR. Translated from Khimiya Prirodnykh Soedinenii, No. 3, pp. 387–389, May–June, 1974.     

An investigation of the venom of Renard's viperVipera ursini renardi. III. Isolation of phospholipases A2

Two phospholipases A₂ with molecular weights of 15,600 and 13,200 and pI values of 7.57 and 6.73, respectively, have been isolated from viper venom in the pure state with the aid of gel filtration on Sephadex G-75 followed by ion-exchange chromatography on CM-cellulose and SE-Sephadex C-25.Institute of Biochemistry, Academy of Sciences of the Uzbek SSR, Tashkent. Translated from Khimiya Prirodnykh Soedinenii, No. 3, pp. 389–392, May–June, 1980.     

Coupling techniques in speciation analysis

Summary Direct coupling techniques in speciation analysis lead to reproducible results with lower risks of trace contamination or losses of analytes in a shorter period of time. The direct coupling mode seems to be most promising for liquid chromatographic separation methods — especially gel permeation chromatography — and for some electrophoretic methods — as flow-through and capillary techniques. Promising detection methods are Flame-AAS and ICP-OES but also hydride-generation AAS and chemical reaction detectors in continuous-flow technique.     

Structures of two new flavanones fromVexibia alopecuroides

Two new flavanones — vexibinol and vexibidin — have been isolated from the chloroform fraction of an ethanolic extract of the roots ofVexibia alopecuroides by column chromatography on silica gel. Their structures have been established on the basis of chemical and spectral characteristics. Their IR, UV,¹H, and¹³C NMR, and mass spectra are given.Institute of the Chemistry of Plant Substances, Academy of Sciences of the Uzbek SSR, Tashkent. I. P. Pavlov Samarkand State Medical Institute. Translated from Khimiya Prirodnykh Soedinenii, No. 1, pp. 35–41, January–February, 1985.     

Polysaccharides ofEremurus. X. Characteristics of the polysaccharides ofEremurus lactiflorus andE. luteus

Water-soluble polysaccharides have been isolated from two species ofEremurus — E. lactiflorus andE. luteus — with yields of 13.5% and 20.5%, respectively. They contained mainly glucose and mannose in ratios of 1:5 and 1:3.1. The polysaccharides ofE. lactiflorus were separated from a column of DEAE-cellulose. The yield of neutral fraction was 10.3%. Gel filtration of the polysaccharides on Sephadex G-200 showed their polydispersity. Homogeneous fractions were obtained by fractional precipitation with ethanol. They have been characterized with respect to monosaccharide composition, molecular weight, and IR spectra.Institute of the Chemistry of Plant Substances, Academy of Sciences of the Uzbek SSR, Tashkent. Translated from Khimiya Prirodnykh Soedinenii, No. 5, pp. 604–608, September–October, 1979.     

Auftrennung der Proteine der Alge Chlorella

Zusammenfassung Die wasserlöslichen Proteine aus der einzelligen süßwasseralge Chlorella pyrenoidosa wurden nach Homogenisierung der Zellen erhalten. Die Proteine wurden dann einerseits durch Chromatographie auf DEAE-Zellulose, gefolgt von Elektrophorese in Polyacrylamid-Gel, und anderseits durch Gel-Filtration in Sephadex G-200 fraktioniert. Man findet, daß die Proteine nach elektrischer Ladung und Molekulargewicht aus vielen Komponenten bestehen. Einige dieser Komponenten lassen sich jedoch durch die Methode der kombinierten Chromatographie und Elektrophorese charakterisieren. Wesentliche Gehalte an Nukleinsäure wurden bei der Chromatographie nur in der letzten — am schwersten eluierbaren — Fraktion gefunden. Dagegen ist die Nukleinsäure bei der Gel-Filtration über mehrere Fraktionen verteilt.

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The water soluble proteins of the unicellular freshwater alga Chlorella pyrenoidosa were obtained after homogenization of the cells. The proteins were then fractionated either by DEAE chromatography, followed by electrophoresis in polyacrylamide gel, or by gel filtration in Sephadex G-200. The proteins were found to consist of many components differing in electric charge and molecular size. Some of these components may be characterized by combined chromatography and electrophoresis. Appreciable amounts of nucleic acid have been found in chromatography only in the fraction eluted last. In contrast, in gel filtration nucleic acid is distributed over several fractions.

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