Effect of arabinose substitution on the material properties of arabinoxylan films

This study was undertaken to investigate the effect of arabinose content on film properties. The substrate used was a rye arabinoxylan that had an Ara/Xyl ratio of 0.52 and an average number molecular weight of 305 kDa. Oxalic acid was used to attempt selective removal of the arabinose substituents on the xylan main chain. The debranching of the polymer was coupled with a decrease in molecular weight. The effect of reaction conditions on the decrease in arabinose content and loss of molecular weight was investigated. Optimal conditions were selected using an experimental design. Treatment at lower temperature for longer period of times resulted in debranching with less degradation of the main chain. As the Ara/Xyl ratio was lowered, aggregates began to form in an interval of the Ara/Xyl ratio between 0.31 and 0.23 in a water solution. Precipitation occurred below an Ara/Xyl ratio of 0.1. Thus, removal of arabinose substituents results in a gradual association of unsubstituted chains. There is a linear correlation between arabinose substitution and the moisture content of arabinoxylan at 98% RH. A decrease in arabinose content resulted in the loss of a plasticizing effect, as determined by dynamic mechanical analysis, which is correlated to water binding capacity.     

Experimental study of the antibacterial activity and chemotherapeutic efficacy of the novel glycopeptide eremomycin

The study of antibacterial and chemotherapeutic activity of eremomycin, a novel glycopeptide antibiotic showed that it inhibited the growth of gram positive and acid fast microbes. The antibacterial spectrum of the novel glycopeptide was close to that of ristomycin and vancomycin. However, the in vitro antibacterial activity of eremomycin was 2-10 times higher than that of ristomycin and vancomycin. It also inhibited the growth of oxacillin resistant microbes at concentrations 5.20 times lower than those of vancomycin and ristomycin. By the therapeutic efficacy in albino mice with staphylococcal or streptococcal sepsis eremomycin was 2-4 times as superior as vancomycin and ristomycin and by the chemotherapeutic indices it was more than 10 times as superior.     

Quantitative amino acid makeup and the characteristics of the amino groups of ristomycin and the products of its partial acid hydrolysis]

The quantitative amino acid composition of ristomycin A, a glycopeptide antibiotic, peptides I-IV (from partial acid hydrolysis of the antibiotic) and their dinitrophenylic derivatives was determined. It was shown that both ristomycin and free peptides I-IV contained one residue of ristomycinic acid and one residue of actinoidinic acid, diamino-dicarbonic amino acids of the glycylphenolic type. Peptides I-IV had close molecular weights, i.e. 1100-1200 and differed from each other in the gradually increasing numbers of NH2- and COON- groups, from one in peptide I to four in peptide IV. The quantitative amino acid analysis of the dinitrophenylic derivatives of ristomycin and peptides I-IV showed that the free NH2-group in peptide I belonged to ristomycinic acid, the same as in the antibiotic, while in peptides III-IV at least one of the free NH2-groups belonged to ristomycinic acid and the other belonged to actinoidinic acid.     

Therapeutic effectiveness of ristomycin in the visceral form of actinomycosis (experimental research)

Therapeutic efficacy of ristomycin in visceral actinomycosis, a severe disease difficult for diagnosis requiring long-term complex therapy was studied on 24 rabbits. The animals were divided into 4 groups. A model of thoracal actinomycosis was developed. 7 rabbits were treated with ristomycin. The other 7 rabbits were treated with ristomycin in combination with actinolysate, a specific immune preparation. 5 rabbits were treated with actinolysate alone and the other 5 rabbits received no treatment. alone and especially in combination with actinolysate was an efficient therapeutic agent for treatment of severe visceral actinomycosis. This allowed to recommend ristomycin for clinical trials in treatment of the disease.     

Ability of vancomycin-group antibiotics to induce or to inhibit thrombocyte agglutination

The effect of 4 vancomycin antibiotics on factor VIII-dependent agglutination of thrombocytes was studied. Significant similarity, both quantitative and qualitative, between ristocetin and ristomycin was found. In this connection ristomycin may be used for determination of the so-called ristocetin cofactor. Actinoidin and vancomycin inhibited agglutination of platelets induced by ristocetin or ristomycin in platelet-enriched plasma with citrate or EDTA the same as in the system contaning platelets treated with formalin and did not inhibit agglutination induced by the bovine factor VIII. The 4 antibiotics induced precipitation of the plasma protein. Vancomycin was most active and actinoidin ws lest active in this respect. Ristocetin and ristomycin also possessed such capacity, the effect of the latter being higher. Actinoidin and vancomycin did not prevent the immediate effect of light absorption increasing due to addition of ristocetin or ristomycin to fixed platelets in concentrations completely inhibiting agglutination of platelets in the presence of the protein cofactor. Inhibition of this direct effect of ristocetin and ristomycin was observd only at higher concentrations, which indicated that this effect was not probably associated with agglutination. The results of the study on various ristomycin derivatives showed that methylated carboxylic groups and free hydroxyls of phenol may play the main role in ristomycin binding with the thrombocytic membrane and/or protein cofactor.     

Effect of various carbon and nitrogen sources on the biosynthesis of ristomycin, protease and pigments by a culture of Nocardia fructiferi var. ristomycini

The effect of various sources of carbon and nitrogen on the biosynthesis of ristomycin, protease and pigments by Nocardia fructiferi was studied. It was shown that the carbon sources had the most significant effect on the biosynthesis of the antibiotic. The maximum biosynthetic activity of the Nocardia was observed in the medium containing 1-2 per cent of soybean meal and 2 per cent of glycerol. Under such conditions all the three biologically active substances formed. The contents of ristomycin, protease and pigments amounted to 562-649 microgram/ml, 26-30 PU/ml and 0.45-0.63 conditional units, respectively.     

Permeability of an actinomycoma for ristomycin (experimental studies)

In vitro studies showed that ristomycin was the most active against actinomycetes causing actinomycosis as compared to benzylpenicillin, ampicillin, methicillin and lincomycin. The growth of the test microbes was inhibited by ristomycin in concentrations of 61--122 mg/ml. Since ristomycin was the most active against actinomycetes, its levels in the blood, parenchymatous organs, capsule and pus of actinomycomas of 5 rabbits infected wtih actino nycosis in the submaxillary area were determined. In the control healthy rabbit, the ristomycin levels were determined in the blood and organs. Ristomycin was administered intravenously in a single dose of 7000 mg/kg. Its concentrations in the animals were determined in 2.5 hours. The results of the experiments showed that ristomycin penetrated in therapeutic concentrations into the connective tissue capsule of actinomycoma. As for the other antibiotics tested earlier, they failed to penetrate this barrier. In 3 infected rabbits, ristomycin penetrated even the pus contained in actinomycoma. Ristomycin provides therapeutic concentrations in the disease focus and may produce a satisfactory therapeutic effect in treatment of actinomycosis.     

Study of the sorption of glycopeptide antibiotics on an ultradisperse carbon sorbent

The sorption capacity of a novel ultradisperse carbon sorbent (UDCS) towards antibiotics of the glycopeptide group, namely, eremomycin, vancomycin, ristomycin A, and teicoplanin A2, has been studied. The conditions for maximum sorption of the antibiotics from solutions have been determined, and a mathematical model of the sorption of the antibiotics of the above named group has been put forward.     

Chemical modification of ristomycin A with bifunctional reagents

Various bifunctional reagents by the free NH2 group of ristomycinic acid of ristomycin A were used for selective chemical modification of the antibiotic. The bifunctional reagents were the following: di-N-hydroxysuccinimide ether of suberic acid and 4,4'-difluoro-3,3'-dinitrodiphenylsulfone. Bis-N,N'-derivatives of ristomycin A were prepared using these reagents. The derivatives inhibited the growth of Bac. subtilis but the concentrations required for the inhibition were 2-4 times higher than those of ristomycin A. It was noted that the MIC of the bis-N,N'-derivatives depended on the length and flexibility of the "binding foot". The MIC of the bis-N,N'-derivative prepared with using suberic acid was 2 times higher than that of the derivative prepared with the use of 4,4'-difluoro-3,3'-dinitrodiphenylsulfone.     

Effect of ristomycin on the electrokinetic properties of Staphylococcus aureus cells

The electrophoretic mobility of the cells of Staph. aureus cultivated on the medium with ristomycin is markedly decreased at pH 3.0-5.0. This indicates that ristomycin added to the cultivation medium lowers the number of the phosphate groups in teichoic acids of the staphylococcal cell wall. The effect of ristomycin on the cells of the staphylococci was studied in vitro during the process of their incubation. It was shown that almost within the first minutes of the incubation ristomycin interacts immediately with the ionogenic groups of the cell wall and first of all with the phosphate groups of teichoic acids.     

Optimization of acid hydrolysis from the hemicellulosic fraction of Eucalyptus grandis residue using response surface methodology

Biotechnological conversion of biomass into fuels and chemicals requires hydrolysis of the polysaccharide fraction into monomeric sugars. Hydrolysis can be performed enzymatically and with dilute or concentrate mineral acids. The present study used dilute sulfuric acid as a catalyst for hydrolysis of Eucalyptus grandis residue. The purpose of this paper was to optimize the hydrolysis process in a 1.4 l pilot-scale reactor and investigate the effects of the acid concentration, temperature and residue/acid solution ratio on the hemicellulose removal and consequently on the production of sugars (xylose, glucose and arabinose) as well as on the formation of by-products (furfural, 5-hydroxymethylfurfural and acetic acid). This study was based on a model composition corresponding to a 2(3) orthogonal factorial design and employed the response surface methodology (RSM) to optimize the hydrolysis conditions, aiming to attain maximum xylose extraction from hemicellulose of residue. The considered optimum conditions were: H(2)SO(4) concentration of 0.65%, temperature of 157 degrees C and residue/acid solution ratio of 1/8.6 with a reaction time of 20 min. Under these conditions, 79.6% of the total xylose was removed and the hydrolysate contained 1.65 g/l glucose, 13.65 g/l xylose, 1.55 g/l arabinose, 3.10 g/l acetic acid, 1.23 g/l furfural and 0.20 g/l 5-hydroxymethylfurfural.     

Practical synthesis of a tetrasaccharide derivative corresponding to ristomycin A and ristocetin A

A practical synthesis of fully benzoylated tetrasaccharide, whose free form is indispensable to the antibiotic ristomycin A for the process of dimerization and binding to the cell wall, was achieved via sequential assembly of the building blocks, allyl 3,4-di-O-benzoyl-alpha-D-glucopyranoside, 2,3,4-tri-O-benzoyl-alpha-L-rhamnopyranosyl trichloroacetimidate, 2-O-acetyl-3,4,6-tri-O-benzoyl-alpha-D-mannopyranosyl trichloroacetimidate, and 2,3,5-tri-O-benzoyl-alpha-D-arabinofuranosyl trichloroacetimidate. A one-pot preparation of allyl 3,4-di-O-benzoyl-2-O-tert-butyldimethylsilyl-6-O-triphenylmethyl-alpha-D-glucopyranoside is described, and regioselective glycosylation is carried out using perbenzoylated sugar trichloroacetimidates as glycosyl donors in the presence of a catalytic amount of trimethylsilyl trifluoromethanesulfonate (TMSOTf).     

Growth of Neocallimastix sp. Strain R1 on Italian Ryegrass Hay: Removal of Neutral Sugars from Plant Cell Walls

The anaerobic fungus Neocallimastix sp. strain R1 was grown for up to 5 days on a medium containing autoclaved Italian ryegrass hay as the carbon source. Culture supernatants and digested cell walls were harvested at 12-h intervals. Supernatants were analyzed for the fermentation products formate and acetate, and residual cell walls were analyzed for dry-matter and neutral-sugar losses. Fungal growth was accompanied by the digestion of plant cell walls and the accumulation of fermentation products in culture media. Dry-matter losses were accounted for by removal of four major neutral sugars (arabinose, galactose, glucose, and xylose) from the plant cell walls. First-order reaction kinetics could be used to describe the loss of each sugar. All cell wall sugars, including arabinose and galactose, which are not fermented by Neocallimastix sp. strain R1 were removed simultaneously. Although the rates of removal of individual sugars were similar, there were significant differences in their extents of removal: the extent of removal of arabinose exceeded that of the other three sugars, and xylose was the least digestible. This study provides the first account of simultaneous (nonpreferential) removal of neutral sugars from plant cell walls by an anaerobic fungus. Although in vitro techniques were used, these results indicate a potentially significant role for the anaerobic fungi as fiber digesters in the rumen.     

Experimental study of histamine-liberating properties of eremomycin

Eremomycin, a new glycopeptide antibiotic showed the same as ristomycin, polymyxins B and M and kanamycin histamine liberating properties. Liberation of endogenic histamine induced destructive lesions on the mucosa of the gastrointestinal tract. The most pronounced lesions were observed after intraperitoneal administration of the antibiotic. When eremomycin was administered intravenously or subcutaneously the affection of the gastrointestinal tract epithelium was less pronounced. After subplantar administration of the antibiotics pad edema in albino rats was observed. The most pronounced edema was after the use of ristomycin and the polymyxins. After the use of eremomycin and kanamycin it was at the average 2 times less pronounced. Preliminary administration of dimedrol decreased intensity of the pad edema induced by the antibiotics.     

Microbial synthesis of 3-dehydroshikimic acid: a comparative analysis of D-xylose, L-arabinose, and D-glucose carbon sources.

3-Dehydroshikimic acid is a hydroaromatic precursor to chemicals ranging from L-phenylalanine to adipic acid. The concentration and yield of 3-dehydroshikimic acid microbially synthesized from various carbon sources has been examined under fed-batch fermentor conditions. Examined carbon sources included D-xylose, L-arabinose, and D-glucose. A mixture consisting of a 3:3:2 molar ratio of glucose/xylose/arabinose was also evaluated as a carbon source to model the composition of pentose streams potentially resulting from the hydrolysis of corn fiber. Escherichia coli KL3/pKL4.79B, which overexpresses feedback-insensitive DAHP synthase, synthesizes higher concentrations and yields of 3-dehydroshikimic acid when either xylose, arabinose, or the glucose/xylose/arabinose mixture is used as a carbon source relative to when glucose alone is used as a carbon source. E. coli KL3/pKL4.124A, which overexpresses transketolase and feedback-insensitive DAHP synthase, synthesizes higher concentrations and yields of 3-dehydroshikimic acid when the glucose/xylose/arabinose mixture is used as the carbon source relative to when either xylose or glucose is used as a carbon source. Observed high-titer, high-yielding synthesis of 3-dehydroshikimic acid from the glucose/xylose/arabinose mixture carries significant ramifications relevant to the employment of corn fiber in the microbial synthesis of value-added chemicals.     

Microbial Synthesis of Antiviral Nucleosides Using Escherichia coli BL21 as Biocatalyst

Experimental conditions such as shaking (aeration) rate, concentration of reagents and extent of culture growth for the optimal synthesis of adenosine using Escherichia coli BL21 as biocatalyst were assessed, achieving 95% yield in 30 min of reaction using microorganisms harvested from late exponential phase. The ability of E. coli BL21 to synthesise purine nucleosides containing sugar residues such as 2'-deoxyribose, 2',3'-dideoxyribose and arabinose was also verified. 2'-Deoxyribo- and arabinonucleosides could be prepared in high yield, while the results obtained with 2',3'-dideoxyribonucleosides were not satisfactory. In the case of 2'-deoxyadenosine, using thymidine as a starting material, a yield of 94% was achieved at 45°C.     

Effect of antibiotics on indices of immunologic reactivity in mice

The effect of ristomycin, chloramphenicol, kanamycin, benzylpenicillin, streptomycin, and cephaloridine on the indices of cellular and humoral immunity was studied comparatively on intact animals and on animals with secondary immune deficiency. The study of the antibiotic effect on the count of rosette-forming lymphocytes (RFL) and the total count of lymphocytes showed that all the antibiotics except streptomycin induced a significant decrease in the count of RFL. The most active was kanamycin. It lowered the count of RFL 5-fold as compared to the control. The total count of lymphocytes was lowered after administration of ristomycin, chloramphenicol and kanamycin. In the animals with immune deficiency induced by cyclophosphamide benzylpenicillin potentiated the inhibitory effect of cyclophosphamide on the weight of the lymphoid organs, while streptomycin lowered the effect of cyclophosphamide. No such effect was observed with the use of the other antibiotics. The data indicated the necessity of taking into account the effect of various antibiotics on the immune system, especially under conditions of immune deficiency.     

Fermentation of corn fibre sugars by an engineered xylose utilizing Saccharomyces yeast strain

The ability of a recombinant Saccharomyces yeast strain to ferment the sugars glucose, xylose, arabinose and galactose which are the predominant monosaccharides found in corn fibre hydrolysates has been examined. Saccharomyces strain 1400 (pLNH32) was genetically engineered to ferment xylose by expressing genes encoding a xylose reductase, a xylitol dehydrogenase and a xylulose kinase. The recombinant efficiently fermented xylose alone or in the presence of glucose. Xylose-grown cultures had very little difference in xylitol accumulation, with only 4 to 5g/l accumulating, in aerobic, micro-aerated and anaerobic conditions. Highest production of ethanol with all sugars was achieved under anaerobic conditions. From a mixture of glucose (80g/l) and xylose (40g/l), this strain produced 52g/l ethanol, equivalent to 85% of theoretical yield, in less than 24h. Using a mixture of glucose (31g/l), xylose (15.2g/l), arabinose (10.5g/l) and galactose (2g/l), all of the sugars except arabinose were consumed in 24h with an accumulation of 22g ethanol/l, a 90% yield (excluding the arabinose in the calculation since it is not fermented). Approximately 98% theoretical yield, or 21g ethanol/l, was achieved using an enzymatic hydrolysate of ammonia fibre exploded corn fibre containing an estimated 47.0g mixed sugars/l. In all mixed sugar fermentations, less than 25% arabinose was consumed and converted into arabitol.     

Esterification of ferulic acid with polyols using a ferulic acid esterase from Aspergillus niger

Commercially available enzyme preparations were screened for enzymes that have a high ability to catalyze direct ester-synthesis of ferulic acid with glycerol. Only a preparation, Pectinase PL "Amano" produced by Aspergillus niger, feruloylated glycerol under the experimental conditions. The enzyme responsible for the esterification was purified and characterized. This enzyme, called FAE-PL, was found to be quite similar to an A. niger ferulic acid esterase (FAE-III) in terms of molecular mass, pH and temperature optima, substrate specificity on synthetic substrates, and the N-terminal amino acid sequence. FAE-PL highly catalyzed direct esterification of ferulic acid and sinapinic acid with glycerol. FAE-PL could feruloylate monomeric sugars including arabinose, fructose, galactose, glucose, and xylose. We determined the suitable conditions for direct esterification of ferulic acid with glycerol to be as follows: 1% ferulic acid in the presence of 85% glycerol and 5% dimethyl sulfoxide at pH 4.0 and 50 degrees C. Under these conditions, 81% of ferulic acid could be converted to 1-glyceryl ferulate, which was identified by (1)H-NMR. The ability of 1-glyceryl ferulate to scavenge 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals was higher than that of the anti-oxidant butyl hydroxytoluene.     

Changes in the microflora of the intestine after administration of antibiotics to animals maintained conventionally

A series of experiments were performed on mice and rats subjected to total decontamination by oral administration of 3 antibiotics: gentamicin, ristomycin and nystatin. The results of experiments are summarized. It was shown that under the conventional conditions the decontamination was effective, when the intervals between the experiments were 3-5 months. When the intervals were less or the experiments were performed in the same room without intervals, the efficacy of the decontamination markedly decreased or it was ineffective. The results are discussed from the standpoints of the necessity of observing definite requirements for performance of such procedures in hospitals and laboratories.