Study of some physico-chemical characteristics of a Saccharomyces cerevisiae endopolygalacturonase: a possible use in beverage industry

Zymograms of a crude protein extract from S. cerevisiae strain SCPP containing endopolygalacturonase were studied and compared to the purified enzyme by determining their physico-chemical properties. The results obtained with crude extract were similar to those of the purified enzyme. The endopolygalacturonase from both sources displayed a pH optimum between 3.0 and 4.0, and was active at temperatures between 4 and 50°C on a large panel of substrates. These characteristics make this S. cerevisiae endopolygalacturonase an attractive tool for the beverage industry. Journal of Industrial Microbiology & Biotechnology (2000) 24, 296–300. Received 24 September 1999/ Accepted in revised form 29 January 2000     

Sodium stibogluconate (SSG) & paromomycin combination compared to SSG for visceral leishmaniasis in East Africa: a randomised controlled trial

Background

Alternative treatments for visceral leishmaniasis (VL) are required in East Africa. Paromomycin sulphate (PM) has been shown to be efficacious for VL treatment in India.

Methods

A multi-centre randomized-controlled trial (RCT) to compare efficacy and safety of PM (20 mg/kg/day for 21 days) and PM plus sodium stibogluconate (SSG) combination (PM, 15 mg/kg/day and SSG, 20 mg/kg/day for 17 days) with SSG (20 mg/kg/day for 30 days) for treatment of VL in East Africa. Patients aged 4–60 years with parasitologically confirmed VL were enrolled, excluding patients with contraindications. Primary and secondary efficacy outcomes were parasite clearance at 6-months follow-up and end of treatment, respectively. Safety was assessed mainly using adverse event (AE) data.

Findings

The PM versus SSG comparison enrolled 205 patients per arm with primary efficacy data available for 198 and 200 patients respectively. The SSG & PM versus SSG comparison enrolled 381 and 386 patients per arm respectively, with primary efficacy data available for 359 patients per arm. In Intention-to-Treat complete-case analyses, the efficacy of PM was significantly lower than SSG (84.3% versus 94.1%, difference = 9.7%, 95% confidence interval, CI: 3.6 to 15.7%, p = 0.002). The efficacy of SSG & PM was comparable to SSG (91.4% versus 93.9%, difference = 2.5%, 95% CI: −1.3 to 6.3%, p = 0.198). End of treatment efficacy results were very similar. There were no apparent differences in the safety profile of the three treatment regimens.

Conclusion

The 17 day SSG & PM combination treatment had a good safety profile and was similar in efficacy to the standard 30 day SSG treatment, suggesting suitability for VL treatment in East Africa.

Clinical Trials Registration

www.clinicaltrials.gov {"type":"clinical-trial","attrs":{"text":"NCT00255567","term_id":"NCT00255567"}}NCT00255567     

Growth, D-glucose utilization and malolactic fermentation by Leuconostoc œnos strains in 18 media deficient in one amino acid

Six Leuconostoc œnos strains were used to study the effect of the deficiency of one amino acid on growth, heterofermentative pathway and malolactic fermentation. All strains had an absolute requirement for four amino acids (isoleucine, glutamic acid, tryptophan and arginine) and needed six other amino acids (valine, methionine, cysteine, leucine, aspartic acid and histidine) for optimum growth. Each deficiency in one amino acid had a particular effect on D-glucose utilization. Overproduction and underproduction of D-lactic acid were observed. The rate of L-malic acid consumption in media deficient in one amino acid was lower than in the complete medium with all amino acids.
Although some deficiencies (glycine, phenylalanine, proline or tyrosine) had no influence on the growth, they noticeably limited the malolactic fermentation.     

Recovery of microalgal biomass and metabolites: process options and economics

Commercial production of intracellular microalgal metabolites requires the following: (1) large-scale monoseptic production of the appropriate microalgal biomass; (2) recovery of the biomass from a relatively dilute broth; (3) extraction of the metabolite from the biomass; and (4) purification of the crude extract. This review examines the options available for recovery of the biomass and the intracellular metabolites from the biomass. Economics of monoseptic production of microalgae in photobioreactors and the downstream recovery of metabolites are discussed using eicosapentaenoic acid (EPA) recovery as a representative case study.     

Characterization of Virion RNAs of Southern Bean Mosaic Virus by Electron Microscopy

Electron microscopy of denatured RNA of southern bean mosaic virus (SBMV) shows two principal linear components, 0.31 ± 0.08 μm (subgenomic RNAs, MW 0.51 × 10⁶) and 0.80 ± 0.17 μm (genomic RNA, MW 1.3 × 10⁶ 25S). Nondenatured RNA (～ 32S) from heat-inactivated virions measure 1.0 ± 0.20 μm (MW 1.64 × 10⁶) but is poorly-infectious. Upon denaturation the 325 RNA disaggregates into components of lengths typical of the genomic and subgenornic RNAs and infectivity is restored.     

Cloning and analysis of the gene for the major outer membrane lipoprotein from Pseudomonas aeruginosa

The gene for the Pseudomonas aeruginosa outer membrane lipoprotein I was isolated from a genomic library in the phage lambda EMBL3 vector and subsequently subcloned in the low copy-number, wide host-range plasmid vector, pKT240. The cloned gene was highly expressed, resulting in the production of a low molecular-weight protein (8 kD) that was found to be associated with the outer membrane. Sequence analysis showed an open reading frame of 83 amino acids with a putative N-terminal hydrophobic signal peptide of 19 residues immediately followed by the lipoprotein consensus sequence, GLY-CYS-SER-SER (residues 19-22). The predicted amino acid composition of the mature polypeptide and that of the purified lipoprotein I of P. aeruginosa (Mizuno and Kageyama, 1979) were identical. In contrast with other Gram-negative outer membrane lipoproteins, conformation predictions suggested that the mature protein was a single alpha helix.     

Rapid simultaneous lipid extraction and transesterification for fatty acid analyses

An improved adaptation of the direct transesterification method of Lepage and Roy (J. Lipid Res. 25, 1391–96, 1984) for the preparation of fatty acid methyl esters allows notable saving of time and reagents. The material being analysed is heated for 10 minutes with methanol, acetyl chloride and hexane. © Rapid Science Ltd. 1998     

Use of a new gelling agent (Eladium©) as an alternative to agar-agar and its adaptation to screen biofilm-forming yeasts

The incidence of yeast-induced infections has increased in the last decade, mainly because of the increasing number of immunodeficient patients. Since biofilm production is believed to be responsible for fungal virulence, we propose screening yeasts of various genera in order to determine their ability to form biofilms. This is an important issue because yeast cells that form biofilms are particularly resistant to anti-fungal agents used in human patients. For screening, we used Eladium©, a new polysaccharide produced by a Rhizobium sp., as an alternative gelling agent to agar. We also established the conditions necessary to detect biofilm formation. The adapted medium provides the missing link between liquid and solid media. Its advantages include enhancement of growth of microorganisms and facilitation of quick and easy monitoring of biofilm formation.     

Morphological and molecular taxonomy of Pythium longisporangium sp. nov. isolated from the Burgundian region of France

During the course of an investigation on the Pythiaceous oomycetes occurring in the Burgundian vineyards, some species of Pythium possessing mainly hypogynous antheridia were found. These had been classified as oomycetes belonging to the "Pythium rostratum" group for a long time. Three of these isolates, having similar structures and growth, are very closely related to a recently described species, Pythium bifurcatum Paul. A close look at these, however, underlines some fundamental differences with the latter. Not all of them produce zoospores but have very large sporangia. The type specimen is F-1200 (B 76a) which is a medium-slow growing saprophyte. The sequence of the ITS region of the rDNA also shows a very close relationship with P. bifurcatum. On the basis of morphological and molecular analysis, we now describe this species as Pythium longisporangium sp. nov. Morphological features of this new species, the sequences of the ITS region of its nuclear ribosomal DNA, and its comparison with related species are discussed.