Scanning electron microscopy investigations on bis(2-ethylhexyl)phthalate treated Mycobacterium cells.

Comparative investigation of steroid transforming activity and ultrastructural changes of bis(2-ethylhexyl)phthalate (BEHP, phthalate) treated Mycobacterium sp. NRRL B-3805 cells was carried out. Transformation of beta-sitosterol into androstenedione (AD) and androstadienedione (ADD) was performed in phthalate medium by resting cells preincubated in the organic solvent for a period from 3 to 24 h. It was observed that a preincubation greater than 12 h leads to the development of dense formations on the cells surface, reduction in the cell turgor, disruption in the cell walls, and formation of zones with reduced electron density. The preincubation for 24 h causes deeper changes in the cell ultrastructure but the treated cells retain their steroid transforming activity, allowing up to 80% of the substrate to be converted into AD and ADD. A preincubation of the resting Mycobacterium cells in BEHP for 6 h might be recommended as it leads to an achievement of stoichiometrical transformation of the substrate into AD and ADD and slightly higher initial rate of the reaction performed.

Stoichiometry of microbial continuous-flow purification of toluene-contaminated air.

The applicability of a recently published modification of the chemostat, named "titrostat", for microbial continuous-flow purification of toluene-contaminated air is discussed. This article describes the operative range and the toluene elimination efficiency of a 2-l titrostat running with a mixed bacterial culture dominated by two Acinetobacter species: A. calcoaceticus and A. radioresistens. The study focuses on the kinetics and stoichiometry of the process. Special attention is paid to the peculiarities of toluene as an unconventional growth substrate having high carbon and energy content. Removal productivity as high as 2.24 g l(-1) h(-1) with 99.9% elimination efficiency was observed at air flow rate 60 l h(-1), temperature 32 degrees C, pH 6.2, toluene concentration in the inlet air 37.4 mg l(-1) and titrant solution containing NH3 at 1.87 g l(-1). The maximum biomass yield from assimilated toluene, Ysm=0.880+/-0.011, and a rate of substrate expenditures for cell maintenance, ms=0.022+/-0.002 h(-1), were estimated.

Titrostat: a reliable technique for continuous cultivation of microorganisms utilizing liquid or gaseous water insoluble substrates.

We describe a method for continuous cultivation of microorganisms utilizing liquid or gaseous water insoluble substrates as a single source of carbon and energy. The water insoluble substrate, which is also the growth-limiting factor, enters the cultivation space as a manually adjustable single-compound material flow. All nonlimiting nutrients (with the exception of oxygen) enter the cultivation space as ingredients of titrant solution which feed rate is reliably coupled to the rate of substrate addition by means of the system titrator. The method provides mild starting conditions appropriate for primary isolation of microorganisms utilizing substrates with growth-inhibiting properties such as BTEX (benzene, toluene, ethylbenzene, and xylene). The sound control over the microbial specific growth rate makes it suitable for precise kinetic studies as well. We provide a detailed description of both the principles of the method and the equipment used. The dependence of the systems operative range on the concentration of titrant solution is illustrated in the case of continuous cultivation of a mixed bacterial culture on toluene.