Purification and Characterization of an Aminopeptidase from Lactococcus lactis subsp. cremoris AM2

An aminopeptidase was purified from cell extracts of Lactococcus lactis subsp. cremoris AM2 by ion-exchange chromatography. After electrophoresis of the purified enzyme in the presence or absence of sodium dodecyl sulfate, one protein band was detected. The enzyme was a 300-kilodalton hexamer composed of identical subunits not linked by disulfide bridges. Activity was optimal at 40 degrees C and pH 7 and was inhibited by classical thiol group inhibitors. The aminopeptidase hydrolyzed naphthylamide-substituted amino acids, as well as dipeptides and tripeptides. Longer protein chains such as the B chain of insulin were hydrolyzed, but at a much slower rate. The Michaelis constant (K(m)) and the maximal rate of hydrolysis (V(max)) were, respectively, 4.5 mM and 3,600 pkat/mg for the substrate l-histidyl-beta-naphthylamide. Amino acid analysis showed that the enzyme contained low levels of hydrophobic residues. The partial N-terminal sequence of the first 19 residues of the mature enzyme was determined. Polyclonal antibodies were obtained from the purified enzyme, and after immunoblotting, there was no cross-reaction between these antibodies and other proteins in the crude extract.     

Infectious disease and the Unesco basic education initiative

As part of a Basic Education for All campaign, Unesco is exploring ways to improve health and nutrition in school-age children in order to help them take advantage of the only schooling they may ever receive. At the first international Unesco working meeting on health, nutrition and school performance, intestinal helminths were identified as more appropriate immediate targets than malaria and other infectious diseases. Prevalence and intensity of infection in school-age children, the schools as vehicles of intervention, and the availability of interventions contributed to this choice. However, there are still many gaps in our knowledge of the effects of infectious diseases in schoolchildren. Through the improvement of understanding in this area, the parasitology community may be able to contribute to the Unesco project.     

RAMARIA SUBGENERA RAMARIA AND LAETICOLORA IN YUNNAN

Three taxa of Ramaria subgenus Ramaria, and 18 of subg. Laeticolora are reported from Yunnan Province. Of these 13 are considered new to science. Only one taxon (R. formosa) is considered of widespread distribution, the others being known olay from the Indo-Asian subcontinent. Wide distribution patterns, based on phenetic similarity, seem to be at the infraspecific to species complex level, but only rarely contaxie. New taxa are as follows: subg. Ramaria: R. eryuanensis; subg. Laeticolora: R. brunneipes, R. distinctissima, R. ephemeroderma, R. hilaris, R. lacviformosoides, R. linearioides, R. linearis, R. nanispora, R. rubri-attenuipcs, R. sino-conjunctipes, R. rubricarnata var. laeta, R. botrytoides var. microspora, and R. neoformosa var. sinensis. Keys to Yunnan taxa of Ramaria subgenera Ramarla and Laeticolora are furnished.     

Mathematical model for microbial oxidation of pure lead sulfide by Thiobacillus ferrooxidans

A shrinking-core mathematical model describing bioleaching of lead sulfide is developed considering the deposition of insoluble bio-oxidation products on metal sulfide particle surfaces. Variations in particle size are considered as it affects diffusion limitations.     

Sliding controller design for a nonlinear fermentation system

The design of a sliding controller for a continuous fermentation process is presented. The results obtained by simulation have proved the control scheme to be very robust. Regulation of substrate concentration at its optimal value has been achieved even though process parameters change their nominal values. Chattering effects are decreased by introducing a minor modification of the control variable around the sliding surface.     

Reactive extraction of penicillin G from mycel-containing broth in a countercurrent extraction decanter

Penicillin was recovered from mycel-containing fermentation broth by direct reactive extraction into a counter-current extraction decanter, Type CA 226-290 of the Westfalia Separator Co., at room temperature via steady state operation. Penicillin concentrations in the feed varied from 3 to 41 g L(-1), Amberlite LA-2 carrier concentrations from 7 to 20 g L(-1)and/or DITDA carrier concentrations from 7.2 to 84 g L(-1), the LA-2-to-penicillin mole concentration ratio from 4 to 6.4, and/or the DITDA-to-penicillin mole concentration ratio was maintained at 2. The throughputs of the fermentation broth (520 to 880 L h(-1)) of the solvent phase (200 to 860 L h(-1)) and the over all throughput (800 to 1750 L h(-1)) were high. Extraction degrees of 72 to 96% were achieved between pH 4.6 and 5.1. Without carriers in the same pH range, extraction degrees of only 17 to 19% were attained. By reducing the pH to 2.3 and in the absence of carriers, the degree of extraction was increased to 61%. However, during the extraction, 6.5% of the penicillin decomposed. At these high throughputs, the steady state was attained within 1 to 4 min. Through the mechanical stress, the length of the hyphae was reduced and the protein content of the broth was increased by 50 to 100%. However, this protein content had no appreciable influence on the phase separation.     

Mathematical descriptions of hybridoma culture kinetics: II. The relationship between thiol chemistry and the degradation of serum activity

The growth rate of the hybridoma cell line ATCC-CRL-1606 in low serum medium declines rapidly with time after inoculation. To characterize this phenomenon, the stability of the growth-promoting activity of serum was investigated. The activity of serum was found to de grade with time, and was stabilized by alterations in the medium formulation that acted to lower the oxidation/reduction potential. This included both the addition of thiols and the elimination of disulfides from the medium. Additionally, cysteine and other thiols were shown to stimulate growth in low serum, low cell density cultures, suggesting that thiols may be rate-limiting in low serum medium. Stimulation of growth by thiol addition was less significant at high cell levels, implying that the cells themselves may be acting to reduce their environment. A hypothesis is presented based on these results which suggests that the actual rate-limiting moieties in low serum cultures may be dithiols.