Biomedical subjects
Publications and source records attributed to J Barrios-González.
Amylomyces rouxii eliminated 85% of initial pentachlorophenol (PCP) at 12.5 mg l(-1) when grown with 0.1 g tyrosine l(-1), but only 55% without tyrosine. Addition of tyrosine in the culture medium increased the monophenolase activity by 1.8-fold. Tyrosinase is thus indicated to be the phenoloxidase involved in PCP degradation by A. rouxii .
An earlier work showed that when the bagasse content (BC) of the solid medium was decreased within a wide range of values, penicillin production by solid-state fermentation was always increased. Respiration studies were performed to understand how BC controls the secondary metabolism in this culture system. CO2 production of solid cultures with different compositions was monitored. In cultures of series A, the initial moisture content was increased and this variation was compensated by decreasing the nutrient and BC of the medium. In series B the initial moisture content was increased while BC was decreased and the nutrient content increased. In addition, penicillin production and respiration was also studied in extreme media (dry and concentrated and humid and diluted), with high and low BC. Criteria for the interpretation of respiration kinetics of the idiophase were established for the first time in this work. For the cumulative form (total CO2/g dry matter vs t) as well as for derivative (CO2/g dm/h vs t) respiration kinetics, the CO2 production rate (Q(CO2)) was determined by calculating the slope of the cumulative curve. Results indicate that Q(CO2) of the tropho- and idiophases was directly related to the BC of the solid medium (and inversely related to penicillin yields). These conclusions were confirmed by analysis of the derivative form, the results of which indicate that a lower but stable metabolic activity is essential for obtaining high penicillin yields in solid-state fermentation (SSF). The results indicate that the derivative CO2 production kinetics proved to be a more precise and sensitive indicator of the culture metabolic activity during idiophase than the cumulative respiration kinetics.
A novel method for selecting overproducing strains of rifamycin B was developed. This technique involves the use of lysozyme and the effect of barbital on the growth of A. mediterranei. Complete medium added with glycine and barbital was inoculated with mutagenized mycelium, incubated for 48 hours and treated with lysozyme. The lysozyme resistant mycelium was washed with dilute detergent. Complete medium with glycine but without barbital was inoculated with the washed mycelium. Protoplasts were obtained and regenerated and the colonies were picked and seeded on Bennet agar plates with and without barbital. Two selected mutants were sensitive to 0.5% barbital producing 200% more rifamycin than the parental strain. In addition, 30 barbital resistant mutants were isolated and their production level was lower than the one observed with the parental strain. These results suggest that the effect of barbital on secondary metabolism (rifamycin production) is related to its effect on primary metabolism.
Microbial secondary metabolites are useful high value products that are normally produced by liquid culture; but could be advantageously produced by solid-state fermentation (SSF). Particularly if SSF could benefit from a deeper understanding of microbial physiology in a solid environment. Recent research indicates that different kind of secondary metabolites can be produced by SSF: antibiotics, phytohormones, food grade pigments, alkaloids, etc. Physiology in SSF shows several similarities with physiology in liquid medium, so similar strategies must be adapted for efficient processes. However, there are certain particularities of idiophase in solid medium which dictate the need for special strains.
Penicillin production with an industrial strain and 4 strains of P. chrysogenum, in solid state fermentation (SSF) and liquid submerged fermentation (LSF), was determined. Their ability to produce the antibiotic in SSF in relation to their capacity to do so in LSF was evaluated. this was done by calculating the ratio PS/PL (production in SSF/production in LSF), which was called relative production. Clones were isolated from each strain and evaluated in a similar way. The strains presented different relative productions (from 1.4 to 2.5). Within the clones, a much wider range of relative productions was observed (0.6 to 16.7). On the other hand, the highest-producing strains in LSF were also the highest producers in SSF. This indicates that the production potential of a strain is an important factor in its production level in SSF. Moreover, the highest penicillin producing ciones (9,500 to 10,500 microg of penicillin/g were generated from high-yielding strains (P2 and ASP-78). However, the higher-producing strains (in LSF) showed lower relative performance, suggesting that higher producing strains tend to express less efficiently their potential in SSF. In this study, several overproducing clones, particularly suited for SSF, were obtained by the procedures followed. Production increases of 500 to 600 %, in this culture system, were achieved.
The use of a large particle size (14 mm) support (sugar cane bagasse) increased penicillin production by solid state fermentation by 37 %, however this effect was due to a higher sugar concentration in this bagasse fraction. Cultures with closer packing densities (0.35) produced 20 % more penicillin. Agitation did not have a negative effect on production if moisture loss during the operation is restituted.