Statistical medium optimization of an alkaline protease from Pseudomonas aeruginosa MTCC 10501, its characterization and application in leather processing.

Proteases are shown to have greener mode of application in leather processing for dehairing of goat skins and cow hides. Production of protease by submerged fermentation with potent activity is reported using a new isolate P. aeruginosa MTCC 10501. The production parameters were optimized by statistical methods such as Plackett-Burman and response surface methodology. The optimized production medium contained (g/L); tryptone, 2.5; yeast extract, 3.0; skim milk 30.0; dextrose 1.0; inoculum concentration 4%: initial pH 6.0; incubation temperature 30 °C and optimum production at 48 h with protease activity of 7.6 U/mL. The protease had the following characteristics: pH optima, 9.0; temperature optima 50 °C; pH stability between 5.0-10.0 and temperature stability between 10-40 °C. The protease was observed to have high potential for dehairing of goat skins in the pre- tanning process comparable to that of the chemical process as evidenced by histology. The method offers cleaner processing using enzyme only instead of toxic chemicals in the pre-tanning process of leather manufacture.

Response surface optimization of effective medium constituents for the production of alkaline protease from a newly isolated strain of Pseudomonas aeruginosa.

Optimization of the fermentation medium for maximum alkaline protease production was carried out with a new strain of Pseudomonas aeruginosa (B-2). Replacing the protein source/inducer (albumin in place of casein) brought about significant increase in yield after 48 hr of inoculation. Three most effective medium constituents identified by initial screening method of Plackett-Burman were albumin, (NH4)2SO4 and glucose. Central Composite Design (CCD) and Response Surface Methodology (RSM) were used in the design of the experiment and in the analysis of the results. Optimum levels of the effective medium constituents were albumin (6.586%); (NH4)2SO4, 0.164%; and glucose, 6.72%. The alkaline protease production increased from 533460 to 793492 Ul(-1).

Dislipidemia associada à terapia anti-retroviral em pacientes com AIDS / Dyslipidemia associated with antiretroviral therapy in AIDS patients

A disponibilidade da terapia anti-retroviral tem resultado em um enorme declínio nas taxas de morbidade e mortalidade dos pacientes infectados pelo vírus da imunodeficiência humana. entretanto, tal terapia tem sido associada a efeitos metabólicos adversos, como a dislipidemia; sendo assim, indivíduos com doença arterial coronariana devem ser identificados e tratados. Este artigo faz uma revisão das anormalidades dos lipídios e das lipoproteínas associadas ao uso dos inibidores de proteases, sobre o possível mecanismo da dislipidemia associada a estes inibidores e usa o protocolo do programa de educação para tratamento do colesterol em adultos(National Cholesterol Education Program Adult Treatment Panel III Guidelines) para o acompanhamento do paciente com dislipidemia.

Freqüência e atividade enzimática de Candida albicans isoladas da mucosa bucal de crianças de uma creche da prefeitura de Fortaleza / Frequency and enzymatic activity of Candida albicans isolated from the buccal mucosa of children of a day-care center of the city hall of Fortaleza, Ceará, Brazil

As candidíases bucais (também chamadas sapinhos) que ocorrem em crianças são causadas por uma deficiência imunológica, bem como por outros fatores tais como má higiene bucal e esterilização inadequada dos utensílios utilizados pelas mesmas, que potencializam a ocorrência dessa infecção fúngica. Considerando esse fato, foram avaliadas a freqüência e a atividade enzimática de Candida sp. isoladas em crianças de uma creche pública (Aprisco) na cidade de Fortaleza, Ceará. Foram coletadas amostras da mucosa bucal de 364 alunos de 1 a 5 anos de idade. Elas foram semeadas em ágar Sabouraud dextrose com cloranfenicol, incubadas por 72 horas a 37ºC e identificadas por testes micológicos. Verificou-se que 67 (18 por cento) apresentaram leveduras do gênero Candida. A Candida albicans foi a mais freqüente, com 30 isolados (45 por cento), seguida pelas C. tropicalis (31 por cento), C. guilliermondii (17 por cento), C. glabrata (4,5 por cento) e C. stellatoidea (1,5 por cento). Com relação às atividades enzimáticas das cepas de Candida albicans, 20 por cento produziram a enzima proteinase e 33 por cento, a fosfolipase. As Candida albicans isoladas da mucosa bucal de crianças dessa creche da prefeitura apresentaram uma fraca atividade enzimática. Assim, conclui-se que essas cepas parecem ter uma baixa virulência.

Influence of nutritional supplements on keratinolysis by Amycolatopsis keratinophila.

Keratinolytic potential of A. keratinophila (DSM 44409T), a newly described Amycolatopsis sp. isolated from cultivated soil in Kuwait, was demonstrated using keratinazure as the sole source of carbon and nitrogen as estimated by gel diffusion assay. Effects of 12 various nutritional supplements on the keratinolytic and azocollytic activities were determined. NH4H2PO4 and KNO3 in the medium supported a significantly higher keratinolytic activity than other supplements. However, azocollytic activities in all the supplemented media and the control were same. Best combination of carbon and nitrogen supplements (galactose and NH4H2PO4 respectively) used to evaluate the dynamics of growth and enzymes (keratinase and protease) activities of the isolate revealed a luxuriant growth with optimal keratinolytic activity occurring during the log phase. Other parameters of the fermentation medium, including pH, biomass accumulation, total protein and free amino acid concentrations were also studied.

Optimization of some additives to improve protease production under SSF.

In a locally isolated Rhizopus oryzae strain highest-production of protease (388.54/g wheat bran) was observed in presence of Tween-80 and dioctyl sodium sulfosuccinate individually at 40mg/g wheat bran concentration. Under solid state fermentation biotin (0.0025mg/g wheat bran); Ca2+ (0.05mg/g wheat bran) and 1-Naphthyl acetic acid (0.01mg/g wheat bran) also showed some inducing effect on the synthesis of the enzyme protease by solid state fermentation.

Environmental and nutritional parameters controlling the biosynthesis of thermostable alkaline protease [TAP] from Bacillus sterrothurmophilos, S-WN 1616 B isolated from Wady El - Natroon lakes in Egypt

A special study was undertaken concerning the productivity of thermo- alkaline protease[s] [TAP] by the proteolytic thermoalkalophilic Bacillus stearothermophilus S-WN1616B isolated from Wady El-Natroon in Egypt. Optimal parameters controlling TAP productivity were inoculum size [0.23x109 CFU/ml], incubation period [30 days], incubation temperature [55C], pH [10-10.2] and NaCl 3%. Although L-threonine, Ca-nitrate, gelatin, peptone pantothenic acid and Zn++ enhanced [TAP] formation while added separately to gelatin-production medium, yet their elimination in addition to the absence of all ingredients of medium gave evidence that only gelatin in tap water stimulated the highest yield of [TAP] under other optimal specified conditions. Data emphasized the inducing role of gelatin on TAP-biosynthesis at relatively high temperature and pH value

Protease production during growth and autolysis of submerged Metarhizium anisopliae cultures

The growth and autolysis of two strains of the entomopathogenic deuteromycete fungus Metarhizium anisopliae var.anisopliae were evaluated in medium containing casein or glucose as carbon source. Parameters such as economic coefficient and degree of autolysis were determined for each strain. Protease production was determined throughout the growth and autolysis phases of the cultures on medium under conditions of proteaseinduction (in the presence of casein as sole source of carbon and nitrogen). The fungus was shown to utilize casein as a carbon/energy source in a more efficient manner than glucose. The autolysis shown by the strains was intense under both types of growth conditions, reaching up to 62,7 (per cent) of the dry mass produced and started soon after the depletion of the exogenous carbon source. The relationship between the proteolytic activities of the two strains evaluated varied significantly (a maximum of 19.78 on the 5th day and a minimum of 2.03 on the 16th day of growth) during the various growth and autolysis phases, clearly showing that the difference between the growth curves and the difference in the kinetics of enzyme production may decisively affect the process of strain selection for protease production.

Toxinas killer e produção de enzimas por Candida albicans isoladas da mucosa bucal de pacientes com câncer / Killer toxin and enzyme production by Candida albicans isolated from buccal mucosa in patients with cancer

Opportunistic infections of the oral cavity are primarily caused by Candida and frequently occur in patients with cancer who are undergoing chemotherapy and antibiotic treatment. Of the specimens received from the oral mucosa of 44 patients with cancer, 25 (56.8%) yielded Candida on culture in Sabouraud agar. Twenty four of these isolates were identified as C. albicans (96%) and 1 as C. krusei (4%). The phenotypic characteristics of these isolates showed that all of them were strongly proteolytic, had a high ability to produce phospholipase, and presented the byotypes characterized as 811 (95.8%) and 511 (4.2%) in terms of susceptibility to killer toxins.

Infecções oportunistas da cavidade bucal são primariamente causadas por fungos do gênero Candida e freqüentemente ocorrem em pacientes com câncer que estão sobtratamento quimioterápico e antibacteriano. De 44 amostras coletadas da mucosa oral de pacientes com câncer, observou-se o isolamento de 25 leveduras do gênero Candida em cultivo realizado em ágar Sabouraud-dextrose. Foram identificados Candida albicans em 24 (96%) isolados e C. krusei em 1 (4%). As características fenotípicas das amostras de Candida albicans mostraram que todos os isolados foram fortemente proteolíticos, capazes de produzir fosfolipases e possuíam os biotipos caracterizados como 811(95,8%) e 511 (4,2%) em relação a susceptibilidade às toxinas killer.

Production and some properties of proteinase by fungi utilizing whey

Ten fungal strains were screened on different kinds of whey and permeate for their ability to produce proteinase enzymes. Rhizopus sp. NRC 5 was selected as a new isolate from acid whey characterized by its high proteolytic productivity when grown in submerged culture of acid whey for 5 days at 30C +/- 2. Addition of CaC12 to the fermentation medium [acid whey] stimulated the enzymes formation, while MgC12 inhibited the enzymes biosynthesis. The produced enzymes gave high proteolytic activity at acidic and alkaline pH [5.5 and 8]. Optimum reaction temperature and time for acidic and alkaline proteinase enzymes were 40C for 15 and 25 min, respectively. Maximum acidic and alkaline proteolytic enzymes activity was obtained at enzyme concentrations of 0.54 mg/ml besides substrate concentration of 2.0 g%

Production of extracellular proteases and amylases by some acidophilic and alkalophilic bacteria.

Twelve bacteria were isolated from two effluent sources of Shaw-Wallace Gelatins, Jabalpur. Six bacteria from dicalcium phosphate plant effluent (pH-5) and six from main drain of the factory (pH-10) were isolated. Two facultatively acidophilic and two facultatively alkalophilic bacteria were selected and tentatively identified as Plesiomonas shigelloides, Aeromonas hydrophilla, Klebsiella pneumoniae and Staphylococcus saprophyticus respectively. Acidic amylases were produced in higher amounts on 4th day of incubation by Plesiomonas shigelloides and on 6th day by Aeromonas hydrophilla. Alkaline amylases were produced in higher amounts on 4th day of incubation by Klebsiella pneumoniae and on 8th day by Staphylococcus saprophyticus in vitro.

Production of extracellular protease by Streptomyces fradiae.

A new strain of Streptomyces fradiae, isolated and identified in our laboratory, was found to be a potential producer of protease enzyme. An enzyme yield of 285 P.U./ml. was obtained prior to optimization in a synthetic medium containing potato starch, potassium nitrate and trace salts by shake flask method. The carbon and nitrogen compounds play prominent role in the protease production. The maximum enzyme yield of 930 P.U./ml. (about 3-fold increase) was obtained with all optimum parameters such as pH. 7.0, with 3% jowar starch, 0.5% tryptone, 28 degrees C incubation temperature, with 1:20 medium to flask volume ratio and with 48 hrs. inoculum.

Serotyping, proteinase and phospholipase production associated with the pathogenicity of Candida albicans strains isolated from frog meat

Quinze amostras de Candida albicans isoladas de coxas de räs (Rana catesbeiana, Shaw, 1802) comercializadas no Brasil, foram estudadas quanto ao sorotipo e fatores de virulência "in vitro" e "in vivo". A sorotipagem revelou predominância do sorotipo B. Todas as amostras estudadas foram produtoras de fosfolipases e 66,6 por cento produziram proteinases. No estudo da patogenicidade foram observados dois tipos de alteraçöes renais: uma do tipo agudo com abscessos corticais e outra do tipo crônico com pielonefrite e envolvimento unilateral

Proteolytic activity of saccharomyces cerevisiae as affected by cultural conditions. II. Effect of some nutritional requirements

Supplementation of the fermentation medium with different nitrogen and carbon sources and their effects on growth and proteinase production by S. cerevisiae was studied. Organic complex nitrogen compounds enhanced yeast growth and stimulated the process of proteinase production. Similarly, medium fortified with molasses produced the highest growth and proteinase yields. Effect of different concentrations of peptone as the best nitrogen source - on growth and proteinase production by the yeast cells was also studied. 0.5% was the optimum concentration of peptone at which maximum growth and proteinase production were yielded. Effect of different concentrations of molasses - as the best carbon source - was studied. 1.0% was the optimum concentration of molasses at which maximal biomass yield was gained. Increasing molasses concentration in the medium resulted in a concomitant decrease of the proteolytic activity of the organism

Evidence for the participation of proteinases released by Candida albicans in the early killing of peritoneal macrophages in vitro

We have investigated the possibility that proteinases released by Candida albicans participate in the early killing of three types of mice peritoneal macrophage (resident, thioglycollate-elicited, or Con-A-activated)in vitro. Phagocytic assays were performed by incubation of macrophages and C. albicans together at a 1:10 ratio for 30 min at 37 grade C in RPMI medium buffered to pH 7.0 with 12 mM Hepes without serum. With no albumin added to the medium, the medium, the macrophages were 85% to 100% damaged and unviable, to a greater extent than expected from the proportion of phagocytic cells containing germ tubes. When 10 mg/ml of albumin was added to the medium, however, 90% of the macrophages remained viable for the 2 hours of the phagocytic assay, suggesting that albumin may have acted as a substrate for or inhibitor of proteinases released by C. albicans, thereby protecting the macrophage from the proteolytic action of the proteinases. The phagocytosis of IgG-coacted erythrocytes was reduced to 43% when IgG was preincubated with the supernatant from C. albicans cultures, but the addition of 10 mg/ml albuumin or of 5 ug/ml pepstatin (an inhibitor of C. albicans acid proteinases) to the same supernatans prevented the effect on phagocytosis of IgG-coated erythrocytes. These results suggest that proteinases released from C. albicans are involved in the early killing of macrophages