Cyclodextrin glycosyltransferase from Bacillus licheniformis: Optimization of production and its properties

Cyclodextrin glycosyltransferase (EC 2.4.1.19) is an enzyme that produces cyclodextrins from starch via an intramolecular transglycosylation reaction. An alkalophilic Bacillus strain, isolated from cassava peels, was identified as Bacillus licheniformis. CGTase production by this strain was better when potato starch was used as carbon source, followed by cassava starch and amylopectin. Glucose and amylose, on the other hand, acted as synthesis repressors. When the cultivation was supplemented with sodium ions and had the pH adjusted between 6.0 and 9.0, the microorganism maintained the growth and enzyme production capacity. This data is interesting because it contradicts the concept that alkalophilic microorganisms do not grow in this pH range. After ultrafiltration-centrifugation, one protein of 85.2 kDa with CGTase activity was isolated. This protein was identified in plates with starch and phenolphthalein. Determination of the optimum temperature showed higher activities at 25 degrees C and 55 degrees C, indicating the possible presence of more than one CGTase in the culture filtrate. Km and Vmax values were 1.77 mg/mL and 0.0263 U/mg protein, respectively, using potato starch as substrate.

PRODUCTION OF MICROBIAL ALKALINE CELLULASE AND STUDIES OF THEIR CHARACTERISTICS

In order to obtain cellulases that improve the detergency of laundry detergent products, two alkalophilic microorganims, Bacillus sp B38-2 and Streptomyces sp S36-2, were isolated from soil and compost by incubating samples in enrichment culture medium containing CMC and Na2CO3 at pH9.6. It was found that they secrete a constitutive extracellular alkaline carboxymethyl cellulase (CMCase) in high quantity. The maximum enzyme activity was observed between 48hr to 72 hr at 30-degrees-C for the Streptomyces and between 72hr to 96hr at 35-degrees-C for the Bacillus. The optimum pH and temperature of the crude enzyme activities ranged from 6.0 to 7.0 at 55-degrees-C for the Streptomyces and 7.0 to 8.0 at 60-degrees-C for the Bacillus. Two crude CMCases activities were termostable at 45-degrees-C for 1hr and the both crude enzyme activities of the Bacillus as of the Streptomyces were stable at pH 5.0 to 9.0 after pH treatments in various buffer solutions at 30-degrees-C for 24hr.

Organic acids and/or compound with defined microorganisms to control Salmonella enterica serovar Enteritidis experimental infection in chickens

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Retention of oral microorganisms on conventional and resin-modified glass-ionomer cements

A cárie secundária representa problema de saúde pública e socioeconômico no mundo. A restauração de dentes acometidos por cárie pode criar condições favoráveis à proliferação microbiana na superfície do material restaurador ou na interface dente/restauração, criando ambiente propício para o estabelecimento de cárie secundária. O objetivo deste estudo foi avaliar a capacidade de retenção de placa bacteriana em cimentos de ionômero de vidro convencionais (Chelon-Fil e Vidrion R) e modificados por resina (Vitremer e Fuji II LC) e de resina composta híbrida (Z100), utilizada como controle. Nos testes de retenção de microrganismos, in situ, 12 voluntários utilizaram, por 7 dias, placa de Hawley contendo corpos-de-prova de todos os materiais. A seguir, os corpos-de-prova foram transferidos para tubos contendo 2,0 ml de Ringer-PRAS e os microrganismos presentes em sua superfície foram cultivados em placa com ágar-sangue e ágar Mitis Salivarius Bacitracina, os quais foram incubados, a 37ºC, em anaerobiose (90% N2, 10% CO2), por 10 e 2 dias, respectivamente. Os ionômeros modificados por resina retiveram quantidade de bactérias similar àquela mostrada pela resina testada. Os ionômeros modificados por resina também apresentaram menor número de estreptococos do grupo mutans do que a resina e os cimentos ionoméricos convencionais. Os ionômeros de vidro convencionais apresentaram menor número de estreptococos do grupo mutans que a resina, sendo que essa diferença não foi estatisticamente significativa.

Photodynamic inactivation of microorganisms present on complete dentures. A clinical investigation

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Purification and characterization of two xylanases from alkalophilic and thermophilic Bacillus licheniformis 77-2

The alkalophilic bacteria Bacillus licheniformis 77-2 produces significant quantities of thermostable cellulase-free xylanases. The crude xylanase was purified to apparent homogeneity by gel filtration (G-75) and ionic exchange chromatography (carboxymethyl sephadex, Q sepharose, and Mono Q), resulting in the isolation of two xylanases. The molecular masses of the enzymes were estimated to be 17 kDa (X-I) and 40 kDa (X-II), as determined by SDS-PAGE. The K(m) and V(max) values were 1.8 mg/mL and 7.05 U/mg protein (X-I), and 1.05 mg/mL and 9.1 U/mg protein (X-II). The xylanases demonstrated optimum activity at pH 7.0 and 8.0-10.0 for xylanase X-I and X-II, respectively, and, retained more than 75% of hydrolytic activity up to pH 11.0. The purified enzymes were most active at 70 and 75 degrees C for X-I and X-II, respectively, and, retained more than 90% of hydrolytic activity after 1 h of heating at 50 degrees C and 60 degrees C for X-I and X-II, respectively. The predominant products of xylan hydrolysates indicated that these enzymes were endoxylanases.

Contribution of the Malpighian tubules for the maintenance of symbiotic microorganisms in cephalotes ants

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Ileum of the Cephalotes ants: A specialized structure to harbor symbionts microorganisms

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Production and characteristics comparison of crude beta-glucosidases produced by microorganisms Thermoascus aurantiacus e Aureobasidium pullulans in agricultural wastes

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Endophytic microorganisms from coffee tissues as plant growth promoters and biocontrol agents of coffee leaf rust

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Action of soil microorganisms on PCL and PHBV blend and films

Poly(hydroxybutyrate-co-valerate) (PHBV) and poly(epsilon-caprolactone) (PCL) PCL/PHBV (4:1) blend films were prepared by melt-pressing. The biodegradation of the films in response to burial in soil for 30 days was investigated by Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), differential scanning calorimetry (DSC) and thermogravimetry (TG). The PHBV film was the most susceptible to microbial attack, since it was rapidly biodegraded via surface erosion in 15 days and completely degraded in 30 days. The PCL film also degraded but more slowly than PHBV. The degradation of the PCL/PHBV blend occurred in the PHBV phase, inducing changes in the PCL phases (interphase) and resulting in an increase of its crystalline fraction.

Culture-dependent and culture-independent characterization of microorganisms associated with Aedes aegypti (Diptera: Culicidae) (L.) and dynamics of bacterial colonization in the midgut

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Antimicrobial activity of Calendula officinalis, Camellia sinensis and chlorhexidine against the adherence of microorganisms to sutures after extraction of unerupted third molars

Objective: The objective of this study was to compare the antimicrobial effect of mouthwashes containing Calendula officinalis L., Camellia sinensis (L.) Kuntze and 0.12% chlorhexidine digluconate on the adherence of microorganisms to suture materials after extraction of unerupted third molars. Material and Methods: Eighteen patients with unerupted maxillary third molars indicated for extraction were selected (n=6 per mouthwash). First, the patients were subjected to extraction of the left tooth and instructed not to use any type of antiseptic solution at the site of surgery (control group). After 15 days, the right tooth was extracted and the patients were instructed to use the Calendula officinalis, Camellia sinensis or chlorhexidine mouthwash during 1 week (experimental group). For each surgery, the sutures were removed on postoperative day 7 and placed in sterile phosphate-buffered saline. Next, serial dilutions were prepared and seeded onto different culture media for the growth of the following microorganisms: blood agar for total microorganism growth; Mitis Salivarius bacitracin sucrose agar for mutans group streptococci; mannitol agar for Staphylococcus spp.; MacConkey agar for enterobacteria and Pseudomonas spp., and Sabouraud dextrose agar containing chloramphenicol for Candida spp. The plates were incubated during 24-48 h at 37 degrees C for microorganism count (CFU/nnL). Results: The three mouthwashes tested reduced the number of microorganisms adhered to the sutures compared to the control group. However, significant differences between the control and experimental groups were only observed for the mouthwash containing 0.12% chlorhexidine digluconate. Conclusions: Calendula officinalis L. and Camellia sinensis (L.) Kuntze presented antimicrobial activity against the adherence of microorganisms to sutures but were not as efficient as chlorhexidine digluconate.