Denitrifying phosphate accumulating organisms and its mechanism of nitrogen and phosphorus removal / 生物工程学报

Water eutrophication poses great threats to protection of water environment. Microbial remediation of water eutrophication has shown high efficiency, low consumption and no secondary pollution, thus becoming an important approach for ecological remediation. In recent years, researches on denitrifying phosphate accumulating organisms and their application in wastewater treatment processes have received increasing attention. Different from the traditional nitrogen and phosphorus removal process conducted by denitrifying bacteria and phosphate accumulating organisms, the denitrifying phosphate accumulating organisms can simultaneously remove nitrogen and phosphorus under alternated anaerobic and anoxic/aerobic conditions. It is worth noting that microorganisms capable of simultaneously removing nitrogen and phosphorus absolutely under aerobic conditions have been reported in recent years, but the mechanisms remain unclear. This review summarizes the species and characteristics of denitrifying phosphate accumulating organisms and the microorganisms capable of performing simultaneous nitrification-denitrification and phosphorous removal. Moreover, this review analyzes the relationship between nitrogen removal and phosphorus removal and the underlying mechanisms, discusses the challenges of denitrifying phosphorus removal, and prospects future research directions, with the aim to facilitate process improvement of denitrifying phosphate accumulating organisms.

Enhanced nitrogen removal by bioelectrochemical coupling anammox and characteristics of microbial communities / 生物工程学报

To investigate the bioelectrochemical enhanced anaerobic ammonia oxidation (anammox) nitrogen removal process, a bioelectrochemical system with coupled anammox cathode was constructed using a dual-chamber microbial electrolysis cell (MEC). Specifically, a dark incubation batch experiment was conducted at 30 ℃ with different influent total nitrogen concentrations under an applied voltage of 0.2 V, and the enhanced denitrification mechanism was investigated by combining various characterization methods such as cyclic voltammetry, electrochemical impedance spectroscopy and high-throughput sequencing methods. The results showed that the total nitrogen removal rates of 96.9%±0.3%, 97.3%±0.4% and 99.0%±0.3% were obtained when the initial total nitrogen concentration was 200, 300 and 400 mg/L, respectively. In addition, the cathode electrode biofilm showed good electrochemical activity. High-throughput sequencing results showed that the applied voltage enriched other denitrifying functional groups, including Denitratisoma, Limnobacter, and ammonia oxidizing bacteria SM1A02 and Anaerolineaceae, Nitrosomonas europaea and Nitrospira, besides the anammox bacteria. These electrochemically active microorganisms comprised of ammonium oxidizing exoelectrogens (AOE) and denitrifying electrotrophs (DNE). Together with anammox bacteria Candidatus Brocadia, they constituted the microbial community structure of denitrification system. Enhanced direct interspecies electron transfer between AOE and DNE was the fundamental reason for the further improvement of the total nitrogen removal rate of the system.

Functional analysis of functional membrane microdomains in the biosynthesis of menaquinone-7 / 生物工程学报

Functional membrane microdomains (FMMs) that are mainly composed of scaffold proteins and polyisoprenoids play important roles in diverse cellular physiological processes in bacteria. The aim of this study was to identify the correlation between MK-7 and FMMs and then regulate the MK-7 biosynthesis through FMMs. Firstly, the relationship between FMMs and MK-7 on the cell membrane was determined by fluorescent labeling. Secondly, we demonstrated that MK-7 is a key polyisoprenoid component of FMMs by analyzing the changes in the content of MK-7 on cell membrane and the changes in the membrane order before and after destroying the integrity of FMMs. Subsequently, the subcellular localization of some key enzymes in MK-7 synthesis was explored by visual analysis, and the intracellular free pathway enzymes Fni, IspA, HepT and YuxO were localized to FMMs through FloA to achieve the compartmentalization of MK-7 synthesis pathway. Finally, a high MK-7 production strain BS3AT was successfully obtained. The production of MK-7 reached 300.3 mg/L in shake flask and 464.2 mg/L in 3 L fermenter.

Mechanism of trehalose-enhanced metabolism of heterotrophic nitrification-aerobic denitrification community under high-salt stress / 生物工程学报

Heterotrophic nitrification-aerobic denitrification (HN-AD) bacteria are aerobic microorganisms that can remove nitrogen under high-salt conditions, but their performance in practical applications are not satisfactory. As a compatible solute, trehalose helps microorganisms to cope with high salt stress by participating in the regulation of cellular osmotic pressure, and plays an important role in promoting the nitrogen removal efficiency of microbial populations in the high-salt environment. We investigated the mechanism of exogenous-trehalose-enhanced metabolism of HN-AD community under high-salt stress by starting up a membrane aerobic biofilm reactor (MABR) to enrich HN-AD bacteria, and designed a C150 experimental group with 150 μmol/L trehalose addition and a C0 control group without trehalose. The reactor performance and the community structure showed that NH4+-N, total nitrogen (TN) and chemical oxygen demand (COD) removal efficiency were increased by 29.7%, 28.0% and 29.1%, respectively. The total relative abundance of salt-tolerant HN-AD bacteria (with Acinetobacter and Pseudofulvimonas as the dominant genus) in the C150 group reached 66.8%, an 18.2% increase compared with that of the C0 group. This demonstrated that trehalose addition promoted the enrichment of salt-tolerant HN-AD bacteria in the high-salt environment to enhance the nitrogen removal performance of the system. In-depth metabolomics analysis showed that the exogenous trehalose was utilized by microorganisms to improve proline synthesis to increase resistance to high-salt stress. By regulating the activity of cell proliferation signaling pathways (cGMP-PKG, PI3K-Akt), phospholipid metabolism pathway and aminoacyl-tRNA synthesis pathway, the abundances of phosphoethanolamine, which was one of the glycerophospholipid metabolites, and purine and pyrimidine were up-regulated to stimulate bacterial aggregation and cell proliferation to promote the growth of HN-AD bacteria in the high-salt environment. Meanwhile, the addition of trehalose accelerated the tricarboxylic acid (TCA) cycle, which might provide more electron donors and energy to the carbon and nitrogen metabolisms of HN-AD bacteria and promote the nitrogen removal performance of the system. These results may facilitate using HN-AD bacteria in the treatment of high-salt and high-nitrogen wastewater.

Media development and process parameter optimization using statistical experimental designs for the production of nonribosomal peptides in Escherichia coli

BACKGROUND: Nonribosomal peptide synthases (NRPS) can synthesize functionally diverse bioactive peptides by incorporating nonproteinogenic amino acids, offering a rich source of new drug leads. The bacterium Escherichia coli is a well-characterized production host and a promising candidate for the synthesis of nonribosomal peptides, but only limited bioprocess engineering has been reported for such molecules. We therefore developed a medium and optimized process parameters using the design of experiments (DoE) approach. RESULTS: We found that glycerol is not suitable as a carbon source for rhabdopeptide production, at least for the NRPS used for this study. Alternative carbon sources from the tricarboxylic acid cycle achieved much higher yields. DoE was used to optimize the pH and temperature in a stirred-tank reactor, revealing that optimal growth and optimal production required substantially different conditions. CONCLUSIONS: We developed a chemically defined adapted M9 medium matching the performance of complex medium (lysogeny broth) in terms of product concentration. The maximum yield in the reactor under optimized conditions was 126 mg L-1, representing a 31-fold increase compared to the first shaking-flask experiments with M9 medium and glycerol as the carbon source. Conditions that promoted cell growth tended to inhibit NRPS productivity. The challenge was therefore to find a compromise between these factors as the basis for further process development.

Ammonia determines transcriptional profile of microorganisms in anaerobic digestion

ABSTRACT Anaerobic digestion is important for the management of livestock manure with high ammonia level. Although ammonia effects on anaerobic digestion have been comprehensively studied, the molecular mechanism underlying ammonia inhibition still remains elusive. In this study, based on metatranscriptomic analysis, the transcriptional profile of microbial community in anaerobic digestion under low (1500 mg L-1) and high NH4 + (5000 mg L-1) concentrations, respectively, were revealed. The results showed that high NH4 + concentrations significantly inhibited methane production but facilitated the accumulations of volatile fatty acids. The expression of methanogenic pathway was significantly inhibited by high NH4 + concentration but most of the other pathways were not significantly affected. Furthermore, the expressions of methanogenic genes which encode acetyl-CoA decarbonylase and methyl-coenzyme M reductase were significantly inhibited by high NH4 + concentration. The inhibition of the co-expressions of the genes which encode acetyl-CoA decarbonylase was observed. Some genes involved in the pathways of aminoacyl-tRNA biosynthesis and ribosome were highly expressed under high NH4 + concentration. Consequently, the ammonia inhibition on anaerobic digestion mainly focused on methanogenic process by suppressing the expressions of genes which encode acetyl-CoA decarbonylase and methyl-coenzyme M reductase. This study improved the accuracy and depth of understanding ammonia inhibition on anaerobic digestion.

Optimizing culture conditions for production of intra and extracellular inulinase and invertase from Aspergillus niger ATCC 20611 by response surface methodology (RSM)

Abstract The aim of this study was obtain a model that maximizes growth and production of inulinase and invertase by Aspergillus niger ATCC 20611, employing response surface methodology (RSM). The RSM with a five-variable and three-level central composite design (CCD) was employed to optimize the medium composition. Results showed that the experimental data could be appropriately fitted into a second-order polynomial model with a coefficient of determination (R2) more than 0.90 for all responses. This model adequately explained the data variation and represented the actual relationships between the parameters and responses. The pH and temperature value of the cultivation medium were the most significant variables and the effects of inoculum size and agitation speed were slightly lower. The intra-extracellular inulinase, invertase production and biomass content increased 10-32 fold in the optimized medium condition (pH 6.5, temperature 30 °C, 6% (v/v), inoculum size and 150 rpm agitation speed) by RSM compared with medium optimized through the one-factor-at-a-time method. The process development and intensification for simultaneous production of intra-extracellular inulinase (exo and endo inulinase) and invertase from A. niger could be used for industrial applications.

Effects of oxygen transfer coefficient on dihydroxyacetone production from crude glycerol

Abstract The principal objective of this study was to evaluate the kinetics of dihydroxyacetone production by Gluconobacter frateurii CGMCC 5397 under different oxygen volumetric mass transfer coefficient (kLa) conditions in submerged bioreactors using biodiesel-derived crude glycerol as the carbon source. kLa is a key fermentation parameter for the production of dihydroxyacetone. Cultivations were conducted in baffled- and unbaffled-flask cultures (the kLa values were 24.32 h−1 and 52.05 h−1, respectively) and fed-batch cultures (the kLa values were held at 18.21 h−1, 46.03 h−1, and 82.14 h−1) to achieve high dihydroxyacetone concentration and productivity. The results showed that a high kLa could dramatically increase dihydroxyacetone concentrations and productivities. The baffled-flask culture (with a kLa of 52.05 h−1) favored glycerol utilization and dihydroxyacetone production, and a dihydroxyacetone concentration as high as 131.16 g/L was achieved. When the kLa was set to 82.14 h−1 in the fed-batch culture, the dihydroxyacetone concentration, productivity and yield were 175.44 g/L, 7.96 g/L/h and 0.89 g/g, respectively, all of which were significantly higher than those in previous studies and will benefit dihydroxyacetone industrial production.

Enhancing inulinase yield by irradiation mutation associated with optimization of culture conditions

A new inulinase-producing strain was isolated from rhizosphere soils of Jerusalem artichoke collected from Shihezi (Xinjiang, China) using Jerusalem artichoke power (JAP) as sole carbon source. It was identified as an Aspergillus niger strain by analysis of 16S rRNA. To improve inulinase production, this fungus was subjected to mutagenesis induced by ⁶⁰Co γ-irradiation. A genetically stable mutant (designated E12) was obtained and it showed 2.7-fold higher inulinase activity (128 U/mL) than the parental strain in the supernatant of a submerged culture. Sequential methodology was used to optimize the inulinase production of stain E12. A screening trial was first performed using Plackett-Burman design and variables with statistically significant effects on inulinase bio-production were identified. These significant factors were further optimized by central composite design experiments and response surface methodology. Finally, it was found that the maximum inulinase production (185 U/mL) could be achieved under the optimized conditions namely pH 7.0, yeast extract concentration of 5.0 g/L, JAP concentration of 66.5 g/L, peptone concentration of 29.1 g/L, solution volume of 49.4 mL in 250-mL shake flasks, agitation speed of 180 rpm, and fermentation time of 60 h. The yield of inulinase under optimized culture conditions was approximately 1.4-fold of that obtained by using basal culture medium. These findings are of significance for the potential industrial application of the mutant E12.

Bioleaching of gold, copper and nickel from waste cellular phone PCBs and computer goldfinger motherboards by two Aspergillus nigerstrains

In an effort to develop alternate techniques to recover metals from waste electrical and electronic equipment (WEEE), this research evaluated the bioleaching efficiency of gold (Au), copper (Cu) and nickel (Ni) by two strains of Aspergillus niger in the presence of gold-plated finger integrated circuits found in computer motherboards (GFICMs) and cellular phone printed circuit boards (PCBs). These three metals were analyzed for their commercial value and their diverse applications in the industry. Au-bioleaching ranged from 42 to 1% for Aspergillus niger strain MXPE6; with the combination of Aspergillus niger MXPE6 + Aspergillus niger MX7, the Au-bioleaching was 87 and 28% for PCBs and GFICMs, respectively. In contrast, the bioleaching of Cu by Aspergillus niger MXPE6 was 24 and 5%; using the combination of both strains, the values were 0.2 and 29% for PCBs and GFICMs, respectively. Fungal Ni-leaching was only found for PCBs, but with no significant differences among treatments. Improvement of the metal recovery efficiency by means of fungal metabolism is also discussed.

Aerobic cyanide degradation by bacterial isolates from cassava factory wastewater

Ten bacterial strains that utilize cyanide (CN) as a nitrogen source were isolated from cassava factory wastewater after enrichment in a liquid media containing sodium cyanide (1 mM) and glucose (0.2% w/v). The strains could tolerate and grow in cyanide concentrations of up to 5 mM. Increased cyanide levels in the media caused an extension of lag phase in the bacterial growth indicating that they need some period of acclimatisation. The rate of cyanide removal by the strains depends on the initial cyanide and glucose concentrations. When initial cyanide and glucose concentrations were increased up to 5 mM, cyanide removal rate increased up to 63 and 61 per cent by Bacillus pumilus and Pseudomonas putida. Metabolic products such as ammonia and formate were detected in culture supernatants, suggesting a direct hydrolytic pathway without an intermediate formamide. The study clearly demonstrates the potential of aerobic treatment with cyanide degrading bacteria for cyanide removal in cassava factory wastewaters.

Anaerobic degradation of anionic surfactants by indigenous microorganisms from sediments of a tropical polluted river in Brazil / Degradación anaeróbica de tensioactivos aniónicos en microorganismos autóctonos de los sedimentos de un río tropical contaminado en Brasil

Linear alkylbenzene sulfonate (LAS) is widely used in the formulation of domestic and industrial cleaning products, the most synthetic surfactants used worldwide. These products can reach water bodies through the discharge of untreated sewage or non-effective treatments. This study evaluates the ability of the microorganisms found in the Tietê river sediment to degrade this synthetic surfactant. The experiment was conducted in a bioreactor, operated in batch sequences under denitrifying conditions, with cycles of 24 hours and stirring at 150rpm, using 430mL of sediments and 1 070mL of a synthetic substrate consisting of yeast extract, soluble starch, sodium bicarbonate and sucrose. LAS was added at different concentrations of 15mg/L and 30mg/L. The reactor operation was divided into the biomass adaptation to the synthetic substrate without LAS and three experimental conditions: a) addition of 15mg/L of LAS; b) 50% reduction the co-substrate concentration and 15mg/L of LAS, and c) addition of 30mg/L of LAS and 100% co-substrate concentration. The results showed that the degradation efficiency of LAS was directly related to the addition of co-substrates and the population of denitrifying bacteria. The removal of LAS and nitrate can be achieved simultaneously in wastewater with low organic loads. The reduction in the co-substrates concentration was directly influenced by the number of denitrifying bacteria (2.2x10(13) to 1.0x10(8)MPN/gTVS), and consequently, LAS degradation (60.1 to 55.4%). The sediment microorganisms in the Tietê river can be used as an alternative inoculum in the treatment of wastewater with nitrate and LAS contamination.

El alquilbenceno sulfonato lineal (LAS) es el tensoactivo sintético más usado en todo el mundo en los produtos de limpeza domestica e industrial y puede llegar a las masas de agua a través de la descarga de aguas residuales sin tratamiento o con un tratamiento ineficaz. El objetivo del estudio consistió en evaluar la capacidad de la microbiota presente en el sedimento del río Tietê en la degradación del tensoactivo anionico - LAS. El experimento se llevó a cabo en un bioreactor de lotes secuenciales en condiciones de desnitrificación con ciclos de 24 horas, agitación de 150rpm, usando 430mL de sedimento y 1 070mL de sustrato sintético constituido por extracto de levadura, almidón soluble, bicarbonato de sodio y sacarosa. El LAS fue añadido a diferentes concentraciones de 15mg/L y 30mg/L. El funcionamiento del bioreactor se dividió en la adaptación de la biomasa con sustrato sintético sin LAS y tres condiciones experimentales: A) adición de 15mg/L de LAS; B) 15mg/L de LAS y reducción del 50% de la concentración del co-sustrato y C) 30mg/L de LAS y la concentración de 100% de co-substrato. Los resultados obtenidos muestran que la eficiencia en la degradación del LAS está directamente relacionada con la población de bacterias desnitrificadoras y que el sedimento del río Tietê se puede utilizar como inóculo en el tratamiento de LAS en condiciones desnitrificadoras. La población de bacterias fue capaz de degradar el LAS independiente de la fuente de carbón adicionada. La remoción de LAS y de nitrato se puede lograr simultáneamente en aguas residuales con una baja carga orgánica. La reducción de la concentración del co-sustrato fue influenciado directamente por la población de bacterias desnitrificantes (2.2x10(13) a 1.0x10(8)MNP/gTVS) y por lo tanto la degradación de LAS (60.1-55.4%). Los microorganismos en el sedimento del río Tietê se pueden usar como inóculo alternativo para el tratamiento de efluentes contaminados con nitrato y LAS.

Microbial dynamics during azo dye degradation in a UASB reactor supplied with yeast extract

The present work aimed to investigate the microbial dynamics during the anaerobic treatment of the azo dye blue HRFL in bench scale upflow anaerobic sludge bed (UASB) reactor operated at ambient temperature. Sludge samples were collected under distinct operational phases, when the reactor were stable (low variation of color removal), to assess the effect of glucose and yeast extract as source of carbon and redox mediators, respectively. Reactors performance was evaluated based on COD (chemical oxygen demand) and color removal. The microbial dynamics were investigated by PCR-DGGE (Polimerase Chain Reaction - Denaturing Gradient of Gel Electrophoresis) technique by comparing the 16S rDNA profiles among samples. The results suggest that the composition of microorganisms changed from the beginning to the end of the reactor operation, probably in response to the presence of azo dye and/or its degradation byproducts. Despite the highest efficiency of color removal was observed in the presence of 500 mg/L of yeast extract (up to 93%), there were no differences regarding the microbial profiles that could indicate a microbial selection by the yeast extract addition. On the other hand Methosarcina barkeri was detected only in the end of operation when the best efficiencies on color removal occurred. Nevertheless the biomass selection observed in the last stages of UASB operation is probably a result of the washout of the sludge in response of accumulation of aromatic amines which led to tolerant and very active biomass that contributed to high efficiencies on color removal.

Obtención de sobrenadante de cultivo aglutinador de hematíes portadores del antígeno de grupo sanguíneo B / Obtention of cell culture supernatant for erythrocyte agglutination of blood group B antigen

Introducción: la obtención de anticuerpos monoclonales en líquido ascítico ha ido decayendo paulatinamente por la aparición de alternativas de producción in Vitro, que permiten alcanzar mayores volúmenes y un control más riguroso del proceso productivo, lo que incrementa la reproducibilidad de procesos y la calidad de los productos. Objetivo: evaluar dos métodos de producción de sobrenadante de cultivo rico en una inmunoglobulina de ratón del tipo IgG2b, aglutinadora de hematíes humanos portadores del antígeno del grupo sanguíneo B, según el sistema ABO; la cual es secretada por el hibridoma C6G4. Métodos: se evaluaron dos métodos de producción del anticuerpo con el empleo de un biorreactor CELLine, útil como modelo para la obtención de anticuerpos monoclonales en cultivos de alta densidad celular. Los métodos se diferenciaron esencialmente en la densidad celular de siembra en el biorreactor y en la duración del periodo de fermentación entre la siembra y la cosecha del caldo de cultivo rico en anticuerpos. Para cada método se determinó la concentración específica de anticuerpos y la potencia de aglutinación del sobrenadante, así como la densidad y la viabilidad celular del cultivo alcanzadas en el momento de la cosecha. Resultados: se observó que ambos métodos generaron sobrenadantes de cultivo con una potencia de aglutinación similar, a pesar de que se encontraron diferencias en el resto de las variables medidas. Si bien uno de los métodos produjo una mayor concentración de anticuerpos en el sobrenadante, no se observaron diferencias en la potencia de aglutinación de los sobrenadantes obtenidos por ambas alternativas. Conclusiones: los dos métodos estudiados permitieron obtener volúmenes semejantes de sobrenadante anti-B con diferentes concentraciones de anticuerpos, pero con una potencia de aglutinación similar. La principal diferencia residió en que uno de los métodos permitió obtener el mismo volumen del producto en un tiempo sensiblemente menor...

Introduction : the obtention of monoclonal antibodies in ascite fluid has been declining gradually due to the appearance of alternative in vitro production that achieve higher volumes and a more precise monitoring of the production process, which increases the reproducibility of processes and the quality of products. Objective : to evaluate two methods to make cell culture supernatant rich in murine monoclonal IgG2b type, with agglutinating activity against human red cell of blood group antigen B (ABO system), which is secreted by murine hybridoma C6G4. Methods : two methods were evaluated for antibody production in cell culture supernatant using as model a CELLine bioreactor for the production of monoclonal antibodies in high cell density culture. Both methods essentially differed in the seeding cell density in the bioreactor and the fermentation period between seeding and harvesting of the culture broth rich in antibodies. The specific antibody concentration and potency of agglutination was determined in the obtained supernatant and also the cell density and cell viability of the culture reached at the time of harvest. Results : both methods generated culture supernatants with similar agglutination strength despite differences found in the rest of the variables measured. Even when one of the methods produced a higher antibody concentration in the supernatants, no differences in potency of the supernatants agglutination obtained by both alternatives were observed. Conclusions : both methods generated supernatant anti-B with different concentrations of antibodies but similar potency of agglutination. The main difference was that with one of the methods the same volume of the product was obtained in a considerably minor time...

Volatile fatty acids influence on the structure of microbial communities producing PHAs

Polyhydroxyalkanoates (PHAs) can be produced by microorganisms and are a biodegradable alternative to fossil-fuel based plastics. Currently, the focus is on reducing production costs by exploring alternative substrates for PHAs production, and on producing copolymers which are less brittle than monomers. Accordingly, this study used a substrate consisting of wastewater from waste-glycerol fermentation, supplemented with different amounts of acetic and propionic acids. These substrates were used to feed mixed microbial communities enriched from activated sludge in a sequencing batch reactor. A reactor supplemented with 2 mL of acetic acid produced 227.8 mg/L of a homopolymer of hydroxybutyrate (3HB); 4 mL of acetic acid produced 279.8 mg/L 3HB; whereas 4 mL of propionic acid produced 673.0 mg/L of a copolymer of 3HB and 3HV (hydroxyvalerate). Ribosomal Intergenic Spacer Analysis (RISA) was used to show the differences between the communities created in the reactors. Thauera species predominated in biomass that produced 3HB; Paracoccus denitrificans in the biomass that produced 3HB-co-3HV. Because P. denitrificans produced the more desirable copolymer, it may be advantageous to promote its growth in PHAs-producing reactors by adding propionate.

Canthaxanthin biosynthesis by Dietzia natronolimnaea HS-1: effects of inoculation and aeration rate

The interest in production of natural colorants by microbial fermentation has been currently increased. The effects of D-glucose concentration (3.18-36.82 g/L), inoculum size (12.5 x 10(9)-49.5 x 10(9) cfu cells/mL) and air-flow rate (1.95-12.05 L/L min) on the biomass, total carotenoid and canthaxanthin (CTX) accumulation of Dietzia natronolimnaea HS-1 in a batch bioreactor was scrutinized using a response surface methodology-central composite rotatable design (RSM-CCRD). Second-order polynomial models with high R² values ranging from 0.978 to 0.990 were developed for the studied responses using multiple linear regression analysis. The models showed the maximum cumulative amounts of biomass (7.85 g/L), total carotenoid (5.48 mg/L) and CTX (4.99 mg/L) could be achieved at 23.38 g/L of D-glucose, 31.2 x 10(9) cfu cells/mL of inoculation intensity and air-flow rate of 7.85 L/L min. The predicted values for optimum conditions were in good agreement with experimental data.

Application of statistical experimental design for optimisation of bioinsecticides production by sporeless Bacillus thuringiensis strain on cheap medium

In order to overproduce bioinsecticides production by a sporeless Bacillus thuringiensis strain, an optimal composition of a cheap medium was defined using a response surface methodology. In a first step, a Plackett-Burman design used to evaluate the effects of eight medium components on delta-endotoxin production showed that starch, soya bean and sodium chloride exhibited significant effects on bioinsecticides production. In a second step, these parameters were selected for further optimisation by central composite design. The obtained results revealed that the optimum culture medium for delta-endotoxin production consists of 30 g L-1 starch, 30 g L-1 soya bean and 9g L-1 sodium chloride. When compared to the basal production medium, an improvement in delta-endotoxin production up to 50% was noted. Moreover, relative toxin yield of sporeless Bacillus thuringiensis S22 was improved markedly by using optimised cheap medium (148.5 mg delta-endotoxins per g starch) when compared to the yield obtained in the basal medium (94.46 mg delta-endotoxins per g starch). Therefore, the use of optimised culture cheap medium appeared to be a good alternative for a low cost production of sporeless Bacillus thuringiensis bioinsecticides at industrial scale which is of great importance in practical point of view.

Semi-scale production of PHAs from waste frying oil by Pseudomonas fluorescens S48

The present study aimed at developing a strategy to improve the volumetric production of PHAs by Pseudomonas fluorescens S48 using waste frying oil (WFO) as the sole carbon source. For this purpose, several cultivations were set up to steadily improve nutrients supply to attain high cell density and high biopolymer productivity. The production of PHAs was examined in a 14 L bioreactor as one-stage batch, two-stage batch, and high-cell-density fed-batch cultures. The highest value of polymer content in one-stage bioreactor was obtained after 60 h (33.7%). Whereas, the two-stage batch culture increased the polymer content to 50.1% after 54 h. High-cell-density (0.64 g/L) at continuous feeding rate 0.55 mL/l/h of WFO recorded the highest polymer content after 54 h (55.34%). Semi-scale application (10 L working volume) increased the polymer content in one-stage batch, two-stage batch and high cell density fed-batch cultures by about 12.3%, 5.8% and 11.3%, respectively, as compared with that obtained in 2 L fermentation culture. Six different methods for biopolymer extraction were done to investigate their efficiency for optimum polymer recovery. The maximum efficiency of solvent recovery of PHA was attained by chloroform-hypochlorite dispersion extraction. Gas chromatography (GC) analysis of biopolymer produced by Pseudomonas fluorescens S48 indicated that it solely composed of 3-hydrobutyric acid (98.7%). A bioplastic film was prepared from the obtained PHB. The isolate studied shares the same identical sequence, which is nearly the complete 16S rRNA gene. The identity of this sequence to the closest pseudomonads strains is about 98-99%. It was probably closely related to support another meaningful parsiomony analysis and construction of a phylogenetic tree. The isolate is so close to Egyptian strain named EG 639838.

Yeast biomass production: a new approach in glucose-limited feeding strategy

The aim of this work was to implement experimentally a simple glucose-limited feeding strategy for yeast biomass production in a bubble column reactor based on a spreadsheet simulator suitable for industrial application. In biomass production process using Saccharomyces cerevisiae strains, one of the constraints is the strong tendency of these species to metabolize sugars anaerobically due to catabolite repression, leading to low values of biomass yield on substrate. The usual strategy to control this metabolic tendency is the use of a fed-batch process in which where the sugar source is fed incrementally and total sugar concentration in broth is maintained below a determined value. The simulator presented in this work was developed to control molasses feeding on the basis of a simple theoretical model in which has taken into account the nutritional growth needs of yeast cell and two input data: the theoretical specific growth rate and initial cell biomass. In experimental assay, a commercial baker's yeast strain and molasses as sugar source were used. Experimental results showed an overall biomass yield on substrate of 0.33, a biomass increase of 6.4 fold and a specific growth rate of 0.165 h-1 in contrast to the predicted value of 0.180 h-1 in the second stage simulation.

Tratamento de compostos orgânicos odoríferos tóxicos por biorreatores / Treatment of odorous toxic organic compounds by bioreactor

Introdução É importante o desenvolvimento de sistemas de controle de poluição do ar que sejam eficientes, além de aplicáveis à condição nacional e pra proteção da saúde humana, uma vez que os compostos do grupo BTEX são tóxicos. Objetivo - Avaliar o desempenho de sistema de tratamento biológico para vapores de BTEX e investigar as melhores condições de operação para os critérios de projeto adotados. Métodos. Trata-se de trabalho experimental com utilização de unidade piloto constituída de coluna de vidro (diâmetro interno de 80 mm e altura total de 1,2 m) tendo no seu interior um meio filtrante composto vegetal e anéis de Pall - que serviram de suporte para os microrganismos e onde se realizou a biodegradação. Foram monitorados parâmetros como temperatura, perda de carga, vazão, concentração dos gases na entrada e na saída, que constituíram a base para desenvolver intervenções e melhorar seu desempenho. A análise dos gases foi feita por fotoionização (PID) em aparelho portátil. Conclusões - Conclui-se que é viável o tratamento biológico para remoção do BTEX de efluentes gasosos, nas condições operacionais adotadas, com eficiência máxima de remoção em torno de 90 por cento . A máxima eficiência foi obtida para tempo de retenção de 2,4 min., carga superficial do gás de 11,9 m3/m2xh, carga mássica no leito de 67 g/m3xh e capacidade de eliminação de 4 g/m3xh. O uso de anéis de Pall misturados ao composto evitou que valores elevados de perda de carga. Foi relevante a participação da adsorção. A utilização de composto mostrou-se viável como alternativa para a biodegradação do BTEX, fortalecendo seu uso com essa prática ambiental.