Lactic acid production by new Lactobacillus plantarum LMISM6 grown in molasses: optimization of medium composition

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

The Use of Response Surface Methodology in Optimization of Lactic Acid Production: Focus on Medium Supplementation, Temperature and pH Control

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

Response surface optimization of D(-)-lactic acid production by Lactobacillus SMI8 using corn steep liquor and yeast autolysate as an alternative nitrogen source

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

A systems biology framework for pathway level culture media engineering: pplication to Pichia pastoris cultures

Dissertação para obtenção do Grau de Doutor em Engenharia Química e Bioquímica

Development of process control strategies exploiting knowledge from systems biology: application to MDCK suspension cells

Madine Darby Canine Kidney (MDCK) cell lines have been extensively evaluated for their potential as host cells for influenza vaccine production. Recent studies allowed the cultivation of these cells in a fully defined medium and in suspension. However, reaching high cell densities in animal cell cultures still remains a challenge. To address this shortcoming, a combined methodology allied with knowledge from systems biology was reported to study the impact of the cell environment on the flux distribution. An optimization of the medium composition was proposed for both a batch and a continuous system in order to reach higher cell densities. To obtain insight into the metabolic activity of these cells, a detailed metabolic model previously developed by Wahl A. et. al was used. The experimental data of four cultivations of MDCK suspension cells, grown under different conditions and used in this work came from the Max Planck Institute, Magdeburg, Germany. Classical metabolic flux analysis (MFA) was used to estimate the intracellular flux distribution of each cultivation and then combined with partial least squares (PLS) method to establish a link between the estimated metabolic state and the cell environment. The validation of the MFA model was made and its consistency checked. The resulted PLS model explained almost 70% of the variance present in the flux distribution. The medium optimization for the continuous system and for the batch system resulted in higher biomass growth rates than the ones obtained experimentally, 0.034 h-1 and 0.030 h-1, respectively, thus reducing in almost 10 hours the duplication time. Additionally, the optimal medium obtained for the continuous system almost did not consider pyruvate. Overall the proposed methodology seems to be effective and both proposed medium optimizations seem to be promising to reach high cell densities.

Halocin SH10 production by an extreme haloarchaeon Natrinema sp. BTSH10 isolated from salt pans of South India

Halobacteria, members of the domain Archaea that live under extremely halophilic conditions, are often considered as dependable source for deriving novel enzymes, novel genes, bioactive compounds and other industrially important molecules. Protein antibiotics have potential for application as preserving agents in food industry, leather industry and in control of infectious bacteria. Halocins are proteinaceous antibiotics synthesized and released into the environment by extreme halophiles, a universal characteristic of halophilic bacteria. Herein, we report the production of halocin (SH10) by an extremely halophilic archeon Natrinema sp. BTSH10 isolated from salt pan of Kanyakumari, Tamilnadu, India and optimization of medium for enhanced production of halocin. It was found that the optimal conditions for maximal halocin production were 42 C, pH 8.0, and 104 h of incubation at 200 rpm with 2% (V/V) inoculum concentration in Zobell’s medium containing 3 M NaCl, Galactose, beef extract, and calcium chloride as additional supplements. Results indicated scope for fermentation production of halocin for probable applications using halophilic archeon Natrinema sp. BTSH10

Otimização do meio de cultura para a produção de quitosanase por metarhizium anisopliae em cultivo descontínuo submerso

In this work a Plackett-Burman Design with 8 factors and 12 trials in 2 levels with 3 repetitions at the center point was used in order to investigate the influence of the concentration of chitosan, peptone, yeast extract, NaNO3, K2HPO4, KCl, MgSO4.7H2O and FeSO4 on chitosanase production by Metarhizium anisopliae. Runs were carried out using submerged discontinuous cultivation for enzyme production. The results of the Plackett & Burman Design showed that only two factors, chitosan concentration as well as FeSO4 had influence on chitosanolytic activity, while the increase in concentration of other factors not contributed significantly to the quitosanolítica activity. Cultivation medium optimization for enzyme production was carried out using a Composite Central Design, with the most important factors for chitosanolytic activity (chitosan and FeSO4), in accordance with Plackett & Burman Design, and keeping the other nutrients in their minimum values. On this other design, it was taken the highest limit in Plackett & Burman Design as the lowest limit (-1) to FeSO4 factor. The results showed that the enzyme production was favoured by increasing the chitosan concentration and by decreasing FeSO4. Maximum production for chitosanolytic activity was about 70.0 U/L and was reached in only 18 h of fermentation, a result about twenty-eight times greater than a former study using the same microorganism (about 2.5 U/L at 48 h)

d(-)-Lactic Acid Production by Leuconostoc mesenteroides B512 Using Different Carbon and Nitrogen Sources

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

In vitro evaluation of antimicrobial activity of sealers and pastes used in endodontics

The antimicrobial activity of four root canal sealers (AH Plus, Sealapex, Ketac Endo, and Fill Canal), two calcium hydroxide pastes (Calen and Calasept), and a zinc oxide paste was evaluated. Seven bacterial strains were used, six of them standard; Micrococcus luteus ATCC 9341, Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 27853, Staphylococcus epidermidis ATCC 12228, Escherichia coli ATCC 25922, and Enterococcus faecalis ATCC 10541. There was a wild strain of Streptococcus mutans isolated from saliva obtained in an adult dental clinic. Activity was evaluated using the agar diffusion method with Brain Heart Infusion agar and Müller Hinton medium seeded by pour plate. Calcium hydroxide-based sealers and pastes were either placed directly into 4.0 × 4.0 mm wells or by using absorbent paper points. The plates were kept at room temperature for 2 hr for diffusion. After incubation at 37°C for 24 hr, the medium was optimized with 0.05 g% TTC gel and inhibition haloes were measured. All bacterial strains were inhibited by all materials using the well method. However, when the materials were applied with absorbent paper points, Enterococcus faecalis was not inhibited by zinc oxide, and Pseudomonas aeruginosa was not inhibited by AH Plus, Fill Canal, and the zinc oxide-based paste. We conclude that sealers and pastes presented antimicrobial activity in vitro and culture medium optimization with 0.05 g% TTC gel facilitated observation of the inhibition haloes. Copyright © 2000 by The American Association of Endodontists.

Optimization model for medium term natural gas commitment

In this paper we study the optimal natural gas commitment for a known demand scenario. This study implies the best location of GSUs to supply all demands and the optimal allocation from sources to gas loads, through an appropriate transportation mode, in order to minimize total system costs. Our emphasis is on the formulation and use of a suitable optimization model, reflecting real-world operations and the constraints of natural gas systems. The mathematical model is based on a Lagrangean heuristic, using the Lagrangean relaxation, an efficient approach to solve the problem. Computational results are presented for Iberian and American natural gas systems, geographically organized in 65 and 88 load nodes, respectively. The location model results, supported by the computational application GasView, show the optimal location and allocation solution, system total costs and suggest a suitable gas transportation mode, presented in both numerical and graphic supports.

Cooling system optimization in medium-speed engines

Tässä työssä optimoidaan keskinopean Wärtsilä 32 -dieselmoottorin jäähdytysjärjestelmää ja tutkitaan taajuusmuuttajien käyttömahdollisuutta kiertopumppujen yhteydessä niin, että järjestelmässä saataisiin kiertämään vain kulloinkin tarvittava määrä vettä. Tutkimuksen mallinnus on toteutettu laatimalla aiemmin käytössä olleista yksinkertaisista simulointimalleista yksi malli, johon on sisällytetty sekä virtauksen että lämmönsiirron laskenta, jotka on aiemmin mallinnettu erillisillä ohjelmilla. Diplomityö on osa projektia, joka on tehty Sähkötekniikan osaston tutkijan Mikko Pääkkösen kanssa yhteistyössä. Tämän diplomityö keskittyy lähinnä virtausteknisiin ja lämmönsiirtoon liittyviin asioihin, kun taas sähkötekniikan osuus on esitetty Mikko Pääkkösen raportissa. Tulosten perustella voidaan sanoa, että taajuusmuuttajakäyttö kannattaa kiertopumppujen yhteydessä. Käyttämällä pumppujen virtaussäätöä voidaan jäähdytysjärjestelmästä jättää monia komponentteja, kuten termostaattiventtiilejä pois. Mallinnetut yksinkertaiset piiriratkaisut näyttävät toimivan ainakin yleisellä tasolla. Tutkimusta pumppujen säädöstä ja tässä projektissa luoduista jäähdytysjärjestelmäkonfiguraatioista kannattaa jatkaa.

Optimization of medium for the production of a novel aquaculture probiotic, Micrococcus MCCB 104 using central composite design

A marine isolate of jáÅêçÅçÅÅìë MCCB 104 has been identified as an aquaculture probiotic antagonistic to sáÄêáç. In the present study different carbon and nitrogen sources and growth factors in a mineral base medium were optimized for enhanced biomass production and antagonistic activity against the target pathogen, sáÄêáç=Ü~êîÉóá, following response surface methodology (RSM). Accordingly the minimum and maximum limits of the selected variables were determined and a set of fifty experiments programmed employing central composite design (CCD) of RSM for the final optimization. The response surface plots of biomass showed similar pattern with that of antagonistic activity, which indicated a strong correlation between the biomass and antagonism. The optimum concentration of the carbon sources, nitrogen sources, and growth factors for both biomass and antagonistic activity were glucose (17.4 g/L), lactose (17 g/L), sodium chloride (16.9 g/L), ammonium chloride (3.3 g/L), and mineral salts solution (18.3 mL/L). © KSBB

Antibiosis associated to Lactobacillus acidophilus growth .2. Optimization of some culture parameters in whey and soy milk medium

Whey supplemented with soy milk has been used as a low-cost alternative in the growth of Lactobacillus acidophilus for the production of antimicrobial compounds. Response Surface Metodology has been employed in order to study the effects of initial pH, incubation temperature and soy milk rate for supplementation in the production of antimicrobial substance. It has been observed that both tested microrganisms used (S. aureus and E. coli) were inhibited by antimicrobial substance produced by L. acidophilus. The results obtained with E. coli inhibition did not follow the employed statistical model. on the other hand, when the tested microorganism S. aureus was used, the best inhibition results have been obtained when L. acidophilus was incubated at 36.80 degrees C in whey with 5.6 initial pH and 31,90% (v/v) rate supplemented with soy milk. The analysed antimicrobial substances were nor acids neither hidrogen peroxid.

Thermoplastic polyesters and Co-polyesters derived from vegetable oil: synthesis and optimization of melt polycondensation for medium and long chain poly(omega-hydroxyfatty acid)s and their ester derivatives

The synthesis of a series of omega-hydroxyfatty acid (omega-OHFA) monomers and their methyl ester derivatives (Me-omega-OHFA) from mono-unsaturated fatty acids and alcohols via ozonolysis-reduction/crossmetathesis reactions is described. Melt polycondensation of the monomers yielded thermoplastic poly(omega-hydroxyfatty acid)s [-(CH2)(n)-COO-](x) with medium (n = 8 and 12) and long (n = 17) repeating monomer units. The omega-OHFAs and Me-omega-OHFAs were all obtained in good yield (>= 80%) and purity (>= 97%) as established by H-1 NMR, Fourier Transform infra-red spectroscopy (FT-IR), mass spectroscopy (ESI-MS) and high performance liquid chromatography (HPLC) analyses. The average molecular size (M-n) and distribution (PDI) of the poly(omega-hydroxyfatty acid)s (P(omega-OHFA)s) and poly(omega-hydroxyfatty ester) s (P(Me-omega-OHFA) s) as determined by GPC varied with organo-metallic Ti(IV) isopropoxide [Ti(OiPr)(4)] polycondensation catalyst amount, reaction time and temperature. An optimization of the polymerization process provided P(omega-OHFA) s and P(Me-omega-OHFA) s with M-n and PDI values desirable for high end applications. Co-polymerization of the long chain (n = 12) and medium chain (n = 8) Me-omega-OHFAs by melt polycondensation yielded poly(omega-hydroxy tridecanoate/omega-hydroxy nonanoate) random co-polyesters (M-n = 11000- 18500 g mol(-1)) with varying molar compositions.