Optimal Conditions for Biomass and Recombinant Glycerol Kinase Production Using the Yeast Pichia pastoris

The extracellular glycerol kinase gene from Saccharomyces cerevisiae (GUT]) was cloned into the expression vector pPICZ alpha. A and integrated into the genome of the methylotrophic yeast Pichia pastoris X-33. The presence of the GUT1 insert was confirmed by PCR analysis. Four clones were selected and the functionality of the recombinant enzyme was assayed. Among the tested clones, one exhibited glycerol kinase activity of 0.32 U/mL, with specific activity of 0.025 U/mg of protein. A medium optimized for maximum biomass production by recombinant Pichia pastoris in shaker cultures was initially explored, using 2.31 % (by volume) glycerol as the carbon source. Optimization was carried out by response surface methodology (RSM). In preliminary experiments, following a Plackett-Burman design, glycerol volume fraction (phi(Gly)) and growth time (t) were selected as the most important factors in biomass production. Therefore, subsequent experiments, carried out to optimize biomass production, followed a central composite rotatable design as a function of phi(Gly) and time. Glycerol volume fraction proved to have a significant positive linear effect on biomass production. Also, time was a significant factor (at linear positive and quadratic levels) in biomass production. Experimental data were well fitted by a convex surface representing a second order polynomial model, in which biomass is a function of both factors (R(2)=0.946). Yield and specific activity of glycerol kinase were mainly affected by the additions of glycerol and methanol to the medium. The optimized medium composition for enzyme production was: 1 % yeast extract, 1 % peptone, 100 mM potassium phosphate buffer, pH=6.0, 1.34 % yeast nitrogen base (YNB), 4.10(-5) % biotin, 1 %, methanol and 1 %, glycerol, reaching 0.89 U/mL of glycerol kinase activity and 14.55 g/L of total protein in the medium after 48 h of growth.

Harmonic, reactive and unbalance compensation by means of CPT framework

This paper presents possible selective current compensation strategies based on the Conservative Power Theory (CPT). This recently proposed theory, introduces the concept of complex power conservation under non-sinusoidal conditions. Moreover, the related current decompositions results in several current terms, which are associated with a specific physical phenomena (power absorption P, energy storage Q, voltage and current distortion D). Such current components are used in this work for the definition of different current compensators, which can be selective in terms of minimizing particular disturbing effects. The choice of one or other current component for compensation directly affects the sizing and cost of active and/or passive devices and it will be demonstrated that it can be done to attend predefined limits for harmonic distortion, unbalances and/or power factor. Single and three-phase compensation strategies will be discussed by means of the CPT Framework. Simulation and experimental results will be demonstrated in order to validate their performance. © 2009 IEEE.

Development of a DC-DC converter for DC bus voltage control of series connected device

The purpose of this work is to study voltage control and energy balance of a split DC bus topology within a power electronics equipment connected to the AC mains, such as UPS systems, wind power generators, active filters and FACTS devices. A typical configuration in such equipment has two mains connected converters sharing a common DC bus, one series connected and the other parallel connected. The DC bus is usually composed by a battery or a capacitor bank. In the proposed topology, the DC bus is divided in two sides, interconnected with a buck-boost converter, which controls power flow and DC voltage on both sides. © 2009 IEEE.

Dynamic voltage restorer development and testing

The growing use of sensitive loads in the electric power system, especially in industrial applications, increases voltage sags related production losses considerably, stimulating a demand for power electronics' based solutions to mitigate the effects of such problems. This paper shows the implementation and some industrial certification tests of a power equipment prototype designed to correct sags and swells, a dynamic voltage restorer, which is one of the many possible solutions for voltage sags and swells problems Experimental results of a 75kVA prototype are shown both in laboratory and full load conditions, in a certification institution (IEE-USP). © 2011 IEEE.

Reference generator for voltage controlled power conditioners

Voltage reference generation is an important issue on electronic power conditioners or voltage compensators connected to the electric grid. Several equipments, such as Dynamic Voltage Restorers (DVR), Uninterruptable Power Supplies (UPS) and Unified Power Quality Conditioners (UPQC) need a proper voltage reference to be able to compensate electric network disturbances. This work presents a new reference generator's algorithm, based on vector algebra and digital filtering techniques. It is particularly suited for the development of voltage compensators with energy storage, which would be able to mitigate steady state disturbances, such as waveform distortions and unbalances, and also transient disturbances, like voltage sags and swells. Simulation and experimental results are presented for the validation of the proposed algorithm. © 2011 IEEE.

Shunt active compensation based on the Conservative Power Theory current's decomposition

Considering the operation of shunt active compensators, such as active power filters, this paper proposes possible compensation strategies by means of the recent formulation of the Conservative Power Theory (CPT). The CPT current's decomposition results in several current components, which are associated with specific load characteristics (power transfer, energy storage, unbalances and/or non linearities). These current components are used for the definition of different compensation strategies, which can be selective in terms of minimizing particular disturbing effects. In order to validate the applicability of these new compensation strategies, simulation and experimental results for three-phase four-wire systems are presented. © 2011 IEEE.

Algoritmos para geração de referências, inicialização e proteção de um condicionador unificado de qualidade de energia

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Fatores que influenciam a produção de biomassa e glicerol quinase pela levedura recombinante Pichia pastoris

Pós-graduação em Alimentos e Nutrição - FCFAR

Estudo do potencial genotóxico da Gutiferona A em diferentes células de camundongos in vitro

Pós-graduação em Biologia Geral e Aplicada - IBB

Benzophenone guttiferone A from Garcinia achachairu Rusby (Clusiaceae) Presents Genotoxic Effects in Different Cells of Mice

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

Análise de parâmetros nutricionais e metabólicos de jogadores de futebol suplementados com suco de laranja

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Using Pichia pastoris to produce recombinant glycerol kinase

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