O acido indolacético (IAA) é uma auxina natural e um dos principais reguladores do crescimento de plantas superiores. A quantidade de ácido indolacético extracelular produzida pela cultura de microalga verde Scenedesmus obliquus foi estabelecida por cromatografia líquida de alta eficiência (HPLC). Avaliou-se o efeito de três variáveis de processo na produção de IAA extracelular nestas culturas usando planejamento fatorial 2 3 para uma metodologia de screening. As variáveis de planejamento experimental foram agitação, iluminância (intensidade do fluxo luminoso) e fonte de carvão (mistura CO 2 /air v/v (%)) em três diferentes níveis. A concentração de IAA e a taxa de crescimento específico foram usadas como variáveis de resposta. As análises de HPLC mostraram que em condições experimentais, a quantidade de IAA que foi liberada no meio pela alga S. obliquus depende somente da variável iluminância. Os efeitos da luz e as concentrações de carvão no meio de cultura foram significantes (p < 0.05) para uma taxa de crescimento específico.The indoleacetic acid (IAA) is a natural auxin and one of the main growth regulators in higher plants. The extracellular indoleacetic acid (IAA) amount produced by the green microalga Scenedesmus obliquus culture was detected by high performance liquid chromatography (HPLC). It was evaluated the effect of three process variables in the extracellular IAA production in these cultures using 2 3 factorial design for screening methodology. The experimental design variables were agitation, illuminance (intensity of luminous flux) and carbon source (CO 2 /air v/v (%) mixture) at three different levels. The IAA concentration and the specific growth rate were used as response variables. The HPLC analyses showed that under experimental conditions, the IAA amount that is released to the medium by S. obliquus depends only on the illuminance variable. The light effects and the carbon concentrations in culture medium were significant (p < 0.05) for specific growth rate.

This study aimed to express heterologously the lipase LipA from Pseudomonas aeruginosa PSA01 obtained from palm fruit residues. In previous approaches, LipA was expressed in Escherichia coli fused with its signal peptide and without its disulfide bond, displaying low activity. We cloned the mature LipA with its truncated chaperone Lif in a dual plasmid and overexpressed the enzyme in two E. coli strains: the traditional BL21 (DE3) and the SHuffle® strain, engineered to produce stable cytoplasmic disulfide bonds. We evaluated the effect of the disulfide bond on LipA stability using molecular dynamics. We expressed LipA successfully under isopropyl β-d-1-thio-galactopyranoside (IPTG) and slow autoinducing conditions. The SHuffle LipA showed higher residual activity at 45 °C and a greater hyperactivation after incubation with ethanol than the enzyme produced by E. coli BL21 (DE3). Conversely, the latter was slightly more stable in methanol 50% and 60% (t½: 49.5 min and 9 min) than the SHuffle LipA (t½: 31.5 min and 7.4 min). The molecular dynamics simulations showed that removing the disulfide bond caused some regions of LipA to become less flexible and some others to become more flexible, significantly affecting the closing lid and partially exposing the active site at all times.

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