Producción de plásticos biodegradables a partir de bacterias de hábitats salinos aisladas de la Laguna de Ayarza / Production of biodegradable plastics from saline habitats bacteria isolated from Laguna de Ayarza

La contaminación por plásticos petroquímicos es una grave amenaza para el medio ambiente que requiere im-plementar alternativas como los bioplásticos para lograr un desarrollo sostenible. Los polihidroxialcanoatos (PHA) son polímeros utilizados para la producción de plásticos biodegradables y que han llamado la atención como sustitutos de los plásticos de base fósil. Sin embargo, el costo de producción de los PHA constituye una barrera para su producción industrial a gran escala. Las de bacterias de hábitats salinos son microorganismos prometedores para la síntesis de PHA debido a sus características tales como altos requisitos de salinidad que previenen la contaminación microbiana, la alta presión osmótica intracelular que permite una fácil lisis celular para purificar los PHA y la capacidad para usar un amplio espectro de sustratos. La presente investigación planteó determinar las cepas nativas de bacterias halófilas y halotolerantes de la Laguna de Ayarza capaces de producir PHA, establecer la capacidad que tienen de utilizar residuos agrícolas para la producción de PHA y determinar su eficiencia. Esto se logró a través de la inoculación de las cepas productoras de PHA en medios de fermentación con pulpa de café, cáscaras de plátanos y salvado de trigo lo que permitió determinar las cepas más eficientes. Se encontró que las bacterias productoras de PHA pertenecen a las especies: Alcaligenes faecalis, Bacillus idriensis, Bacillus megaterium, Exiguobacterium acetylicum, E. aurantiacum, Pseudomonas cuatrocienegasensis y Sta-phylococcus capitis y que las cepas AP21-14, AP21-10 y AP21-03 mostraron los mejores resultados que podrían ser prometedores para la producción a nivel industrial.

Pollution by petrochemical plastics is a serious threat to the environment that requires the implementation of al-ternatives such as bioplastics to achieve sustainable development. Polyhydroxyalkanoates (PHAs) are polymers used for the production of biodegradable plastics and have drawn attention as substitutes for fossil-based plastics. However, the cost of producing PHAs constitutes a barrier to their large-scale industrial production. Bacteria from saline environments bacteria are promising microorganisms for PHA synthesis due to their characteristics such as high salinity requirements that prevent microbial contamination, high intracellular osmotic pressure that allows easy cell lysis to purify PHAs, and the ability to use a broad spectrum of substrates. This research project aimed to determine the native strains of halophilic and halotolerant bacteria from Laguna de Ayarza capable of producing PHA, establish their ability to use agricultural residues for the production of PHA, and determine their efficiency. This was achieved through the inoculation of the PHA-producing strains in fermentation media with coffee pulp, banana peels and wheat bran, which allowed determining the most efficient strains. It was found that the PHA-producing bacteria belong to the species: Alcaligenes faecalis, Bacillus idriensis, Bacillus mega-terium, Exiguobacterium acetylicum, E. aurantiacum, Pseudomonas cuatrocienegasensis and Staphylococcus capitis and that the strains AP21-14, AP21-10 and AP21-03 showed the best results that could be promising for production at an industrial level.

Agro-industrial residues and starch for growth and co-production of Polyhydroxyalkanoate copolymer and alfa-amylase by Bacillus. SP CFR67îen

Polyhydroxyalkanoates (PHA) and α-amylase (α-1,4 glucan-4-glucanohydrolase, E.C. 3.2.1.1) were co-produced by Bacillus sp. CFR-67 using unhydrolysed corn starch as a substrate. Bacterial growth and polymer production were enhanced with the supplementation of hydrolysates of wheat bran (WBH) or rice bran (RBH) individually or in combination (5-20 g L-1, based on weight of soluble substrates-SS). In batch cultivation, a mixture of WBH and RBH (1:1, 10 g L-1 of SS) along with ammonium acetate (1.75 g L-1) and corn starch (30 g L-1) produced maximum quantity of biomass (10 g L-1) and PHA (5.9 g L-1). The polymer thus produced was a copolymer of polyhydroxybutyrate-co-hydroxyvalerate of 95:5 to 90:10 mol%. Presence of WBH and corn starch (10-50 g L-1) in the medium enhanced fermentative yield of α-amylase (2-40 U mL-1 min-1). The enzyme was active in a wide range of pH (4-9) and temperature (40-60ºC). This is the first report on simultaneous production of copolymer of bacterial PHA and α-amylase from unhydrolysed corn starch and agro-industrial residues as substrates.

New PHA products using unrelated carbon sources

Polyhydroxyalkanoates (PHA) are natural polyesters stored by a wide range of bacteria as carbon source reserve. Due to its chemical characteristics and biodegradability PHA can be used in chemical, medical and pharmaceutical industry for many human purposes. Over the past years, few Burkholderia species have become known for production of PHA. Aside from that, these bacteria seem to be interesting for discovering new PHA compositions which is important to different industrial applications. In this paper, we introduce two new strains which belong either to Burkholderia cepacia complex (Bcc) or genomovar-type, Burkholderia cepacia SA3J and Burkholderia contaminans I29B, both PHA producers from unrelated carbon sources. The classification was based on 16S rDNA and recA partial sequence genes and cell wall fatty acids composition. These two strains were capable to produce different types of PHA monomers or precursors. Unrelated carbon sources were used for growth and PHA accumulation. The amount of carbon source evaluated, or mixtures of them, was increased with every new experiment until it reaches eighteen carbon sources. As first bioprospection experiments staining methods were used with colony fluorescent dye Nile Red and the cell fluorescent dye Nile Blue A. Gas chromatography analysis coupled to mass spectrometry was used to evaluate the PHA composition on each strain cultivated on different carbon sources. The synthesized polymers were composed by short chain length-PHA (scl-PHA), especially polyhydroxybutyrate, and medium chain length-PHA (mcl-PHA) depending on the carbon source used.

Production and characterization of PHA from recombinant E. coli harbouring phaC1 gene of indigenous Pseudomonas sp. LDC-5 using molasses

Polyhydroxyalkanoates (PHA) are biodegradable and biocompatible green thermoplastics, synthesized by wide variety of bacteria as an intracellular carbon and energy storage intermediate. They are used as an alternative to nonrenewable petroleum derived plastics. The current interest in these biopolyesters is stimulated by the search for cost-effective capitalized production. This paper attempts to achieve maximized production rate from recombinant system using inexpensive substrate. Molasses from agro-industrial waste was used to produce PHA from recombinant E.coli in batch culture. PHA yield in molasses (3.06g/L ± 0.05-75.5 percent) was higher than that of sucrose (2.5g/L ± 0.05 - 65.1 percent). Properties of the polymer produced from molasses and sucrose were analyzed by DSC, TGA, DTA, GC/MS, TLC and optical rotation studies. The findings suggested that molasses enhanced PHA production in recombinant E.coli.