Verónica Leticia Colin

Actinobacteria: Current research and perspectives for bioremediation of pesticides and heavy metals

Actinobacteria exhibit cosmopolitan distribution since their members are widely distributed in aquatic and terrestrial ecosystems. In the environment they play relevant ecological roles including recycling of substances, degradation of complex polymers, and production of bioactive molecules. Biotechnological potential of actinobacteria in the environment was demonstrated by their ability to remove organic and inorganic pollutants. This ability is the reason why actinobacteria have received special attention as candidates for bioremediation, which has gained importance because of the widespread release of contaminants into the environment. Among organic contaminants, pesticides are widely used for pest control, although the negative impact of these chemicals in the environmental balance is increasingly becoming apparent. Similarly, the extensive application of heavy metals in industrial processes lead to highly contaminated areas worldwide. Several studies focused in the use of actinobacteria for cleaning up the environment were performed in the last 15 years. Strategies such as bioaugmentation, biostimulation, cell immobilization, production of biosurfactants, design of defined mixed cultures and the use of plant-microbe systems were developed to enhance the capabilities of actinobacteria in bioremediation. In this review, we compiled and discussed works focused in the study of different bioremediation strategies using actinobacteria and how they contributed to the improvement of the already existing strategies. In addition, we discuss the importance of omic studies to elucidate mechanisms and regulations that bacteria use to cope with pollutant toxicity, since they are still little known in actinobacteria. A brief account of sources and harmful effects of pesticides and heavy metals is also given.

The Role of Synthetic Biology in the Design of Microbial Cell Factories for Biofuel Production

Insecurity in the supply of fossil fuels, volatile fuel prices, and major concerns regarding climate change have sparked renewed interest in the production of fuels from renewable resources. Because of this, the use of biodiesel has grown dramatically during the last few years and is expected to increase even further in the future. Biodiesel production through the use of microbial systems has marked a turning point in the field of biofuels since it is emerging as an attractive alternative to conventional technology. Recent progress in synthetic biology has accelerated the ability to analyze, construct, and/or redesign microbial metabolic pathways with unprecedented precision, in order to permit biofuel production that is amenable to industrial applications. The review presented here focuses specifically on the role of synthetic biology in the design of microbial cell factories for efficient production of biodiesel.

Production and partial characterization of bioemulsifier from a chromium-resistant actinobacteria

Surface-active compounds such as synthetic emulsifiers have been used for several decades, both for the degradation of hydrocarbons and increasing desorption of soil-bound metals. However, due to their high toxicity, low degradability, and production costs unaffordable for use in larger ecosystems, synthetic emulsifiers have been gradually replaced by those derived from natural sources such as plants or microbes. In previous studies, the bacterium Streptomyces sp. MC1 has shown the ability to reduce and/or accumulate Cr(VI), a highly promising advance in the development of methods for environmental clean-up of sites contaminated with chromium. Here, new studies on the production of emulsifier from this strain are presented. The cultivation factors that have a significant influence on emulsifier biosynthesis, as well as the interactions among them, were studied by factorial design. Based upon optimization studies, maximum bioemulsifier production was detected in the culture medium having an initial pH of 8 with phosphate 2.0 g L(-1) and Ca(+2) 1.0 g L(-1) added, with an emulsification index about 3.5 times greater compared to the basal value. Interestingly, in the presence of 5.0 g L(-1) Cr(VI), Streptomyces sp. MC1 retained about 65% of its emulsifier production ability. Partially purified emulsifier presented high thermo-stability and partial water solubility. These findings could have promising future prospects for the remediation of organic- and metal-contaminated sites.

Potential application of a bioemulsifier-producing actinobacterium for treatment of vinasse

Vinasse is a complex effluent created during production of ethyl alcohol, which can present serious pollution hazard in areas where it is discharged. A variety of technologies, many based upon recovery of the effluent via microbial pathways, are continually being evaluated in order to mitigate the pollution potential of vinasse. The present work reports on initial advances related to the effectiveness of the actinobacterium Streptomyces sp. MC1 for vinasse treatment. Alternative use of raw vinasse as a substrate for producing metabolites of biotechnological interest such as bioemulsifiers, was also evaluated. The strain was able to grow at very high vinasse concentrations (until 50% v/v) and remove over 50% of the biodegradable organic matter in a time period as short as 4 d. Potentially toxic metals such as Mn, Fe, Zn, As, and Pb were also effectively removed during bacterial growth. Decrease in the pollution potential of treated vinasse compared to raw effluent, was reflected in a significant increase in the vigour index of Lactuca sativa (letucce) used as bioremediation indicator. Finally, significant bioemulsifier production was detected when this strain was incubated in a vinasse-based culture medium. These results represent the first advances on the recovery and re-valuation of an actual effluent, by using an actinobacterium from our collection of cultures.

Production of bioemulsifiers by Amycolatopsis tucumanensis DSM 45259 and their potential application in remediation technologies for soils contaminated with hexavalent chromium.

In recent years, increasing interest has been shown in the use of bioemulsifiers as washing agents that can enhance desorption of soil-bound metals. However, high production costs derived from the use of expensive substrates for formulation of the fermentation media represent the main challenge for full, large-scale implementation of bioemulsifiers. This work reports on a first study of bioemulsifier production by the actinobacterium Amycolatopsis tucumanensis DSM 45259 using different carbon and nitrogen sources. Preliminary results on the potential use of these compounds as washing agents for soils contaminated with Cu(II) and Cr(VI) are also presented. The best specific production was detected using glycerol and urea as carbon and nitrogen substrates, respectively. However, with all of the substrates used during the batch assay, the bioemulsifiers showed high levels of stability at extreme conditions of pH, temperature, and salt concentration. Under the current assay conditions, the bioemulsifiers were not effective in removing Cu(II) from soil. However, they were able to mediate Cr(VI) recovery, with the removal percentage doubled compared to that seen when using deionized water. These findings appear promising for the development of remediation technologies for hexavalent chromium compounds based upon direct use of these microbial emulsifiers.

Effect of environmental conditions on extracellular lipases production and fungal morphology from Aspergillus niger MYA 135

Under the current assay conditions, lipase production in mineral medium was only detected in the presence of vegetable oils, reaching the highest specific activity with olive oil. In this way, effect of different environmental conditions on fungal morphology and olive oil‐induced extracellular lipases production from Aspergillus niger MYA 135 was studied. It was observed that addition of 1.0 g l–1 FeCl3to the medium encouraged filamentous growth and increased the specific activity 6.6 fold after 4 days of incubation compared to the control. However, major novelty of this study was the satisfactory production of an acidic lipase at initial pH 3 of the culture medium (1.74 ± 0.06 mU μg–1), since its potencial applications in food and pharmaceutical industry are highly promising. (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)

Mycelium-bound lipase production from Aspergillus niger MYA 135, and its potential applications for the transesterification of ethanol

The potential biotecnological applications of both constitutive and inducible lipase sources from Aspergillus niger MYA 135 were evaluated. To this end, the effect of environmental conditions on mycelium‐bound lipase production from this strain was studied, when cultured either in the absence or presence of 2% olive oil. It was previously reported that mycelium‐bound lipase from Aspergillus niger MYA 135 possess high stability in reaction mixtures containing ethanol; which could be especially important for their use in biodiesel synthesis. In this connection, the performance of the lipase sources produced in the transesterification of ethanol using p‐nitrophenyl palmitate as acyl donor was also explored.

Tailoring fungal morphology of Aspergillus niger MYA 135 by altering the hyphal morphology and the conidia adhesion capacity: biotechnological applications

Current problems of filamentous fungi fermentations and their further successful developments as microbial cell factories are dependent on control fungal morphology. In this connection, this work explored new experimental procedures in order to quantitatively check the potential of some culture conditions to induce a determined fungal morphology by altering both hyphal morphology and conidia adhesion capacity. The capacity of environmental conditions to modify hyphal morphology was evaluated by examining the influence of some culture conditions on the cell wall lytic potential of Aspergillus niger MYA 135. The relative value of the cell wall lytic potential was determined by measuring a cell wall lytic enzyme activity such as the mycelium-bound β-N-acetyl-D-glucosaminidase (Mb-NAGase). On the other hand, the quantitative value of conidia adhesion was considered as an index of its aggregation capacity. Concerning microscopic morphology, a highly negative correlation between the hyphal growth unit length (lHGU) and the specific Mb-NAGase activity was found (r = -0.915, P < 0.001). In fact, the environment was able to induce highly branched mycelia only under those culture conditions compatible with specific Mb-NAGase values equal to or higher than 190 U gdry.wt-1. Concerning macroscopic morphology, a low conidia adhesion capacity was followed by a dispersed mycelial growth. In fact, this study showed that conidia adhesion units per ml equal to or higher than 0.50 were necessary to afford pellets formation. In addition, it was also observed that once the pellet was formed the lHGU had an important influence on its final diameter. Finally, the biotechnological significance of such results was discussed as well.

Selection of an actinobacteria mixed culture for chlordane remediation. Pesticide effects on microbial morphology and bioemulsifier production

Chlordane bioremediation using actinobacteria mixed culture is an attractive clean‐up technique. Their ability to produce bioemulsifiers could increase the bioavailability of this pesticide. In order to select a defined actinobacteria mixed culture for chlordane remediation, compatibility assays were performed among six Streptomyces strains. The strains did not show growth inhibition, and they were assayed for chlordane removal, either as pure or as mixed cultures. In pure cultures, all of the strains showed specific dechlorination activity (1.42–24.20 EU mg−1) and chlordane removal abilities (91.3–95.5%). The specific dechlorination activity was mainly improved with cultures of three or four microorganisms. The mixed culture consisting of Streptomyces sp. A2‐A5‐A13 was selected. Their ability to produce bioemulsifiers in the presence of glucose or chlordane was tested, but no significant differences were observed (p > 0.05). However, the stability of the emulsions formed was linked to the carbon source used. Only in chlordane presence the emulsions retained 100% of their initial height. Finally, the selected consortium showed a high degree of sporulation in the pesticide presence. This is the first study on the effects that chlordane exerts on microbe morphology and emulsifier production for a defined mixed culture of Streptomyces with ability to remediate the pesticide.

Surface-Active Compounds of Microbial Origin and Their Potential Application in Technologies of Environmental Remediation

The world is now confronted with serious problems of environmental contamination. Industrial and mining activities represent the main sources of heavy metal contamination, which provide unique challenges for their remediation, as they cannot be degraded into innocuous products. On the other hand, extensive production and use of hydrocarbons and pesticides in diverse fields has resulted in widespread environmental contamination by these chemicals. A variety of remediation technologies that include physicochemical methods are available to address contamination with organic and inorganic pollutants. However, these technologies have high cost and the risk of secondary environmental pollution. The microbial remediation includes the use of certain microorganisms or products derived to them to address the problems of environmental pollution. In this connection, the synthesis and release of exopolymers with surfactants activity is one of the strategies of the first microbial cell defense line against diverse toxics. These compounds can be successfully used to release hydrocarbons and other pollutants of low solubility from soils matrix. There are also reports regarding the release of heavy metals from soils and sediments using compounds of biological origin as surfactants microbial. This chapter considers the current advances on this topic, emphasizing on the properties and potential applications of microbial surfactants in environmental remediation technologies.