An interlaboratory comparison of physiological and genetic properties of four Saccharomyces cerevisiae strains

To select a Saccharomyces cerevisiae reference strain amenable to experimental techniques used in (molecular) genetic, physiological and biochemical engineering research, a variety of properties were studied in four diploid, prototrophic laboratory strains. The following parameters were investigated: 1) maximum specific growth rate in shake-flask cultures; 2) biomass yields on glucose during growth on defined media in batch cultures and steady-state chemostat cultures under controlled conditions with respect to pH and dissolved oxygen concentration; 3) the critical specific growth rate above which aerobic fermentation becomes apparent in glucose-limited accelerostat cultures; 4) sporulation and mating efficiency; and 5) transformation efficiency via the lithium-acetate, bicine, and electroporation methods. On the basis of physiological as well as genetic properties, strains from the CEN.PK family were selected as a platform for cell-factory research on the stoichiometry and kinetics of growth and product formation.     

A novel mode of bathing behavior of hummingbirds recorded in the Brazilian ruby Heliodoxa rubricauda and allies (Aves: Trochilidae)

acta ethologica - The Brazilian ruby, Heliodoxa rubricauda, is a forest species of hummingbird endemic to the Atlantic Forest. It belongs to an Andean clade of birds with robust and strong legs and...     

Adaptation of acidogenic sludge to increasing glycerol concentrations for biohydrogen production

Hydrogen is a promising alternative as an energetic carrier and its production by dark fermentation from wastewater has been recently proposed, with special attention to crude glycerol as potential substrate. In this study, two different feeding strategies were evaluated for replacing the glucose substrate by glycerol substrate: a one-step strategy (glucose was replaced abruptly by glycerol) and a step-by-step strategy (progressive decrease of glucose concentration and increase of glycerol concentration from 0 to 5 g L^?1), in a continuous stirred tank reactor (12 h of hydraulic retention time (HRT), pH 5.5, 35 °C). While the one-step strategy led to biomass washout and unsuccessful H₂ production, the step-by-step strategy was efficient for biomass adaptation, reaching acceptable hydrogen yields (0.4?±?0.1 mol_H2?mol^?1 _{glycerol consumed}) around 33 % of the theoretical yield independently of the glycerol concentration. Microbial community structure was investigated by single-strand conformation polymorphism (SSCP) and denaturing gradient gel electrophoresis (DGGE) fingerprinting techniques, targeting either the total community (16S ribosomal RNA (rRNA) gene) or the functional Clostridium population involved in H₂ production (hydA gene), as well as by 454 pyrosequencing of the total community. Multivariate analysis of fingerprinting and pyrosequencing results revealed the influence of the feeding strategy on the bacterial community structure and suggested the progressive structural adaptation of the community to increasing glycerol concentrations, through the emergence and selection of specific species, highly correlated to environmental parameters. Particularly, this work highlighted an interesting shift of dominant community members (putatively responsible of hydrogen production in the continuous stirred tank reactor (CSTR)) according to the gradient of glycerol proportion in the feed, from the family Veillonellaceae to the genera Prevotella and Clostridium sp., putatively responsible of hydrogen production in the CSTR.     

The challenge of small and rare species in marine biodiversity surveys: microgastropod diversity in a complex tropical coastal environment

Although molluscs feature prominently in the semi-popular and academic literature on marine biodiversity, field surveys largely ignore the small and rare species that form the majority of marine molluscan diversity. As a result of a massive effort to sample the benthic molluscs of a complex tropical coastal environment, 23,238 gastropod specimens representing 259 species of Triphoridae—a family with most adult species ranging from 2 to 10 mm—were obtained from a 45,000 hectares study area off the island of Espiritu Santo, Vanuatu. Most species are represented by fewer than 20 specimens and, despite the intensity of the sampling effort, 13% of the species are unique singletons. Spatial heterogeneity was high: out of 416 sampling events, 187 contained triphorids, and 42% of the species occurred at fewer than 5 stations. Most species were small (68% below 5 mm) or very small (22% below 3 mm). A faunal turnover was documented at around 10 m, and another at around 60 m, at the onset of the “twilight zone” that is particularly difficult to sample. On the order of 70% of the species are probably new to science. When dealing with taxonomically difficult groups a morphospecies segregation approach is operationally appropriate to detect patterns of richness, rarity and spatial turn-over. Very few, if any, conservation surveys have the human and funding resources to carry out baseline surveys of the intensity that generated the results presented here. However, as species numbers are often used to promote the conservation interest of a reef, a bay or a stretch of coast, it is essential to know how the numbers were generated: absolute numbers of species are meaningless unless sampling effort and techniques, area surveyed, and size classes targeted are described. This is very rarely the case, even in the academic literature.     

Phosphorus retention in lab and field-scale subsurface-flow wetlands treating plant nursery runoff

Constructed wetland systems built to handle nutrient contaminants are often efficient at removing nitrogen, but ineffective at reducing phosphorus (P) loads. Incorporating a clay-based substrate can enhance P removal in subsurface-flow constructed wetland systems. We evaluated the potential of crushed brick, a recycled building product, and two particle sizes of a palygorskite–bentonite industrial mineral aggregate (calcined clay) to sorb P from simulated nutrient-rich plant nursery effluent. The three substrates were screened for P sorbing behavior using sorption, desorption, and equilibration experiments. We selected one substrate to evaluate in an 8-month field trial to compare field sorption capacity with laboratory sorption capacity. In the laboratory, coarse calcined clay average sorption capacity was 497 mg kg⁻¹ and it sorbed the highest percentage of P supplied (76%), except at exposure concentrations >100 mg L⁻¹ where the increased surface area of fine calcined clay augmented its P sorption capacity. Subsurface-flow mesocosms were filled with coarse calcined clay and exposed to a four and seven day hydraulic retention time treatment. Phosphorus export was reduced by 60 to 74% for both treatments until substrate P-binding sites began to saturate during month seven. During the eight month experiment, the four and seven day treatments fixed 1273 ± 22 mg kg⁻¹ P and 937 ± 16 mg kg⁻¹ P, respectively. Sequential extractions of the P saturated clay indicated that P could desorb slowly over time from various pools within the calcined clay; thus, if the calcined clay were recycled as a soil amendment, most P released would be slowly available for plant uptake and use. This study demonstrated the viability of using coarse calcined clay as a root bed substrate in subsurface-flow treatment wetlands remediating phosphorus from plant nursery runoff.     

Isolation and characterization of novel bacterial strains exhibiting ligninolytic potential

Background

The number of biotransformations that use nicotinamide recycling systems is exponentially growing. For this reason one of the current challenges in biocatalysis is to develop and optimize more simple and efficient cofactor recycling systems. One promising approach to regenerate NAD⁺ pools is the use of NADH-oxidases that reduce oxygen to hydrogen peroxide while oxidizing NADH to NAD⁺. This class of enzymes may be applied to asymmetric reduction of prochiral substrates in order to obtain enantiopure compounds.

Results

The NADH-oxidase (NOX) presented here is a flavoenzyme which needs exogenous FAD or FMN to reach its maximum velocity. Interestingly, this enzyme is 6-fold hyperactivated by incubation at high temperatures (80°C) under limiting concentrations of flavin cofactor, a change that remains stable even at low temperatures (37°C). The hyperactivated form presented a high specific activity (37.5 U/mg) at low temperatures despite isolation from a thermophile source. Immobilization of NOX onto agarose activated with glyoxyl groups yielded the most stable enzyme preparation (6-fold more stable than the hyperactivated soluble enzyme). The immobilized derivative was able to be reactivated under physiological conditions after inactivation by high solvent concentrations. The inactivation/reactivation cycle could be repeated at least three times, recovering full NOX activity in all cases after the reactivation step. This immobilized catalyst is presented as a recycling partner for a thermophile alcohol dehydrogenase in order to perform the kinetic resolution secondary alcohols.

Conclusion

We have designed, developed and characterized a heterogeneous and robust biocatalyst which has been used as recycling partner in the kinetic resolution of rac-1-phenylethanol. The high stability along with its capability to be reactivated makes this biocatalyst highly re-useable for cofactor recycling in redox biotransformations.     

Influence of physico-chemical parameters of the aquatic medium on germination of Eichhornia crassipes seeds

The germination of seeds of Eichhornia crassipes in locations distant from the foci of infestation may be a means of dispersal of this invasive plant. Nonetheless, no modern studies have examined the influence of single components on germination, although the influence of temperature, oxygen and redox potential was examined in earlier studies. The effects of pH, conductivity, nitrate, phosphate, potassium, calcium, iron and boron on germination of E. crassipes were determined in seed germination tests. The results showed that external input of nutrients influences germination, since neither pH nor conductivity alone had any effect, but there was a significant increase in germination in growth media containing phosphorus at 3.95 mg·l(-1) (P/L; 95.5%) and boron at 10 mg·l(-1) (B/L; 97.5%); at higher concentrations, the latter was toxic. These findings contribute to knowledge of factors controlling the germination of E. crassipes seeds. Consequently, E. crassipes seeds would find very good conditions for germination in water classified as hypereutrophic, which may play a decisive role in expansion of this plant.