Fermentation Methods for Protein Enrichment of Cassava and Corn with Candida tropicalis

Candida tropicalis grows on soluble starch, corn, and cassava powders without requiring that these substrates be previously hydrolyzed. C. tropicalis possesses the enzyme needed to hydrolyze starch, namely, an α-amylase. That property has been used to develop a fermentation process whereby C. tropicalis can be grown directly on corn or cassava powders so that the resultant mixture of biomass and residual corn or cassava contains about 20% protein, which represents a balanced diet for either animal fodder or human food. The fact that no extra enzymes are required to hydrolyze starch results in a particularly efficient way of improving the nutritional value of amylaceous products, through a single-step fermentation process.     

Life cycle assessment of soybean-based biodiesel in Argentina for export

Background, aim and scope

Regional specificities are a key factor when analyzing the environmental impact of a biofuel pathway through a life cycle assessment (LCA). Due to different energy mixes, transport distances, agricultural practices and land use changes, results can significantly vary from one country to another. The Republic of Argentina is the first exporter of soybean oil and meal and the third largest soybean producer in the world, and therefore, soybean-based biodiesel production is expected to significantly increase in the near future, mostly for exportation. Moreover, Argentinean biodiesel producers will need to evaluate the environmental performances of their product in order to comply with sustainability criteria being developed. However, because of regional specificities, the environmental performances of this biofuel pathway can be expected to be different from those obtained for other countries and feedstocks previously studied. This work aims at analyzing the environmental impact of soybean-based biodiesel production in Argentina for export. The relevant impact categories account for the primary non-renewable energy consumption (CED), the global warming potential (GWP), the eutrophication potential (EP), the acidification potential (AP), the terrestrial ecotoxicity (TE), the aquatic ecotoxicity (AE), the human toxicity (HT) and land use competition (LU). The paper tackles the feedstock and country specificities in biodiesel production by comparing the results of soybean-based biodiesel in Argentina with other reference cases. Emphasis is put on explaining the factors that contribute most to the final results and the regional specificities that lead to different results for each biodiesel pathway.

Materials and methods

The Argentinean (AR) biodiesel pathway was modelled through an LCA and was compared with reference cases available in the ecoinvent® 2.01 database, namely, soybean-based biodiesel production in Brazil (BR) and the United States (US), rapeseed-based biodiesel production in the European Union (EU) and Switzerland (CH) and palm-oil-based biodiesel production in Malaysia (MY). In all cases, the systems were modelled from feedstock production to biodiesel use as B100 in a 28 t truck in CH. Furthermore, biodiesel pathways were compared with fossil low-sulphur diesel produced and used in CH. The LCA was performed according to the ISO standards. The life cycle inventory and the life cycle impact assessment (LCIA) were performed in Excel spreadsheets using the ecoinvent® 2.01 database. The cumulative energy demand (CED) and the GWP were estimated through the CED for fossil and nuclear energy and the IPCC 2001 (climate change) LCIA methods, respectively. Other impact categories were assessed according to CML 2001, as implemented in ecoinvent. As the product is a fuel for transportation (service), the system was defined for one vehicle kilometre (functional unit) and was divided into seven unit processes, namely, agricultural phase, soybean oil extraction and refining, transesterification, transport to port, transport to the destination country border, distribution and utilisation.

Results

The Argentinean pathway results in the highest GWP, CED, AE and HT compared with the reference biofuel pathways. Compared with the fossil reference, all impact categories are higher for the AR case, except for the CED. The most significant factor that contributes to the environmental impact in the Argentinean case varies depending on the evaluated category. Land provision through deforestation for soybean cultivation is the most impacting factor of the AR biodiesel pathway for the GWP, the CED and the HT categories. Whilst nitrogen oxide emissions during the fuel use are the main cause of acidification, nitrate leaching during soybean cultivation is the main factor of eutrophication. LU is almost totally affected by arable land occupation for soybean cultivation. Cypermethrin used as pesticide in feedstock production accounts for almost the total impact on TE and AE.

Discussion

The sensitivity analysis shows that an increase of 10% in the soybean yield, whilst keeping the same inputs, will reduce the total impact of the system. Avoiding deforestation is the main challenge to improve the environmental performances of soybean-based biodiesel production in AR. If the soybean expansion can be done on marginal and set-aside agricultural land, the negative impact of the system will be significantly reduced. Further implementation of crops’ successions, soybean inoculation, reduced tillage and less toxic pesticides will also improve the environmental performances. Using ethanol as alcohol in the transesterification process could significantly improve the energy balance of the Argentinean pathway.

Conclusions

The main explaining factors depend on regional specificities of the system that lead to different results from those obtained in the reference cases. Significantly different results can be obtained depending on the level of detail of the input data, the use of punctual or average data and the assumptions made to build up the LCA inventory. Further improvement of the AR biodiesel pathways should be done in order to comply with international sustainability criteria on biofuel production.

Recommendations and perspectives

Due to the influence of land use changes in the final results, more efforts should be made to account for land use changes others than deforestation. More data are needed to determine the part of deforestation attributable to soybean cultivation. More efforts should be done to improve modelling of interaction between variables and previous crops in the agricultural phase, future transesterification technologies and market prices evolution. In order to assess more accurately the environmental impact of soybean-based biodiesel production in Argentina, further considerations should be made to account for indirect land use changes, domestic biodiesel consumption and exportation to other regions, production scale and regional georeferenced differentiation of production systems.     

Growth and Pigment Production by Hairy Root Cultures of Beta Vulgaris L. in a Bubble Column Reactor

Beet hairy root cultures established from red and yellow varieties were grown in a 2 L bubble column reactor. The yellow clone showed profuse root hairs and a predominance of betaxanthin pigment with the red clone showed fewer root hairs and both betaxanthin and betacyanin pigments. The cultures displayed different ionic and sugar yields: 2.1 mg dry wt / mS.mL and 0.361 g dry wt / g sugar for the yellow clone and 2.3 mg dry wt / mS.mL and 0.375 g dry wt / g sugar for the red one. Both cultures grew at the same specific growth rate of 0.22 d^-1in the bubble column, as compared to 0.32 d^-1in shake flasks, indicating mass transfer limitations for growth in reactors.     

Nature of the interactions involved in the lipid-protein complexes of the Escherichia coli N-acetylmuramoyl-L-alanine amidase

Depending on its concentration, phosphatidylglycerol, one of the three main Escherichia coli phospholipid species, is able to activate or inactivate the E. coli murein amidase (N-acetylmuramoyl-L-alanine amidase, EC 3.5.1.28) (Vanderwinkel, E. and De Vlieghere, M. (1985) Biochim. Biophys. Acta 838, 54-59). The mechanisms underlying the modulation of this enzyme activity were studied by analyzing the effects of cations, polycationic molecules, various surfactants and amphiphilic water-soluble compounds. K+, Mg2+ and polyamines were all able to prevent completely the enzyme inactivation produced by millimolar order concentration of phosphatidylglycerol. The efficiencies of the ionic species tested were in the order K+ less than Mg2+ = putrescine less than spermidine less than spermine. The kinetics of the counteraction processes were all sigmoidal. By contrast, the activation of the murein amidase produced by phosphatidylglycerol in micromolar concentration appeared to be insensitive to the ionic strength of the medium. Surfactants and amphiphilic molecules differing in their polar head and hydrophobic tail were found to activate the enzyme at various degrees for concentrations below their critical micellar concentration. The non-ionic surfactants were the most potent activators and remarkably mimicked the phosphatidylglycerol activation. The enzyme activation process appeared to require only a hydrophobic solvation shell around the protein. All kinetic data supported our previous interpretation of the phosphatidylglycerol-enzyme interactions in terms of multisite non-allosteric theory.     

Genome-Wide Association Study of Metabolic Traits Reveals Novel Gene-Metabolite-Disease Links

Metabolic traits are molecular phenotypes that can drive clinical phenotypes and may predict disease progression. Here, we report results from a metabolome- and genome-wide association study on ¹H-NMR urine metabolic profiles. The study was conducted within an untargeted approach, employing a novel method for compound identification. From our discovery cohort of 835 Caucasian individuals who participated in the CoLaus study, we identified 139 suggestively significant (P<5×10⁻⁸) and independent associations between single nucleotide polymorphisms (SNP) and metabolome features. Fifty-six of these associations replicated in the TasteSensomics cohort, comprising 601 individuals from São Paulo of vastly diverse ethnic background. They correspond to eleven gene-metabolite associations, six of which had been previously identified in the urine metabolome and three in the serum metabolome. Our key novel findings are the associations of two SNPs with NMR spectral signatures pointing to fucose (rs492602, P = 6.9×10⁻⁴⁴) and lysine (rs8101881, P = 1.2×10⁻³³), respectively. Fine-mapping of the first locus pinpointed the FUT2 gene, which encodes a fucosyltransferase enzyme and has previously been associated with Crohn''s disease. This implicates fucose as a potential prognostic disease marker, for which there is already published evidence from a mouse model. The second SNP lies within the SLC7A9 gene, rare mutations of which have been linked to severe kidney damage. The replication of previous associations and our new discoveries demonstrate the potential of untargeted metabolomics GWAS to robustly identify molecular disease markers.