Construction of a Stable α-Galactosidase-Producing Baker's Yeast Strain

Molasses is widely used as a substrate for commercial yeast production. The complete hydrolysis of raffinose, which is present in beet molasses, by Saccharomyces strains requires the secretion of α-galactosidase, in addition to the secretion of invertase. Raffinose is not completely utilized by commercially available yeast strains used for baking, which are Mel⁻. In this study we integrated the yeast MEL1 gene, which codes for α-galactosidase, into a commercial mel⁰ baker's yeast strain. The Mel⁺ phenotype of the new strain was stable. The MEL1 gene was expressed when the new Mel⁺ baker's yeast was grown in molasses medium under conditions similar to those used for baker's yeast production at commercial factories. The α-galactosidase produced by this novel baker's yeast strain hydrolyzed all the melibiose that normally accumulates in the growth medium. As a consequence, additional carbohydrate was available to the yeasts for growth. The new strain also produced considerably more α-galactosidase than did a wild-type Mel⁺ strain and may prove useful for commercial production of α-galactosidase.     

Fructose metabolism in Zymomonas mobilis

Summary In the metabolism of fructose by Zymomonas, the ethanol yield is decreased due to the formation of dihydroxyacetone, mannitol and glycerol. The reduction of fructose to mannitol by an NADPH-dependent mannitol dehydrogenase is apparently coupled to the oxidation of glucose-6-phosphate by glucose-6-phosphate dehydrogenase, which exhibits higher activity with NADP than with NAD as cofactor. The relatively low cell yield on fructose can partly be explained by the loss of ATP in the formation of dihydroxyacetone and glycerol and partly by the toxic effect of dihydroxyacetone and acetaldehyde on the growth of the organism.     

Production of cyclomaltodextrin glucanotransferase of Bacillus circulans var. alkalophilus ATCC21783 in B. subtilis

Summary The cyclomaltodextrin glucanotransferase (CGTase, E.C. 2.4.1.19) gene from an alkalophilic Bacillus circulans var. alkalophilus ATCC21783 was cloned into Escherichia coli and B. subtilis. When cloned from E. coli to B. subtilis, the entire insert containing the CGTase gene was, depending on the plasmid construction, either unstable or the recombinant B. subtilis did not secrete the enzyme in significant amounts. To achieve efficient enzyme production in B. subtilis, the gene was placed under the control of the B. amyloliquefaciens -amylase promoter. In one of the constructions, both the promoter and the signal sequence of the gene were replaced with those of B. amyloliquefaciens, whereas in another construction only the promoter area was exchanged. The recombinant B. subtilis clones transformed with these plasmid constructions secreted CGTase into the culture medium 14 times as much as did the parental strain in shake flask cultures. In fermentor cultures in an industrially feasible medium the enzyme production was substantially higher, yielding 1.2 g/l of CGTase, which is about 33 times the amount of the enzyme produced by the parental strain in corresponding fermentations. Both of the plasmid constructions were stable when grown over 50 generations without antibiotic selection.     

Core concepts of spatial prioritisation in systematic conservation planning

Systematic conservation planning (SCP) is a field of conservation biology concerned with delivering on‐the‐ground actions that achieve conservation goals. It describes a set of operational models that cover both design and implementation of conservation, with a strong focus on mobilising the collective action typically required to implement conservation. SCP, as it was originally described, was composed of six different stages: collection of data, identification of conservation goals, evaluation of the existing protected area network, design of expansions, implementation of conservation action, and long‐term maintenance of biodiversity in the network. Since then, the operational model has been expanded into several different variants. Conservation actions applied inside SCP include establishment and expansion of reserve networks and allocation of habitat restoration and management. Within the broader context of SCP, there is a fundamental biogeographic‐economic analysis frequently called spatial conservation prioritisation or conservation assessment, which is used for identifying where important areas for biodiversity are and how conservation goals might be achieved efficiently. Here, we review the usage and meaning of the 12 biogeographic‐economic core concepts of SCP: adequacy, complementarity, comprehensiveness, effectiveness, efficiency, flexibility, irreplaceability, replacement cost, representation, representativeness, threat, and vulnerability. Some of the concepts have clear definitions whereas others may have alternative and possibly conflicting definitions. With a comprehensive literature review literature, we elucidate the historical backgrounds of these concepts, the first definitions and usages, alternative later definitions, key applications, and prior reviews. This review reduces linguistic uncertainty in the application of SCP. Since SCP is a global activity with a multitude of different stakeholders involved, it is vital that those involved can speak the same language. Through these concepts, this review serves as a source of information about the historical development of SCP. It provides a comprehensive review for anyone wishing to understand the key concepts of spatial prioritisation within SCP.     

Maximum shear strain-based algorithm can predict proteoglycan loss in damaged articular cartilage

Post-traumatic osteoarthritis (PTOA) is a common disease, where the mechanical integrity of articular cartilage is compromised. PTOA can be a result of chondral defects formed due to injurious loading. One of the first changes around defects is proteoglycan depletion. Since there are no methods to restore injured cartilage fully back to its healthy state, preventing the onset and progression of the disease is advisable. However, this is problematic if the disease progression cannot be predicted. Thus, we developed an algorithm to predict proteoglycan loss of injured cartilage by decreasing the fixed charge density (FCD) concentration. We tested several mechanisms based on the local strains or stresses in the tissue for the FCD loss. By choosing the degeneration threshold suggested for inducing chondrocyte apoptosis and cartilage matrix damage, the algorithm driven by the maximum shear strain showed the most substantial FCD losses around the lesion. This is consistent with experimental findings in the literature. We also observed that by using coordinate system-independent strain measures and selecting the degeneration threshold in an ad hoc manner, all the resulting FCD distributions would appear qualitatively similar, i.e., the greatest FCD losses are found at the tissue adjacent to the lesion. The proposed strain-based FCD degeneration algorithm shows a great potential for predicting the progression of PTOA via biomechanical stimuli. This could allow identification of high-risk defects with an increased risk of PTOA progression.

     

Genotoxicity of gliotoxin,a secondary metabolite of Aspergillus fumigatus,in a battery of short-term test systems

The genotoxic effects of gliotoxin, a known fungal secondary metabolite, were studied. Gliotoxin was purified from cultivation medium of Aspergillus fumigatus isolated from the indoor air of a moisture problem house. The genotoxicity of gliotoxin was assessed both in bacterial test systems including bacterial repair assay, Ames Salmonella assay and SOS-chromotest, and in mammalian cells using single cell gel (SCG) electrophoresis assay and sister-chromatid exchange (SCE) test. Gliotoxin was found to be genotoxic in the bacterial repair assay but, not in the Salmonella test or SOS-chromotest. A dose-related increase in DNA damage was observed in mouse RAW264.7 macrophages exposed to gliotoxin for 2h in plain medium in the SCG assay. In contrast to the positive response in the SCG assay, gliotoxin did not induce any clear, dose-related increase in SCEs in Chinese hamster ovary (CHO) cells.     

Prioritizing multiple-use landscapes for conservation: methods for large multi-species planning problems

Across large parts of the world, wildlife has to coexist with human activity in highly modified and fragmented landscapes. Combining concepts from population viability analysis and spatial reserve design, this study develops efficient quantitative methods for identifying conservation core areas at large, even national or continental scales. The proposed methods emphasize long-term population persistence, are applicable to both fragmented and natural landscape structures, and produce a hierarchical zonation of regional conservation priority. The methods are applied to both observational data for threatened butterflies at the scale of Britain and modelled probability of occurrence surfaces for indicator species in part of Australia. In both cases, priority landscapes important for conservation management are identified.     

Reserve selection using nonlinear species distribution models

Reserve design is concerned with optimal selection of sites for new conservation areas. Spatial reserve design explicitly considers the spatial pattern of the proposed reserve network and the effects of that pattern on reserve cost and/or ability to maintain species there. The vast majority of reserve selection formulations have assumed a linear problem structure, which effectively means that the biological value of a potential reserve site does not depend on the pattern of selected cells. However, spatial population dynamics and autocorrelation cause the biological values of neighboring sites to be interdependent. Habitat degradation may have indirect negative effects on biodiversity in areas neighboring the degraded site as a result of, for example, negative edge effects or lower permeability for animal movement. In this study, I present a formulation and a spatial optimization algorithm for nonlinear reserve selection problems in grid-based landscapes that accounts for interdependent site values. The method is demonstrated using habitat maps and nonlinear habitat models for threatened birds in the Netherlands, and it is shown that near-optimal solutions are found for regions consisting of up to hundreds of thousands grid cells, a landscape size much larger than those commonly attempted even with linear reserve selection formulations.     

The role of niche breadth, resource availability and range position on the life history of butterflies

We analysed the relationship between three life history characteristics (mobility, length of flight period and body size) and niche breadth (larval host plant specificity and adult habitat breadth), resource availability (distribution and abundance of host plants) and range position (distance between the northernmost distribution record and southernmost point of Finland) of the butterfly fauna of Finland. The data is based on literature and questionnaires. Often in across species studies phylogeny may create spurious relationships between life-history and ecological variables. We took the phylogenetic relatedness of butterfly species into account by analysing the data with phylogenetically independent contrasts (CAIC method). Butterfly mobility was positively related to the niche breadth, resource availability and range position. The length of the flight period was negatively related to the range position, indicating that the species at the northern edge of their distribution range have shorter flight period than species which are further way from the range edge. After controlling for the phylogenetic relatedness we found no significant correlations between body size and niche breadth, resource availability or range position. We suggest that the relationship between the length of the flight period and range position may arise as a consequence of lower hatching asynchrony in edge species as a result of lower environmental variance in larval growth conditions. Our results on the mobility suggest that there is selection pressure towards lower migration rate in species that have restricted niche breadth, low resource availability and in species that are on the northern edge of their geographical distribution range. In such species, selection against mobile individuals is likely to result from the decreased probability of finding another habitat patch suitable for egg laying.