| 1. |
- Blomqvist, Johanna, et al.
(författare)
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Fermentation of lignocellulosic hydrolysate by the alternative industrial ethanol yeast Dekkera bruxellensis
- 2011
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Ingår i: Letters in Applied Microbiology. - Malden, USA : Wiley-Blackwell. - 0266-8254 .- 1472-765X. ; 53:1, s. 73-78
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Tidskriftsartikel (refereegranskat)abstract
- Aim: Testing the ability of the alternative ethanol production yeast Dekkera bruxellensis to produce ethanol from lignocellulose hydrolysate and comparing it to Saccharomyces cerevisiae.Methods and Results: Industrial isolates of D. bruxellensis and S. cerevisiae were cultivated in small-scale batch fermentations of enzymatically hydrolysed steam exploded aspen sawdust. Different dilutions of hydrolysate were tested. None of the yeasts grew in undiluted or 1 : 2 diluted hydrolysate [final glucose concentration always adjusted to 40 g l(-1) (0.22 mol l(-1))]. This was most likely due to the presence of inhibitors such as acetate or furfural. In 1 : 5 hydrolysate, S. cerevisiae grew, but not D. bruxellensis, and in 1 : 10 hydrolysate, both yeasts grew. An external vitamin source (e.g. yeast extract) was essential for growth of D. bruxellensis in this lignocellulosic hydrolysate and strongly stimulated S. cerevisiae growth and ethanol production. Ethanol yields of 0 42 +/- 0 01 g ethanol (g glucose)(-1) were observed for both yeasts in 1 : 10 hydrolysate. In small-scale continuous cultures with cell recirculation, with a gradual increase in the hydrolysate concentration, D. bruxellensis was able to grow in 1 : 5 hydrolysate. In bioreactor experiments with cell recirculation, hydrolysate contents were increased up to 1 : 2 hydrolysate, without significant losses in ethanol yields for both yeasts and only slight differences in viable cell counts, indicating an ability of both yeasts to adapt to toxic compounds in the hydrolysate.Conclusions: Dekkera bruxellensis and S. cerevisiae have a similar potential to ferment lignocellulose hydrolysate to ethanol and to adapt to fermentation inhibitors in the hydrolysate.Significance and Impact of the study: This is the first study investigating the potential of D. bruxellensis to ferment lignocellulosic hydrolysate. Its high competitiveness in industrial fermentations makes D. bruxellensis an interesting alternative for ethanol production from those substrates.
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| 2. |
- Feng, X. M., et al.
(författare)
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Rhizopus oligosporus and yeast co-cultivation during barley tempeh fermentation-Nutritional impact and real-time PCR quantification of fungal growth dynamics
- 2007
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Ingår i: Food microbiology (Print). - London, United Kingdom : Elsevier BV. - 0740-0020 .- 1095-9998. ; 24:4, s. 393-402
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Tidskriftsartikel (refereegranskat)abstract
- Barley tempeh was produced by fermenting barley kernels with Rhizopus oligosporus. The potential of the yeasts Saccharomyces cerevisiae (three strains), S. boulardii (one strain), Pichia anomala (one strain) and Kluyveromyces lactis (one strain) to grow together with R. oligosporus during barley tempeh fermentation was evaluated. All yeast strains grew during the fermentation and even during cold storage of tempeh (P < 0.01). The growth of yeasts slightly increased the ergosterol contents, but did not influence amino acid contents and compositions, and did not reduce phytate contents. Slight increases of vitamins B6 and niacinamide, and slight decreases of B1 and biotin were observed. Quantification of fungal growth is difficult during mixed species fermentations because ergosterol is found in all fungal species, and colony-forming-unit (cfu) estimations are not reliable for R. oligosporus and other sporulating fungi. Therefore, we developed a quantitative real-time PCR method for individually quantifying S. cerevisiae and R. oligosporus growth in barley tempeh. The PCR results were highly correlated with the ergosterol content of R. oligosporus and with the number of cfu of S. cerevisiae. Thus, real-time PCR is a rapid and selective method to quantify yeasts and R. oligosporus during mixed species fermentation of inhomogenous substrate such as barley tempeh. © 2006 Elsevier Ltd. All rights reserved.
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| 3. |
- Ballerini, Lucia, et al.
(författare)
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Comparison of histomorphometrical data obtained with two different image analysis methods
- 2007
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Ingår i: Journal of materials science. Materials in medicine. - : Springer. - 0957-4530 .- 1573-4838. ; 18:8, s. 1471-1479
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Tidskriftsartikel (refereegranskat)abstract
- A common way to determine tissue acceptance of biomaterials is to perform histomorphometrical analysis on histologically stained sections from retrieved samples with surrounding tissue, using various methods. The “time and money consuming” methods and techniques used are often “in house standards”. We address light microscopic investigations of bone tissue reactions on un-decalcified cut and ground sections of threaded implants. In order to screen sections and generate results faster, the aim of this pilot project was to compare results generated with the in-house standard visual image analysis tool (i.e., quantifications and judgements done by the naked eye) with a custom made automatic image analysis program. The histomorphometrical bone area measurements revealed no significant differences between the methods but the results of the bony contacts varied significantly. The raw results were in relative agreement, i.e., the values from the two methods were proportional to each other: low bony contact values in the visual method corresponded to low values with the automatic method. With similar resolution images and further improvements of the automatic method this difference should become insignificant. A great advantage using the new automatic image analysis method is that it is time saving—analysis time can be significantly reduced.
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| 4. |
- Basheer, Shabana, et al.
(författare)
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A membrane protein based biosensor: Use of a phosphate - H+ symporter membrane protein (Pho84) in the sensing of phosphate ions.
- 2011
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Ingår i: Biosensors & bioelectronics. - : Elsevier. - 0956-5663 .- 1873-4235. ; 27:1, s. 58-63
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Tidskriftsartikel (refereegranskat)abstract
- A label free biosensor for direct detection of inorganic phosphate based on potential-step capacitance measurements has been developed. The high-affinity Pho84 plasma membrane phosphate/proton symporter of Saccharomyces cerevisiae was used as a sensing element. Heterologously expressed and purified Pho84 protein was immobilized on a self-assembled monolayer (SAM) on a capacitance electrode. Changes in capacitance were recorded upon exposure to phosphate compared to the control substance, phosphate analogue methylphosphonate. Hence, even without the explicit use of lipid membranes, the Pho84 membrane protein could retain its capacity of selective substrate binding, with a phosphate detection limit in the range of the apparent in vivo K(m). A linear increase in capacitance was monitored in the phosphate concentration range of 5-25 mu M. The analytical response of the capacitive biosensor is in agreement with that the transporter undergoes significant conformational changes upon exposure to inorganic phosphate, while exposure to the analogue only causes minor responses.
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| 5. |
- Cicortas Gunnarsson, Lavinia, et al.
(författare)
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Evolution of a carbohydrate binding module into a protein-specific binder
- 2006
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Ingår i: Biomolecular Engineering. - : Elsevier BV. - 1389-0344 .- 1878-559X. ; 23:2-3, s. 111-117
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Tidskriftsartikel (refereegranskat)abstract
- A carbohydrate binding module, CBM4-2, derived front the xylanase (Xyn 10A) of Rhodothermus marinus has been used as a scaffold for molecular diversification. Its binding specificity has been evolved to recognise a quite different target, a human monoclonal IgG4. In order to understand the basis for this drastic change in specificity we have further investigated the target recognition of the IgG4-specific CBMs. Firstly, we defined that the structure target recognised by the selected CBM-variants was the protein and not the carbohydrates attached to the glycoprotein. We also identified key residues involved in the new specificity and/or responsible for the swap in specificity, from xylan to human IgG4. Specific changes present in all these CBMs included mutations not introduced in the design of the library from which the specific clones were selected. Reversion of such mutations led to a complete loss of binding to the target molecule, suggesting that they are critical for the recognition of human IgG4. Together with the mutations introduced at will, they had transformed the CBM scaffold into a protein binder. We have thus shown that the scaffold of CBM4-2 is able to harbour molecular recognition for either carbohydrate or protein structures. (c) 2005 Elsevier B.V. All rights reserved.
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| 6. |
- Cicortas Gunnarsson, Lavinia, et al.
(författare)
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Molecular engineering of a thermostable carbohydrate-binding module
- 2006
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Ingår i: Biocatalysis and Biotransformation. - : Informa UK Limited. - 1029-2446 .- 1024-2422. ; 24:1-2, s. 31-37
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Konferensbidrag (refereegranskat)abstract
- Structure-function studies are frequently practiced on the very diverse group of natural carbohydrate-binding modules in order to understand the target recognition of these proteins. We have taken a step further in the study of carbohydrate-binding modules and created variants with novel binding properties by molecular engineering of one such molecule of known 3D-structure. A combinatorial library was created from the sequence encoding a thermostable carbohydrate-binding module, CBM4-2 from a Rhodothermus marinus xylanase, and phage-display technology was successfully used for selection of variants with specificity towards different carbohydrate polymers (birchwood xylan, Avicel (TM), ivory nut mannan and recently also xyloglucan), as well as towards a glycoprotein (human IgG4). Our work not only generated a number of binders with properties that would suite a range of biotechnological applications, but analysis of selected binders also helped us to identify residues important for their specificities.
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| 7. |
- Gunnarsson, Lavinia Cicortas, 1977-, et al.
(författare)
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A carbohydrate binding module as a diversity-carrying scaffold
- 2004
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Ingår i: Protein Engineering Design & Selection. - : Oxford University Press. - 1741-0126 .- 1741-0134. ; 17:3, s. 213-221
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Tidskriftsartikel (refereegranskat)abstract
- The growing field of biotechnology is in constant need of binding proteins with novel properties. Not just binding specificities and affinities but also structural stability and productivity are important characteristics for the purpose of large-scale applications. In order to find such molecules, libraries are created by diversifying naturally occurring binding proteins, which in those cases serve as scaffolds. In this study, we investigated the use of a thermostable carbohydrate binding module, CBM4-2, from a xylanase found in Rhodothermus marinus, as a diversity-carrying scaffold. A combinatorial library was created by introducing restricted variation at 12 positions in the carbohydrate binding site of the CBM4-2. Despite the small size of the library (1.6 x 10(6) clones), variants specific towards different carbohydrate polymers (birchwood xylan, Avicel and ivory nut mannan) as well as a glycoprotein (human IgG4) were successfully selected for, using the phage display method. Investigated clones showed a high productivity (on average 69 mg of purified protein/l shake flask culture) when produced in Escherichia coli and they were all stable molecules displaying a high melting transition temperature (75.7 +/- 5.3 degrees C). All our results demonstrate that the CBM4-2 molecule is a suitable scaffold for creating variants useful in different biotechnological applications.
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| 8. |
- Håkansson, Torbjörn, et al.
(författare)
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Sulphate reducing bacteria to precipitate mercury after electrokinetic soil remediation
- 2008
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Ingår i: International journal of environmental science and technology. - 1735-1472 .- 1735-2630. ; 5:2, s. 267-274
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Tidskriftsartikel (refereegranskat)abstract
- Combined treatment with electroremediation and sulphate reducing bacteria (SRB) was tested in laboratory and pilot scale. The contaminated soil came from a chlor-alkali factory and contained about 100 mg/kg Hg. Iodide/iodine complexing agent was used to mobilize mercury. Mercury iodide complexes were moved to the anode solution using an electric field. The anode solution was then mixed with hydrogen sulphide (H2S) containing water, causing precipitation of mercury sulphide. The H2S was produced at site by a SRB reactor. Precipitation problems arising from the nature of the anode solution were expected, since this solution is highly acidic, very oxidised and may contain iodide/iodine that strongly complexes mercury and can hinder mercury sulphide precipitation. Mercury concentrations in the anode solution were up to 65.7 mg/L (field) and 15.4 mg/L (lab. scale). Reduction of mercury in the water was >93% at all times. Iodide did not hinder the process: Nonetheless, in the lab system, iodide concentration was high in the anode solution but mercury reduction was > 99.9%. The redox potential was sufficiently low for HgS precipitation during the experiments, except for a short period, when the mercury removal decreased to 94%. Sulphate reducing bacteria are shown as a viable tool for the treatment of mercury contaminated, acidic, oxidative, iodide containing water, such as that produced by electrokinetic remediation. A second SRB step or other water treatment is required to reduce the mercury concentration to environmentally acceptable levels. Redox. potential is the most sensitive factor in the system.
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| 9. |
- Lacayo, Martha, et al.
(författare)
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A toxaphene-degrading bacterium related to Enterobacter cloacae, strain D1 isolated from aged contaminated soil in Nicaragua
- 2005
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Ingår i: Systematic and Applied Microbiology. - : Elsevier BV. - 0723-2020 .- 1618-0984. ; 28:7, s. 632-639
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Tidskriftsartikel (refereegranskat)abstract
- Enterobacter sp. strain D1 is a facultative anaerobic, Gram-negative heterotrophic bacterium isolated from toxaphene-contaminated soil. This organism was identified and characterized through phylogenetic and taxonomic studies. Based on 16S rDNA analysis, the strain D1 was clustered closely with the species Enterobacter cloacae subsp. dissolvens (LMG 2683) and E. cloacae (ATCC 13047T). Strain D1 resembled these E. cloacae strains with respect to various biochemical and nutritional characteristics, but also exhibited differences. Moreover, strain D1 is able to grow and survive with toxaphene supplied in the medium in the range 3-96 mg/L. Amongst the chemical components of toxaphene, octachlorocamphenes, nonachlorobornanes and decachlorobornanes were seen to be rapidly metabolized, although levels of hexachlorocamphenes and heptachlorobornanes were found to be slowly degraded, and subsequently accumulated during the last stage of the cultivation.
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| 10. |
- Lacayo, Martha, et al.
(författare)
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Degradation of toxaphene by Bjerkandera sp. strain BOL13 using waste biomass as a cosubstrate
- 2006
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Ingår i: Applied Microbiology and Biotechnology. - : Springer Science and Business Media LLC. - 0175-7598 .- 1432-0614. ; 71:4, s. 549-554
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Tidskriftsartikel (refereegranskat)abstract
- The white-rot fungus Bjerkandera sp. strain BOL13 was capable of degrading toxaphene when supplied with wood chips, wheat husk or cane molasses as cosubstrates in batch culture experiments. Approximately 85% of toxaphene was removed when wheat husk was the main substrate. The production of lignin peroxidase was only stimulated when wheat husk was present in the liquid medium. Although xylanase was always detected, wheat husk supported the highest xylanase production. A negligible amount of beta-glucosidase and cellulase were found in the batch culture medium. To the best of our knowledge, this is the first reported case of toxaphene degradation by white-rot fungi.
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