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Category Archives: Biotechnology
Impact of high biomass loading on ionic liquid pretreatment
Background:
Ionic liquid (IL) pretreatment has shown great potential as a novel pretreatment technology with high sugar yields. To improve process economics of pretreatment, higher biomass loading is desirable. The goal of this work is to establish, the impact of high biomass loading of switchgrass on IL pretreatment in terms of viscosity, cellulose crystallinity, chemical composition, saccharification kinetics, and sugar yield.
Results:
The pretreated switchgrass/IL slurries show frequency dependent shear thinning behavior. The switchgrass/IL slurries show a crossover from viscous behavior at 3 wt% to elastic behavior at 10 wt%. The relative glucan content of the recovered solid samples is observed to decrease with increasing levels of lignin and hemicelluloses with increased biomass loading. The IL pretreatment led to a transformation of cellulose crystalline structure from I to II for 3, 10, 20 and 30 wt% samples, while a mostly amorphous structure was found for 40 and 50 wt% samples.
Conclusions:
IL pretreatment effectively reduced the biomass recalcitrance at loadings as high as 50 wt%. Increased shear viscosity and a transition from 'fluid' like to 'solid' like behavior was observed with increased biomass loading. At high biomass loadings shear stress produced shear thinning behavior and a reduction in viscosity by two orders of magnitude, thereby reducing the complex viscosity to values similar to lower loadings. The rheological properties and sugar yields indicate that 10 to 50 wt% may be a reasonable and desirable target for IL pretreatment under certain operating conditions.Source:
http://www.biotechnologyforbiofuels.com/content/6/1/52
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Land-use change and greenhouse gas emissions from corn and cellulosic ethanol
Background:
The greenhouse gas (GHG) emissions that may accompany land-use change (LUC) from increased biofuel feedstock production are a source of debate in the discussion of drawbacks and advantages of biofuels. Estimates of LUC GHG emissions focus mainly on corn ethanol and vary widely. Increasing the understanding of LUC GHG impacts associated with both corn and cellulosic ethanol will inform the on-going debate concerning their magnitudes and sources of variability.
Results:
In our study, we estimate LUC GHG emissions for ethanol from four feedstocks: corn, corn stover, switchgrass, and miscanthus. We use new computable general equilibrium (CGE) results for worldwide LUC. U.S. domestic carbon emission factors are from state-level modelling with a surrogate CENTURY model and U.S. Forest Service data. This paper investigates the effect of several key domestic lands carbon content modelling parameters on LUC GHG emissions. International carbon emission factors are from the Woods Hole Research Center. LUC GHG emissions are calculated from these LUCs and carbon content data with Argonne National Laboratory's Carbon Calculator for Land Use Change from Biofuels Production (CCLUB) model. Our results indicate that miscanthus and corn ethanol have the lowest (-10 g CO2e/MJ) and highest (7.6 g CO2e/MJ) LUC GHG emissions under base case modelling assumptions. The results for corn ethanol are lower than corresponding results from previous studies. Switchgrass ethanol base case results (2.8 g CO2e/MJ) were the most influenced by assumptions regarding converted forestlands and the fate of carbon in harvested wood products. They are greater than miscanthus LUC GHG emissions because switchgrass is a lower-yielding crop. Finally, LUC GHG emissions for corn stover are essentially negligible and insensitive to changes in model assumptions.
Conclusions:
This research provides new insight into the influence of key carbon content modelling variables on LUC GHG emissions associated with the four bioethanol pathways we examined. Our results indicate that LUC GHG emissions may have a smaller contribution to the overall biofuel life cycle than previously thought. Additionally, they highlight the need for future advances in LUC GHG emissions estimation including improvements to CGE models and aboveground and belowground carbon content data.Source:
http://www.biotechnologyforbiofuels.com/content/6/1/51
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Development of an electrotransformation protocol for genetic manipulation of Clostridium pasteurianum
Background:
Reducing the production cost of, and increasing revenues from, industrial biofuels will greatly facilitate their proliferation and co-integration with fossil fuels. The cost of feedstock is the largest cost in most fermentation bioprocesses and therefore represents an important target for cost reduction. Meanwhile, the biorefinery concept advocates revenue growth through complete utilization of by-products generated during biofuel production. Taken together, the production of biofuels from low-cost crude glycerol, available in oversupply as a by-product of bioethanol production, in the form of thin stillage, and biodiesel production, embodies a remarkable opportunity to advance affordable biofuel development. However, few bacterial species possess the natural capacity to convert glycerol as a sole source of carbon and energy into value-added bioproducts. Of particular interest is the anaerobe Clostridium pasteurianum, the only microorganism known to convert glycerol alone directly into butanol, which currently holds immense promise as a high-energy biofuel and bulk chemical. Unfortunately, genetic and metabolic engineering of C. pasteurianum has been fundamentally impeded due to lack of an efficient method for deoxyribonucleic acid (DNA) transfer.
Results:
This work reports the development of an electrotransformation protocol permitting high-level DNA transfer to C. pasteurianum ATCC 6013 together with accompanying selection markers and vector components. The CpaAI restriction-modification system was found to be a major barrier to DNA delivery into C. pasteurianum which we overcame by in vivo methylation of the recognition site (5'-CGCG-3') using the M.FnuDII methyltransferase. With proper selection of the replication origin and antibiotic-resistance marker, we initially electroporated methylated DNA into C. pasteurianum at a low efficiency of 2.4 x 101 transformants mug-1 DNA by utilizing conditions common to other clostridial electroporations. Systematic investigation of various parameters involved in the cell growth, washing and pulse delivery, and outgrowth phases of the electrotransformation procedure significantly elevated the electrotransformation efficiency, up to 7.5 x 104 transformants mug-1 DNA, an increase of approximately three order of magnitude. Key factors affecting the electrotransformation efficiency include cell-wall-weakening using glycine, ethanol-mediated membrane solubilization, field strength of the electric pulse, and sucrose osmoprotection.
Conclusions:
C. pasteurianum ATCC 6013 can be electrotransformed at a high efficiency using appropriately methylated plasmid DNA. The electrotransformation method and tools reported here should promote extensive genetic manipulation and metabolic engineering of this biotechnologically important bacterium.Source:
http://www.biotechnologyforbiofuels.com/content/6/1/50
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Biotechnology Project Mrs Nangle 2013 – Video
Biotechnology Project Mrs Nangle 2013
By: Bonnie Bower
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Biotechnology Project Mrs Nangle 2013 - Video
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Detailed analysis of metagenome datasets obtained from biogas-producing microbial communities residing in biogas reactors does not indicate the presence of putative pathogenic microorganisms
Background:
In recent years biogas plants in Germany have been supposed to be involved in amplification and dissemination of pathogenic bacteria causing severe infections in humans and animals. In particular, biogas plants are discussed to contribute to the spreading of Escherichia coli infections in humans or chronic botulism in cattle caused by Clostridium botulinum. Metagenome datasets of microbial communities from an agricultural biogas plant as well as from anaerobic lab-scale digesters operating at different temperatures and conditions were analyzed for the presence of putative pathogenic bacteria and virulence determinants by various bioinformatic approaches.
Results:
All datasets featured a low abundance of reads that were taxonomically assigned to the genus Escherichia or further selected genera comprising pathogenic species. Higher numbers of reads were taxonomically assigned to the genus Clostridium. However, only very few sequences were predicted to originate from pathogenic clostridial species. Moreover, mapping of metagenome reads to complete genome sequences of selected pathogenic bacteria revealed that not the pathogenic species itself, but only species that are more or less related to pathogenic ones are present in the fermentation samples analyzed. Likewise, known virulence determinants could hardly be detected. Only a marginal number of reads showed similarity to sequences described in the Microbial Virulence Database MvirDB such as those encoding protein toxins, virulence proteins or antibiotic resistance determinants.
Conclusions:
Findings of this first study of metagenomic sequence reads of biogas producing microbial communities suggest that the risk of dissemination of pathogenic bacteria by application of digestates from biogas fermentations as fertilizers is low, because obtained results do not indicate the presence of putative pathogenic microorganisms in the samples analyzed.Source:
http://www.biotechnologyforbiofuels.com/content/6/1/49
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Biotechnology Review Song – Video
Biotechnology Review Song
This is an AP Biology Review Song to the tune of "Wobble" by VIC. Enjoy!
By: jchvatal1
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Biotechnology Review Song - Video
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