Microbial genetic engineering approach to replace shark livering for squalene

Squalene is generally sourced from the liver oil of deep sea sharks (Squalus spp.), in which it accounts for 40–70% of liver mass. To meet the growing demand for squalene because of its beneficial effects for human health, three to six million deep sea sharks are slaughtered each year, profoundly endangering marine ecosystems. To overcome this unsustainable practice, microbial sources of squalene might offer a viable alternative to plant- or animal-based squalene, although only a few microorganisms have been found that are capable of synthesizing up to 30% squalene of dry biomass by native biosynthetic pathways. These squalene biosynthetic pathways, on the other hand, can be genetically manipulated to transform microorganisms into \u27cellular factories\u27 for squalene overproduction

Development of Thermophilic Tailor-Made Enzyme Mixtures for the Bioconversion of Agricultural and Forest Residues

Even though the main components of all lignocellulosic feedstocks include cellulose, hemicellulose, as well as the protective lignin matrix, there are some differences in structure, such as in hardwoods and softwoods, which may influence the degradability of the materials. Under this view, various types of biomass might require a minimal set of enzymes that has to be tailor-made. Partially defined complex mixtures that are currently commercially used are not adapted to efficiently degrade different materials, so novel enzyme mixtures have to be customized. Development of these cocktails requires better knowledge about the specific activities involved, in order to optimize hydrolysis. The role of filamentous fungus Myceliophthora thermophila and its complete enzymatic repertoire for the bioconversion of complex carbohydrates has been widely proven. In this study, four core cellulases (MtCBH7, MtCBH6, MtEG5 and MtEG7), in the presence of other four accessory enzymes (mannanase, lytic polyssacharide monooxygenase MtGH61, xylanase, MtFae1a) and β-glucosidase MtBGL3, were tested as a 9-component cocktail against one model substrate (phosphoric acid swollen cellulose) and four hydrothermally pretreated natural substrates (wheat straw as an agricultural waste, birch and spruce biomass, as forest residues). Synergistic interactions among different enzymes were determined using a suitable design of experiments methodology. The results suggest that for the hydrolysis of the pure substrate (PASC), high proportions of MtEG7 are needed for efficient yields. MtCBH7 and MtEG7 are enzymes of major importance during the hydrolysis of pretreated wheat straw, while MtCBH7 plays a crucial role in case of spruce. Cellobiohydrolases MtCBH6 and MtCBH7 act in combination and are key enzymes for the hydrolysis of the hardwood (birch). Optimum combinations were predicted from suitable statistical models which were able to further increase hydrolysis yields, suggesting that tailor-made enzyme mixtures targeted towards a particular residual biomass can help maximize hydrolysis yields. The present work demonstrates the change from ‘one cocktail for all’ to ‘tailor-made cocktails’ that are needed for the efficient saccharification of targeted feed stocks prior to the production of biobased products through the biorefinery concept

Carbonic anhydrase to boost CO2 sequestration : Improving carbon capture utilization and storage (CCUS)

CO2 Capture Utilization and Storage (CCUS) is a fundamental strategy to mitigate climate change, and carbon sequestration, through absorption, can be one of the solutions to achieving this goal. In nature, carbonic anhydrase (CA) catalyzes the CO2 hydration to bicarbonates. Targeting the development of novel biotechnological routes which can compete with traditional CO2 absorption methods, CA utilization has presented a potential to expand as a promising catalyst for CCUS applications. Driven by this feature, the search for novel CAs as biocatalysts and the utilization of enzyme improvement techniques, such as protein engineering and immobilization methods, has resulted in suitable variants able to catalyze CO2 absorption at relevant industrial conditions. Limitations related to enzyme recovery and recyclability are still a concern in the field, affecting cost efficiency. Under different absorption approaches, CA enhances both kinetics and CO2 absorption yields, besides reduced energy consumption. However, efforts directed to process optimization and demonstrative plants are still limited. A recent topic with great potential for development is the CA utilization in accelerated weathering, where industrial residues could be re-purposed towards becoming carbon sequestrating agents. Furthermore, research of new solvents has identified potential candidates for integration with CA in CO2 capture, and through techno-economic assessments, CA can be a path to increase the competitiveness of alternative CO2 absorption systems, offering lower environmental costs. This review provides a favorable scenario combining the enzyme and CO2 capture, with possibilities in reaching an industrial-like stage in the future

High concentrations of dried sorghum stalks as a biomass feedstock for single cell oil production by Rhodosporidium toruloides

Environmental crisis and concerns for energy security have made the research for renewable fuels that will substitute the usage of fossil fuels an important priority. Biodiesel is a potential substitute for petroleum, but its feasibility is hindered by the utilization of edible vegetable oil as raw material, which is responsible for a large fraction of the production cost and fosters the food versus fuel competition. Microbial oils are an interesting alternative as they do not compete with food production, and low cost renewable materials could serve as raw materials during cultivation of microorganisms. Sweet sorghum is an excellent candidate as substrate for microbial oil production, as it possesses high photosynthetic activity yielding high amounts of soluble and insoluble carbohydrates, and does not require high fertilization and irrigation rates8FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPsem informaçã

Structural and Molecular Characterization of Squalene Synthase Belonging to the Marine Thraustochytrid Species Aurantiochytrium limacinum Using Bioinformatics Approach

The marine microorganisms thraustochytrids have been explored for their potential in the production of various bioactive compounds, such as DHA, carotenoids, and squalene. Squalene is a secondary metabolite of the triterpenoid class and is known for its importance in various industrial applications. The bioinformatic analysis for squalene synthase (SQS) gene (the first key enzyme in the triterpenoid synthesis pathway), that is prevailing among thraustochytrids, is poorly investigated. In-silico studies combining sequence alignments and bioinformatic tools helped in the preliminary characterization of squalene synthases found in Aurantiochytrium limacinum. The sequence contained highly conserved regions for SQS found among different species indicated the enzyme had all the regions for its functionality. The signal peptide sequence and transmembrane regions were absent, indicating an important aspect of the subcellular localization. Secondary and 3-D models generated using appropriate templates demonstrated the similarities with SQS of the other species. The 3-D model also provided important insights into possible active, binding, phosphorylation, and glycosylation sites

Characterization of Organosolv Birch Lignins: Toward Application-Specific Lignin Production

Organosolv pretreatment represents one of the most promising biomass valorization strategies for renewable carbon-based products; meanwhile, there is an overall lack of holistic approach to how extraction conditions affect the suitable end-usages. In this context, lignin extracted from silver birch (Betula pendula L.) by a novel hybrid organosolv/steam-explosion treatment at varying process conditions (EtOH %; time; catalyst %) was analyzed by quantitative NMR (1H-13C HSQC; 13C NMR; 31P NMR), gel permeation chromatography, Fourier transform infrared (FT-IR), Pyr-gas chromatography-mass spectroscopy (GC/MS), and thermogravimetric analysis, and the physicochemical characteristics of the lignins were discussed regarding their potential usages. Characteristic lignin interunit bonding motifs, such as beta-O-4', beta-beta', and beta-5', were found to dominate in the extracted lignins, with their abundance varying with treatment conditions. Low-molecular-weight lignins with fairly unaltered characteristics were generated via extraction with the highest ethanol content potentially suitable for subsequent production of free phenolics. Furthermore, beta-beta' and beta-5' structures were predominant at higher acid catalyst contents and prolonged treatment times. Higher acid catalyst content led to oxidation and ethoxylation of side-chains, with the concomitant gradual disappearance of p-hydroxycinnamyl alcohol and cinnamaldehyde. This said, the increasing application of acid generated a broad set of lignin characteristics with potential applications such as antioxidants, carbon fiber, nanoparticles, and water remediation purposes

Green conversion of municipal solid wastes into fuels and chemicals

Presently, the society is facing a serious challenge for the effective management of the increasing amount of produced municipal solid wastes. The accumulated waste has caused a series of environmental problems such as uncontrolled release of greenhouse gases. Moreover, the increasing amount of wastes has resulted in a shortage of areas available for waste disposal, resulting in a nonsustainable waste management. These problems led to serious public concerns, which in turn resulted in political actions aiming to reduce the amount of wastes reaching the environment. These actions aim to promote sustainable waste management solutions. The main objective of these policies is to promote the recycling of municipal solid waste and the conversion of waste to energy and valuable chemicals. These conversions can be performed using either biological (e.g., anaerobic digestion) or thermochemical processes (e.g., pyrolysis). Research efforts during the last years have been fruitful, and many publications demonstrated the effective conversation of municipal solid waste to energy and chemicals. These processes are discussed in the current review article together with the change of the waste policy that was implemented in the EU during the last years

Isolation and modification of nano-scale cellulose from organosolv-treated birch through the synergistic activity of LPMO and endoglucanases

Nanocellulose isolation fromlignocellulose is a tedious and expensive processwith high energy and harsh chemical requirements, primarily due to the recalcitrance of the substrate, which otherwise would have been costeffective due to its abundance. Replacing the chemical steps with biocatalytic processes offers opportunities to solve this bottleneck to a certain extent due to the enzymes substrate specificity and mild reaction chemistry. In this work, we demonstrate the isolation of sulphate-free nanocellulose from organosolv pretreated birch biomass using different glycosyl-hydrolases, along with accessory oxidative enzymes including a lytic polysaccharide monooxygenase (LPMO). The suggested process produced colloidal nanocellulose suspensions (zeta-potential-19.4 mV) with particles of 7-20 nm diameter, high carboxylate content and improved thermostability (T-o= 301 degrees C, T-max= 337 degrees C). Nanocelluloseswere subjected to post-modification using LPMOs of different regioselectivity. The sample from chemical route was the least favorable for LPMO to enhance the carboxylate content, while that from the C1-specific LPMO treatment showed the highest increase in carboxylate content. (c) 2021 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http:// creativecommons.org/licenses/by/4.0/)

optimization of enzymatic synthesis of l arabinose ferulate catalyzed by feruloyl esterases from myceliophthora thermophila in detergentless microemulsions and assessment of its antioxidant and cytotoxicity activities

The feruloyl esterases FaeA1, FaeA2, FaeB1, FaeB2 from Myceliophthora thermophila C1 and MtFae1a from M. thermophila ATCC 42464 were used as biocatalysts for the transesterification of vinyl ferula ..

Lignin-first biomass fractionation using a hybrid organosolv – Steam explosion pretreatment technology improves the saccharification and fermentability of spruce biomass

For a transition to a sustainable society, fuels, chemicals, and materials should be produced from renewable resources. Lignocellulosic biomass constitutes an abundant and renewable feedstock; however, its successful application in a biorefinery requires efficient fractionation into its components; cellulose, hemicellulose and lignin. Here, we demonstrate that a newly established hybrid organosolv – steam explosion pretreatment can effectively fractionate spruce biomass to yield pretreated solids with high cellulose (72% w/w) and low lignin (delignification up to 79.4% w/w) content. The cellulose-rich pretreated solids present high saccharification yields (up to 61% w/w) making them ideal for subsequent bioconversion processes. Moreover, under high-gravity conditions (22% w/w) we obtained an ethanol titer of 61.7 g/L, the highest so far reported for spruce biomass. Finally, the obtained high-purity lignin is suitable for various advanced applications. In conclusion, hybrid organosolv pretreatment could offer a closed-loop biorefinery while simultaneously adding value to all biomass components