Pretreatment strategies for microbial valorization of bio‐oil fractions produced by fast pyrolysis of ash‐rich lignocellulosic biomass

This work evaluates a biorefinery approach for microbial valorization of bio‐oil fractions produced by fast pyrolysis of ash‐rich lignocellulosic biomass. Different methods are presented for the pretreatment of the low‐sugar complex bio‐oil consisting of organic condensate (OC) and aqueous condensate (AC) to overcome their strong inhibitory effects and unsuitability for common analytical methods. Growth of Pseudomonas putida KT2440, which was chosen as a reference system, on untreated bio‐oil fractions was only detectable using solid medium with OC as sole carbon source. Utilization of a pretreated OC which was filtered, autoclaved, neutralized and centrifuged enabled growth in liquid medium with significant remaining optical instability. By subjecting the pretreated fractions to solid phase extraction, more stable and less inhibitory bio‐oil fractions could be obtained enabling the appliance of common analytical methods. Furthermore, this pretreatment facilitated growth of the applied reference organism Pseudomonas putida KT2440. As there is currently no convincing strategy for reliable application of bio‐oil as a sole source of carbon in industrial biotechnology, the presented work depicts a first step toward establishing bio‐oil as a future sustainable feedstock for a bio‐based economy.     

A brief review of extrusion‐based tissue scaffold bio‐printing

Extrusion‐based bio‐printing has great potential as a technique for manipulating biomaterials and living cells to create three‐dimensional (3D) scaffolds for damaged tissue repair and function restoration. Over the last two decades, advances in both engineering techniques and life sciences have evolved extrusion‐based bio‐printing from a simple technique to one able to create diverse tissue scaffolds from a wide range of biomaterials and cell types. However, the complexities associated with synthesis of materials for bio‐printing and manipulation of multiple materials and cells in bio‐printing pose many challenges for scaffold fabrication. This paper presents an overview of extrusion‐based bio‐printing for scaffold fabrication, focusing on the prior‐printing considerations (such as scaffold design and materials/cell synthesis), working principles, comparison to other techniques, and to‐date achievements. This paper also briefly reviews the recent development of strategies with regard to hydrogel synthesis, multi‐materials/cells manipulation, and process‐induced cell damage in extrusion‐based bio‐printing. The key issue and challenges for extrusion‐based bio‐printing are also identified and discussed along with recommendations for future, aimed at developing novel biomaterials and bio‐printing systems, creating patterned vascular networks within scaffolds, and preserving the cell viability and functions in scaffold bio‐printing. The address of these challenges will significantly enhance the capability of extrusion‐based bio‐printing.     

Maßgeschneiderte Mikroorganismen

Tailor‐made microorganisms Microbial diversity provides unlimited resources for the development of novel industrial processes and products. Since the beginning of the 20th century microorganisms have been successfully applied for the large scale production of bio‐based products. In recent years, modern methods of strain development and Synthetic Biology have enabled biotech engineers to design even more sophisticated and tailor‐made microorganisms. These microbes serve industrial processes for the production of bulk chemicals, enzymes, polymers, biofuels as well as plant‐derived ingredients such as Artemisinin in an ecologically and economically sustainable and attractive fashion. In the future, production of advanced biofuels, microbial fuel cells, CO₂ as feedstock and microbial cellulose are research topics as well as challenges of global importance. Continuous efforts in microbiology and biotechnology research will be pivotal for white biotechnology to gain more momentum in transforming the chemical industry towards a knowledge based bio‐economy.     

Bio‐Nanotechnology in High‐Performance Supercapacitors

The use of bio‐nanotechnology for the fabrication of diverse functional nanomaterials with precisely controlled morphologies and microstructures is attracting considerable attention due to its sustainability and renewability. As one of the key energy storage devices, supercapacitor (SC) requires the active electrode material to have high specific surface area, interconnected porous structure, excellent electronic conductivity, and appropriate heteroatom doping for promoting the transfer of electrons and electrolyte ions. The combination of bio‐technology and SC will open up a new avenue for the large‐scale fabrication of high performance functional energy storage devices. In this review, the most state‐of‐the‐art research progress in bio‐nanotechnological fabrication of different nanomaterials, including carbon materials, metal oxides, conducting polymers, and their corresponding composites are reviewed with the following three bio‐nanotechnical approaches covered: (1) biomass carbonization technologies; (2) bio‐template methods; and (3) bio‐complex technologies, while also highlighting their applications as functional SC electrodes.     

Next‐Generation Industrial Biotechnology‐Transforming the Current Industrial Biotechnology into Competitive Processes

The chemical industry has made a contribution to modern society by providing cost‐competitive products for our daily use. However, it now faces a serious challenge regarding environmental pollutions and greenhouse gas emission. With the rapid development of molecular biology, biochemistry, and synthetic biology, industrial biotechnology has evolved to become more efficient for production of chemicals and materials. However, in contrast to chemical industries, current industrial biotechnology (CIB) is still not competitive for production of chemicals, materials, and biofuels due to their low efficiency and complicated sterilization processes as well as high‐energy consumption. It must be further developed into “next‐generation industrial biotechnology” (NGIB), which is low‐cost mixed substrates based on less freshwater consumption, energy‐saving, and long‐lasting open continuous intelligent processing, overcoming the shortcomings of CIB and transforming the CIB into competitive processes. Contamination‐resistant microorganism as chassis is the key to a successful NGIB, which requires resistance to microbial or phage contaminations, and available tools and methods for metabolic or synthetic biology engineering. This review proposes a list of contamination‐resistant bacteria and takes Halomonas spp. as an example for the production of a variety of products, including polyhydroxyalkanoates under open‐ and continuous‐processing conditions proposed for NGIB.     

A review of thermochemical upgrading of pyrolysis bio‐oil: Techno‐economic analysis,life cycle assessment,and technology readiness

Technologies for upgrading fast pyrolysis bio‐oil to drop‐in fuels and coproducts are under development and show promise for decarbonizing energy supply for transportation and chemicals markets. The successful commercialization of these fuels and the technologies deployed to produce them depend on production costs, scalability, and yield. To meet environmental regulations, pyrolysis‐based biofuels need to adhere to life cycle greenhouse gas intensity standards relative to their petroleum‐based counterparts. We review literature on fast pyrolysis bio‐oil upgrading and explore key metrics that influence their commercial viability through life cycle assessment (LCA) and techno‐economic analysis (TEA) methods together with technology readiness level (TRL) evaluation. We investigate the trade‐offs among economic, environmental, and technological metrics derived from these methods for individual technologies as a means of understanding their nearness to commercialization. Although the technologies reviewed have not attained commercial investment, some have been pilot tested. Predicting the projected performance at scale‐up through models can, with industrial experience, guide decision‐making to competitively meet energy policy goals. LCA and TEA methods that ensure consistent and reproducible models at a given TRL are needed to compare alternative technologies. This study highlights the importance of integrated analysis of multiple economic, environmental, and technological metrics for understanding performance prospects and barriers among early stage technologies.     

Approaches for Addressing Life Cycle Assessment Data Gaps for Bio‐based Products

There is an increasing need for life cycle data for bio‐based products, which becomes particularly evident with the recent drive for greenhouse gas reporting and carbon footprinting studies. Meeting this need is challenging given that many bio‐products have not yet been studied by life cycle assessment (LCA), and those that have are specific and limited to certain geographic regions. In an attempt to bridge data gaps for bio‐based products, LCA practitioners can use either proxy data sets (e.g., use existing environmental data for apples to represent pears) or extrapolated data (e.g., derive new data for pears by modifying data for apples considering pear‐specific production characteristics). This article explores the challenges and consequences of using these two approaches. Several case studies are used to illustrate the trade‐offs between uncertainty and the ease of application, with carbon footprinting as an example. As shown, the use of proxy data sets is the quickest and easiest solution for bridging data gaps but also has the highest uncertainty. In contrast, data extrapolation methods may require extensive expert knowledge and are thus harder to use but give more robust results in bridging data gaps. They can also provide a sound basis for understanding variability in bio‐based product data. If resources (time, budget, and expertise) are limited, the use of averaged proxy data may be an acceptable compromise for initial or screening assessments. Overall, the article highlights the need for further research on the development and validation of different approaches to bridging data gaps for bio‐based products.     

Nature Driven Bio‐Piezoelectric/Triboelectric Nanogenerator as Next‐Generation Green Energy Harvester for Smart and Pollution Free Society

Electronics wastes (e‐wastes) are the major concern in the rapid expansion of smart/wearable/portable electronics in modern high‐tech society. Informal processing and enormous gathering of e‐wastes can lead to adverse human/animal health effects and environmental pollution worldwide. Currently, these issues are a big headache and require the scientific community to develop effective green energy harvesting technologies using biodegradable/biocompatible materials. Piezoelectric/triboelectric nanogenerators (PNGs/TNGs) are considered one of the most promising renewable green energy sources for the conversion of mechanical/biomechanical energies into electricity. However, organic/inorganic material based PNGs/TNGs are very much incompatible, and considered e‐wastes for their non‐biodegradability. This review covers potential uses of biodegradable/biocompatible materials which are wasted every day as nature driven material based bio‐nanogenerators with a particular focus on their applications in flexible PNGs/TNGs fabrication. Structural investigation and possible working principles are described first in order to outline the basic mechanism of bio‐inspired materials behind energy harvesting. Then, energy harvesting abilities and the mechanical sensing of bio‐inspired integrated flexible devices are discussed under various mechanical/biomechanical activities. Finally, their potential applications in various flexible, wearable, and portable electronic fields are demonstrated. These bio‐inspired energy harvesting devices can make huge changes in fields as diverse as portable electronics, in vitro/in vivo biomedical applications, and many more.     

A framework for the identification of promising bio‐based chemicals

Recent progress in metabolic engineering and synthetic biology enables the use of microorganisms for the production of chemicals—“bio‐based chemicals.” However, it is still unclear which chemicals have the highest economic prospect. To this end, we develop a framework for the identification of such promising ones. Specifically, we first develop a genome‐scale constraint‐based metabolic modeling approach, which is used to identify a candidate pool of 209 chemicals (together with the estimated yield, productivity, and residence time for each) from the intersection of the high‐production‐volume chemicals and the KEGG and MetaCyc databases. Second, we design three screening criteria based on a chemical’s profit margin, market volume, and market size. The total process cost, including the downstream separation cost, is systematically incorporated into the evaluation. Third, given the three aforementioned criteria, we identify 32 products as economically promising if the maximum yields can be achieved, and 22 products if the maximum productivities can be achieved. The breakeven titer that renders zero profit margin for each product is also presented. Comparisons between extracellular and intracellular production, as well as Escherichia coli and Saccharomyces cerevisiae systems are also discussed. The proposed framework provides important guidance for future studies in the production of bio‐based chemicals. It is also flexible in that the databases, yield estimations, and criteria can be modified to customize the screening.     

Revealing the Environmental Advantages of Industrial Symbiosis in Wood‐Based Bioeconomy Networks: An Assessment From a Life Cycle Perspective

The German government has recently initiated funding schemes that incentivize strategies for wood‐based bioeconomy regions. Regional wood and chemical industries have been encouraged to act symbiotically, that is, share pilot plant facilities, couple processes where feasible, and cascade woody feedstock throughout their process networks. However, during the planning stages of these bioeconomy regions, options need to be assessed for sustainably integrating processes and energy integration between the various industries that produce bio‐based polymers and engineered wood products. The aim of this paper is to identify the environmental sustainability of industrial symbiosis for producing high‐value‐added, bio‐based products in the wood‐based bioeconomy region of Central Germany. An analysis was conducted of three possible future scenarios with varying degrees of symbiosis in the bioeconomy network. A life cycle assessment (LCA) approach was used to compare these three scenarios to a traditional fossil‐based production system. Eleven environmental impact categories were considered. The results show that, in most cases, the bioeconomy network outperformed the fossil‐based production system, mitigating environmental impacts by 25% to 130%.     

Separation of bio‐based chemicals from fermentation broths by salting‐out extraction

The possibility of creating a biorefinery using inexpensive biomass has attracted a great deal of attention, which is mainly focused on the improvement of strains and fermentation, whereas few resources have been spent on downstream processing. Bio‐based chemical downstream processing can become a bottleneck in industrial production because so many impurities are introduced into the fermentation broth. This review introduces a technique referred to as salting‐out extraction, which is based on the partition difference between chemicals in two phases consisting of salts and polymers or hydrophilic solvents, hydrophobic solvents, and amphipathic chemicals. The effects of solvents and salts on the formation of two phases were discussed, as was the use of this method to recover bio‐based chemicals. This review focused on the separation of hydrophilic chemicals (1,3‐propanediol, 2,3‐butanediol, acetoin, and lactic acid) from fermentation broths. Diols could be recovered at a high yield from fermentation broths without pretreatment especially with a hydrophilic solvent‐based system, whereas the recovery of organic acids was slightly lower. Most of the impurities (cells and proteins) were removed during the same step. Extractive fermentations were also used for polymer‐based aqueous two‐phase systems.     

Hydrothermal liquefaction of Malaysia's algal biomass for high‐quality bio‐oil production

Currently, fossil materials form the majority of our energy and chemical source. Many global concerns force us to rethink about our current dependence on the fossil energy. Limiting the use of these energy sources is a key priority for most countries that pledge to reduce greenhouse gas emissions. The application of biomass, as substitute fossil resources for producing biofuels, plastics and chemicals, is a widely accepted strategy for sustainable development. Aquatic plants including algae possess competitive advantages as biomass resources compared to the terrestrial plants in this current global situation. Bio‐oil production from algal biomass is technically and economically viable, cost competitive, requires no capacious lands and minimal water use and reduces atmospheric carbon dioxide. The aim of this paper is to review the potential of converting algal biomass, as an aquatic plant, into high‐quality crude bio‐oil through applicable processes in Malaysia. In particular, bio‐based materials and fuels from algal biomass are considered as one of the reliable alternatives for clean energy. Currently, pyrolysis and hydrothermal liquefaction (HTL) are two foremost processes for bio‐oil production from biomass. HTL can directly convert high‐moisture algal biomass into bio‐oil, whereas pyrolysis requires feedstock drying to reduce the energy consumption during the process. Microwave‐assisted HTL, which can be conducted in aqueous environment, is suitable for aquatic plants and wet biomass such as algae.     

Production of oncolytic adenovirus and human mesenchymal stem cells in a single‐use,Vertical‐Wheel bioreactor system: Impact of bioreactor design on performance of microcarrier‐based cell culture processes

Anchorage‐dependent cell cultures are used for the production of viruses, viral vectors, and vaccines, as well as for various cell therapies and tissue engineering applications. Most of these applications currently rely on planar technologies for the generation of biological products. However, as new cell therapy product candidates move from clinical trials towards potential commercialization, planar platforms have proven to be inadequate to meet large‐scale manufacturing demand. Therefore, a new scalable platform for culturing anchorage‐dependent cells at high cell volumetric concentrations is urgently needed. One promising solution is to grow cells on microcarriers suspended in single‐use bioreactors. Toward this goal, a novel bioreactor system utilizing an innovative Vertical‐Wheel? technology was evaluated for its potential to support scalable cell culture process development. Two anchorage‐dependent human cell types were used: human lung carcinoma cells (A549 cell line) and human bone marrow‐derived mesenchymal stem cells (hMSC). Key hydrodynamic parameters such as power input, mixing time, Kolmogorov length scale, and shear stress were estimated. The performance of Vertical‐Wheel bioreactors (PBS‐VW) was then evaluated for A549 cell growth and oncolytic adenovirus type 5 production as well as for hMSC expansion. Regarding the first cell model, higher cell growth and number of infectious viruses per cell were achieved when compared with stirred tank (ST) bioreactors. For the hMSC model, although higher percentages of proliferative cells could be reached in the PBS‐VW compared with ST bioreactors, no significant differences in the cell volumetric concentration and expansion factor were observed. Noteworthy, the hMSC population generated in the PBS‐VW showed a significantly lower percentage of apoptotic cells as well as reduced levels of HLA‐DR positive cells. Overall, these results showed that process transfer from ST bioreactor to PBS‐VW, and scale‐up was successfully carried out for two different microcarrier‐based cell cultures. Ultimately, the data herein generated demonstrate the potential of Vertical‐Wheel bioreactors as a new scalable biomanufacturing platform for microcarrier‐based cell cultures of complex biopharmaceuticals. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:1600–1612, 2015     

All‐Solid‐State Printed Bipolar Li–S Batteries

Despite their potential advantages over currently widespread lithium‐ion batteries, lithium–sulfur (Li–S) batteries are not yet in practical use. Here, for the first time bipolar all‐solid‐state Li–S batteries (ASSLSBs) are demonstrated that exhibit exceptional safety, flexibility, and aesthetics. The bipolar ASSLSBs are fabricated through a solvent‐drying‐free, ultraviolet curing‐assisted stepwise printing process at ambient conditions, without (high‐temperature/high‐pressure) sintering steps that are required for inorganic electrolyte‐based all‐solid‐state batteries. Two thermodynamically immiscible and nonflammable gel electrolytes based on ethyl methyl sulfone (EMS) and tetraethylene glycol dimethyl ether (TEGDME) are used to address longstanding concerns regarding the grain boundary resistance of conventional inorganic solid electrolytes, as well as the polysulfide shuttle effect in Li–S batteries. The EMS gel electrolytes embedded in the sulfur cathodes facilitate sulfur utilization, while the TEGDME gel composite electrolytes serve as polysulfide‐repelling separator membranes. Benefiting from the well‐designed cell components and printing‐driven facile processability, the resulting bipolar ASSLSBs exhibit unforeseen advancements in bipolar cell configuration, safety, foldability, and form factors, which lie far beyond those achievable with conventional Li–S battery technologies.     

Microbial‐based motor fuels: science and technology

The production of biofuels via microbial biotechnology is a very active field of research. A range of fuel molecule types are currently under consideration: alcohols, ethers, esters, isoprenes, alkenes and alkanes. At the present, the major alcohol biofuel is ethanol. The ethanol fermentation is an old technology. Ongoing efforts aim to increase yield and energy efficiency of ethanol production from biomass. n‐Butanol, another microbial fermentation product, is potentially superior to ethanol as a fuel but suffers from low yield and unwanted side‐products currently. In general, biodiesel fuels consist of fatty acid methyl esters in which the carbon derives from plants, not microbes. A new biodiesel product, called microdiesel, can be generated in engineered bacterial cells that condense ethanol with fatty acids. Perhaps the best fuel type to generate from biomass would be biohydrocarbons. Microbes are known to produce hydrocarbons such as isoprenes, long‐chain alkenes and alkanes. The biochemical mechanisms of microbial hydrocarbon biosynthesis are currently under study. Hydrocarbons and minimally oxygenated molecules may also be produced by hybrid chemical and biological processes. A broad interest in novel fuel molecules is also driving the development of new bioinformatics tools to facilitate biofuels research.     

Structure‐Based Site‐Specific PEGylation of Fibroblast Growth Factor 2 Facilitates Rational Selection of Conjugate Sites

Polyethylene glycol modification (PEGylation) can enhance the pharmacokinetic properties of therapeutic proteins by the attachment of polyethylene glycol (PEG) to the surface of a protein to shield the protein surface from proteolytic degradation and limit aggregation. However, current PEGylation strategies often reduce biological activity, potentially as a result of steric hindrance of PEG. Overall, there are no structure‐based guidelines for selection of conjugate sites that retain optimal biological activity with improved pharmacokinetic properties. In this study, site‐specific PEGylation based on the FGF2‐FGFR1‐heparin complex structure is performed. The effects of the conjugate sites on protein function are investigated by measuring the receptor/heparin binding affinities of the modified proteins and performing assays to measure cell‐based bio‐activity and in vivo stability. Comprehensive analysis of these data demonstrates that PEGylation of FGF2 that avoids the binding sites for fibroblast growth factor receptor 1 (FGFR1) and heparin provides optimal pharmacokinetic enhancement with minimal losses to biological activity. Animal experiments demonstrate that PEGylated FGF2 exhibits greater efficacy in protecting against traumatic brain injury‐induced brain damage and neurological functions than the non‐modified FGF2. This rational structure‐based PEGylation strategy for protein modification is expected to have a major impact in the area of protein‐based therapeutics.     

Allogeneic cell therapy bioprocess economics and optimization: Single‐use cell expansion technologies

For allogeneic cell therapies to reach their therapeutic potential, challenges related to achieving scalable and robust manufacturing processes will need to be addressed. A particular challenge is producing lot‐sizes capable of meeting commercial demands of up to 10⁹ cells/dose for large patient numbers due to the current limitations of expansion technologies. This article describes the application of a decisional tool to identify the most cost‐effective expansion technologies for different scales of production as well as current gaps in the technology capabilities for allogeneic cell therapy manufacture. The tool integrates bioprocess economics with optimization to assess the economic competitiveness of planar and microcarrier‐based cell expansion technologies. Visualization methods were used to identify the production scales where planar technologies will cease to be cost‐effective and where microcarrier‐based bioreactors become the only option. The tool outputs also predict that for the industry to be sustainable for high demand scenarios, significant increases will likely be needed in the performance capabilities of microcarrier‐based systems. These data are presented using a technology S‐curve as well as windows of operation to identify the combination of cell productivities and scale of single‐use bioreactors required to meet future lot sizes. The modeling insights can be used to identify where future R&D investment should be focused to improve the performance of the most promising technologies so that they become a robust and scalable option that enables the cell therapy industry reach commercially relevant lot sizes. The tool outputs can facilitate decision‐making very early on in development and be used to predict, and better manage, the risk of process changes needed as products proceed through the development pathway. Biotechnol. Bioeng. 2014;111: 69–83. © 2013 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc.     

Cell wall biochemical alterations during Agrobacterium‐mediated expression of haemagglutinin‐based influenza virus‐like vaccine particles in tobacco

Influenza virus‐like particles (VLPs) have been shown to induce a safe and potent immune response through both humoral and cellular responses. They represent promising novel influenza vaccines. Plant‐based biotechnology allows for the large‐scale production of VLPs of biopharmaceutical interest using different model organisms, including Nicotiana benthamiana plants. Through this platform, influenza VLPs bud from the plasma membrane and accumulate between the membrane and the plant cell wall. To design and optimize efficient production processes, a better understanding of the plant cell wall composition of infiltrated tobacco leaves is a major interest for the plant biotechnology industry. In this study, we have investigated the alteration of the biochemical composition of the cell walls of N. benthamiana leaves subjected to abiotic and biotic stresses induced by the Agrobacterium‐mediated transient transformation and the resulting high expression levels of influenza VLPs. Results show that abiotic stress due to vacuum infiltration without Agrobacterium did not induce any detectable modification of the leaf cell wall when compared to non infiltrated leaves. In contrast, various chemical changes of the leaf cell wall were observed post‐Agrobacterium infiltration. Indeed, Agrobacterium infection induced deposition of callose and lignin, modified the pectin methylesterification and increased both arabinosylation of RG‐I side chains and the expression of arabinogalactan proteins. Moreover, these modifications were slightly greater in plants expressing haemagglutinin‐based VLP than in plants infiltrated with the Agrobacterium strain containing only the p19 suppressor of silencing.