Novel applications for glycosylphosphatidylinositol-anchored proteins in pharmaceutical and industrial biotechnology

Glycosylphosphatidylinositol (GPI)-anchored proteins have been regarded as typical cell surface proteins found in most eukaryotic cells from yeast to man. They are embedded in the outer plasma membrane leaflet via a carboxy-terminally linked complex glycolipid GPI structure. The amphiphilic nature of the GPI anchor, its compatibility with the function of the attached protein moiety and the capability of GPI-anchored proteins for spontaneous insertion into and transfer between artificial and cellular membranes initially suggested their potential for biotechnological applications. However, these expectations have been hardly fulfilled so far. Recent developments fuel novel hopes with regard to: (i) Automated online expression, extraction and purification of therapeutic proteins as GPI-anchored proteins based on their preferred accumulation in plasma membrane lipid rafts, (ii) multiplex custom-made protein chips based on GPI-anchored cell wall proteins in yeast, (iii) biomaterials and biosensors with films consisting of sets of distinct GPI-anchored binding-proteins or enzymes for sequential or combinatorial catalysis, and (iv) transport of therapeutic proteins across or into relevant tissue cells, e.g., enterocytes or adipocytes. Latter expectations are based on the demonstrated translocation of GPI-anchored proteins from plasma membrane lipid rafts to cytoplasmic lipid droplets and eventually further into microvesicles which upon release from donor cells transfer their GPI-anchored proteins to acceptor cells. The value of these technologies, which are all based on the interaction of GPI-anchored proteins with membranes and surfaces, for the engineering, production and targeted delivery of biomolecules for a huge variety of therapeutic and biotechnological purposes should become apparent in the near future.     

Winter oilseed production for biofuel in the US Corn Belt: opportunities and limitations

Interest from the US commercial aviation industry and commitments established by the US Navy and Air Force to use renewable fuels has spurred interest in identifying and developing crops for renewable aviation fuel. Concern regarding greenhouse gas emissions associated with land‐use change and shifting land grown for food to feedstock production for fuel has encouraged the concept of intensifying current prominent cropping systems through various double cropping strategies. Camelina (Camelina sativa L.) and field pennycress (Thlaspi arvense L.) are two winter oilseed crops that could potentially be integrated into the corn (Zea mays L.)–soybean [(Glycine max (L.) Merr.] cropping system, which is the prominent cropping system in the US Corn Belt. In addition to providing a feedstock for renewable aviation fuel production, integrating these crops into corn–soybean cropping systems could also potentially provide a range of ecosystem services. Some of these include soil protection from wind and water erosion, soil organic C (SOC) sequestration, water quality improvement through nitrate reduction, and a food source for pollinators. However, integration of these crops into corn–soybean cropping systems also carries possible limitations, such as potential yield reductions of the subsequent soybean crop. This review identifies and discusses some of the key benefits and constraints of integrating camelina or field pennycress into corn–soybean cropping systems and identifies generalized areas for potential adoption in the US Corn Belt.     

Functionalized nanocompartments (Synthosomes): limitations and prospective applications in industrial biotechnology

Synthosomes are mechanically stable vesicles with a block copolymer membrane and an engineered transmembrane protein acting as selective gate. The polymer vesicles are nanometer-sized (50-1000 nm) and functionalized by loading them with enzymes for bioconversions or encapsulating charged macromolecules for selective compound recovery/release. The Synthosome system might become a novel technology platform for biocatalysis and selective product recovery. Progress in Synthosome research comprises employed block copolymers, transmembrane channel engineering, and functionalizations, which are discussed here in detail. The challenges in transmembrane protein engineering, as well as cost-effective production, in block copolymer design and the state of the art in Synthosome characterization comprising quantification of encapsulated protein, translocation efficiency, number of transmembrane channels per vesicle, and enzyme kinetics are also presented and discussed. An assessment of the Synthosome technology platform for prospective applications in industrial (white) biotechnology concludes this review.     

ATP in current biotechnology: Regulation,applications and perspectives

Adenosine tri-phosphate (ATP), the most important energy source for metabolic reactions and pathways, plays a vital role in the growth of industrial strain and the production of target metabolites. In this review, current advances in manipulating ATP in industrial strains, including altering NADH availability, and regulating NADH oxidation pathway, oxygen supply, proton gradient, the electron transfer chain activity and the F₀F₁-ATPase activity, are summarized and discussed. By applying these strategies, optimal product concentrations, yields and productivity in industrial biotechnology have been achieved. Furthermore, the mechanisms by which ATP extends the substrate utilization spectra and enhances the ability to challenge harsh environmental stress have been elucidated. Finally, three critical issues related to ATP manipulation have been addressed.     

Harvesting for food versus feed: a review of Peruvian fisheries in a global context

Peru is the top exporter of fishmeal and fish oil (FMFO) worldwide and is responsible for half and a third of global production, respectively. Landings of “anchoveta” (Engraulis ringens) are used nearly exclusively for FMFO production, despite a proactive national food policy aimed at favoring the direct human consumption of this inexpensive species. It may be surprising that in a country where malnutrition and caloric deficit constitute major issues, a low-priced and highly nutritious fish such as anchovy does not have stronger domestic demand as a food fish. Here, we review and assess eight potential politico-socio-economic processes that can explain this situation. The main explanation are dietary habits, the preference for broiler and the higher profit from anchovy sold as feed fish compared to its use as a food fish due to historically high FMFO prices, boosted by an increasing demand for aquaculture in a context of finite forage and trash fish resources. In addition, the recent introduction of an individual quota system has shifted bargaining power from processors to fishers, thereby increasing competition for the raw material. This competition results in an increase in anchovy prices offered by the feed fish industry due to its onshore processing overcapacity, which is detrimental to the food fish industry. In the end, although the dominant use of anchovy for fish feed is largely explained by integrating these market mechanisms and other minor ones, this use raises other issues, such as rent redistribution through public policies, employment, equitability and utility (low social costs), and resource management (threats to ecosystems or global change). Different policy scenarios are proposed in relation to these issues.     

Review: Feed demand landscape and implications of food-not feed strategy for food security and climate change

The food-feed competition is one of the complex challenges, and so are the ongoing climate change, land degradation and water shortage for realizing sustainable food production systems. By 2050 the global demand for animal products is projected to increase by 60% to 70%, and developing countries will have a lion’s share in this increase. Currently, ~800 million tonnes of cereals (one-third of total cereal production) are used in animal feed and by 2050 it is projected to be over 1.1 billion tonnes. Most of the increase in feed demand will be in developing countries, which already face many food security challenges. Additional feed required for the projected increased demand of animal products, if met through food grains, will further exacerbate the food insecurity in these countries. Furthermore, globally, the production, processing and transport of feed account for 45% of the greenhouse gas emissions from the livestock sector. This paper presents approaches for addressing these challenges in quest for making livestock sector more sustainable. The use of novel human-inedible feed resources such as insect meals, leaf meals, protein isolates, single cell protein produced using waste streams, protein hydrolysates, spineless cactus, algae, co-products of the biofuel industry, food wastes among others, has enormous prospects. Efficient use of grasslands also offers possibilities for increasing carbon sequestration, land reclamation and livestock productivity. Opportunities also exist for decreasing feed wastages by simple and well proven practices such as use of appropriate troughs, increase in efficiency of harvesting crop residues and their conversion to complete feeds especially in the form of densified feed blocks or pellets, feeding as per the nutrient requirements, among others. Available evidence have been presented to substantiate arguments that: (a) for successful and sustained adoption of a feed technology, participation of the private sector and a sound business plan are required, (b) for sustainability of the livestock production systems, it is also important to consider the consumption of animal products and a case has been presented to assess future needs of animal source foods based on their requirements for healthy living, (c) for dairy animals, calculation of Emission Intensity based on the lifetime lactation rather than one lactation may also be considered and (d) for assessment of the efficiency of livestock production systems a holistic approach is required that takes into consideration social dimensions and net human-edible protein output from the system in addition to carbon and water footprints.     

Ionic liquids in biotechnology: applications and perspectives for biotransformations

Ionic liquids are considered as an alternative to organic solvents for catalysis. The literature in this field is reviewed with focus on advantageous use of ionic liquids in biocatalysis and biotransformations. The overview reveals that the exploration and mapping of ionic liquids with respect to biocatalysis is still sketchy. It is apparent that advantages can be gained in view of activity, stability and selectivity. Furthermore, integration of reaction and separation has a high potential in the field. The review presents quantitative data on the productivities, space–time yields, as well as stability as far as they can be extracted from the literature. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Axenic cultures for microalgal biotechnology: Establishment,assessment, maintenance,and applications

The impact of microalgae (including blue-green algae or cyanobacteria) on human life can be both beneficiary and deleterious. While microalgae can be cultivated and used as feedstocks for the production of bioenergy and high value-added products in nutraceuticals, pharmaceuticals, and aquaculture feeds, some microalgae cause harmful algal blooms (HABs) that cause large-scale mortality in aquatic environments around the world. Thus, with the development of microalgal biotechnology and increasing concern about HABs, research on microscopic algae has increased significantly. However, this growth of academic research and application fields has been hindered by difficulties in obtaining axenic cultures. Therefore, this review provides a brief explanation of diverse establishment techniques, along with their strengths and weaknesses, with the hope of facilitating successful axenic cultures. A compilation of research fields and relevant important findings is also presented to clarify the importance of pure algal cultures. Finally, several controversial and sometimes overlooked issues related to the establishment, maintenance, and utilization of axenic cultures are discussed.     

Disposable bioreactors: the current state-of-the-art and recommended applications in biotechnology

Disposable bioreactors have increasingly been incorporated into preclinical, clinical, and production-scale biotechnological facilities over the last few years. Driven by market needs, and, in particular, by the developers and manufacturers of drugs, vaccines, and further biologicals, there has been a trend toward the use of disposable seed bioreactors as well as production bioreactors. Numerous studies documenting their advantages in use have contributed to further new developments and have resulted in the availability of a multitude of disposable bioreactor types which differ in power input, design, instrumentation, and scale of the cultivation container. In this review, the term “disposable bioreactor” is defined, the benefits and constraints of disposable bioreactors are discussed, and critical phases and milestones in the development of disposable bioreactors are summarized. An overview of the disposable bioreactors that are currently commercially available is provided, and the domination of wave-mixed, orbitally shaken, and, in particular, stirred disposable bioreactors in animal cell-derived productions at cubic meter scale is reported. The growth of this type of reactor system is attributed to the recent availability of stirred disposable benchtop systems such as the Mobius CellReady 3 L Bioreactor. Analysis of the data from computational fluid dynamic simulation studies and first cultivation runs confirms that this novel bioreactor system is a viable alternative to traditional cell culture bioreactors at benchtop scale.     

Green leaf volatiles: biosynthesis,biological functions and their applications in biotechnology

Plants have evolved numerous constitutive and inducible defence mechanisms to cope with biotic and abiotic stresses. These stresses induce the expression of various genes to activate defence‐related pathways that result in the release of defence chemicals. One of these defence mechanisms is the oxylipin pathway, which produces jasmonates, divinylethers and green leaf volatiles (GLVs) through the peroxidation of polyunsaturated fatty acids (PUFAs). GLVs have recently emerged as key players in plant defence, plant–plant interactions and plant–insect interactions. Some GLVs inhibit the growth and propagation of plant pathogens, including bacteria, viruses and fungi. In certain cases, GLVs released from plants under herbivore attack can serve as aerial messengers to neighbouring plants and to attract parasitic or parasitoid enemies of the herbivores. The plants that perceive these volatile signals are primed and can then adapt in preparation for the upcoming challenges. Due to their ‘green note’ odour, GLVs impart aromas and flavours to many natural foods, such as vegetables and fruits, and therefore, they can be exploited in industrial biotechnology. The aim of this study was to review the progress and recent developments in research on the oxylipin pathway, with a specific focus on the biosynthesis and biological functions of GLVs and their applications in industrial biotechnology.     

Exploiting the peptidoglycan-binding motif,LysM, for medical and industrial applications

The lysin motif (LysM) was first identified by Garvey et al. in 1986 and, in subsequent studies, has been shown to bind noncovalently to peptidoglycan and chitin by interacting with N-acetylglucosamine moieties. The LysM sequence is present singly or repeatedly in a large number of proteins of prokaryotes and eukaryotes. Since the mid-1990s, domains containing one or more of these LysM sequences originating from different LysM-containing proteins have been examined for purely scientific reasons as well as for their possible use in various medical and industrial applications. These studies range from detecting localized binding of LysM-containing proteins onto bacteria to actual bacterial cell surface analysis. On a more applied level, the possibilities of employing the LysM domains for cell immobilization, for the display of peptides, proteins, or enzymes on (bacterial) surfaces as well as their utility in the development of novel vaccines have been scrutinized. To serve these purposes, the chimeric proteins containing one or more of the LysM sequences have been produced and isolated from various prokaryotic and eukaryotic expression hosts. This review gives a succinct overview of the characteristics of the LysM domain and of current developments in its application potential.     

Catalytic antibodies and their applications in biotechnology: state of the art

Catalytic antibodies are immunoglobulins endowed with enzymatic properties. Discovered in the second part of the 1980s, the enthusiasm they initially aroused was counterbalanced by the difficulty of their production and their low catalytic rates. Nevertheless, improvements in expression systems and engineering technologies, combined with various studies suggesting that catalytic antibodies play a role in the immune system, have opened the way to new applications for these proteins. Herein we review catalytic antibodies from a biotechnological point of view, focusing our study on the different production methods, expression systems and their potential clinical applications dedicated to these proteins.