Advances in biochemical engineering/biotechnology最新文献

The eCell technology is a recently introduced, specialized protein production platform with uses in a multitude of biotechnological applications. This chapter summarizes the use of eCell technology in four selected application areas. Firstly, for detecting heavy metal ions, specifically mercury, in an in vitro protein expression system. Results show improved sensitivity and lower limit of detection compared to comparable in vivo systems. Secondly, eCells are semipermeable, stable, and can be stored for extended periods of time, making them a portable and accessible technology for bioremediation of toxicants in extreme environments. Thirdly and fourthly, applications of eCell technology are shown to facilitate expression of correctly folded disulfide-rich proteins and incorporate chemically interesting derivatives of amino acids into proteins which are toxic to in vivo protein expression. Overall, eCell technology presents a cost-effective and efficient method for biosensing, bioremediation, and protein production.

Metalloproteins, proteins containing metal atoms or clusters within their structures, are critical for various biological functions across all domains of life. More than hundreds of different types have been discovered, which conduct various roles such as transportation of O₂, catalyzing chemical reactions, sensing environmental changes, and relaying electrons. Metalloprotein molecules incorporate a variety of metal atoms, coordinated to specific amino acid residues that affect their conformation and functionality. The process of metal incorporation typically occurs during or post-protein folding, often requiring chaperones for metal ion delivery and quality control. Progress in understanding metal incorporation and metalloprotein functionality has been enhanced by cell-free protein synthesis (CFPS) methods that offer direct control over the synthesis environment. This chapter reviews the diverse applications of CFPS methods in metalloprotein research, encompassing structure-function studies, protein engineering, and creation of artificial metalloproteins. Examples demonstrating the utility and advances brought about by CFPS in synthetic biology, electrochemistry, and drug discovery are highlighted. Despite remarkable progress, challenges remain in optimizing and advancing the CFPS methods, underscoring the need for future explorations in this transformative approach to metalloprotein study and engineering.

Medicinal mushrooms are higher fungi that consist of ascomycetes, basidiomycetes, and imperfect fungi. They have been long used as tonic and traditional medicine in East Asia, Europe, and Africa. Contemporary pharmacological researches have revealed that they possess a wide spectrum of bioactivity due to their production of a variety of bioactive compounds. Some of them have entered into the market; some are ready for industrial trials and further commercialization, while others are in various stages of development. According to the purpose of usage, a variety of medicinal mushroom-based products have been developed, which could be roughly divided into three general categories, i.e., nutraceuticals/functional foods, nutriceuticals/dietary supplements, and pharmaceuticals. Accordingly, the downstream processing of medicinal mushroom products varies greatly. Indeed, a major characteristic of medicinal mushroom is the wide variety of secondary metabolites, due to which a broad spectrum of separation techniques must be employed. In this chapter we will present an overview of the achievements in downstream processing technology for medicinal mushroom products. Examples of separation of products such as bioactive high-molecular-weight products like polysaccharides and low-molecular-weight products like triterpenoids are given. The application of some special separation strategy, e.g., chemical reaction-assisted separation for tackling some analogs with similar physicochemical properties from medicinal mushroom, is also described.

Production of mushroom fruit bodies using farming technology could hardly meet the increasing demand of the world market. During the last several decades, there have been various basic and applied studies on fungal physiology, metabolism, process engineering, and (pre)clinical studies. The fundamental aspects of solid-state cultivation of various kinds of medicinal mushroom mycelia in various types of bioreactors were established. Solid-state cultivation of medicinal mushrooms for their biomass and bioactive metabolites production appear very suitable for veterinary use. Development of comprehensive submerged technologies using stirred tank and airlift bioreactors is the most promising technology for fast and large-scale production of medicinal fungi biomass and their pharmaceutically active products for human need. The potentials initiate the development of new drugs and some of the most attractive over-the-counter human and veterinary remedies. This article is to overview the engineering achievements in solid state and submerged cultivations of medicinal mushrooms in bioreactors.

Since most of the medicinal mushrooms are rare in nature, production of fungal fruiting bodies is hardly covering the food market and the production of pharmaceutically active products, so artificial cultivation of fruiting bodies in a form of farming has been intensively established during the last 40 years. Various cultivation technologies are presented, including traditional farming of fruiting bodies on wood logs and beds, and also on other substrate-based media, such as cultivation in bags, bottles, and others. The advantage of farming is a cheap but time-consuming large-scale production. Agriculture, wood, and food industry wastes represent the main substrates that are in this process delignified and enriched in proteins and highly valuable pharmaceutically active compounds. The present article presents an overview of achievements in artificial cultivation of fruiting bodies, including the most relevant medicinal mushroom species, such as Ganoderma lucidum, Grifola frondosa, Pleurotus ostreatus, Agaricus brasiliensis, and Lentinula edodes.

Cell-free protein synthesis (CFPS) with flexibility and controllability can provide a powerful platform for high-throughput screening of biomolecules, especially in the evolution of peptides or proteins. In this chapter, the emerging strategies for enhancing the protein expression level using different source strains, energy systems, and template designs in constructing CFPS systems are summarized and discussed in detail. In addition, we provide an overview of the ribosome display, mRNA display, cDNA display, and CIS display in vitro display technologies, which can couple genotype and phenotype by forming fusion complexes. Moreover, we point out the trend that improving the protein yields of CFPS itself can offer more favorable conditions for maintaining library diversity and display efficiency. It is hoped that the novel CFPS system can accelerate the development of protein evolution in biotechnological and medical applications.

Anthropogenic global warming is driven by the increasing energy demand and the still dominant use of fossil energy carriers to meet these needs. New carbon-neutral energy sources are urgently needed to solve this problem. Biophotovoltaics, a member of the so-called bioelectrochemical systems family, will provide an important piece of the energy puzzle. It aims to harvest the electrons from sunlight-driven water splitting using the natural oxygenic photosystem (e.g., of cyanobacteria) and utilize them in the form of, e.g., electricity or hydrogen. Several key aspects of biophotovoltaics have been intensively studied in recent years like physicochemical properties of electrodes or efficient wiring of microorganisms to electrodes. Yet, the exact mechanisms of electron transfer between the biocatalyst and the electrode remain unresolved today. Most research is conducted on microscale reactors generating small currents over short time-scales, but multiple experiments have shown biophotovoltaics great potential with lab-scale reactors producing currents over weeks to months. Although biophotovoltaics is still in its infancy with many open research questions to be addressed, new promising results from various labs around the world suggest an important opportunity for biophotovoltaics in the decades to come. In this chapter, we will introduce the concept of biophotovoltaics, summarize its recent key progress, and finally critically discuss the potentials and challenges for future rational development of biophotovoltaics.

Cyanobacteria are highly interesting microbes with the capacity for oxygenic photosynthesis. They fulfill an important purpose in nature but are also potent biocatalysts. This chapter gives a brief overview of this diverse phylum and shortly addresses the functions these organisms have in the natural ecosystems. Further, it introduces the main topics covered in this volume, which is dealing with the development and application of cyanobacteria as solar cell factories for the production of chemicals including potential fuels. We discuss cyanobacteria as industrial workhorses, present established chassis strains, and give an overview of the current target products. Genetic engineering strategies aiming at the photosynthetic efficiency as well as approaches to optimize carbon fluxes are summarized. Finally, main cultivation strategies are sketched.

The human uses of mushrooms and cultured mycelium products for nutrition and medicine are detailed and supported by available human studies, which in many cases are clinical trials published in peer-reviewed journals. The major medically active immunomodulating compounds in the cell walls-chitin, beta-glucans, and glycoproteins, as well as lower weight molecules-nitrogen-containing compounds, phenolics, and terpenes-are discussed in relation to their current clinical uses. The nutritional content and foods derived from mushrooms, particularly related to their medical benefits, are discussed. High-quality major nutrients such as the high amounts of complete protein and prebiotic fibers found in edible and medicinal fungi and their products are presented. Mushrooms contain the highest amount of valuable medicinal fiber, while dried fruiting bodies of some fungi have up to 80% prebiotic fiber. These fibers are particularly complex and are not broken down in the upper gut, so they can diversify the microbiome and increase the most beneficial species, leading to better immune regulation and increasing normalizing levels of crucial neurotransmitters like serotonin and dopamine. Since the growth of medicinal mushroom products is expanding rapidly worldwide, attention is placed on reviewing important aspects of mushroom and mycelium cultivation and quality issues relating to adulteration, substitution, and purity and for maximizing medicinal potency. Common questions surrounding medicinal mushroom products in the marketplace, particularly the healing potential of fungal mycelium compared with fruiting bodies, extraction methods, and the use of fillers in products, are all explored, and many points are supported by the literature.