Critical Reviews in Biotechnology最新文献

Gluconobacter oxydans have been widely used in industrial compound production for their incomplete oxidation ability. However, they are often subjected to a wide variety of severe environmental stresses, such as extreme pH, high temperature, osmotic pressure, and organic solvents, which greatly repress microbial growth viability and productivity. As typical biocatalysis chassis cells with a high tolerance to external environmental stresses, it is extremely important to construct highly tolerant chassis cells and understand the tolerance mechanisms of G. oxydans and how different stresses interact with the cell: membranes, phospholipid bilayers, transporters, and chaperone proteins. In this review, we discuss and summarize the mechanisms of environmental stress tolerance in G. oxydans, and the promising strategies that can be used to further construct tolerant strains are prospected.

Nicotinamide mononucleotide (NMN) presents significant therapeutic potential against aging-related conditions, such as Alzheimer's disease, due to its consistent and strong pharmacological effects. Aside from its anti-aging effect, NMN is also an emerging noncanonical cofactor for orthogonal metabolic pathways in the field of biomanufacturing. This has significant advantages in the field of metabolic engineering, allowing cells to produce unnatural chemicals without disrupting the natural cellular processes. NMN is produced through both the chemical and biological methods, with the latter being more environmentally sustainable. The primary biological production pathway centers on the enzyme nicotinamide phosphoribosyltransferase, which transforms nicotinamide and phosphoribosyl pyrophosphate to NMN. Efforts to increase NMN production have been explored in microorganisms, such as: Escherichia coli, Bacillus subtilis, and yeast, serving as biocatalysts, by rewiring their metabolic processes. Although most researchers are focusing on genetically and metabolically manipulating microorganisms to act as biocatalysts, a growing number of studies on cell-free synthesis are emerging as a promising strategy for producing NMN. This review explores the different biological production techniques of NMN employing microorganisms. This article, in particular, is essential to those who are working on NMN production using microbial strain engineering and cell-free systems.

CRISPR-based diagnostics (CRISPR/Dx) have revolutionized the field of molecular diagnostics. It enables home self-test, field-deployable, and point-of-care testing (POCT). Despite the great potential of CRISPR/Dx in diagnoses of biologically complex diseases, preamplification of the template often is required for the sensitive detection of low-abundance nucleic acids. Various amplification-free CRISPR/Dx systems were recently developed to enhance signal detection at sufficient sensitivity. Broadly, these amplification-free CRISPR/Dx systems are classified into five groups depending on the signal enhancement strategies employed: CRISPR/Cas12a and/or CRISPR/Cas13a are integrated with: (1) other catalytic enzymes (Cas14a, Csm6, Argonaute, duplex-specific nuclease, nanozyme, or T7 exonuclease), (2) rational-designed oligonucleotides (multivalent aptamer, tetrahedral DNA framework, RNA G-quadruplexes, DNA roller machine, switchable-caged guide RNA, hybrid locked RNA/DNA probe, hybridized cascade probe, or "U" rich stem-loop RNA), (3) nanomaterials (nanophotonic structure, gold nanoparticle, micromotor, or microbeads), (4) electrochemical and piezoelectric plate biosensors (SERS nanoprobes, graphene field-effect transistor, redox probe, or primer exchange reaction), or (5) cutting-edge detection technology platforms (digital bioanalysis, droplet microfluidic, smartphone camera, or single nanoparticle counting). Herein, we critically discuss the advances, pitfalls and future perspectives for these amplification-free CRISPR/Dx systems in nucleic acids detection. The continued refinement of these CRISPR/Dx systems will pave the road for rapid, cost-effective, ultrasensitive, and ultraspecific on-site detection without resorting to target amplification, with the ultimate goal of establishing CRISPR/Dx as the paragon of diagnostics.

The development and commercialization of bio-based and biodegradable polyhydroxyalkanoates (PHAs) biopolymers could be crucial for the transition toward a sustainable circular economy. However, despite potential traditional and novel applications in the packaging, textiles, agriculture, automotive, electronics, and biomedical industries, the commercialization of PHAs is limited by their current market competitiveness. This review provides the first critical assessment of the current pure culture pilot-scale PHA literature, which could be crucial in translating promising laboratory-scale developments into industrial-scale commercial PHA production. It will also complement reviews of mixed microbial cultures currently dominating pilot-scale PHA literature. Pure culture fermentations could provide advantages, such as ease of characterizing microbial producers' behavior, higher PHA productivities, and better alignment with existing PHA commercialization and industrial biotechnology approaches. Key aspects, including producer organisms, fermentation volumes and schemes, control schemes, optimization, and properties of the polymers produced, are discussed in-depth, to elucidate important trends, achievements, and knowledge gaps. Furthermore, specific ways for future pilot-scale studies to help address current PHA commercialization challenges are also identified. The insights, and recommendations provided will be extremely beneficial for the future development of PHA production, at both pilot and commercial scales, whilst also being beneficial to the production of other microbial polymers and industrial biotechnology as a whole.

Vibrational spectroscopy is a nondestructive analysis technique that depends on the periodic variations in dipole moments and polarizabilities resulting from the molecular vibrations of molecules/atoms. These methods have important advantages over conventional analytical techniques, including (a) their simplicity in terms of implementation and operation, (b) their adaptability to on-line and on-farm applications, (c) making measurement in a few minutes, and (d) the absence of dangerous solvents throughout sample preparation or measurement. Food safety is a concept that requires the assurance that food is free from any physical, chemical, or biological hazards at all stages, from farm to fork. Continuous monitoring should be provided in order to guarantee the safety of the food. Regarding their advantages, vibrational spectroscopic methods, such as Fourier-transform infrared (FTIR), near-infrared (NIR), and Raman spectroscopy, are considered reliable and rapid techniques to track food safety- and food authenticity-related issues throughout the food chain. Furthermore, coupling spectral data with chemometric approaches also enables the discrimination of samples with different kinds of food safety-related hazards. This review deals with the recent application of vibrational spectroscopic techniques to monitor various hazards related to various foods, including crops, fruits, vegetables, milk, dairy products, meat, seafood, and poultry, throughout harvesting, transportation, processing, distribution, and storage.

Statins are the most prescribed drug for regulating the high cholesterol level in the blood, which can lead to severe complications, such as cardiovascular diseases and other health complications. A wide range of analytical techniques have been employed for the quantification of statins from various origins, including fermentation derived (lovastatin, pravastatin, and compactin), semi-synthetic (simvastatin), and synthetic (atorvastatin, rosuvastatin, and fluvastatin) routes. The presence of more than one structural form and structural analogue generated in the biosynthesis pathway, as well as reaction intermediates and macromolecules in the clinical sample, complicates the quantification of statins. Furthermore, significant concentrations of statins in environmental samples pose serious health and ecology hazards, and estimating statins in those diluted samples is extremely difficult. On the other hand, the: cost, accurate estimation of the desired one from other structural forms, sample complexity, time, limits of detection and quantification, were major criteria distinguishing the usability of each technique. As a result, the current manuscript focuses on analytical techniques such as molecular spectroscopy (normal and derivatives UV-Visible spectrophotometer), chromatography (TLC, HP-TLC, HPLC, GC, swing column, micellar, and supercritical fluid), mass spectroscopy (HPLC-MS/MS and GC-MS/MS), sequential flow injection, capillary electrophoresis, and cyclic voltammetry, as well as their: optimal operating conditions, limits of detection and quantification, advancements, and limitations. Furthermore, various online and offline sample preparations (precipitation, solid phase extraction, liquid-liquid extraction, and micellar extraction) have been highlighted as an essential pretreatment technique to avoid the interference caused by structural analogues and other macromolecules. The greener and more sustainable concepts used in analytical approaches for the quantification statins are also highlighted with current advancements.

Peptide-based medications hold immense potential in addressing a wide range of human disorders and discomforts. However, their widespread utilization encounters two major challenges: preservation and production efficiency. Cyclotides, a class of ribosomally synthesized and post-translationally modified peptides (RiPPs), exhibit unique characteristics, such as a cyclic backbone and cystine knot, enhancing their stability and contributing to a wide range of pharmacological properties exhibited by these compounds. Cyclotides are efficient in the biomedical (e.g., antitumor, antidiabetic, antimicrobial, antiviral) and agrochemical fields by exhibiting activity against pests and plant diseases. Furthermore, their structural attributes make them suitable as molecular scaffolds for grafting and drug delivery. Notably, the mutated variant of kalata B1 cyclotide ([T20K] kalata B1) has recently entered phase 1 of human clinical trials for multiple sclerosis, building upon the success observed in animal trials. To enable large-scale production of cyclotides, it is crucial to further explore their remarkable structural and bioactive properties. This necessitates extensive research focused on enhancing the efficiency of the processes required for their production. This study provides a comprehensive review of the biological synthesis methods of cyclotides, with particular emphasis on various expression systems, namely bacteria, plants, yeast, and cell-free systems. By investigating these expression systems, it becomes possible to design production systems that are adaptable, economically viable, and efficient for generating active and pure cyclotides at an industrial scale. The advantages of biological synthesis over chemical synthesis are thoroughly explored, highlighting the potential of these expression systems in meeting the demands of large-scale cyclotide production.

Agricultural byproducts generally contain abundant bioactive compounds (e.g., cellulose/hemicellulose, phenolic compounds (PCs), and dietary fibers (DFs)), but most of them are neglected and underutilized. Owing to the complicated and rigid structures of agricultural byproducts, a considerable amount of bioactive compounds are entrapped in the polymer matrix, impeding their further development and utilization. In recent years, the prominent performance of cellulolytic fungi to grow and degrade agricultural byproducts has been applied to achieve efficient biotransformation of byproducts to high-value compounds, which is a green and sustainable strategy for the reutilization of agricultural byproducts. This review comprehensively summarizes recent progress in the value-added biotransformation of agricultural byproducts by cellulolytic fungi, including (1) direct utilization of agricultural byproducts for biochemicals and bioethanol production via a consolidated bioprocessing, (2) recovery and biotransformation of bounded PCs from agricultural byproducts for higher bioactive properties, as well as (3) modification and conversion of insoluble DF from agricultural byproducts to produce functional soluble DF. The functional enzymes, potential mechanisms, and metabolic pathways involved are emphasized. Moreover, promising advantages and current bottlenecks using cellulolytic fungi have also been elucidated, shedding further perspectives for sustainable and efficient reutilization of agricultural byproducts by cellulolytic fungi.

Millets, often overlooked as food crops, have regained potential as promising stable food sources of bioactive compounds to regulate blood sugar levels in the diabetic populace. This comprehensive review delves into various millet varieties, processing methods, and extraction techniques aimed at isolating bioactive compounds. The review elucidates the inhibitory effects of millet-derived bioactive compounds on key enzymes involved in carbohydrate metabolism, such as α-amylase and α-glucosidase. It further explores the relationship between the antibacterial activity of phenols, flavonoids, and anthocyanins in millets and their role in amylase inhibition. In particular, phenols, flavonoids, and proteins found in millets play pivotal roles in inhibiting enzymes responsible for glucose digestion and absorption. However, processing methods can either enhance or reduce the bioactive compounds, thereby influencing enzyme inhibition capacity. Studies underscore the presence of phenolic compounds with notable inhibitory activity in: foxtail, finger, barnyard, and pearl millet varieties. Furthermore, extraction techniques, such as Soxhlet and ultrasonic-assisted extraction, emerge as efficient methods for isolating bioactive compounds, thus enhancing their therapeutic efficacy. This review highlights the challenges in preserving the inhibitory activity of millets during processing and optimizing processing methods to ensure better retention of bioactive compounds. It also emphasizes the utilization of millet as a natural dietary supplement or functional food to manage diabetes and promote overall well-being.

Cyanobacteria, the only oxygenic photoautotrophs among prokaryotes, are developing as both carbon building blocks and energetic self-supported chassis for the generation of various bioproducts. However, one of the challenges to optimize it as a more sustainable platform is how to release intracellular bioproducts for an easier downstream biorefinery process. To date, the major method used for cyanobacterial cell lysis is based on mechanical force, which is energy-intensive and economically unsustainable. Phage-mediated bacterial cell lysis is species-specific and highly efficient and can be conducted under mild conditions; therefore, it has been intensively studied as a bacterial cell lysis weapon. In contrast to heterotrophic bacteria, biological cell lysis studies in cyanobacteria are lagging behind. In this study, we reviewed cyanobacterial cell envelope features that could affect cell strength and elicited a thorough presentation of the necessary phage lysin components for efficient cell lysis. We then summarized all bioengineering manipulated pipelines for lysin component optimization and further revealed the challenges for each intent-oriented application in cyanobacterial cell lysis. In addition to applied biotechnology usage, the significance of phage-mediated cyanobacterial cell lysis could also advance sophisticated biochemical studies and promote biocontrol of toxic cyanobacteria blooms.