A rapid and ultrasensitive CRISPR/Cas12a-based assay for the accurate identification of T-even type phages.

Phage contamination poses a significant threat to industrial fermentation, leading to substantial economic losses. Virulent T-even type phages (T2/T4/T6) represent particularly concerning biological hazards in fermentation systems. This paper developed a novel CRISPR/Cas12a-based system integrated with recombinase polymerase amplification (RPA), enabling ultrasensitive identification of T-even type phages.

Temperature-Modulation Categorization of Argonaute-Based Bioreactions Coupled with Amplification Methods for Nucleic Acid Detection.

Argonaute (Ago), a programmable nuclease, has recently emerged as a promising tool for nucleic acid detection. To achieve higher sensitivity, a nucleic acid amplification step is often introduced into the Ago-based detection system, leading the process suffered from the issues of temperature compatibility and device complexity. To address these limitations, integrating multiple reactions within a portable temperature control device shows significant potential to streamline the development and practical application of these systems.

Reusable combinatorial libraries with high diversity for efficient multi-gene expression optimization in Escherichia coli.

Efficient multi-gene expression in Escherichia coli is critical for advancing metabolic engineering and synthetic biology. However, existing strategies for combinatorial optimization remain labor-intensive and low-throughput. In addressing this challenge, a high-throughput platform was developed, encompassing the engineering of standardized genetic elements (promoters and 5' UTRs) with fluorescent reporters (e.

Efficient whole-cell biosynthesis of S-adenosyl-L-methionine by the engineered with high ATP regenerating system.

S-adenosyl-L-methionine (SAM) is an important intermediate metabolite and widely used in the treatment of liver disease, arthritis, and depression. In this work, a whole-cell catalysis strategy was employed to enhance SAM production by combining the SAM biosynthetic pathway with an adenosine triphosphate (ATP) regeneration system in Specifically, the , , and genes were previously introduced into the genome of the host strain. We then confirmed the availability of the ATP regeneration system under the condition of adding adenosine monophosphate (AMP) and acetyl phosphate (ACP) as supplements.

Enhancing terpenoid production in Saccharomyces cerevisiae via cell morphology engineering.

There is a growing interest in establishing yeast as a cell factory for terpenoid production. Given the hydrophobicity of many terpenoids, expanding the intracellular storage pool is an effective strategy to improve titers. In this study, the storage capacity was enhanced by increasing yeast cell size through the knockout of EST1 and SSN8.

Highly efficient loop cleavage for human papillomavirus detection with a novel thermophilic Argonaute from Thermus brockianus.

Argonaute proteins (Agos), endowed with the capacity to cleave DNA or RNA under the guidance of small nucleic acid guides, have emerged as versatile biotechnological tools. This study endeavored to characterize a novel thermophilic Argonaute protein from Thermus brockianus (TbAgo), revealing its proficiency as a DNA-guided DNA endonuclease. Demonstrating high catalytic efficiency and precision at 65 °C, TbAgo possessed compatibility with loop-mediated isothermal amplification (LAMP) method, whose optimal temperature is also around 65 °C.

DNAzyme-Triggered Equilibrium Transfer with Self-Activated CRISPR-Cas12a Biosensor Enables One-Pot Diagnosis of Nucleic Acids.

Integrating recombinase-polymerase amplification (RPA) with CRISPR-Cas12a holds significant potential to simplify and improve nucleic acid diagnostic procedures. However, current strategies face limitations, such as complexity, reduced efficiency, and potential compromises in Cas12a activity. In response, we developed a NAzyme-triggered quilibrium transfer with a elf-activated SPR-Cas12a iosnso (DESCRIBER) for integrated nucleic acid detection.

Transportation engineering for enhanced production of plant natural products in microbial cell factories.

Plant natural products (PNPs) exhibit a wide range of biological activities and have essential applications in various fields such as medicine, agriculture, and flavors. Given their natural limitations, the production of high-value PNPs using microbial cell factories has become an effective alternative in recent years. However, host metabolic burden caused by its massive accumulation has become one of the main challenges for efficient PNP production.

Intein-mediated temperature control for complete biosynthesis of sanguinarine and its halogenated derivatives in yeast.

While sanguinarine has gained recognition for antimicrobial and antineoplastic activities, its complex conjugated structure and low abundance in plants impede broad applications. Here, we demonstrate the complete biosynthesis of sanguinarine and halogenated derivatives using highly engineered yeast strains. To overcome sanguinarine cytotoxicity, we establish a splicing intein-mediated temperature-responsive gene expression system (SIMTeGES), a simple strategy that decouples cell growth from product synthesis without sacrificing protein activity.

Metabolic engineering of Pichia pastoris for overproduction of cis-trans nepetalactol.

Monoterpene indole alkaloids (MIAs) are a group of plant-derived natural products with high-value medicinal properties. However, their availability for clinical application is limited due to challenges in plant extraction. Microbial production has emerged as a promising strategy to meet the clinical demands for MIAs.

Development of a Pichia pastoris cell factory for efficient production of germacrene A: a precursor of β-elemene.

β-Elemene, an active ingredient found in medicinal plants like turmeric and zedoary, is a sesquiterpene compound with antitumor activity against various cancers. However, its current mode of production through plant extraction suffers from low efficiency and limited natural resources. Recently, there has been an increased interest in establishing microbial cell factories to produce germacrene A, which can be converted to β-elemene by a one-step reaction in vitro.

Efficient production of l-glutathione by whole-cell catalysis with ATP regeneration from adenosine.

l-glutathione (GSH) is an important tripeptide compound with extensive applications in medicine, food additives, and cosmetics industries. In this work, an innovative whole-cell catalytic strategy was developed to enhance GSH production by combining metabolic engineering of GSH biosynthetic pathways with an adenosine-based adenosine triphosphate (ATP) regeneration system in Escherichia coli. Concretely, to enhance GSH production in E.

Characterization and engineering of peroxisome targeting sequences for compartmentalization engineering in Pichia pastoris.

Peroxisomal compartmentalization has emerged as a highly promising strategy for reconstituting intricate metabolic pathways. In recent years, significant progress has been made in the peroxisomes through harnessing precursor pools, circumventing metabolic crosstalk, and minimizing the cytotoxicity of exogenous pathways. However, it is important to note that in methylotrophic yeasts (e.

Decoupling tourism growth from carbon emissions: Evidence from Chengdu, China.

The fast growth increases in energy use and greenhouse gas emissions in China's tourism industry. This article assessed the carbon emissions of tourist Chengdu's traffic patterns and how they are expected to change between 2005 and 2021 and explains why tourism carbon decoupling is important for sustainable tourism development. In order to investigate how carbon emissions from tourism may expand without necessarily increasing in tandem with GDP and the variables that influence it, the Tapir model and the LMDI (logarithmic mean Divisia index) method were used.

De Novo Biosynthesis of Vindoline and Catharanthine in .

Vinblastine has been used clinically as one of the most potent therapeutics for the treatment of several types of cancer. However, the traditional plant extraction method suffers from unreliable supply, low abundance, and extremely high cost. Here, we use synthetic biology approach to engineer for de novo biosynthesis of vindoline and catharanthine, which can be coupled chemically or biologically to vinblastine.

Magnetofluid-integrated biosensors based on DNase-dead Cas12a for visual point-of-care testing of HIV-1 by an up and down chip.

The CRISPR Cas system, as a novel nucleic acid detection tool, is often hindered by cumbersome experimental procedures, complicated reagent transfer processes, and associated aerosol pollution risks. In this study, an integrated nucleic acid detection platform named "up and down chip" was developed, which combined RT-RAA technology for nucleic acid amplification, DNase-dead Cas12a-modified magnetic beads for specific recognition of target nucleic acid, and HRP-TMB chromogenic reaction for signal output in different chambers of a single microfluidic chip. The magnetic beads were migrated in an up-and-down manner between different chambers through magnetic driving, achieving a "sample-in, result-out" detection mode.

Retron-mediated multiplex genome editing and continuous evolution in Escherichia coli.

While there are several genome editing techniques available, few are suitable for dynamic and simultaneous mutagenesis of arbitrary targeted sequences in prokaryotes. Here, to address these limitations, we present a versatile and multiplex retron-mediated genome editing system (REGES). First, through systematic optimization of REGES, we achieve efficiency of ∼100%, 85 ± 3%, 69 ± 14% and 25 ± 14% for single-, double-, triple- and quadruple-locus genome editing, respectively.

The greenhouse effect of antibiotics: The influence pathways of antibiotics on methane release from freshwater sediment.

The impact of antibiotics on methane (CH) release from sediment involves both CH production and consumption processes. However, most relevant studies lack a discussion of the pathways by which antibiotics affect CH release and do not highlight the role played by the sediment chemical environment in this influence mechanism. Here, we collected field surface sediments and grouped them with various antibiotic combination concentration gradients (50, 100, 500, 1000 ng g) under a 35-day indoor anaerobic constant temperature incubation.

CRISPR Cas12a-enabled biosensors coupled with commercial pregnancy test strips for the visible point-of-care testing of SARS-CoV-2.

The rapid spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has promoted the development of nucleic acid diagnosis technology. Several platforms with isothermal amplification methods have achieved sensitive and specific detection of SARS-CoV-2. However, they still suffer from complicated operations, delicate instruments, and unintuitive signal output modes.

Ecotoxicological effects of soil lithium on earthworm Eisenia fetida: Lethality, bioaccumulation, biomarker responses, and histopathological changes.

Lithium is an emerging environmental contaminant in the current low-carbon economy, but little is known about its influences on soil invertebrates. In this work, earthworm Eisenia fetida was exposed to soils treated with different levels of lithium for 7 d, and multiple ecotoxicological parameters were evaluated. The results showed that mortality was dose-dependent and lithium's median lethal content (LC) to earthworm was respectively 865.

Metabolic Engineering of for the Production of Triacetic Acid Lactone.

Triacetic acid lactone (TAL) is a promising renewable platform polyketide with broad biotechnological applications. In this study, we constructed an engineered strain for the production of TAL. We first introduced a heterologous TAL biosynthetic pathway by integrating the 2-pyrone synthase encoding gene from ().