Engineering the Methylerythritol Phosphate Pathway and Using a Temporal Promoter for Enhanced Lycopene Production in HY01.

HY01 is a high-yield strain for industrial production of coenzyme Q (Q), indicating its potential for producing other terpenoids. However, the production of Q substantially depletes isoprene precursors, nearly eliminating other terpenoids like spheroidene and spheroidenone commonly found in wild-type . Lycopene was used as an example to demonstrate its potential for terpenoid biosynthesis.

[Developing a curriculum cluster in "Synthetic Biology" to cultivate inter-disciplinary and innovative talents for biomanufacturing].

Synthetic Biology, as an emerging discipline, has gained widespread attention and is developing rapidly, profoundly impacting the fields of life sciences and biotechnology. Concurrently, as emerging engineering education programs take shape, accelerated cultivation of multifaceted innovative talents represents a new mission and imperative for higher education in China. In the context of the flourishing development of Synthetic Biology, East China University of Science and Technology has established a curriculum cluster in Synthetic Biology, focusing on microbiological drug discovery and biomanufacturing.

Unleashing the potential: type I CRISPR-Cas systems in actinomycetes for genome editing.

Covering: up to the end of 2023Type I CRISPR-Cas systems are widely distributed, found in over 40% of bacteria and 80% of archaea. Among genome-sequenced actinomycetes (particularly spp.), 45.

A thermostable type I-B CRISPR-Cas system for orthogonal and multiplexed genetic engineering.

Thermophilic cell factories have remarkably broad potential for industrial applications, but are limited by a lack of genetic manipulation tools and recalcitrance to transformation. Here, we identify a thermophilic type I-B CRISPR-Cas system from Parageobacillus thermoglucosidasius and find it displays highly efficient transcriptional repression or DNA cleavage activity that can be switched by adjusting crRNA length to less than or greater than 26 bp, respectively, without ablating Cas3 nuclease. We then develop an orthogonal tool for genome editing and transcriptional repression using this type I-B system in both thermophile and mesophile hosts.

An in vitro CRISPR-Cas12a-mediated protocol for direct cloning of large DNA fragments.

Large biosynthetic gene cluster (BGC) cloning is important for discovering natural product-based drugs and remains challenging in high GC content microorganisms (e.g., Actinobacteria).

In vitro allosteric transcription factor-based biosensing.

A biosensor is an analytical device that converts a biological response into a measurable output signal. Bacterial allosteric transcription factors (aTFs) have been utilized as a novel class of recognition elements for in vitro biosensing, which circumvents the limitations of aTF-based whole-cell biosensors (WCBs) and helps to meet the increasing requirement of small-molecule biosensors for diverse applications. In this review, we summarize the recent advances related to the configuration of aTF-based biosensors in vitro.

A rapid nucleic acid detection platform based on phosphorothioate-DNA and sulfur binding domain.

Nucleic acid detection plays a key role in diverse diagnosis and disease control. Currently available nucleic acid detection techniques are challenged by trade-offs among speed, simplicity, precision and cost. Here, we described a novel method, designated SENSOR (Sulfur DNA mediated nucleic acid sensing platform), for rapid nucleic acid detection.

Activating cryptic biosynthetic gene cluster through a CRISPR-Cas12a-mediated direct cloning approach.

Direct cloning of biosynthetic gene clusters (BGCs) from microbial genomes facilitates natural product-based drug discovery. Here, by combining Cas12a and the advanced features of bacterial artificial chromosome library construction, we developed a fast yet efficient in vitro platform for directly capturing large BGCs, named CAT-FISHING (CRISPR/Cas12a-mediated fast direct biosynthetic gene cluster cloning). As demonstrations, several large BGCs from different actinomycetal genomic DNA samples were efficiently captured by CAT-FISHING, the largest of which was 145 kb with 75% GC content.

Screening and engineering of high-activity promoter elements through transcriptomics and red fluorescent protein visualization in .

The versatile photosynthetic α-proteobacterium , has recently been extensively engineered as a novel microbial cell factory (MCF) to produce pharmaceuticals, nutraceuticals, commodity chemicals and even hydrogen. However, there are no well-characterized high-activity promoters to modulate gene transcription during the engineering of . In this study, several native promoters from JDW-710 (JDW-710), an industrial strain producing high levels of co-enzyme Q (Q) were selected on the basis of transcriptomic analysis.

Integrating PCR-free amplification and synergistic sensing for ultrasensitive and rapid CRISPR/Cas12a-based SARS-CoV-2 antigen detection.

Antigen detection provides particularly valuable information for medical diagnoses; however, the current detection methods are less sensitive and accurate than nucleic acid analysis. The combination of CRISPR/Cas12a and aptamers provides a new detection paradigm, but sensitive sensing and stable amplification in antigen detection remain challenging. Here, we present a PCR-free multiple trigger dsDNA tandem-based signal amplification strategy and a designed dual aptamer synergistic sensing strategy.

Genome-guided investigation of anti-inflammatory sesterterpenoids with 5-15 trans-fused ring system from phytopathogenic fungi.

Fungal terpenoids catalyzed by bifunctional terpene synthases (BFTSs) possess interesting bioactive and chemical properties. In this study, an integrated approach of genome mining, heterologous expression, and in vitro enzymatic activity assay was used, and these identified a unique BFTS sub-clade critical to the formation of a 5-15 trans-fused bicyclic sesterterpene preterpestacin I (1). The 5-15 bicyclic BFTS gene clusters were highly conserved but showed relatively wide phylogenetic distribution across several species of the diverged fungal classes Dothideomycetes and Sordariomycetes.

Polyketide pesticides from actinomycetes.

Natural product derived pesticides have increased in popularity worldwide because of their high efficacy, eco-friendly nature and favorable safety profile. The development of polyketide pesticides from actinomycetes reflects this increase in popularity in the past decades. These pesticides, which include avermectins, spinosyns, polynactins, tetramycin and their analogues, have been successfully applied in crop protection.

[Exploration of an integrated competency development model for undergraduates training by participating the international Genetic Engineering Machine Competition].

Starting from participating the high-level professional competition, our school has built a talent training system with the spirit of "biomaker" and an innovative practical ability training system. Such system takes the interest of student as the starting point, and relies on the strong scientific research and teaching infrastructure. The programme gives full play to students' initiatives and enhances the scientific research literacy and comprehensive ability of undergraduates majoring in biotechnology.

A versatile biosensing platform coupling CRISPR-Cas12a and aptamers for detection of diverse analytes.

Rapid and sensitive detection of various analytes is in high demand. Apart from its application in genome editing, CRISPR-Cas also shows promises in nucleic acid detection applications. To further exploit the potential of CRISPR-Cas for detection of diverse analytes, we present a versatile biosensing platform that couples the excellent affinity of aptamers for broad-range analytes with the collateral single-strand DNA cleavage activity of CRISPR-Cas12a.

Engineering thermophilic Geobacillus thermoglucosidasius for riboflavin production.

The potential advantages for fermentation production of chemicals at high temperatures are attractive, such as promoting the rate of biochemical reactions, reducing the risk of contamination and the energy consumption for fermenter cooling. In this work, we de novo engineered the thermophile Geobacillus thermoglucosidasius to produce riboflavin, since this bacterium can ferment diverse carbohydrates at an optimal temperature of 60°C with a high growth rate. We first introduced a heterogeneous riboflavin biosynthetic gene cluster and enabled the strain to produce detectable riboflavin (28.

Publisher Correction: Harnessing the intracellular triacylglycerols for titer improvement of polyketides in Streptomyces.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Harnessing the intracellular triacylglycerols for titer improvement of polyketides in Streptomyces.

Pharmaceutically important polyketides such as avermectin are mainly produced as secondary metabolites during the stationary phase of growth of Streptomyces species in fermenters. The source of intracellular metabolites that are funneled into polyketide biosynthesis has proven elusive. We applied multi-omics to reveal that intracellular triacylglycerols (TAGs), which accumulates in primary metabolism, are degraded during stationary phase.

A CRISPR-Cas12a-derived biosensing platform for the highly sensitive detection of diverse small molecules.

Besides genome editing, CRISPR-Cas12a has recently been used for DNA detection applications with attomolar sensitivity but, to our knowledge, it has not been used for the detection of small molecules. Bacterial allosteric transcription factors (aTFs) have evolved to sense and respond sensitively to a variety of small molecules to benefit bacterial survival. By combining the single-stranded DNA cleavage ability of CRISPR-Cas12a and the competitive binding activities of aTFs for small molecules and double-stranded DNA, here we develop a simple, supersensitive, fast and high-throughput platform for the detection of small molecules, designated CaT-SMelor (CRISPR-Cas12a- and aTF-mediated small molecule detector).

Harnessing a previously unidentified capability of bacterial allosteric transcription factors for sensing diverse small molecules in vitro.

A plethora of bacterial allosteric transcription factors (aTFs) have been identified to sense a variety of small molecules. Introduction of a novel aTF-based approach to sense diverse small molecules in vitro will signify a broad series of detection applications. Here, we found that aTFs could interact with their nicked DNA binding sites.

Learn from microbial intelligence for avermectins overproduction.

Microbial strains are amazingly clever by homeostasis of their own survival and optimization for the overproduction of a desired phenotype, for example drugable secondary metabolites through coordination of key genes overexpression and media optimizations. Besides their pesticide activities, avermectins (AVMs) are identified as potent antibiotic agents for a wide range of drug-resistant pathogens by a high-throughput synergy screening strategy. To rewire the genetic circuitry controlling low yields, we summarized the work on balancing the biological chassis with functional parts, and optimized their dynamical process, as well as predicted favorable effective overproduction of AVMs by 5Ms strategy.