A5024332257- Enzyme Catalysis and Immobilization
- Enzyme function and inhibition
- Enzyme Production and Characterization
- Pancreatic function and diabetes
- Bacterial Genetics and Biotechnology
- Bacteriophages and microbial interactions
- Microbial Metabolites in Food Biotechnology
- CRISPR and Genetic Engineering
- RNA and protein synthesis mechanisms
- Phytase and its Applications
- Protein Structure and Dynamics
- Electrochemical sensors and biosensors
- Studies on Chitinases and Chitosanases
- Protein purification and stability
- Biochemical and Molecular Research
- Animal Genetics and Reproduction
- Biofuel production and bioconversion
- Enzyme Structure and Function
- RNA modifications and cancer
- Viral Infectious Diseases and Gene Expression in Insects
- Diabetes Treatment and Management
- Protein Hydrolysis and Bioactive Peptides
- Chemical Synthesis and Analysis
- Microbial Metabolic Engineering and Bioproduction
- Gene Regulatory Network Analysis
Jiangnan University
2015-2025
Bacillus subtilis is an all-important Gram-positive bacterium of valuable biotechnological utility that has been widely used to over-produce industrially and pharmaceutically relevant proteins. There are a variety expression systems in terms types transcriptional patterns, among which the auto-inducible growth-phase-dependent promoters gaining increasing favor due their inducer-independent feature, allowing for potential scale-up. To expand applicability system, novel auto-regulatory system...
Promoter evolution by synthetic promoter library (SPL) is a powerful approach to development of functional promoters biology. However, it requires much tedious and time-consuming screenings because the plethora different variants in SPL. Actually, large proportion mutants SPL are significantly lower strength, which contributes only fabrication with continuum strength. Thus, effectively obtain evolved exhibiting higher essential develop novel strategies construct mutant targeting pivotal...
The narrow substrate scope limits the wide industrial application of enzymes. Here, we successfully broadened a nitrile hydratase (NHase) through mutation two tunnel entrance residues based on rational calculation. Two variants, with increased specific activity, especially toward bulky substrates, were obtained. Crystal structure analysis revealed that mutations led to expansion entrance, which might be conducive entry. More importantly, molecular dynamics simulations illustrated introduced...
Cinnamamide and its derivatives are the most common important building blocks widely present in natural products. Currently, nitrile hydratase (NHase, EC 4.2.1.84) has been used large-scale industrial production of nicotinamide acrylamide, while catalytic activity is extremely low or inactive for bulky substrates such as cinnamonitrile. Therefore, beneficial variant βF37P/L48P/F51N were obtained from PtNHase Pseudonocardia thermophila JCM3095 by reshaping substrate access tunnel binding...
Synthetic riboswitches have been increasingly used to control and tune gene expression in diverse organisms. Although a set of theophylline-responsive developed for bacteria, fully functional elements mediated by synthetic Bacillus subtilis are rarely because the host-dependent compatibility between promoters riboswitches.A novel genetic element composed promoter P43 theophylline-riboswitch was characterized B. subtilis. When combined with (P43'-riboE1), successfully switched constitutive...
High thermostability and catalytic activity are key properties for nitrile hydratase (NHase, EC 4.2.1.84) as a well-industrialized catalyst. In this study, rational design was applied to tailor the of NHase from Pseudonocardia thermophila JCM3095 (PtNHase) by combining FireProt server prediction molecular dynamics (MD) simulation. Site-directed mutagenesis non-catalytic residues provided subsequentially performed. The positive multiple-point mutant, namely, M10...
Protein fusion using a linker plays an important role for protein evolution. However, designing suitable linkers evolution is yet challenging and under-explored. To further clarify the regular pattern of type proteins, one nitrile hydratase (NHase) was used as target subunit strategy carried out to improve its efficient catalysis. Subunit-fused variants with three different types were constructed characterized. All exhibited higher stability than that wild type. The longer was, NHase showed,...
Transcription factor (TF)-based biosensors are expected to serve as powerful tools for the high-throughput screening of biocatalytic systems; however, most them respond ligands in a narrow concentration range, which limits their application. In this study, we constructed heterogenous niacin biosensor using repressive TF BsNadR and its target promoters from Bacillus subtilis. The fine-tunable output was expanded wide range concentrations (0–50 mM) through desensitization engineering, suitable...
Summary Nattokinase ( NK ) is an important serine‐protease with direct fibrinolytic activity involving the prevention of cardiovascular disease as antithrombotic agent. Dozens studies have focused on characterization intrinsic novel promoters and signal peptides to secretory production recombinant proteins in Bacillus subtilis . However, genetic elements several drawbacks, which cannot mediate desired level. In this study, elements, were used overproduce , rationally modified B. a stepwise...
Precise regulation of gene expression is fundamental for tailor-made circuit design in synthetic biology. Current strategies this type development are mainly based on directed evolution beginning with a native promoter template. The performances engineered promoters usually limited by the growth phase because only one recognized sigma factor (σ). Here, we constructed multiple-σ recognizable artificial hybrid (AHPs) composed tandems dual and triple natural minimal (NMPs). These NMPs, which...
Isonicotinamide has important roles and application prospects in pharmaceutical, medical, materials sciences.
Pullulanase is a starch-debranching enzyme that hydrolyzes side chain of starch, oligosaccharides and pullulan. Nevertheless, the limited activities pullulanases constrained their practical application. Herein, hyperthermophilic type II pullulanase from Pyrococcus yayanosii CH1 (PulPY2) was evolved by synergistically engineering substrate-binding pocket active-site lids. The resulting mutant PulPY2-M2 exhibited 5-fold improvement in catalytic efficiency (kcat/Km) compared to PulPY2. utilized...
As self-subunit swapping chaperones or metallochaperones, the activators assist nitrile hydratases to take up metal ions and they are essential for active expression of hydratases. Compared with hydratases, have a low sequence identity. Study activation characteristics relationships between structures functions is great significance understanding maturation mechanism hydratase. We co-expressed low-molecular-mass hydratase (L-NHase) from Rhodococcus rhodochrous J1 four heterologous...