A novel fibrinolytic enzyme from Fusarium sp. CPCC 480097, named Fu-P, was purified to electrophoretic homogeneity using ammonium sulfate precipitation and ion exchange and gel filtration chromatography. Fu-P, a single protein had a molecular weight of 28 kDa, which was determined by SDS-PAGE and gel filtration chromatography. The isoelectric point of Fu-P determined by isoelectric focusing electrophoresis (IEF) was 8.1, and the optimum temperature and pH value were 45 degrees C and 8.5, respectively. Fu-P cleaved the alpha-chain of fibrin (ogen) with high efficiency, and the beta-chain and gamma-gamma (gamma-)-chain with lower efficiency. Fu-P activity was inhibited by EDTA and PMSF, and the enzyme exhibited a high specificity for the chymotrypsin substrate S-2586. Fu-P was therefore identified as a chymotrypsin-like serine metalloprotease. The first 15 amino acids of the N-terminal sequence of Fu-P were Q-A-S-S-G-T-P-A-T-I-R-V-L-V-V and showed no homology with that of other known fibrinolytic enzymes. This protease may have potential applications in thrombolytic therapy and in thrombosis prevention.

Fermentation extracts from 1,075 endophytic fungi were screened by the Markwardt method. The endophytic strain CPCC 480097 had the strongest antithrombotic activity and was identified as Fusarium sp. based on morphologic tests and internal transcriptional spacer sequence analysis. The target of the antithrombotic agent from the endophytic strain CPCC 480097 was identified by analysis of the fibrinogen clotting time, amidolytic activity, and fibrinolytic assay. The results showed that this antithrombotic agent is a 28-kDa single-chain fibrinolytic enzyme. The identification of this fibrinolytic enzyme was performed by liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS). The two internal sequences of this enzyme were obtained, and these showed no homology with those of other known fibrinolytic enzymes.

Biosensors are a class of smart devices fabricated for target analyte detection. A biosensor is commonly made of three basic components: a specific bioreceptor, a physicochemical transducer, and a signal processing device. The bioreceptor part features one of the more crucial technologies of a biosensor to perform specific detection of analytes. They commonly make use of various molecular recognition elements, such as enzymes, natural receptors, antibodies, nucleic acid aptamers, and even synthetic receptors. Then, the molecular recognition events of these bioreceptors can be translated into readable signals in various forms like electrochemical, optical, piezo/pyro electric, etc. Finally, these signals are mathematically processed for quantitative analysis. Nowadays, the great advances in biomimetic materials, in particular the synthetic receptors based on molecularly imprinted polymers (MIPs), promote tremendous development of biosensors. Integration of this field with artificial intelligence enables emerging design of advanced biosensors, which has moved from concept to implementation, meanwhile facing new opportunities and challenges. With no doubt, bioreceptor innovation based on molecular imprinting is an emerging driving force for biosensors development. In this review paper, we provide an overview of MIP‐based biosensors, and also their challenges and opportunities moving forward toward wearable devices are discussed.

Carcinoembryonic antigen (CEA) is an important broad-spectrum tumor marker. For CEA detection, a novel type of metal–organic framework (MOF) was prepared by grafting CEA aptamer-incorporated DNA tetrahedral (TDN) nanostructures into PCN-222 (Fe)-based MOF (referred as CEAapt-TDN-MOF colloid nanorods). The synthesized CEAapt-TDN-MOF is a very stable detection system due to the vertex phosphorylated TDN structure at the interface, possessing a one-year shelf-life. Moreover, it exhibits a significant horseradish peroxidase mimicking activity due to the iron porphyrin ring, which leads to a colorimetric reaction upon binding toward antibody-captured CEA. Using this method, we successfully achieved the highly specific and ultra-sensitive detection of CEA with a limit of detection as low as 3.3 pg/mL. In addition, this method can detect and analyze the target proteins in clinical serum samples, effectively identify the difference between normal individuals and patients with colon cancer, and provide a new method for the clinical diagnosis of tumors, demonstrating a great application potential.