In recent years, the development of chemical biology has been increasing focusing on efficient and mild methods for chemo-selective ligations and site-specific proteins labeling and modifications. These methods have ability to obtain large amount of post translational modifications and artificial proteins, which could not be acquired by using traditional gene cloning and recombinant protein expression strategy. In later 1990s, a new transpeptidase Sortase A was isolated from Staphylococcus Aureus, which can be used to modify proteins bearing a short recognition sequence (most usually as LPXTG or LPAAG). The active-site Cys residue of Sortase A cleaves between LPXT and G residue to produce a thioester intermediate, which can reacts with a nucleophile containing one to five Gly to afford the ligation product. Base on above-mentioned reason, the Sortase-mediated ligation has been successfully applied to many fields such as C-terminals protein modification, labeling and protein semisynthesis with high efficiency recently. Compared to traditional chemical synthesis,Sortase catalyzed semi-chemical synthesis method can preferably address the size problem of protein chemical synthesis. This mini review reports and discusses the recent important development of protein ligations, labeling and modifications by using Sortase mediated ligation method.

Contents
1 Introduction
2 The structure of Sortase and the mechanism of the transpeptidation reaction catalyzed by Sortase
3 Sortases as tools for protein ligations and modifications
3.1 Validation of the in vitro Sortase activity
3.2 Seek for the mutants of Sortase
3.3 Immobilization of Sortase
4 Application of Sortase to protein ligations and modifications
4.1 C terminal modifications
4.2 N terminal modifications
4.3 C and N terminal modifications
4.4 Preparation of unnatural N-N/C-C chimeric proteins by using Sortase
4.5 Protein immobilization
4.6 Preparation of cyclopeptides
4.7 Sortase-tag expressed protein ligation reaction
4.8 Other applications
5 Conclusion and outlook

Both physiological behaviors and pathological processes of living organisms are related to the synergistic co-action of various enzymes. Inspired by this, a new bionic catalytic technology, immobilized multi-enzyme cascade reactor has been proposed for accelerating the bio-reactions including synthesis, substrate degradation, transformation and recombination. By taking advantages of the good stability, reusability and remarkable high efficiency, the immobilized multi-enzyme cascade reactor has attracted more and more attention in many fields including biological sensing, simulation biology and biological transformation and so on. This review focuses on the art-of-state and progress of immobilized multi-enzyme cascade reactor in recent years. The fundamentals, preparation approaches, advantages, factors affecting the efficiency, and applications of the technology are involved. Its trends in future study are also prospected.

Contents
1 Introduction
2 Current application of enzymes
3 Enzyme modification
3.1 Directed evolution
3.2 Glycosylation
3.3 Chemical modification
4 Catalytic engineering of enzymes
4.1 Medium engineering
4.2 Immobilization
4.3 Multi-enzyme complex
5 Conclusion

Contents
1 Introduction
2 Classification and detecting principle of glucose sensors
2.1 Glucose oxidase glucose sensors
2.2 Nonenzymatic glucose sensors
3 Research advances of nonenzymatic glucose sensors
3.1 Nonenzymatic glucose sensors based on metal and metal oxides
3.2 Nonenzymatic glucose sensors based on bimetal
3.3 Nonenzymatic glucose sensors based on carbon materials
4 Conclusion and outlook

Duplex-specific nuclease (DSN) is a type of nuclease, that is isolated from the hepatopancreas of the Kamchatka crab. DSN displays a strong preference for cleaving double-stranded DNA or DNA in DNA-RNA heteroduplexes, and is practically inactive toward single-stranded DNA or RNA. Moreover, this enzyme shows excellent discrimination capability between perfectly and imperfectly matched (up to one mismatch) short duplexes. Owing to its unique feature of cleaving DNA, DSN enzyme is widely applied in the fields of biomedicine and molecular biology, including full-length cDNA library normalization,genomic single-nucleotide polymorphism (SNP) detection and high throughput sequencing. The recent research on DSN are mainly focused on the applications in microRNAs (miRNAs) detection using a DSN-mediated signal amplification strategy. miRNAs are a group of short, endogenous, noncoding RNAs that play vital regulatory roles in physiologic and pathologic processes, including hematopoietic differentiation, cell cycle, regulation, and metabolism. So miRNA is one of the most important biomarkers in individualized treatment, which has great value in terms of improving the diagnosis and treatment of diseases. However, detection of miRNAs is challenging owing to their unique characteristics, including a small size, sequence homology among family members, low abundance in total RNA samples, and susceptibility to degradation in solution. In recent years, isothermal signal amplification and detection of trace miRNA in fluids are reported by many researchers using DSN-mediated biosensors. According to different detection principle of biosensors, DSN-mediated biosensors can be classified as colorimetric, fluorescent, and electrochemical. In this review, we intensively summarize the advantages of DSN in miRNAs detection using DSN-based signal amplification strategy, expanded applications in SNP detection, high throughput sequencing and cDNA library normalization especially when being combined with SMART (Switching Mechanism At 5'end of RNA Transcript) technique.

Contents
1 Introduction
2 The application of DSN enzyme in microRNA detection
2.1 Colorimetric analyses
2.2 Fluorescent analyses
2.3 Electrochemical analyses
3 The application of DSN enzyme in SNP detection
4 Other biological applications of DSN enzyme
4.1 Construction of cDNA Library
4.2 High throughput sequencing
5 Conclusion and outlook

Contents
1 Introduction
2 Research overview of nitrilase
3 Existence range of nitrilase
4 Type of nitrilase catalysts and obtaining manners
4.1 Wild enzyme
4.2 Genetically engineered enzyme
5 The applications in the synthesis of pharmaceutical intermediates
5.1 Picolinic acid
5.2 (R)-Mandelic acid and its derivatives
5.3 Cyanocarboxylic acid
5.4 Pharmaceutical amino acid
5.5 Glycolic acid
6 Conclusion and outlook

Contents
1 Introduction
2 Research of enzyme immobilization on carbon nanotubes
2.1 Hydrolase immobilization on carbon nanotubes
2.2 Oxido-reductase immobilization on carbon nanotubes
2.3 Other kinds of enzymes immobilization on carbon nanotubes
2.4 The application of immobilized enzymes in the biosensors
3 Conclusions

Contents
1 Introduction
2 Relationship between sialic acids and cancers
3 Design and discovery of sialyltransferase inhibitors
3.1 Donor-analog inhibitors
3.2 Acceptor-analog inhibitors
3.3 Bisubstrate-analog inhibitors
3.4 Transition-state-analog of the sialyl donor inhibitors
3.5 Other types of inhibitors
4 Conclusion

Contents
1 Introduction
2 Mechanisms of hydrolases
3 Hydrolases containing no metal ions
3.1 Reuse of natural proteins
3.2 Reconstruction of natural proteins
4 Hydrolases containing metal ions
4.1 Molecular design based on 3-α-helix
4.2 Molecular design based on 4-α-helix
4.3 Molecular design based on zinc-finger protein
5 Hydrolases containing metal complex
5.1 Explore hydrolysis activity of heme proteins
5.2 Tune hydrolysis activity of heme proteins
6 Conclusion and outlook

Contents
1 Introduction
2 Effects of surfactant on enzymatic hydrolysis of pure cellulose and mechanism
2.1 Factors influencing the action of surfactant
2.2 Mechanism for the action of surfactant on pure cellulose hydrolysis
3 Effects of surfactant on enzymatic hydrolysis of pretreated lignocellulose
3.1 Effects of substrate structural features
3.2 Effects of hydrolysis conditions
4 Mechanisms for the action of surfactant on lignocellulose hydrolysis
4.1 Effects of surfactant on substrate structure
4.2 Effects of surfactant on enzyme stability
4.3 Effects of surfactant on the interaction between enzyme and substrate
5 Conclusion

Contents
1 Introduction
2 Reaction types catalyzed by peptide-based artificial enzyme
2.1 Hydrolysis reaction
2.2 Aldol reaction
2.3 Redox reaction
3 Influence factors on the activity of peptide-based artificial enzyme
3.1 Assembly degree
3.2 Microstructure
3.3 Supramolecular structrue
3.4 Active-site microenvironment
3.5 pH
4 Application of peptide-based artificial enzyme
5 Conclusion

The enzymatic probe based on nano-bioconjugate have been studied extensively for many years for their novel electronic, photonic and electrochemical properties. This review summarizes major advances in the gold nanoparticles(GNPs) biocongjugate probes to detect enzyme activity. This is followed by a discussion of the assays in colorimetric, fluorescence and other assays. Throughout the review, a detailed explanation of the unique designs will be presented, and the benefits and shortcomings of these approaches will be highlighted. The review concludes with a brief perspective on future research directions, and remaining barriers that must be overcome for the successful application of these technologies.

Contents
1 Introduction
2 Principle of enzyme activity detection based on GNPs
2.1 Colorimetric assays
2.2 FRET-based assays
2.3 Other assays
3 Neglected enzymes in the field of sensors
4 Conclusion

The telomerase ribonucleoprotein is a reverse transcriptase that plays a critical role in the maintenance of telomere length by synthesizing telomeric DNA repeats using its instinct RNA as the template and thus protects chromosome ends from degradation and fusion. Most normal somatic cells do not express detectable telomerase activity and telomeres shorten progressively with each cell division, while in almost all kinds of cancer cells human telomerase exhibits high activity, suggesting that there is a direct association between telomerase activity and various diseases including cancer and age-related diseases, and that telomerase could be used both as a diagnostic biomarker for the early detection of human cancer and as a potential target for anti-cancer therapy. However, fully understanding the biochemical action of human telomerase is presently a huge challenge since it is a very big complex. Moreover, it is remarkably difficult to get enough amount of heloenzyme for traditional enzyme analysis. Novel and advanced methods therefore should be employed to assay human telomerase and understand enzymatication. Single molecule techniques such as single-molecule fluorescence resonance energy transfer (smFRET) technique and two-color coincidence detection (TCCD) not only can detect dynamic of individual molecule and flexibility of enzyme catalysis, but also can check very little amount of samples in solution or on surface, which can not be performed by bulk experiment. In this review, we summarize the progress and prospect of telomerase research and describe several latest single-molecule methods applied to assaying telomerase structure, function, activity and dynamics.

Contents
1 Introduction
2 Single molecule fluorescence technique
2.1 Development of single molecule fluorescence techniques
2.2 Classification of single molecule fluorescence techniques
2.3 Common single molecule fluorescence techniques
3 Telomere, telomerase and cancer
3.1 Structure and function of telomere and telomerase
3.2 Relationship between telomere, telomerase and cancer
3.3 Telomerase as a diagnostic biomarker and therapeutic target for cancer
4 Single molecule fluorescence studies of human telomerase
4.1 Common methods for telomerase detection
4.2 Single molecule fluorescence techniques applied to telomerase detection
5 Conclusion

Histone deacetylase inhibitor is a novel class of histone deacetylase targeted anti-cancer drug, which takes excellent effects on anti-proliferation, pro-apoptosis, pro-differentiation, cell cycle arrest, anti-angiogenesis and so on. Histone deacetylase inhibitor plays an important role in the development of anticancer drug with its unique anti-tumor mechanism. Among the histone deacetylase inhibitors, the cyclopeptide histone deacetylase inhibitor with the most complicated structure has good antagonism on various types of solid tumors and hematologic cancer. In this review, the structural features of the metal binding region, the surface recognition region and the linker region of natural and synthetic cyclic peptide histone deacetylase inhibitors are summarized, and the inhibitory activity and anti-tumor proliferative activity of various inhibitors are described. Modification of different structure domains can make inhibitors with high efficiency and specificity on different tumor cells. We expect to find the structure rule of the high efficient and low toxic targeted peptide inhibitors through structure-activity relationships, providing new sights for investigation of anti-cancer drugs.

Contents
1 Introduction
2 Histone deacetylase inhibitors and classification
3 Cyclopeptide histone deacetylase inhibitors
3.1 Cyclopeptide histone deacetylase inhibitors containing ketone
3.2 Cyclopeptide histone deacetylase inhibitors containing hydroxamic acid
3.3 Cyclopeptide histone deacetylase inhibitors containing sulfur
3.4 Cyclopeptide histone deacetylase inhibitors containing carboxylic acid or amide
3.5 Other histone deacetylase inhibitors
4 Conclusion