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Über dieses Buch

This brief provides a broad overview of protein-engineering research, offering a glimpse of the most common experimental methods. It also presents various computational programs with applications that are widely used in directed evolution, computational and de novo protein design. Further, it sheds light on the advantages and pitfalls of existing methodologies and future perspectives of protein engineering techniques.



Chapter 1. World of Proteins: Structure-Function Relationships and Engineering Techniques

Proteins are the key biomolecules in almost all the physiological and pathological processes that are occurring in the cell. Functionality of proteins is related to its conformation, which is ultimately dictated by their unique amino acid sequence. In the current chapter, a brief overview will be provided on the nature of amino acids, structural characteristics and functional versatility of proteins. A glimpse on various interactions present in stabilizing the protein fold and the methods for unraveling their atomic level structures are also discussed. Finally, we present the role of engineered synthetic proteins for the welfare of humanity with huge potential in research and industrial sectors including biotechnological and biomedical fields under the emerging concepts of biomolecular/protein engineering.
Krishna Mohan Poluri, Khushboo Gulati

Chapter 2. Expanding the Synthetic Protein Universe by Guided Evolutionary Concepts

The genetic information content of a cell is maintained by the sequence composition of the DNA. The changes in the nucleotide content will potentially alter its transcriptional and translational events thus influencing the characteristics of the newly synthesized proteins. These nature’s alterations can be helpful in the evolution of proteins with novel/improved functionalities or they can contribute to the pathogenesis with loss of native functionalities. Unraveling the logistics of such a molecular evolutionary process is resourceful to strategically implement it for the benefit of the mankind through laboratory techniques. The laboratory process of synthesizing novel proteins in a constructive way through evolutionary guided principles is called “directed evolution”. This chapter will discuss various techniques, their strengths and pitfalls that are developed under the umbrella of directed evolution scheme.
Krishna Mohan Poluri, Khushboo Gulati

Chapter 3. Rational Designing of Novel Proteins Through Computational Approaches

Advances in the computational power has bestowed us with several structural bioinformatics tools that one can utilize them to predict the structures/structural models of the unknown proteins without performing any experimental work. Computational designing of protein involves identification of protein-sequences and predicting their folding to specific conformations. It is broadly classified into rational design and de novo design. Initial Protein designing approaches were mostly based on the primary sequence composition of the proteins and did not account for specific secondary or tertiary interactions. Now the advent of novel molecular force fields, protein threading algorithms and libraries of amino acid conformations etc., pushed the boundaries of in silico designing methods in obtaining structural design and characterization with greater accuracy. In current chapter, we will discuss several of the rational designing computational tools that are capable of obtaining structures of unknown polypeptide chains and characterizing the functional hotspots, thus aid the researchers in designing novel functional motifs with minimal bench work.
Krishna Mohan Poluri, Khushboo Gulati

Chapter 4. Generating the Fancy Protein Basket with De Novo and Combinatorial Approaches

Under the umbrella of computational techniques, de novo approach occupies a unique role as this method is involved in designing the proteins from scratch. In the first part of the current chapter, we will discuss the principles and applications of the de novo approach along with negative designing technique. In the second part, we will elucidate the combinatorial approach of protein engineering, i.e., a hybrid approach to engineer proteins using both the experimental methods such as directed evolution techniques along with rational and de novo computational techniques. Furthermore, we will discuss various examples that glared the field of protein engineering under combinatorial approach.
Krishna Mohan Poluri, Khushboo Gulati

Chapter 5. Biotechnological and Biomedical Applications of Protein Engineering Methods

The fascinating field of protein engineering has provided breakthroughs by producing plethora of specifically engineered/rationally designed proteins with different functionalities and wide-scale applications in industrial, biotechnological, and pharmaceutical sectors. Many protein based therapeutics, vaccines, and scaffolds with greater safety, improved efficacy, reduced immunogenicity and improved delivery have been designed as novel biomedical formulations. In the present chapter, we will discuss the applications of engineered proteins in diversified fields of biotechnological and biomedical sciences including the areas of industrial, environmental, nanotechnology, biosensors, biomaterials, and biologics etc.
Krishna Mohan Poluri, Khushboo Gulati
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