Publisher's Synopsis
Protein engineering is the conception and production of unnatural polypeptides, often through modification of amino acid sequences that are found in nature. Synthetic protein structures and functions can now be designed entirely on a computer or produced through directed evolution in the laboratory. In this book, a chronological review of protein engineering methods and applications is provided. A variety of protein engineering applications have been reported in the literature. These applications range from biocatalysis for food and industry to environmental, medical and nanobiotechnology applications. A broad range of topics are covered by providing a solid foundation in protein engineering and supplies readers with knowledge essential to the design and production of proteins. This title presents in-depth discussions of various methods for protein engineering featuring contributions from leading experts from different countries. Many different protein engineering methods are available today, owing to the rapid development in biological sciences, more specifically, recombinant DNA technology. These methods are chronologically reviewed in this book. The most classical method in protein engineering is the so-called "rational design" approach which involves "site-directed mutagenesis" of proteins. Site-directed mutagenesis allows introduction of specific amino acids into a target gene. There are two common methods for site-directed mutagenesis. One is called the "overlap extension" method. This method involves two primer pairs, where one primer of each primer pair contains the mutant codon with a mismatched sequence. These four primers are used in the first polymerase chain reaction (PCR), where two PCRs take place, and two double-stranded DNA products are obtained. Upon denaturation and annealing of them, two heteroduplexes are formed, and each strand of the heteroduplex involves the desired mutagenic codon. DNA polymerase is then used to fill in the overlapping 3' and 5' ends of each heteroduplex and the second PCR takes place using the nonmutated primer set to amplify the mutagenic DNA. Recently, novel types of proteins have been developed, using combinatorial protein engineering techniques. These binding proteins of non-Ig origin are called "affibody binding proteins". With their high affinity, these proteins have been used in many different applications such as diagnostics, bioseparation, functional inhibition, viral targeting, and in vivo tumor imaging or therapy. Inteins are protein splicing elements that are involved in a variety of applications such as protein purification, protein semisynthesis, in vivo and in vitro protein modifications. The use of intein tags for protein purification in plants with high protein production could potentially enable industrial production of pharmaceutically important proteins. The proteolytic cleavage and ligation activities of inteins have been understood, which resulted in novel intein applications in protein engineering, enzymology, microarray production, target detection and transgene activation in plants. The conversion of inteins into molecular switches was introduced by intein-mediated protein attachment to solid supports for microarray and western blot studies and by linking nucleic acids to proteins and controlled splicing. Recent intein-mediated protein engineering applications like protein purification, ligation, cyclization and selenoprotein production have been discussed in detail lately. Applications of protein engineering in enzymatic biofuel cell design is also becoming increasingly important. Particularly, obtaining biofuels from lignocellulosic resources is a challenge, as the enzyme hydrolysis efficiency of lignocellulose is low which increases the costs of biofuels. Thus, protein engineering methods have been used to improve the performance of lignocellulose-degrading enzymes, and biofuels-synthesizing enzymes. Protein engineering is also applied to obtain an efficient electrical communication between biocatalyst(s) and the electrode by rational design and directed evolution, within the frame of biocatalyst engineering. Protein engineering is the conception and production of unnatural polypeptides, often through modification of amino acid sequences that are found in nature. Synthetic protein structures and functions can now be designed entirely on a computer or produced through directed evolution in the laboratory. In this book, a chronological review of protein engineering methods and applications is provided. A variety of protein engineering applications have been reported in the literature. These applications range from biocatalysis for food and industry to environmental, medical and nanobiotechnology applications. A broad range of topics are covered by providing a solid foundation in protein engineering and supplies readers with knowledge essential to the design and production of proteins. This title presents in-depth discussions of various methods for protein engineering featuring contributions from leading experts from different countries. Many different protein engineering methods are available today, owing to the rapid development in biological sciences, more specifically, recombinant DNA technology. These methods are chronologically reviewed in this book. The most classical method in protein engineering is the so-called "rational design" approach which involves "site-directed mutagenesis" of proteins. Site-directed mutagenesis allows introduction of specific amino acids into a target gene. There are two common methods for site-directed mutagenesis. One is called the "overlap extension" method. This method involves two primer pairs, where one primer of each primer pair contains the mutant codon with a mismatched sequence. These four primers are used in the first polymerase chain reaction (PCR), where two PCRs take place, and two double-stranded DNA products are obtained. Upon denaturation and annealing of them, two heteroduplexes are formed, and each strand of the heteroduplex involves the desired mutagenic codon. DNA polymerase is then used to fill in the overlapping 3' and 5' ends of each heteroduplex and the second PCR takes place using the nonmutated primer set to amplify the mutagenic DNA. Recently, novel types of proteins have been developed, using combinatorial protein engineering techniques. These binding proteins of non-Ig origin are called "affibody binding proteins". With their high affinity, these proteins have been used in many different applications such as diagnostics, bioseparation, functional inhibition, viral targeting, and in vivo tumor imaging or therapy. Inteins are protein splicing elements that are involved in a variety of applications such as protein purification, protein semisynthesis, in vivo and in vitro protein modifications. The use of intein tags for protein purification in plants with high protein production could potentially enable industrial production of pharmaceutically important proteins. The proteolytic cleavage and ligation activities of inteins have been understood, which resulted in novel intein applications in protein engineering, enzymology, microarray production, target detection and transgene activation in plants. The conversion of inteins into molecular switches was introduced by intein-mediated protein attachment to solid supports for microarray and western blot studies and by linking nucleic acids to proteins and controlled splicing. Recent intein-mediated protein engineering applications like protein purification, ligation, cyclization and selenoprotein production have been discussed in detail lately. Applications of protein engineering in enzymatic biofuel cell design is also becoming increasingly important. Particularly, obtaining biofuels from lignocellulosic resources is a challenge, as the enzyme hydrolysis efficiency of lignocellulose is low which increases the costs of biofuels. Thus, protein engineering methods have been used to improve the performance of lignocellulose-degrading enzymes, and biofuels-synthesizing enzymes. Protein engineering is also applied to obtain an efficient electrical communication between biocatalyst(s) and the electrode by rational design and directed evolution, within the frame of biocatalyst engineering. Protein engineering is the conception and production of unnatural polypeptides, often through modification of amino acid sequences that are found in nature. Synthetic protein structures and functions can now be designed entirely on a computer or produced through directed evolution in the laboratory. In this book, a chronological review of protein engineering methods and applications is provided. A variety of protein engineering applications have been reported in the literature. These applications range from biocatalysis for food and industry to environmental, medical and nanobiotechnology applications. A broad range of topics are covered by providing a solid foundation in protein engineering and supplies readers with knowledge essential to the design and production of proteins. This title presents in-depth discussions of various methods for protein engineering featuring contributions from leading experts from different countries. Many different protein engineering methods are available today, owing to the rapid development in biological sciences, more specifically, recombinant DNA technology. These methods are chronologically reviewed in this book. The most classical method in protein engineering is the so-called "rational design" approach which involves "site-directed mutagenesis" of proteins. Site-directed mutagenesis allows introduction of specific amino acids into a target gene. There are two common methods for site-directed mutagenesis. One is called the "overlap extension" method. This method involves two primer pairs, where one primer of each primer pair contains the mutant codon with a mismatched sequence. These four primers are used in the first p