Metabolic engineering has been developed for nearly 20 years since its beginning on 1990s, and it has significantly promoted the improvement of microbial fermentation industry. This review summarized the technology development and their applications in fermentation industry in each of the three important phases during the development of metabolic engineering. Finally, the key issues for future development and solving strategies were discussed.
Biotechnology ; trends ; DNA, Recombinant ; genetics ; Genetic Engineering ; methods ; Industrial Microbiology ; methods ; Metabolism ; Protein Engineering

Databases, Protein ; Information Dissemination ; Internet ; Protein Engineering ; methods ; Protein Structure, Tertiary ; Recombinant Proteins ; biosynthesis ; chemistry

Terpenoids are present in all organisms but are especially abundant in plants, with more than 30,000 compounds. Not only do they play an important role in the life of plant, but also have high commercial values. However, the content of many important terpenoids in plant is very low. Therefore, how to improve the inefficient production of terpenoids is an urgent task. Metabolic engineering has been one of the most potential technologies to improve terpenoids production in recent years, following the study of metabolic pathway and regulation mechanism of terpenoids. Although there are some breakthroughs, metabolic engineering of terpenoids is still full of challenges because of the lack of knowledge on metabolic control of most terpenoids. Functional genomics approaches, including transcriptomics, proteomics and metabolomics, are potential tools for exploring of metabolic engineering. Integrating transcriptomics and metabolomics is an effective way to discover new genes involved in metabolic pathway. In this paper, the representative research outcomes about the metabolic engineering of terpenoids in plant were reviewed concisely and then the application of functional genomics approaches to study metabolic pathway and regulation mechanism of terpenoids and the strategies for metabolic engineering of terpenoids were discussed.
Genomics ; methods ; Metabolomics ; methods ; Plants ; metabolism ; Protein Engineering ; methods ; Proteomics ; methods ; Terpenes ; metabolism

Self-assembly of peptides is ubiquitous in the body of creatures. The molecules of peptides combine with each other to form proteins with different functions through self-assembly. The formation of a specific conformation of one type of protein is owing to the self-assembly of its compositive amino acids. So, researchers can design self-assembly of peptides at the molecular level and can control its formation and configuration. It has the potential for application in the preparation of new medicines and biomaterials. In recent years, self-assembling peptides have been increasingly high-lighted and used to simulate the function of natural biomolecules, to synthesize peptide-medicine, and to serve as the carriers of medicine.
Biocompatible Materials ; chemical synthesis ; Molecular Conformation ; Nanotechnology ; methods ; Peptides ; chemistry ; Protein Engineering ; methods

As an efficient and promising protein engineering strategy, directed evolution includes the construction of mutant libraries and screening of desirable mutants. A rapid and high-throughput screening method has played a critical role in the successful application of directed evolution strategy. We reviewed several high-throughput screening tools which have great potential to be applied in directed evolution. The development of powerful high-throughput screening tools will make great contributions to the advancement of protein engineering.
Directed Molecular Evolution ; methods ; High-Throughput Screening Assays ; methods ; Mutagenesis, Site-Directed ; methods ; Mutant Proteins ; genetics ; Protein Engineering ; methods

Hepcidin is a small cystein-rich cationic peptide produced mainly by the liver. It was initially isolated from human plasma and exhibited antimicrobial activity. Recently, several lines of evidence have suggested that hepcidin is a key regulator of iron metabolism at the whole body level and is relative to inflammation, infection, hypoxia and anemia. Hepcidin, is implicated in duodenal iron absorption and iron mobilization from reticuloendothelial macrophages. The major mechanism of hepcidin function seems to be the regulation of transmembrane iron transport. As both iron deficiency and iron excess are associated with cellular dysfunction, so hepcidin or hepcidin-related therapeutics could find a place in the treatment of various diseases such as hemochromatosis and anemia of chronic disease. To elucidate biological function of hepcidin further and use it for other research, it is necessary to produce enough hepcidin through DNA recombinant technique. As a highly successful system for the production of a variety of heterologous proteins, the methylotrophic Pichia pastoris system has the probability for a high level production of hepcidin. The subject of this paper is to summarize the regulation of hepcidin gene expression and the understanding of functions of hepcidin. At last, giving a prospect of production hepcidin by gene engineer.
Antimicrobial Cationic Peptides ; biosynthesis ; genetics ; physiology ; Hepcidins ; Humans ; Iron ; metabolism ; Protein Engineering ; methods

According to the amino acid sequence of monellin, a single chain 294bp monellin gene was synthesized and inserted into vector pET-22b to yield the recombinant secretion plasmid pETMO. The single-chain monellin gene was designed based on the biased codons of E. coli so that its expression would be then optimized. Under the expressing conditions, monellin was produced accounting for 44.8% of total soluble proteins. The E. coli-expressed single-chain monellin is 3000 times sweeter than sucrose. The thermal-stability and acid-resistance of the protein are higher than the natural monellin.
Escherichia coli ; genetics ; metabolism ; Plant Proteins ; biosynthesis ; genetics ; Protein Engineering ; methods ; Recombinant Proteins ; biosynthesis ; genetics

The developments of recombinant DNA technology and structural biology make it possible to modify enzyme in molecular level. Scientists show growing interests in the evolution or functional fusion of enzymes. Recent advances and applications of the molecular enzyme engineering are reviewed and discussed in this article.
Enzymes ; genetics ; Protein Engineering ; methods ; Recombinant Fusion Proteins ; genetics ; Research Design

As central players in cellular structure and function, proteins have long been central themes in life science research. Analyzing the impact of protein sequence variation on its structure and function is one of the important means to study proteins. In recent years, a technology called deep mutational scanning (DMS) has been widely used in the field of protein research. It introduces thousands of mutations in parallel in specific regions of proteins through high-abundance DNA libraries. After screening, high-throughput sequencing is employed to score each mutation, revealing sequence-function correlations. Due to its high-throughput, fast and easy, and labor-saving features, DMS has become an important method for protein function research and protein engineering. This review briefly summarizes the principle of DMS technology, highlighting its applications in mammalian cells. Moreover, this review analyzes the current technical bottlenecks, aiming to facilitate relevant research.
Animals ; Mutation ; Proteins/chemistry* ; Protein Engineering ; High-Throughput Nucleotide Sequencing/methods* ; Mammals/genetics*

As a novel BioChip technique, LiquiChip technique uses fluorescent polystyrene beads as the carrriers of various probes, thus the related reactions of bio-molecules are in liquid system. LiquiChip technique is of use in the methods to detect bio-macromolecules, e. g. DNA detection, immunoassay, cytokine assay, hormone assay, environmental survey and analysis. In comparison to conventional biochip, LiquiChip presents the advantages of high-throughput, high sensitivity, high accuracy, fine repeatability, wide linear range, etc. Now LiquiChip technique is widely used in biomedical engineering field.
Animals ; Biomedical Engineering ; trends ; Biosensing Techniques ; methods ; Humans ; Microfluidic Analytical Techniques ; methods ; Molecular Imprinting ; Protein Array Analysis ; methods