Metabolites-BOC Sciences

Directed Evolution Service

Directed Evolution Service

BOC Sciences provides directed evolution service for microbial strains. We are capable of library construction for gene variants by performing multiple rounds of mutagenesis with strains’ genes. In addition to our advanced techniques and extensive microbial strain improvement experience, we provide a professional platform for strains’ directed evolution. Our services served as a tool to screen and select desired functional gene variants, thus helping our customers to express and obtain target proteins.

Introduction

Directed evolution is a cyclic process alternative between gene diversifications and gene screening for desired gene variants selection. Directed evolution mimics the Darwinian evolution in a test tube to artificially accelerate biological molecules' natural evolution process. Directed Evolution has become one of the most powerful tools for various metabolic pathways, enzyme and protein generation, and even whole genomes with desired properties. In directed evolution, a single gene is evolved by iterative rounds of mutagenesis, selection or screening. After all, only the beneficial mutations remain, and variants from the previous round served as the template for the next round to achieve stepwise improvements.

Application of Directed Evolution

Directed evolution has been widely used to engineer nucleic acids, proteins, genetic circuits, metabolic pathways, and whole genomes to improved basic functions for applied biological and medical research. Directed evolution is a powerful tool to enhance the catalytic activities and substrate specificity in numerous enzymes. Furthermore, directed evolution can also upregulate protein’s stability at high temperatures and binding affinity with therapeutic antibodies.

Our advantages

  • Various mutagenesis methods for gene variant library construction.
  • Personalized service using the microbial host of customer choice.
  • Comprehensive microbial expression systems cover bacterial and eukaryotic systems, such as E. coli, yeast, Bacillus subtilis, etc.
  • Advanced high-throughput screening techniques: high-throughput mass spectrometry (MS)-based screening, microtiter plate-based screening, fluorescence imaging-based screening, droplet microfluidics, etc.
  • Extensive experience in microbial strains culture aspect.
  • Optimized medium selection, biomass control and product induction.
  • Improved microbial host with positive performance, such as rapid growth, genetic stability, and eliminates by-products.
  • Cost-effective and high-quality products.

Our methods

Directed evolution includes two main steps: 1) constructing a diverse gene variants library by random mutagenesis, gene recombination, etc. 2) screening and selection of improved phenotypes variants.

Random Mutagenesis: mutation rate and mutational spectrum are the most important factors when designing a random mutagenesis experiment. Commonly, mutagenesis methods involve mutagenic chemicals or exposing organisms to UV radiation.

Site saturation mutagenesis (SSM): a polymerase chain reaction (PCR)-based technique. This method introduce mutations during oligonucleotides annealing or PCR with synthetic DNA oligonucleotides containing one or more degenerate codons at the target residues.

Iterative saturation mutagenesis (ISM): performing iterative cycles of saturation mutagenesis at chosen sites rationally may be crucial for enhancing the protein property of interest.

Gene recombination: the exchange of genetic material between different organisms. This can be used to recombine beneficial mutations while removing deleterious mutations in natural evolution.

Selection and screening: by exerting certain selective pressure to the gene mutation library, unwanted variants are directly eliminated, then positive candidates are allowed to undergo subsequent rounds of directed evolution.

Project workflow

  • Customer advisory
  • Project discussion
  • Microbial host selection
  • Microbial strains directed evolution
  • Novel strain evaluation, transform primitive strains into industrializable strains
  • Project delivery

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