Adaptive Laboratory Evolution Service

Probiotics are widely used in a variety of fields, including food, pharmaceuticals, and others. Efforts to remove barriers to industrial development of probiotics and maximize productivity are still ongoing, and adaptive laboratory evolution (ALE)is one of the most effective ways to achieve this goal. Through our professional ALE platform, Creative Enzymes provides strain modification services to help our clients obtain superior strains that are easy to control culture conditions, have short generation times, and are easy to manufacture and preserve.

Introduction of Adaptive Laboratory Evolutionary Technique

The adaptive laboratory evolution technique is an evolutionary engineering approach under artificial conditions that selects strains with better phenotypes by mimicking natural evolution. Since ALE does not require detailed knowledge of various complex and interacting metabolic networks and only requires the simulation of natural environmental conditions in the laboratory to design selection pressures, it has the advantage of being widely adaptable, practical and more convenient for strain transformation. ALE has been widely used to produce more productive probiotics with beneficial mutations. For example, ALEs of probiotics were performed for acid tolerance with Lactobacillus casei and Bificobacterium longum and they exhibited a significant increase of tolerance showing viability at lethal pH.

Fig. 1 Adaptive laboratory evolution (ALE). (Dragosits, 2013)

Applications of ALE in Strain Improvement

Industrial strains are developed to maximize productivity and yield. For industrial process efficiency, ALE of an industrial host can contribute to cut down the cycle time and improve product titer, strain tolerance, and substrate utilization ability. The application of ALE in strain improvement is usually divided into the following categories:

• Optimization of nutrient-specific growth and induction of native nutrients.
• Growing in harsh conditions (non-optimal pH and temperature) or tolerating adverse environments.
• Produce new substances or increase product titers.

Our Methods

The steps of Creative Enzymes to improve strains by adaptive laboratory evolution technique mainly include the following two aspects.

• We construct a large mutational library of genetic diversity by simulating the natural evolutionary process through random mutagenesis or genetic recombination.

Random mutagenesis mainly involves exposing the organism to radiation or mutagens and can be performed in combination with site-saturation mutagenesis and iterative saturation mutagenesis. Genetic recombination allows the exchange of genetic material between organisms to aggregate beneficial mutations and remove deleterious mutations during evolution, resulting in strains with superior traits.

• Rapidly obtain strains of stable form by screening.

By applying some selection pressure to the mutant library, unwanted variants are eliminated and strains with phenotypic or physiological properties adapted to specific conditions are obtained.

Workflow of Creative Enzymes

Fig. 2 Project workflow of Creative Enzymes.

Creative Enzymes is a professional and experienced probiotic supplier and service provider. We offer customized strain directed evolution services that provide powerful tools for changing the evolutionary power of strain phenotype, performance and stability. To get more information, please contact us and we will be happy to serve your research demands.

Reference

1. Dragosits, M.; et al. Adaptive laboratory evolution–principles and applications for biotechnology. Microbial cell factories. 2013, 12(1): 1-17.