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Expression Evaluation and Optimization



The route to efficient and cost effective gene expression and enzyme production is often case-specific. A wise step before trial-scale production is to quickly screen possible expression systems and identify the most target friendly system. In some cases, even after the full-scale production is set up, evaluation of other expression and production systems could help to save the cost and improve the product quality. To serve these purposes, Creative Enzymes provides the custom service of expression evaluation and optimization.

The service quickly tests the most commonly used expression systems, including host cells, vectors, and promoters, and identify the most efficient system. Production optimization is also available, which targets more on the fermentation conditions and processes to further improve the yield and purity of the desired enzyme. The evaluation and optimization service benefits our customers on several aspects: 1) It saves your valuable time with our unique high-throughput screening platform, which is able identify the most suitable expression system within one week; 2) The production yield is assessed and quantitatively compared among systems, eliminating uncertainties such as codon bias, vector construction, and host cell toxicity; 3) The detailed protocol will be delivered covering key parameters for successful expression; and 4) The service cost is lower, and the overall time is further reduced when integrated with other services such as gene cloning and enzyme production.

Our technology evaluates the impacts of most parameters in gene expression and enzyme production at multiple levels, to ensure the accuracy and consistency of the conclusion. The gene expression level in host cells can be measured by the number of mRNA replicates using sequencing and micro-array techniques. Simultaneously, the target enzyme is also identified and quantified. These results will be used to examine if the enzyme is expressed and to what level. In addition, any discrepancy between the data sets could further locate possible issues in each step of translation, secretion, and folding, etc. Subsequently, the enzyme production yield is then evaluated under various fermentation conditions, along with the enzyme properties. Typical factors contributing to production efficiency include molecular weight, disulfide bonds, post-translational modifications, homogeneity, enzyme activities such as proteolysis, promoter design, strain selection, pH, temperature, induction mechanisms, inoculation, and recovery and purification. All these important parameters can be designed into a batch of condition screening, and the optimal condition will be recommended based on the tested conditions and predictions from the test matrix. In addition, any limits on the production cost will be also considered if requested by the customer. As a result, a majority of customers have seen increases in production yield of at least 10 folds after using the optimization service.

Relying on the advanced research and extensive experiences in gene expression and enzyme production, Creative Enzymes would like to help researchers overcome the challenges in obtaining the desired enzymes and reduce the risk of unknown outcomes from expensive and long-term investigation. As one of the first-comers and best-rated service provider in this field, Creative Enzymes delivers reliable and valuable results of the evaluation and optimization service:



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