Before and After IPTG

Purified Enzyme

Custom production: We can custom produce milligram to multiple gram quantities of any recombinant protein or enzyme for research applications. Our custom manufactured proteins may be used for in vitro reactions or for animal use. We do not produce custom enzymes for human clinical use. We have offered this service since 2002. We have experience in making enzymes of all sizes, single unit or multi-subunit enzymes, tagged or untagged enzymes. We have experience in producing molecular biology grade enzymes, at low cost. Production is customized to accommodate the customer's needs.

Four Scales: Our standard fermentation scales are 16 liters, 32 liters, 100 liters, and 400 liters. We can perform other scales to accomodate the customer's requirements.

Yield: Enzyme yield is highly dependent on the individual enzyme. Although we can take every measure to get the highest yield possible, and to minimize the formation of inclusion bodies, yields vary from about 5 mg per liter to about 150 mg per liter. We use a proprietary enriched broth in the fermentation.

Purity: Typical purity is 95% or higher for enzymes which express well in E. coli. Purification is performed under mild chromatographic conditions, ensuring that the purified enzyme product has the maximum enzyme activity. The enzyme is purified according to the customer specifications, which may entail multiple chromatography columns, and other purification methods, to achieve the customer's objectives.

Codon Optimization: For proteins which do not express well in E. coli, we can contruct an expression clone using a synthetic gene optimized for expression in E. coli hosts, such as BL21. Synthetic genes avoid rare codons which hinder translation of the mRNA into the recombinant protein. For genes which do not require synthetic genes, we can construct an expression clone using high fidelity PCR in our proprietary expression vector. Constructed expression clones are confirmed by DNA sequencing prior to use. Alternatively, the customer may provide us with an expression clone for the gene of interest.

Tags: We recommend using polyhistidine tags in order to facilitate enzyme purification using immobilized metal chelate chromatography. We can also produce native untagged proteins, and proteins with different tags, such as GST tagged proteins.

Supplied: Enzymes are supplied in a stabilizing buffer containing glycerol, usually at 25% to 50% concentration. Enzymes are typically supplied at high concentration, such as 10 mg protein/ml to 30 mg protein/ml. This permits the customer to optionally dilute the enzyme into a different buffer prior to use. Bulk quantites of enzyme are usually shipped on dry ice.

Quality control: The final purified enzyme product can be assayed for enzyme activity according to the desired assay procedure recommended by the customers. In addition, we can assay for contaminating activities, such as nuclease, exonuclease, and RNase contamination, which is standard for molecular biology grade enzymes.

Fast Service: The turnaround time for delivery of custom enzymes depends on the scale of the fermentation and whether or not the customer will provide us with an expression clone. For production of the first batch of an enzyme, additional time is required for us to optimize the production process. The turnaround time for a 16 liter scale in which we are provided with the clone is typcially about two to three weeks.