The primary structure of an enzyme, also known as its amino acid sequence, drives the folding and intramolecular bonding of the amino acid chain, which ultimately determines the enzyme's unique three-dimensional shape and corresponding functions. The primary structure of an enzyme can be easily determined by the corresponding gene sequence. In case no DNA sequence information is available, which is not unusual during enzyme discovery, the determination of the enzyme's primary structure needs to start "from scratch".

Creative Enzymes provides comprehensive and accurate solutions for enzyme primary structure determination based on our advanced technology platform and rich expertise. We help our customers to select the most appropriate assay for enzyme structure determination according to their research objectives.

What do we Offer for Enzyme Primary Structure Determination?

Usually, there are four general steps to determine the primary structure of a pre-purified enzyme without a known DNA sequence.

• First, the amino acid chains of the enzyme are separated by reducing disulfide bonds.
• Next, the polypeptide chains are fragmented enzymatically, chemically, or physically, with endopeptidase, cyanogen bromide, or through collision in Mass spectrometry as perspective examples.
• Then, the fragments are sequenced by Edman degradation or Mass spectrometry.
• Finally, the sequences are reconstructed to get the complete primary structure.

Mass spectrometry is a powerful tool in protein determination. In shotgun proteomics, this technology can be even used to sequence a mixture of proteins. However, in most cases, multiple technologies will be used in combination to generate the most reliable results.

Creative Enzymes ensures high efficiency and accuracy in primary structure determination of enzymes. Using mass spectrometry, we can rapidly analyze the primary structure of enzyme molecules, and we provide effective assistance to researchers in accurately determining the primary structure of enzymes and advancing the study of enzyme function and biochemical mechanisms.

Figure 1 Bottom-up- and top-down analysis for the structural characterization of peptides and protein pharmaceuticals by mass spectrometry. (European Journal of Pharmaceutics and Biopharmaceutics 2015)

Significance of Enzyme Primary Structure Determination

• Functional Study of Enzymes. An enzyme's primary structure determines its function and activity, and different amino acid sequences can lead to different enzyme activities with underlying structural information. Therefore enzyme primary structure determination is crucial to explain the biological function of enzymes. Researchers can study the relationship between enzyme structure and function more thoroughly and reveal a number of mechanisms by understanding the basic structure of enzymes. These mechanisms include enzyme-catalyzed biochemical reactions, the operation of the underlying mechanism, and the interaction with substrates or cofactors.
• Drug Design. Understanding the fundamental structure of enzymes can serve as a research foundation for bioengineering and medication design. Researchers can better create specialized enzyme inhibitors for use in medicinal or commercial applications by understanding the basic structure of enzymes.
• Medical Research. Understanding these variations is crucial for early detection and treatment of some genetic disorders since they are linked to changes in the basic structure of enzymes. The association between protein variations and diseases can be successfully ascertained by figuring out the main structure of enzymes, which is of major importance in medical research.

Our Service Workflow

Creative Enzymes provides solid services for amino acid sequencing. Our service is built with sufficient experience and advanced instruments. Rigorous QC processes are in place to ensure that the customer's requirement is met for the specific purpose. Some highlighted services include:

• Amino acid composition determination.
• N-terminal amino acid sequencing.
• N- and C-terminal sequencing.
• Peptide mapping.

If you are interested in us, please feel free to contact us.