Marine Glycoenzyme Screening Service

Marine glycoenzyme screening is a technology that focuses on the Directed Evolution of Glycoenzymes. Directed evolution experiments include three basic steps, namely library construction, screening, and gene amplification. Screening steps, if performed carelessly or inappropriately, result in no positive candidates. CD BioGlyco is committed to providing excellent Marine Glycoenzyme Development Services, of which marine glycoenzyme screening service is one of our core services. We provide suitable and reliable screening methods to identify variants of interest based on biochemical or biophysical readout assays, including but not limited to: agar plates, microtiter plates, fluorescence-activated cell sorting (FACS), chromatography, and mass spectrometry.

Fig.1 Marine glycoenzyme screening service. (CD BioGlyco)

• Agar Plate-based Marine Glycoenzyme Screening
  This technique uses agar plates for the growth of marine organisms and the subsequent screening of glycoenzymes. The colonies produced on the agar plate can be easily observed and isolated for further study and identification of the specified glycoenzymes. It is a simple, inexpensive, and straightforward method.
• Microtiter Plate-based Marine Glycoenzyme Screening
  This technique employs microtiter plates, where each well serves as a separate reaction chamber for high-throughput glycoenzyme screening. Compared to agar plate screening, this is a quicker method and it allows simultaneous screening of numerous samples.
• FACS-based Marine Glycoenzyme Screening
  FACS offers high-speed and high-sensitivity screening for glycoenzymes. It allows real-time analysis of the individual cells and can sort them based on their characteristics.
• Chromatography-based Marine Glycoenzyme Screening
  This screening technique separates the glycoenzymes based on their physicochemical properties using chromatography. It is a powerful tool for the purification and identification of glycoenzymes.
• Glycophage Display System-based Marine Glycoenzyme Screening
  This state-of-the-art method uses bacteriophages to display glycoenzymes on their surfaces. Individual phages with different glycoenzymes are isolated and studied. This method provides high throughput and specificity.

Publication

Technology: A microtiter plate-based screening assay

Journal: Organic & biomolecular chemistry

IF: 3.89

Published: 2017

Results: The authors developed a screening assay based on a microtiter plate that could accurately determine sialyltransferases' activity and regioselectivity. This tool was specifically used by the authors for screening two single-site saturation libraries of the α2-3-sialyltransferase 1 (PmST1) from Pasteurella multocida for any instances of α2-6-sialyltransferase activity, as well as overall sialyltransferase activity. The PmST1 variation that they found most effective in undertaking the role of an α2-6-sialyltransferase was the double mutant P34H/M144L. Compared to the PmST1 found in the wild type, this mutant demonstrated a 50% reduction in donor hydrolysis and the sialidase activity had been brought down by 50-fold. The interesting feature of this mutant was that it maintained the backdrop of promiscuity found in the donor substrate similar to the wild-type enzyme and was engaged efficiently in a one-pot multienzyme (OPME) system. This was specifically done to catalyze selectively the sialylation of the terminal galactose residue present in a multi galactose-containing tetrasaccharide, namely lacto-N-neotetraoside.

Here are some of the results shown in this article:

Fig.2 A high-throughput screening assay for identifying PmST1 mutants. (McArthur, et al., 2017)

Advantages of Marine Glycoenzyme Screening

• Marine glycoenzyme screening is used to quickly and efficiently select enzymes with superior performance from a large number of variants.
• Marine glycoenzyme screening is used to help speed up the enzyme evolution process and improve enzyme performance.
• Marine glycoenzyme screening is used to improve the efficiency of marine glycoenzyme development.

At CD BioGlyco, we quickly and accurately screen out enzymes with excellent performance from a large number of mutant enzymes. We provide professional and efficient services whether it is agar plate-based, microtiter plate-based, or FACS-based screening. We are committed to continuously optimizing the screening process to provide clients with higher-quality services. You are welcome to contact us at any time to learn more about the marine glycoenzyme screening service.