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Cloning, Expression, and Characterization of a Novel Sialidase From Brevibacterium Casei

Cloning, Expression, and Characterization of a Novel Sialidase From Brevibacterium Casei

Xuedong Wang, Hui Long, Danhong Shen, Long Liu

Biotechnol Appl Biochem. 2017 Mar;64(2):195-200.

PMID: 26748450

Abstract:

The sialidase gene from Brevibacterium casei was cloned in pET28a and overexpressed as a histidine-tagged protein in Escherichia coli BL21(DE3). The histidine-tagged sialidase protein was purified and characterized from the crude cell extracts of isopropyl-β-d-thiogalactopyranoside-induced cells using Ni-NTA agarose chromatography. SDS-PAGE using the purified sialidase indicated a single band at 116 kDa. This sialidase showed maximum activity at a pH of 5.5 and temperature of 37 °C. The kinetic parameters Km and Vmax for the artificial substrate 2'-(4-methylumbelliferyl)-α-d-N-acetyl-neuraminic acid sodium salt hydrate were 1.69 × 10-3 mM and 244 mmol·Min-1 ·mg-1 , respectively. The sialidase may catalyze the hydrolysis of terminal sialic acids linked by the α-(2,3) and α-(2,8) linkage of polysialogangliosides, but it does not act on monosialotetrahexosylganglioside (GM1), which offers it a great potential for commercially producing GM1 from polysialogangliosides.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
IAR42411261	2-Keto-3-methylbutyric acid-3-d sodium salt hydrate			Price

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