Characterization of two β-galactosidases LacZ and WspA1 from Nostoc flagelliforme with focus on the latter’s central active region
Artikel i vetenskaplig tidskrift, 2021

The identification and characterization of new β-galactosidases will provide diverse candidate enzymes for use in food processing industry. In this study, two β-galactosidases, Nf-LacZ and WspA1, from the terrestrial cyanobacterium Nostoc flagelliforme were heterologously expressed in Escherichia coli, followed by purification and biochemical characterization. Nf-LacZ was characterized to have an optimum activity at 40 °C and pH 6.5, different from that (45 °C and pH 8.0) of WspA1. Two enzymes had a similar Michaelis constant (Km = 0.5 mmol/liter) against the substrate o-nitrophenyl-β-D-galactopyranoside. Their activities could be inhibited by galactostatin bisulfite, with IC50 values of 0.59 µM for Nf-LacZ and 1.18 µM for WspA1, respectively. Gel filtration analysis suggested that the active form of WspA1 was a dimer, while Nf-LacZ was functional as a larger multimer. WspA1 was further characterized by the truncation test, and its minimum central region was found to be from residues 188 to 301, having both the glycosyl hydrolytic and transgalactosylation activities. Finally, transgenic analysis with the GFP reporter protein found that the N-terminus of WspA1 (35 aa) might play a special role in the export of WspA1 from cells. In summary, this study characterized two cyanobacterial β-galactosidases for potential applications in food industry.

Författare

Xiang Gao

Shaanxi University of Technology

Huazhong Normal University

Litao Liu

Huazhong Normal University

Lijuan Cui

Huazhong Normal University

Tao Zheng

Shaanxi University of Technology

Boyang Ji

Chalmers, Biologi och bioteknik, Systembiologi

Ke Liu

Huazhong Normal University

Scientific Reports

2045-2322 (ISSN)

Vol. 11 1 18448

Ämneskategorier

Biokemi och molekylärbiologi

Strukturbiologi

Biokatalys och enzymteknik

DOI

10.1038/s41598-021-97929-6

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Senast uppdaterat

2021-10-07