Structural and functional characterization of cold-active sialidase isolated from Antarctic fungus Penicillium griseofulvum P29
The fungal strain, Penicillium griseofulvum P29, isolated from a soil sample taken from Terra Nova Bay, Antarctica, was found to be a good producer of sialidase (P29). The present study was focused on the purification and structural characterization of the enzyme. P29 enzyme was purified using a Q-S...
Published in: | Biochemistry and Biophysics Reports |
---|---|
Main Authors: | , , , , , , , , , , , , |
Format: | Article in Journal/Newspaper |
Language: | English |
Published: |
Elsevier
1481
|
Subjects: | |
Online Access: | https://doi.org/10.1016/j.bbrep.2023.101610 https://doaj.org/article/897e11fc99ad4f11b32bcd2d14843cb0 |
id |
ftdoajarticles:oai:doaj.org/article:897e11fc99ad4f11b32bcd2d14843cb0 |
---|---|
record_format |
openpolar |
spelling |
ftdoajarticles:oai:doaj.org/article:897e11fc99ad4f11b32bcd2d14843cb0 2024-01-21T10:00:30+01:00 Structural and functional characterization of cold-active sialidase isolated from Antarctic fungus Penicillium griseofulvum P29 Aleksandar Dolashki Radoslav Abrashev Dimitar Kaynarov Ekaterina Krumova Lyudmila Velkova Rumyana Eneva Stefan Engibarov Yana Gocheva Jeny Miteva-Staleva Vladislava Dishliyska Boryana Spasova Maria Angelova Pavlina Dolashka 2024-03-01T00:00:00Z https://doi.org/10.1016/j.bbrep.2023.101610 https://doaj.org/article/897e11fc99ad4f11b32bcd2d14843cb0 EN eng Elsevier http://www.sciencedirect.com/science/article/pii/S2405580823001917 https://doaj.org/toc/2405-5808 2405-5808 doi:10.1016/j.bbrep.2023.101610 https://doaj.org/article/897e11fc99ad4f11b32bcd2d14843cb0 Biochemistry and Biophysics Reports, Vol 37, Iss , Pp 101610- (2024) Antarctic fungi Sialidase Cold-active enzyme Structural characterization 3D-structure Fluorescence analysis Biology (General) QH301-705.5 Biochemistry QD415-436 article 1481 ftdoajarticles https://doi.org/10.1016/j.bbrep.2023.101610 2023-12-24T01:41:30Z The fungal strain, Penicillium griseofulvum P29, isolated from a soil sample taken from Terra Nova Bay, Antarctica, was found to be a good producer of sialidase (P29). The present study was focused on the purification and structural characterization of the enzyme. P29 enzyme was purified using a Q-Sepharose column and fast performance liquid chromatography separation on a Mono Q column. The determined molecular mass of the purified enzyme of 40 kDa by SDS-PAGE and 39924.40 Da by matrix desorption/ionization mass spectrometry (MALDI-TOF/MS) analysis correlated well with the calculated mass (39903.75 kDa) from the amino acid sequence of the enzyme. P29 sialidase shows a temperature optimum of 37 °C and low-temperature stability, confirming its cold-active nature. The enzyme is more active towards α(2 → 3) sialyl linkages than those containing α(2 → 6) linkages.Based on the determined amino acid sequence and 3D structural modeling, a 3D model of P29 sialidase was presented, and the properties of the enzyme were explained. The conformational stability of the enzyme was followed by fluorescence spectroscopy, and the new enzyme was found to be conformationally stable in the neutral pH range of pH 6 to pH 9. In addition, the enzyme was more stable in an alkaline environment than in an acidic environment. The purified cold-active enzyme is the only sialidase produced and characterized from Antarctic fungi to date. Article in Journal/Newspaper Antarc* Antarctic Antarctica Directory of Open Access Journals: DOAJ Articles Antarctic Terra Nova Bay Biochemistry and Biophysics Reports 37 101610 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Antarctic fungi Sialidase Cold-active enzyme Structural characterization 3D-structure Fluorescence analysis Biology (General) QH301-705.5 Biochemistry QD415-436 |
spellingShingle |
Antarctic fungi Sialidase Cold-active enzyme Structural characterization 3D-structure Fluorescence analysis Biology (General) QH301-705.5 Biochemistry QD415-436 Aleksandar Dolashki Radoslav Abrashev Dimitar Kaynarov Ekaterina Krumova Lyudmila Velkova Rumyana Eneva Stefan Engibarov Yana Gocheva Jeny Miteva-Staleva Vladislava Dishliyska Boryana Spasova Maria Angelova Pavlina Dolashka Structural and functional characterization of cold-active sialidase isolated from Antarctic fungus Penicillium griseofulvum P29 |
topic_facet |
Antarctic fungi Sialidase Cold-active enzyme Structural characterization 3D-structure Fluorescence analysis Biology (General) QH301-705.5 Biochemistry QD415-436 |
description |
The fungal strain, Penicillium griseofulvum P29, isolated from a soil sample taken from Terra Nova Bay, Antarctica, was found to be a good producer of sialidase (P29). The present study was focused on the purification and structural characterization of the enzyme. P29 enzyme was purified using a Q-Sepharose column and fast performance liquid chromatography separation on a Mono Q column. The determined molecular mass of the purified enzyme of 40 kDa by SDS-PAGE and 39924.40 Da by matrix desorption/ionization mass spectrometry (MALDI-TOF/MS) analysis correlated well with the calculated mass (39903.75 kDa) from the amino acid sequence of the enzyme. P29 sialidase shows a temperature optimum of 37 °C and low-temperature stability, confirming its cold-active nature. The enzyme is more active towards α(2 → 3) sialyl linkages than those containing α(2 → 6) linkages.Based on the determined amino acid sequence and 3D structural modeling, a 3D model of P29 sialidase was presented, and the properties of the enzyme were explained. The conformational stability of the enzyme was followed by fluorescence spectroscopy, and the new enzyme was found to be conformationally stable in the neutral pH range of pH 6 to pH 9. In addition, the enzyme was more stable in an alkaline environment than in an acidic environment. The purified cold-active enzyme is the only sialidase produced and characterized from Antarctic fungi to date. |
format |
Article in Journal/Newspaper |
author |
Aleksandar Dolashki Radoslav Abrashev Dimitar Kaynarov Ekaterina Krumova Lyudmila Velkova Rumyana Eneva Stefan Engibarov Yana Gocheva Jeny Miteva-Staleva Vladislava Dishliyska Boryana Spasova Maria Angelova Pavlina Dolashka |
author_facet |
Aleksandar Dolashki Radoslav Abrashev Dimitar Kaynarov Ekaterina Krumova Lyudmila Velkova Rumyana Eneva Stefan Engibarov Yana Gocheva Jeny Miteva-Staleva Vladislava Dishliyska Boryana Spasova Maria Angelova Pavlina Dolashka |
author_sort |
Aleksandar Dolashki |
title |
Structural and functional characterization of cold-active sialidase isolated from Antarctic fungus Penicillium griseofulvum P29 |
title_short |
Structural and functional characterization of cold-active sialidase isolated from Antarctic fungus Penicillium griseofulvum P29 |
title_full |
Structural and functional characterization of cold-active sialidase isolated from Antarctic fungus Penicillium griseofulvum P29 |
title_fullStr |
Structural and functional characterization of cold-active sialidase isolated from Antarctic fungus Penicillium griseofulvum P29 |
title_full_unstemmed |
Structural and functional characterization of cold-active sialidase isolated from Antarctic fungus Penicillium griseofulvum P29 |
title_sort |
structural and functional characterization of cold-active sialidase isolated from antarctic fungus penicillium griseofulvum p29 |
publisher |
Elsevier |
publishDate |
1481 |
url |
https://doi.org/10.1016/j.bbrep.2023.101610 https://doaj.org/article/897e11fc99ad4f11b32bcd2d14843cb0 |
geographic |
Antarctic Terra Nova Bay |
geographic_facet |
Antarctic Terra Nova Bay |
genre |
Antarc* Antarctic Antarctica |
genre_facet |
Antarc* Antarctic Antarctica |
op_source |
Biochemistry and Biophysics Reports, Vol 37, Iss , Pp 101610- (2024) |
op_relation |
http://www.sciencedirect.com/science/article/pii/S2405580823001917 https://doaj.org/toc/2405-5808 2405-5808 doi:10.1016/j.bbrep.2023.101610 https://doaj.org/article/897e11fc99ad4f11b32bcd2d14843cb0 |
op_doi |
https://doi.org/10.1016/j.bbrep.2023.101610 |
container_title |
Biochemistry and Biophysics Reports |
container_volume |
37 |
container_start_page |
101610 |
_version_ |
1788703238078332928 |