Streptococcus agalactiae amylomaltase offers insight into the transglycosylation mechanism and the molecular basis of thermostability among amylomaltases

Abstract Amylomaltase (AM) catalyzes transglycosylation of starch to form linear or cyclic oligosaccharides with potential applications in biotechnology and industry. In the present work, a novel AM from the mesophilic bacterium Streptococcus agalactiae (SaAM), with 18–49% sequence identity to previ...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Suthipapun Tumhom, Pitchanan Nimpiboon, Kittikhun Wangkanont, Piamsook Pongsawasdi
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/603b3c4a035e433ca06bde6359ea7fbf
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:603b3c4a035e433ca06bde6359ea7fbf
record_format	dspace
spelling	oai:doaj.org-article:603b3c4a035e433ca06bde6359ea7fbf2021-12-02T16:35:55ZStreptococcus agalactiae amylomaltase offers insight into the transglycosylation mechanism and the molecular basis of thermostability among amylomaltases10.1038/s41598-021-85769-32045-2322https://doaj.org/article/603b3c4a035e433ca06bde6359ea7fbf2021-03-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-85769-3https://doaj.org/toc/2045-2322Abstract Amylomaltase (AM) catalyzes transglycosylation of starch to form linear or cyclic oligosaccharides with potential applications in biotechnology and industry. In the present work, a novel AM from the mesophilic bacterium Streptococcus agalactiae (SaAM), with 18–49% sequence identity to previously reported AMs, was characterized. Cyclization and disproportionation activities were observed with the optimum temperature of 30 °C and 40 °C, respectively. Structural determination of SaAM, the first crystal structure of small AMs from the mesophiles, revealed a glycosyl-enzyme intermediate derived from acarbose and a second acarbose molecule attacking the intermediate. This pre-transglycosylation conformation has never been before observed in AMs. Structural analysis suggests that thermostability in AMs might be mainly caused by an increase in salt bridges since SaAM has a lower number of salt bridges compared with AMs from the thermophiles. Increase in thermostability by mutation was performed. C446 was substituted with A/S/P. C446A showed higher activities and higher k cat /K m values for starch in comparison to the WT enzyme. C446S exhibited a 5 °C increase in optimum temperature and the threefold increase in half-life time at 45 °C, most likely resulting from H-bonding interactions. For all enzymes, the main large-ring cyclodextrin (LR-CD) products were CD24-CD26 with CD22 as the smallest. C446S produced more CD35-CD42, especially at a longer incubation time.Suthipapun TumhomPitchanan NimpiboonKittikhun WangkanontPiamsook PongsawasdiNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-15 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Suthipapun Tumhom Pitchanan Nimpiboon Kittikhun Wangkanont Piamsook Pongsawasdi Streptococcus agalactiae amylomaltase offers insight into the transglycosylation mechanism and the molecular basis of thermostability among amylomaltases
description	Abstract Amylomaltase (AM) catalyzes transglycosylation of starch to form linear or cyclic oligosaccharides with potential applications in biotechnology and industry. In the present work, a novel AM from the mesophilic bacterium Streptococcus agalactiae (SaAM), with 18–49% sequence identity to previously reported AMs, was characterized. Cyclization and disproportionation activities were observed with the optimum temperature of 30 °C and 40 °C, respectively. Structural determination of SaAM, the first crystal structure of small AMs from the mesophiles, revealed a glycosyl-enzyme intermediate derived from acarbose and a second acarbose molecule attacking the intermediate. This pre-transglycosylation conformation has never been before observed in AMs. Structural analysis suggests that thermostability in AMs might be mainly caused by an increase in salt bridges since SaAM has a lower number of salt bridges compared with AMs from the thermophiles. Increase in thermostability by mutation was performed. C446 was substituted with A/S/P. C446A showed higher activities and higher k cat /K m values for starch in comparison to the WT enzyme. C446S exhibited a 5 °C increase in optimum temperature and the threefold increase in half-life time at 45 °C, most likely resulting from H-bonding interactions. For all enzymes, the main large-ring cyclodextrin (LR-CD) products were CD24-CD26 with CD22 as the smallest. C446S produced more CD35-CD42, especially at a longer incubation time.
format	article
author	Suthipapun Tumhom Pitchanan Nimpiboon Kittikhun Wangkanont Piamsook Pongsawasdi
author_facet	Suthipapun Tumhom Pitchanan Nimpiboon Kittikhun Wangkanont Piamsook Pongsawasdi
author_sort	Suthipapun Tumhom
title	Streptococcus agalactiae amylomaltase offers insight into the transglycosylation mechanism and the molecular basis of thermostability among amylomaltases
title_short	Streptococcus agalactiae amylomaltase offers insight into the transglycosylation mechanism and the molecular basis of thermostability among amylomaltases
title_full	Streptococcus agalactiae amylomaltase offers insight into the transglycosylation mechanism and the molecular basis of thermostability among amylomaltases
title_fullStr	Streptococcus agalactiae amylomaltase offers insight into the transglycosylation mechanism and the molecular basis of thermostability among amylomaltases
title_full_unstemmed	Streptococcus agalactiae amylomaltase offers insight into the transglycosylation mechanism and the molecular basis of thermostability among amylomaltases
title_sort	streptococcus agalactiae amylomaltase offers insight into the transglycosylation mechanism and the molecular basis of thermostability among amylomaltases
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/603b3c4a035e433ca06bde6359ea7fbf
work_keys_str_mv	AT suthipapuntumhom streptococcusagalactiaeamylomaltaseoffersinsightintothetransglycosylationmechanismandthemolecularbasisofthermostabilityamongamylomaltases AT pitchanannimpiboon streptococcusagalactiaeamylomaltaseoffersinsightintothetransglycosylationmechanismandthemolecularbasisofthermostabilityamongamylomaltases AT kittikhunwangkanont streptococcusagalactiaeamylomaltaseoffersinsightintothetransglycosylationmechanismandthemolecularbasisofthermostabilityamongamylomaltases AT piamsookpongsawasdi streptococcusagalactiaeamylomaltaseoffersinsightintothetransglycosylationmechanismandthemolecularbasisofthermostabilityamongamylomaltases
_version_	1718383691774820352

Streptococcus agalactiae amylomaltase offers insight into the transglycosylation mechanism and the molecular basis of thermostability among amylomaltases

Ejemplares similares