A set of simple methods for detection and extraction of laminarinase

Abstract A carefully designed ammonium sulfate precipitation will simplify extraction of proteins and is considered to be a gold standard among various precipitation methods. Therefore, optimization of ammonium sulfate precipitation can be an important functional step in protein purification. The pr...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Ananthamurthy Koteshwara, Nancy V. Philip, Jesil Mathew Aranjani, Raghu Chandrashekhar Hariharapura, Subrahmanyam Volety Mallikarjuna
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/8fe55ed7f95f404283336a7dae8c3df8
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:8fe55ed7f95f404283336a7dae8c3df8
record_format	dspace
spelling	oai:doaj.org-article:8fe55ed7f95f404283336a7dae8c3df82021-12-02T13:57:49ZA set of simple methods for detection and extraction of laminarinase10.1038/s41598-021-81807-22045-2322https://doaj.org/article/8fe55ed7f95f404283336a7dae8c3df82021-01-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-81807-2https://doaj.org/toc/2045-2322Abstract A carefully designed ammonium sulfate precipitation will simplify extraction of proteins and is considered to be a gold standard among various precipitation methods. Therefore, optimization of ammonium sulfate precipitation can be an important functional step in protein purification. The presence of high amounts of ammonium sulphate precludes direct detection of many enzymatically active proteins including reducing sugar assays (e.g. Nelson-Somogyi, Reissig and 3,5-dinitrosalicylic acid methods) for assessing carbohydrases (e.g. laminarinase (β (1–3)-glucanohydrolase), cellulases and chitinases). In this study, a simple method was developed using laminarin infused agarose plate for the direct analysis of the ammonium sulphate precipitates from Streptomyces rimosus AFM-1. The developed method is simple and convenient that can give accurate results even in presence of ammonium sulfate in the crude precipitates. Laminarin is a translucent substrate requiring the use of a stain to visualize the zones of hydrolysis in a plate assay. A very low-cost and locally available fluorescent optical fabric brightener Tinopal CBS-X has been used as a stain to detect the zones of hydrolysis. We also report simple methods to prepare colloidal chitin and cell free supernatant in this manuscript.Ananthamurthy KoteshwaraNancy V. PhilipJesil Mathew AranjaniRaghu Chandrashekhar HariharapuraSubrahmanyam Volety MallikarjunaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ananthamurthy Koteshwara Nancy V. Philip Jesil Mathew Aranjani Raghu Chandrashekhar Hariharapura Subrahmanyam Volety Mallikarjuna A set of simple methods for detection and extraction of laminarinase
description	Abstract A carefully designed ammonium sulfate precipitation will simplify extraction of proteins and is considered to be a gold standard among various precipitation methods. Therefore, optimization of ammonium sulfate precipitation can be an important functional step in protein purification. The presence of high amounts of ammonium sulphate precludes direct detection of many enzymatically active proteins including reducing sugar assays (e.g. Nelson-Somogyi, Reissig and 3,5-dinitrosalicylic acid methods) for assessing carbohydrases (e.g. laminarinase (β (1–3)-glucanohydrolase), cellulases and chitinases). In this study, a simple method was developed using laminarin infused agarose plate for the direct analysis of the ammonium sulphate precipitates from Streptomyces rimosus AFM-1. The developed method is simple and convenient that can give accurate results even in presence of ammonium sulfate in the crude precipitates. Laminarin is a translucent substrate requiring the use of a stain to visualize the zones of hydrolysis in a plate assay. A very low-cost and locally available fluorescent optical fabric brightener Tinopal CBS-X has been used as a stain to detect the zones of hydrolysis. We also report simple methods to prepare colloidal chitin and cell free supernatant in this manuscript.
format	article
author	Ananthamurthy Koteshwara Nancy V. Philip Jesil Mathew Aranjani Raghu Chandrashekhar Hariharapura Subrahmanyam Volety Mallikarjuna
author_facet	Ananthamurthy Koteshwara Nancy V. Philip Jesil Mathew Aranjani Raghu Chandrashekhar Hariharapura Subrahmanyam Volety Mallikarjuna
author_sort	Ananthamurthy Koteshwara
title	A set of simple methods for detection and extraction of laminarinase
title_short	A set of simple methods for detection and extraction of laminarinase
title_full	A set of simple methods for detection and extraction of laminarinase
title_fullStr	A set of simple methods for detection and extraction of laminarinase
title_full_unstemmed	A set of simple methods for detection and extraction of laminarinase
title_sort	set of simple methods for detection and extraction of laminarinase
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/8fe55ed7f95f404283336a7dae8c3df8
work_keys_str_mv	AT ananthamurthykoteshwara asetofsimplemethodsfordetectionandextractionoflaminarinase AT nancyvphilip asetofsimplemethodsfordetectionandextractionoflaminarinase AT jesilmathewaranjani asetofsimplemethodsfordetectionandextractionoflaminarinase AT raghuchandrashekharhariharapura asetofsimplemethodsfordetectionandextractionoflaminarinase AT subrahmanyamvoletymallikarjuna asetofsimplemethodsfordetectionandextractionoflaminarinase AT ananthamurthykoteshwara setofsimplemethodsfordetectionandextractionoflaminarinase AT nancyvphilip setofsimplemethodsfordetectionandextractionoflaminarinase AT jesilmathewaranjani setofsimplemethodsfordetectionandextractionoflaminarinase AT raghuchandrashekharhariharapura setofsimplemethodsfordetectionandextractionoflaminarinase AT subrahmanyamvoletymallikarjuna setofsimplemethodsfordetectionandextractionoflaminarinase
_version_	1718392272989454336

A set of simple methods for detection and extraction of laminarinase

Ejemplares similares