Expression, Purification, and Characteristic of Tibetan Sheep Breast Lysozyme Using Pichia pastoris Expression System

A lysozyme gene from breast of Tibetan sheep was successfully expressed by secretion using a-factor signal sequence in the methylotrophic yeast, Pichia pastoris GS115. An expression yield and specific activity greater than 500 mg/L and 4,000 U/mg was obtained. Results at optimal pH and temperature s...

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Detalles Bibliográficos
Autores principales: Li, Jianbo, Jiang, Mingfeng, Wang, Yong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Asian-Australasian Association of Animal Production Societies (AAAP) and Korean Society of Animal Science and Technology (KSAST) 2014
Materias:
Articles
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4093532/
https://www.ncbi.nlm.nih.gov/pubmed/25049990
http://dx.doi.org/10.5713/ajas.2013.13529
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author Li, Jianbo
Jiang, Mingfeng
Wang, Yong
author_facet Li, Jianbo
Jiang, Mingfeng
Wang, Yong
author_sort Li, Jianbo
collection PubMed
description A lysozyme gene from breast of Tibetan sheep was successfully expressed by secretion using a-factor signal sequence in the methylotrophic yeast, Pichia pastoris GS115. An expression yield and specific activity greater than 500 mg/L and 4,000 U/mg was obtained. Results at optimal pH and temperature showed recombinant lysozyme has higher lytic activity at pH 6.5 and 45°C. This study demonstrates the successful expression of recombinant lysozyme using the eukaryotic host organism P. pastoris paving the way for protein engineering. Additionally, this study shows the feasibility of subsequent industrial manufacture of the enzyme with this expression system together with a high purity scheme for easy high-yield purification.
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spelling pubmed-40935322014-07-21 Expression, Purification, and Characteristic of Tibetan Sheep Breast Lysozyme Using Pichia pastoris Expression System Li, Jianbo Jiang, Mingfeng Wang, Yong Asian-Australas J Anim Sci Articles A lysozyme gene from breast of Tibetan sheep was successfully expressed by secretion using a-factor signal sequence in the methylotrophic yeast, Pichia pastoris GS115. An expression yield and specific activity greater than 500 mg/L and 4,000 U/mg was obtained. Results at optimal pH and temperature showed recombinant lysozyme has higher lytic activity at pH 6.5 and 45°C. This study demonstrates the successful expression of recombinant lysozyme using the eukaryotic host organism P. pastoris paving the way for protein engineering. Additionally, this study shows the feasibility of subsequent industrial manufacture of the enzyme with this expression system together with a high purity scheme for easy high-yield purification. Asian-Australasian Association of Animal Production Societies (AAAP) and Korean Society of Animal Science and Technology (KSAST) 2014-04 /pmc/articles/PMC4093532/ /pubmed/25049990 http://dx.doi.org/10.5713/ajas.2013.13529 Text en Copyright © 2014 by Asian-Australasian Journal of Animal Sciences This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License http://creativecommons.org/licenses/by-nc/3.0/ which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Articles
Li, Jianbo
Jiang, Mingfeng
Wang, Yong
Expression, Purification, and Characteristic of Tibetan Sheep Breast Lysozyme Using Pichia pastoris Expression System
title Expression, Purification, and Characteristic of Tibetan Sheep Breast Lysozyme Using Pichia pastoris Expression System
title_full Expression, Purification, and Characteristic of Tibetan Sheep Breast Lysozyme Using Pichia pastoris Expression System
title_fullStr Expression, Purification, and Characteristic of Tibetan Sheep Breast Lysozyme Using Pichia pastoris Expression System
title_full_unstemmed Expression, Purification, and Characteristic of Tibetan Sheep Breast Lysozyme Using Pichia pastoris Expression System
title_short Expression, Purification, and Characteristic of Tibetan Sheep Breast Lysozyme Using Pichia pastoris Expression System
title_sort expression, purification, and characteristic of tibetan sheep breast lysozyme using pichia pastoris expression system
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4093532/
https://www.ncbi.nlm.nih.gov/pubmed/25049990
http://dx.doi.org/10.5713/ajas.2013.13529
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AT wangyong expressionpurificationandcharacteristicoftibetansheepbreastlysozymeusingpichiapastorisexpressionsystem

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