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Pilot-Scale Production and Thermostability Improvement of the M23 Protease Pseudoalterin from the Deep Sea Bacterium Pseudoalteromonas sp. CF6-2

Pseudoalterin is the most abundant protease secreted by the marine sedimental bacterium Pseudoalteromonas sp. CF6-2 and is a novel cold-adapted metalloprotease of the M23 family. Proteases of the M23 family have high activity towards peptidoglycan and elastin, suggesting their promising biomedical a...

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Autores principales: Yang, Jie, Yu, Yang, Tang, Bai-Lu, Zhong, Shuai, Shi, Mei, Xie, Bin-Bin, Zhang, Xi-Ying, Zhou, Bai-Cheng, Zhang, Yu-Zhong, Chen, Xiu-Lan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6273387/
https://www.ncbi.nlm.nih.gov/pubmed/27869696
http://dx.doi.org/10.3390/molecules21111567
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author Yang, Jie
Yu, Yang
Tang, Bai-Lu
Zhong, Shuai
Shi, Mei
Xie, Bin-Bin
Zhang, Xi-Ying
Zhou, Bai-Cheng
Zhang, Yu-Zhong
Chen, Xiu-Lan
author_facet Yang, Jie
Yu, Yang
Tang, Bai-Lu
Zhong, Shuai
Shi, Mei
Xie, Bin-Bin
Zhang, Xi-Ying
Zhou, Bai-Cheng
Zhang, Yu-Zhong
Chen, Xiu-Lan
author_sort Yang, Jie
collection PubMed
description Pseudoalterin is the most abundant protease secreted by the marine sedimental bacterium Pseudoalteromonas sp. CF6-2 and is a novel cold-adapted metalloprotease of the M23 family. Proteases of the M23 family have high activity towards peptidoglycan and elastin, suggesting their promising biomedical and biotechnological potentials. To lower the fermentive cost and improve the pseudoalterin production of CF6-2, we optimized the fermentation medium by using single factor experiments, added 0.5% sucrose as a carbon source, and lowered the usage of artery powder from 1.2% to 0.6%. In the optimized medium, pseudoalterin production reached 161.15 ± 3.08 U/mL, 61% greater than that before optimization. We further conducted a small-scale fermentation experiment in a 5-L fermenter and a pilot-scale fermentation experiment in a 50-L fermenter. Pseudoalterin production during pilot-scale fermentation reached 103.48 ± 8.64 U/mL, 77% greater than that before the medium was optimized. In addition, through single factor experiments and orthogonal tests, we developed a compound stabilizer for pseudoalterin, using medically safe sugars and polyols. This stabilizer showed a significant protective effect for pseudoalterin against enzymatic thermal denaturation. These results lay a solid foundation for the industrial production of pseudoalterin and the development of its biomedical and biotechnological potentials.
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spelling pubmed-62733872018-12-28 Pilot-Scale Production and Thermostability Improvement of the M23 Protease Pseudoalterin from the Deep Sea Bacterium Pseudoalteromonas sp. CF6-2 Yang, Jie Yu, Yang Tang, Bai-Lu Zhong, Shuai Shi, Mei Xie, Bin-Bin Zhang, Xi-Ying Zhou, Bai-Cheng Zhang, Yu-Zhong Chen, Xiu-Lan Molecules Article Pseudoalterin is the most abundant protease secreted by the marine sedimental bacterium Pseudoalteromonas sp. CF6-2 and is a novel cold-adapted metalloprotease of the M23 family. Proteases of the M23 family have high activity towards peptidoglycan and elastin, suggesting their promising biomedical and biotechnological potentials. To lower the fermentive cost and improve the pseudoalterin production of CF6-2, we optimized the fermentation medium by using single factor experiments, added 0.5% sucrose as a carbon source, and lowered the usage of artery powder from 1.2% to 0.6%. In the optimized medium, pseudoalterin production reached 161.15 ± 3.08 U/mL, 61% greater than that before optimization. We further conducted a small-scale fermentation experiment in a 5-L fermenter and a pilot-scale fermentation experiment in a 50-L fermenter. Pseudoalterin production during pilot-scale fermentation reached 103.48 ± 8.64 U/mL, 77% greater than that before the medium was optimized. In addition, through single factor experiments and orthogonal tests, we developed a compound stabilizer for pseudoalterin, using medically safe sugars and polyols. This stabilizer showed a significant protective effect for pseudoalterin against enzymatic thermal denaturation. These results lay a solid foundation for the industrial production of pseudoalterin and the development of its biomedical and biotechnological potentials. MDPI 2016-11-17 /pmc/articles/PMC6273387/ /pubmed/27869696 http://dx.doi.org/10.3390/molecules21111567 Text en © 2016 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Yang, Jie
Yu, Yang
Tang, Bai-Lu
Zhong, Shuai
Shi, Mei
Xie, Bin-Bin
Zhang, Xi-Ying
Zhou, Bai-Cheng
Zhang, Yu-Zhong
Chen, Xiu-Lan
Pilot-Scale Production and Thermostability Improvement of the M23 Protease Pseudoalterin from the Deep Sea Bacterium Pseudoalteromonas sp. CF6-2
title Pilot-Scale Production and Thermostability Improvement of the M23 Protease Pseudoalterin from the Deep Sea Bacterium Pseudoalteromonas sp. CF6-2
title_full Pilot-Scale Production and Thermostability Improvement of the M23 Protease Pseudoalterin from the Deep Sea Bacterium Pseudoalteromonas sp. CF6-2
title_fullStr Pilot-Scale Production and Thermostability Improvement of the M23 Protease Pseudoalterin from the Deep Sea Bacterium Pseudoalteromonas sp. CF6-2
title_full_unstemmed Pilot-Scale Production and Thermostability Improvement of the M23 Protease Pseudoalterin from the Deep Sea Bacterium Pseudoalteromonas sp. CF6-2
title_short Pilot-Scale Production and Thermostability Improvement of the M23 Protease Pseudoalterin from the Deep Sea Bacterium Pseudoalteromonas sp. CF6-2
title_sort pilot-scale production and thermostability improvement of the m23 protease pseudoalterin from the deep sea bacterium pseudoalteromonas sp. cf6-2
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6273387/
https://www.ncbi.nlm.nih.gov/pubmed/27869696
http://dx.doi.org/10.3390/molecules21111567
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