Preparation and characterization of immobilized mannanase on polyhydroxyalkanoate (PHA)

Abstract Galactomannan oligosaccharides (GMOS), composed of 2–10 mannose units linked with β-1, 4 glycosidic bond as the main chain and galactose linked with α-1, 6 glycosidic bond as the side chain, are crucial for probiotic food synthesis due to their ability to promote the growth and activity of...

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Main Authors: Zhiyue Men, Yafei Zhang, Zhao Pang, Tao Li, Hening Zhang, Yue Zhou, Ning Hao, Yajian Song, Yu Bai
Format: Article
Language:English
Published: SpringerOpen 2025-06-01
Series:Bioresources and Bioprocessing
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Online Access:https://doi.org/10.1186/s40643-025-00886-5
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author Zhiyue Men
Yafei Zhang
Zhao Pang
Tao Li
Hening Zhang
Yue Zhou
Ning Hao
Yajian Song
Yu Bai
author_facet Zhiyue Men
Yafei Zhang
Zhao Pang
Tao Li
Hening Zhang
Yue Zhou
Ning Hao
Yajian Song
Yu Bai
author_sort Zhiyue Men
collection DOAJ
description Abstract Galactomannan oligosaccharides (GMOS), composed of 2–10 mannose units linked with β-1, 4 glycosidic bond as the main chain and galactose linked with α-1, 6 glycosidic bond as the side chain, are crucial for probiotic food synthesis due to their ability to promote the growth and activity of beneficial intestinal microbiota, enhance the host immune system, and improve nutrient digestion. GMOS is usually obtained by hydrolyzing plants such as locust bean gum and guar gum with mannanase. β-mannanase ManA from Alkaliphilic Bacillus sp. N16-5 can hydrolyze β-1, 4 glycosidic bond of galactomannan. In this study, an immobilization system was employed utilizing polyhydroxyalkanoate (PHA) biopolymers, which naturally have an affinity mainly mediated by hydrophobic interaction for PhaP protein. Fusion protein combining ManA with PhaP from Aeromonas hydrophila, was subsequently immobilized on PHA support to form a multi-enzyme complex, facilitating the hydrolysis of locust bean gum to generate GMOS. This immobilized enzyme enhances enzyme stability and reusability, can be reused up to 32 times while maintaining ~ 80% of its activity, offering substantial cost savings through in-situ enzyme and product separation. Additionally, the different PHA forms were developed to hydrolyze locust bean gum to produce GMOS, such as nano PHA particles, PHA electrospun materials, while these preliminary investigations show promise, further research is needed to optimize their performance and practical application. Graphical Abstract
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spelling doaj-art-4e6d06d1817c487bb8191e6eaa00b4fd2025-08-20T03:24:26ZengSpringerOpenBioresources and Bioprocessing2197-43652025-06-011211910.1186/s40643-025-00886-5Preparation and characterization of immobilized mannanase on polyhydroxyalkanoate (PHA)Zhiyue Men0Yafei Zhang1Zhao Pang2Tao Li3Hening Zhang4Yue Zhou5Ning Hao6Yajian Song7Yu Bai8Tianjin Institute of Industrial Biotechnology, Chinese Academy of SciencesTianjin Institute of Industrial Biotechnology, Chinese Academy of SciencesTianjin Institute of Industrial Biotechnology, Chinese Academy of SciencesTianjin Institute of Industrial Biotechnology, Chinese Academy of SciencesTianjin Institute of Industrial Biotechnology, Chinese Academy of SciencesTianjin Institute of Industrial Biotechnology, Chinese Academy of SciencesTianjin Institute of Industrial Biotechnology, Chinese Academy of SciencesCollege of Biological Engineering, Tianjin University of Science & TechnologyHaihe Laboratory of Synthetic BiologyAbstract Galactomannan oligosaccharides (GMOS), composed of 2–10 mannose units linked with β-1, 4 glycosidic bond as the main chain and galactose linked with α-1, 6 glycosidic bond as the side chain, are crucial for probiotic food synthesis due to their ability to promote the growth and activity of beneficial intestinal microbiota, enhance the host immune system, and improve nutrient digestion. GMOS is usually obtained by hydrolyzing plants such as locust bean gum and guar gum with mannanase. β-mannanase ManA from Alkaliphilic Bacillus sp. N16-5 can hydrolyze β-1, 4 glycosidic bond of galactomannan. In this study, an immobilization system was employed utilizing polyhydroxyalkanoate (PHA) biopolymers, which naturally have an affinity mainly mediated by hydrophobic interaction for PhaP protein. Fusion protein combining ManA with PhaP from Aeromonas hydrophila, was subsequently immobilized on PHA support to form a multi-enzyme complex, facilitating the hydrolysis of locust bean gum to generate GMOS. This immobilized enzyme enhances enzyme stability and reusability, can be reused up to 32 times while maintaining ~ 80% of its activity, offering substantial cost savings through in-situ enzyme and product separation. Additionally, the different PHA forms were developed to hydrolyze locust bean gum to produce GMOS, such as nano PHA particles, PHA electrospun materials, while these preliminary investigations show promise, further research is needed to optimize their performance and practical application. Graphical Abstracthttps://doi.org/10.1186/s40643-025-00886-5MannanasePolyhydroxyalkanoatesImmobilization
spellingShingle Zhiyue Men
Yafei Zhang
Zhao Pang
Tao Li
Hening Zhang
Yue Zhou
Ning Hao
Yajian Song
Yu Bai
Preparation and characterization of immobilized mannanase on polyhydroxyalkanoate (PHA)
Bioresources and Bioprocessing
Mannanase
Polyhydroxyalkanoates
Immobilization
title Preparation and characterization of immobilized mannanase on polyhydroxyalkanoate (PHA)
title_full Preparation and characterization of immobilized mannanase on polyhydroxyalkanoate (PHA)
title_fullStr Preparation and characterization of immobilized mannanase on polyhydroxyalkanoate (PHA)
title_full_unstemmed Preparation and characterization of immobilized mannanase on polyhydroxyalkanoate (PHA)
title_short Preparation and characterization of immobilized mannanase on polyhydroxyalkanoate (PHA)
title_sort preparation and characterization of immobilized mannanase on polyhydroxyalkanoate pha
topic Mannanase
Polyhydroxyalkanoates
Immobilization
url https://doi.org/10.1186/s40643-025-00886-5
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