Whole genome sequencing of a novel carrageenan-degrading bacterium Photobacterium rosenbergii and oligosaccharide preparation

IntroductionCarrageenan oligosaccharides are of significant interest due to their diverse bioactivities, necessitating efficient methods for their production. To this day, the discovery and isolation of microorganisms capable of effectively degrading carrageenan is still crucial for the production o...

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Main Authors: Jing Chen, Runmin Chen, Kit-Leong Cheong, Zhuo Wang, Rui Li, Xuejing Jia, Qiaoli Zhao, Xiaofei Liu, Bingbing Song, Saiyi Zhong
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-01-01
Series:Frontiers in Microbiology
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Online Access:https://www.frontiersin.org/articles/10.3389/fmicb.2025.1519074/full
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author Jing Chen
Jing Chen
Runmin Chen
Kit-Leong Cheong
Zhuo Wang
Rui Li
Xuejing Jia
Qiaoli Zhao
Xiaofei Liu
Bingbing Song
Saiyi Zhong
Saiyi Zhong
author_facet Jing Chen
Jing Chen
Runmin Chen
Kit-Leong Cheong
Zhuo Wang
Rui Li
Xuejing Jia
Qiaoli Zhao
Xiaofei Liu
Bingbing Song
Saiyi Zhong
Saiyi Zhong
author_sort Jing Chen
collection DOAJ
description IntroductionCarrageenan oligosaccharides are of significant interest due to their diverse bioactivities, necessitating efficient methods for their production. To this day, the discovery and isolation of microorganisms capable of effectively degrading carrageenan is still crucial for the production of carrageenan oligosaccharides. In addition, there are no current reports of bacteria of the genus Photobacterium capable of secreting κ-carrageenanase or degrading carrageenan.MethodsIn the current study, strain GDSX-4 was obtained from Gracilaria coronopifolia after enrichment culture, primary screening and rescreening and was initially characterized by morphology and 16SrDNA. The pure culture of strain GDSX-4 was further subjected to bacterial genome sequencing assembly and bioinformatic analysis. Specifically, homology group cluster (COG) annotation, CAZy (carbohydrate-active enzyme) database annotation and CAZyme genome clusters (CGCs) annotation were utilized to identify potential polysaccharide degradation functions. Enzymatic activity was assessed under different conditions, including substrate, temperature, pH, and the presence of metal ions. Hydrolysis products were analyzed using thin-layer chromatography (TLC) and electrospray ionization mass spectrometry (ESI-MS).ResultsPhotobacterium rosenbergii GDSX-4 is a Gram-negative bacterium isolated from the red algae, capable of degrading several polysaccharides. The draft genome was predicted to have 6,407,375 bp, 47.55% G+C content and 6,749 genes. Among them, 214 genes encoding carbohydrate enzymes were annotated, including carrageenase, agarose, alginate lyase, and chitinase. GDSX-4 exhibited remarkable carrageenan-degrading activity, with a specific enzyme activity of 46.94 U/mg. Optimal hydrolysis conditions were determined to be 40°C and pH 7.0, with the enzyme retaining 80% of its activity below 30°C and across a pH range of 4.0–10.0. Metal ions such as as K+, Na+, and Ba2+ enhanced enzymatic activity, while Ni2+, Mn2+, and Cu2+ had inhibitory effects. kappa-carrageenan was totally hydrolyzed into oligosaccharides with degrees of polymerization ranging from 2 to 6.DiscussionThese findings highlight the potential of GDSX-4 for the efficient production of carrageenan oligosaccharides, paving the way for applications in the food and agricultural industries. Future studies may focus on the efficient expression of κ-carrageenase and expand its industrial application in the preparation of oligosaccharides.
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spelling doaj-art-7a11902d3b614fc29f4e80ecacf1a8322025-01-23T14:44:37ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2025-01-011610.3389/fmicb.2025.15190741519074Whole genome sequencing of a novel carrageenan-degrading bacterium Photobacterium rosenbergii and oligosaccharide preparationJing Chen0Jing Chen1Runmin Chen2Kit-Leong Cheong3Zhuo Wang4Rui Li5Xuejing Jia6Qiaoli Zhao7Xiaofei Liu8Bingbing Song9Saiyi Zhong10Saiyi Zhong11Shenzhen Research Institute, Guangdong Ocean University, Shenzhen, ChinaGuangdong Provincial Key Laboratory of Aquatic Product Processing and Safety, Guangdong Province Engineering Laboratory for Marine Biological Products, Guangdong Provincial Engineering Technology Research Center of Seafood, College of Food Science and Technology, Guangdong Ocean University, Zhanjiang, ChinaGuangdong Provincial Key Laboratory of Aquatic Product Processing and Safety, Guangdong Province Engineering Laboratory for Marine Biological Products, Guangdong Provincial Engineering Technology Research Center of Seafood, College of Food Science and Technology, Guangdong Ocean University, Zhanjiang, ChinaGuangdong Provincial Key Laboratory of Aquatic Product Processing and Safety, Guangdong Province Engineering Laboratory for Marine Biological Products, Guangdong Provincial Engineering Technology Research Center of Seafood, College of Food Science and Technology, Guangdong Ocean University, Zhanjiang, ChinaGuangdong Provincial Key Laboratory of Aquatic Product Processing and Safety, Guangdong Province Engineering Laboratory for Marine Biological Products, Guangdong Provincial Engineering Technology Research Center of Seafood, College of Food Science and Technology, Guangdong Ocean University, Zhanjiang, ChinaGuangdong Provincial Key Laboratory of Aquatic Product Processing and Safety, Guangdong Province Engineering Laboratory for Marine Biological Products, Guangdong Provincial Engineering Technology Research Center of Seafood, College of Food Science and Technology, Guangdong Ocean University, Zhanjiang, ChinaGuangdong Provincial Key Laboratory of Aquatic Product Processing and Safety, Guangdong Province Engineering Laboratory for Marine Biological Products, Guangdong Provincial Engineering Technology Research Center of Seafood, College of Food Science and Technology, Guangdong Ocean University, Zhanjiang, ChinaGuangdong Provincial Key Laboratory of Aquatic Product Processing and Safety, Guangdong Province Engineering Laboratory for Marine Biological Products, Guangdong Provincial Engineering Technology Research Center of Seafood, College of Food Science and Technology, Guangdong Ocean University, Zhanjiang, ChinaGuangdong Provincial Key Laboratory of Aquatic Product Processing and Safety, Guangdong Province Engineering Laboratory for Marine Biological Products, Guangdong Provincial Engineering Technology Research Center of Seafood, College of Food Science and Technology, Guangdong Ocean University, Zhanjiang, ChinaGuangdong Provincial Key Laboratory of Aquatic Product Processing and Safety, Guangdong Province Engineering Laboratory for Marine Biological Products, Guangdong Provincial Engineering Technology Research Center of Seafood, College of Food Science and Technology, Guangdong Ocean University, Zhanjiang, ChinaShenzhen Research Institute, Guangdong Ocean University, Shenzhen, ChinaGuangdong Provincial Key Laboratory of Aquatic Product Processing and Safety, Guangdong Province Engineering Laboratory for Marine Biological Products, Guangdong Provincial Engineering Technology Research Center of Seafood, College of Food Science and Technology, Guangdong Ocean University, Zhanjiang, ChinaIntroductionCarrageenan oligosaccharides are of significant interest due to their diverse bioactivities, necessitating efficient methods for their production. To this day, the discovery and isolation of microorganisms capable of effectively degrading carrageenan is still crucial for the production of carrageenan oligosaccharides. In addition, there are no current reports of bacteria of the genus Photobacterium capable of secreting κ-carrageenanase or degrading carrageenan.MethodsIn the current study, strain GDSX-4 was obtained from Gracilaria coronopifolia after enrichment culture, primary screening and rescreening and was initially characterized by morphology and 16SrDNA. The pure culture of strain GDSX-4 was further subjected to bacterial genome sequencing assembly and bioinformatic analysis. Specifically, homology group cluster (COG) annotation, CAZy (carbohydrate-active enzyme) database annotation and CAZyme genome clusters (CGCs) annotation were utilized to identify potential polysaccharide degradation functions. Enzymatic activity was assessed under different conditions, including substrate, temperature, pH, and the presence of metal ions. Hydrolysis products were analyzed using thin-layer chromatography (TLC) and electrospray ionization mass spectrometry (ESI-MS).ResultsPhotobacterium rosenbergii GDSX-4 is a Gram-negative bacterium isolated from the red algae, capable of degrading several polysaccharides. The draft genome was predicted to have 6,407,375 bp, 47.55% G+C content and 6,749 genes. Among them, 214 genes encoding carbohydrate enzymes were annotated, including carrageenase, agarose, alginate lyase, and chitinase. GDSX-4 exhibited remarkable carrageenan-degrading activity, with a specific enzyme activity of 46.94 U/mg. Optimal hydrolysis conditions were determined to be 40°C and pH 7.0, with the enzyme retaining 80% of its activity below 30°C and across a pH range of 4.0–10.0. Metal ions such as as K+, Na+, and Ba2+ enhanced enzymatic activity, while Ni2+, Mn2+, and Cu2+ had inhibitory effects. kappa-carrageenan was totally hydrolyzed into oligosaccharides with degrees of polymerization ranging from 2 to 6.DiscussionThese findings highlight the potential of GDSX-4 for the efficient production of carrageenan oligosaccharides, paving the way for applications in the food and agricultural industries. Future studies may focus on the efficient expression of κ-carrageenase and expand its industrial application in the preparation of oligosaccharides.https://www.frontiersin.org/articles/10.3389/fmicb.2025.1519074/fullPhotobacterium rosenbergiicarrageenanoligosaccharidesgenome sequencingpreparation
spellingShingle Jing Chen
Jing Chen
Runmin Chen
Kit-Leong Cheong
Zhuo Wang
Rui Li
Xuejing Jia
Qiaoli Zhao
Xiaofei Liu
Bingbing Song
Saiyi Zhong
Saiyi Zhong
Whole genome sequencing of a novel carrageenan-degrading bacterium Photobacterium rosenbergii and oligosaccharide preparation
Frontiers in Microbiology
Photobacterium rosenbergii
carrageenan
oligosaccharides
genome sequencing
preparation
title Whole genome sequencing of a novel carrageenan-degrading bacterium Photobacterium rosenbergii and oligosaccharide preparation
title_full Whole genome sequencing of a novel carrageenan-degrading bacterium Photobacterium rosenbergii and oligosaccharide preparation
title_fullStr Whole genome sequencing of a novel carrageenan-degrading bacterium Photobacterium rosenbergii and oligosaccharide preparation
title_full_unstemmed Whole genome sequencing of a novel carrageenan-degrading bacterium Photobacterium rosenbergii and oligosaccharide preparation
title_short Whole genome sequencing of a novel carrageenan-degrading bacterium Photobacterium rosenbergii and oligosaccharide preparation
title_sort whole genome sequencing of a novel carrageenan degrading bacterium photobacterium rosenbergii and oligosaccharide preparation
topic Photobacterium rosenbergii
carrageenan
oligosaccharides
genome sequencing
preparation
url https://www.frontiersin.org/articles/10.3389/fmicb.2025.1519074/full
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