Biochemical Characterization and Substrate Degradation Mode of a Novel α-Agarase from <i>Catenovulum agarivorans</i>

Liu, Jie; Liu, Zhen; Jiang, Chen; Mao, Xiangzhao

doi:10.1021/acs.jafc.9b03073

Cited by 25 publications

(13 citation statements)

References 29 publications

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“…Thus, polysaccharide lyases and glycosyl hydrolases were isolated from microorganisms, including fungi and bacteria. The genus Catenovulum have been known as bacteria that can degrade agar and polysaccharide containing uronic acids, such as alginate, pectin, and ulvan (Cui et al, 2014;Li et al, 2015;Lee, Lee & Hong, 2019;Liu et al, 2019a;Xie et al, 2013). Our previous report demonstrated that many genes encoding polysaccharide-degrading enzymes were present in the QB4 genome (Lau et al, 2019).…”

Section: Discussionmentioning

confidence: 99%

Genes for degradation and utilization of uronic acid-containing polysaccharides of a marine bacteriumCatenovulumsp. CCB-QB4

Furusawa

Azami

Teh

2021

PeerJ

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Background Oligosaccharides from polysaccharides containing uronic acids are known to have many useful bioactivities. Thus, polysaccharide lyases (PLs) and glycoside hydrolases (GHs) involved in producing the oligosaccharides have attracted interest in both medical and industrial settings. The numerous polysaccharide lyases and glycoside hydrolases involved in producing the oligosaccharides were isolated from soil and marine microorganisms. Our previous report demonstrated that an agar-degrading bacterium, Catenovulum sp. CCB-QB4, isolated from a coastal area of Penang, Malaysia, possessed 183 glycoside hydrolases and 43 polysaccharide lyases in the genome. We expected that the strain might degrade and use uronic acid-containing polysaccharides as a carbon source, indicating that the strain has a potential for a source of novel genes for degrading the polysaccharides. Methods To confirm the expectation, the QB4 cells were cultured in artificial seawater media with uronic acid-containing polysaccharides, namely alginate, pectin (and saturated galacturonate), ulvan, and gellan gum, and the growth was observed. The genes involved in degradation and utilization of uronic acid-containing polysaccharides were explored in the QB4 genome using CAZy analysis and BlastP analysis. Results The QB4 cells were capable of using these polysaccharides as a carbon source, and especially, the cells exhibited a robust growth in the presence of alginate. 28 PLs and 22 GHs related to the degradation of these polysaccharides were found in the QB4 genome based on the CAZy database. Eleven polysaccharide lyases and 16 glycoside hydrolases contained lipobox motif, indicating that these enzymes play an important role in degrading the polysaccharides. Fourteen of 28 polysaccharide lyases were classified into ulvan lyase, and the QB4 genome possessed the most abundant ulvan lyase genes in the CAZy database. Besides, genes involved in uronic acid metabolisms were also present in the genome. These results were consistent with the cell growth. In the pectin metabolic pathway, the strain had genes for three different pathways. However, the growth experiment using saturated galacturonate exhibited that the strain can only use the pathway related to unsaturated galacturonate.

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Section: Discussionmentioning

confidence: 99%

Genes for degradation and utilization of uronic acid-containing polysaccharides of a marine bacteriumCatenovulumsp. CCB-QB4

Furusawa

Azami

Teh

2021

PeerJ

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“…Agarases play a key role in the preparation of specific agaro-oligosaccharides. According to the glycosidic bonds they cleave in agarose, agarases can be divided into α-agarase of cutting α-1,3-glycosidic linkage and β-agarase of cutting β-1,4-glycosidic linkage to produce agarooligosaccharides (AOS) and neoagarooligosaccharides (NAOS) from hydrolysis of agarose, respectively. , Therein, several α-agarases have been identified and characterized; all of them belong to the glycoside hydrolase (GH) 96 family. − Compared with α-agarases, the β-agarases were explored more exhaustive, so far, over 50 β-agarases has been expressed heterologous. , These characterized β-agarases belong to four GH families, including GH16, GH50, GH86, and GH118. Therein, the β-agarases of GH16 and GH50 were most identified.…”

Section: Introductionmentioning

confidence: 99%

Characterization of a GH50 β-Agarase: A Biotechnological Tool for Preparing Oligosaccharides from Agarose and Porphyran

Jiang

Zhang

et al. 2022

J. Agric. Food Chem.

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Agarase is of vital significance for functional agaro-oligosaccharides production from algal dived agarose. Especially, the exolytic agarases have the advantage of obtaining agaro-oligosaccharides with a specific degree of polymerization. Herein, we cloned and expressed a novel glycoside hydrolase (GH) 50 family β-agarase OUC-PgJC50 from Photobacterium gaetbulicola. The degradation pattern analysis indicated that OUC-PgJC50 not only showed an exolytic activity with main products of neoagarotetraose from hydrolyzing agarose but also show a hydrolytic activity to transform neoagarotetraose into neoagarobiose. This is the first time that the discovery of a neoagarotetraose-producing exolytic GH50 β-agarase possesses the activity to transform neoagarotetraose into neoagarobiose, which provided new insight into the recognition of the degradation mode of agarases. Molecular docking and sequence alignment analysis further revealed the His654 residue in OUC-PgJC50 may play a vital role in forming a strong force with l-AHG residue at −4 subsite that helps to produce neoagarotetraose from catalyzing agarose. Moreover, the catalytic ability of OUC-PgJC50 toward another agar polysaccharide porphyran was also described that could hydrolyze porphyran into sulfated oligosaccharides, in which the LA6S-d-Gal was the main products. This study is of vital significance for developing the application range of GH50 β-agarases.

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“…Among the many agarases reported, most are β-agarases, except some α-agarases isolated from Alteromonas agarilyticus GJ1B [15], Thalassomonas agarivorans JAMB-A33, Catenovulum agarivorans [16], and Caternovulum sediminis WS1-A [17]. Several reports have described that β-agarases hydrolyze agarose to produce various neoagaro-oligosaccharides, such as neoagarobiose, neoagarotetraose, and neoagarohexaose [1].…”

Section: Introductionmentioning

confidence: 99%

A Novel Agarase, Gaa16B, Isolated from the Marine Bacterium Gilvimarinus agarilyticus JEA5, and the Moisturizing Effect of Its Partial Hydrolysis Products

Lee

Eom

et al. 2021

Marine Drugs

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We recently identified a β-agarase, Gaa16B, in the marine bacterium Gilvimarinus agarilyticus JEA5. Gaa16B, belonging to the glycoside hydrolase 16 family of β-agarases, shows less than 70.9% amino acid similarity with previously characterized agarases. Recombinant Gaa16B lacking the carbohydrate-binding region (rGaa16Bc) was overexpressed in Escherichia coli and purified. Activity assays revealed the optimal temperature and pH of rGaa16Bc to be 55 ∘C and pH 6–7, respectively, and the protein was highly stable at 55 ∘C for 90 min. Additionally, rGaa16Bc activity was strongly enhanced (2.3-fold) in the presence of 2.5 mM MnCl2. The Km and Vmax of rGaa16Bc for agarose were 6.4 mg/mL and 953 U/mg, respectively. Thin-layer chromatography analysis revealed that rGaa16Bc can hydrolyze agarose into neoagarotetraose and neoagarobiose. Partial hydrolysis products (PHPs) of rGaa16Bc had an average molecular weight of 88–102 kDa and exhibited > 60% hyaluronidase inhibition activity at a concentration of 1 mg/mL, whereas the completely hydrolyzed product (CHP) showed no hyaluronidase at the same concentration. The biochemical properties of Gaa16B suggest that it could be useful for producing functional neoagaro-oligosaccharides. Additionally, the PHP of rGaa16Bc may be useful in promoting its utilization, which is limited due to the gel strength of agar.

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Biochemical Characterization and Substrate Degradation Mode of a Novel α-Agarase from Catenovulum agarivorans

Cited by 25 publications

References 29 publications

Genes for degradation and utilization of uronic acid-containing polysaccharides of a marine bacteriumCatenovulumsp. CCB-QB4

Genes for degradation and utilization of uronic acid-containing polysaccharides of a marine bacteriumCatenovulumsp. CCB-QB4

Characterization of a GH50 β-Agarase: A Biotechnological Tool for Preparing Oligosaccharides from Agarose and Porphyran

A Novel Agarase, Gaa16B, Isolated from the Marine Bacterium Gilvimarinus agarilyticus JEA5, and the Moisturizing Effect of Its Partial Hydrolysis Products

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