Characterization of an α-agarase from Thalassomonas sp. LD5 and its hydrolysate

Zhang, Wei-Bin; Xu, Jingnan; Liú, Dan; Liu, Huan; Lu, Xin; Yu, Wengong

doi:10.1007/s00253-018-8762-6

Cited by 37 publications

(27 citation statements)

References 27 publications

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“…In contrast, such genes were absent in genome sequences of the nonagarolytic relatives C. psychrerythraea 34H T , C. piezophila Y223G T , and C. chukchiensis BCw111 T (Table S1B). Unique for C. echini was the presence of GH96 ␣-agarases, which are rarely reported and include only a few characterized members (15)(16)(17)(18). Of all the Colwellia genomes analyzed, that of C. echini encoded the largest amount of GH16 CAZymes, comprising ␤-agarases, ␤-porphyranases, laminarinases, and -carrageenases, and only C. echini encoded GH82 -carrageenases.…”

Section: Resultsmentioning

confidence: 99%

A Multifunctional Polysaccharide Utilization Gene Cluster in Colwellia echini Encodes Enzymes for the Complete Degradation of κ-Carrageenan, ι-Carrageenan, and Hybrid β/κ-Carrageenan

Christiansen

Pathiraja

Bech

et al. 2020

mSphere

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Algal cell wall polysaccharides constitute a large fraction in the biomass of marine primary producers and are thus important in nutrient transfer between trophic levels in the marine ecosystem. In order for this transfer to take place, polysaccharides must be degraded into smaller mono-and disaccharide units, which are subsequently metabolized, and key components in this degradation are bacterial enzymes. The marine bacterium Colwellia echini A3 T is a potent enzyme producer since it completely hydrolyzes agar and -carrageenan. Here, we report that the genome of C. echini A3 T harbors two large gene clusters for the degradation of carrageenan and agar, respectively. Phylogenetical and functional studies combined with transcriptomics and in silico structural modeling revealed that the carrageenolytic cluster encodes furcellaranases, a new class of glycoside hydrolase family 16 (GH16) enzymes that are key enzymes for hydrolysis of furcellaran, a hybrid carrageenan containing both ␤and -carrageenan motifs. We show that furcellaranases degrade furcellaran into neocarratetraose-43-O-monosulfate [DA-(␣1,3)-G4S-(␤1,4)-DA-(␣1,3)-G], and we propose a molecular model of furcellaranases and compare the active site architectures of furcellaranases, -carrageenases, ␤-agarases, and ␤-porphyranases. Furthermore, C. echini A3 T was shown to encode -carrageenases, -carrageenases, and members of a new class of enzymes, active only on hybrid ␤/-carrageenan tetrasaccharides. On the basis of our genomic, transcriptomic, and functional analyses of the carrageenolytic enzyme repertoire, we propose a new model for how C. echini A3 T degrades complex sulfated marine polysaccharides such as furcellaran, -carrageenan, and -carrageenan. IMPORTANCE Here, we report that a recently described bacterium, Colwellia echini, harbors a large number of enzymes enabling the bacterium to grow on -carrageenan and agar. The genes are organized in two clusters that encode enzymes for the total degradation of -carrageenan and agar, respectively. As the first, we report on the structure/ function relationship of a new class of enzymes that hydrolyze furcellaran, a partially sulfated ␤/-carrageenan. Using an in silico model, we hypothesize a molecular structure of furcellaranases and compare structural features and active site architectures of furcellaranases with those of other GH16 polysaccharide hydrolases, such as -carrageenases, ␤-agarases, and ␤-porphyranases. Furthermore, we describe a new class of enzymes distantly related to GH42 and GH160 ␤-galactosidases and show that this new class of enzymes is active only on hybrid ␤/-carrageenan oligosaccharides. Finally, we propose a new model for how the carrageenolytic enzyme repertoire enables C. echini to metabolize ␤/-, -, and -carrageenan.

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Section: Resultsmentioning

confidence: 99%

A Multifunctional Polysaccharide Utilization Gene Cluster in Colwellia echini Encodes Enzymes for the Complete Degradation of κ-Carrageenan, ι-Carrageenan, and Hybrid β/κ-Carrageenan

Christiansen

Pathiraja

Bech

et al. 2020

mSphere

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“…However, GH50 enzymes with similar sequences and structures are either endolytic or exolytic (Jung et al., 2017). In contrast to the abundance of β‐agarases, only three α‐agarases have been reported, and they all belong to GH96 (Zhang et al., 2018). NABHs are in family GH117 (Hehemann, Smyth, Yadav, Vocadlo, & Boraston, 2012).…”

Section: Marine‐polysaccharide Degrading Enzymesmentioning

confidence: 99%

Marine‐polysaccharide degrading enzymes: Status and prospects

Sun

Gao

Xue

et al. 2020

Comp Rev Food Sci Food Safe

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Marine-polysaccharide degrading enzymes have recently been studied extensively. They are particularly interesting as they catalyze the cleavage of glycosidic bonds in polysaccharide macromolecules and produce oligosaccharides with low degrees of polymerization. Numerous findings have demonstrated that marine polysaccharides and their biotransformed products possess beneficial properties including antitumor, antiviral, anticoagulant, and anti-inflammatory activities, and they have great value in healthcare, cosmetics, the food industry, and agriculture. Exploitation of enzymes that can degrade marine polysaccharides is in the ascendant, and is important for high-value use of marine biomass resources. In this review, we describe research and prospects regarding the classification, biochemical properties, and catalytic mechanisms of the main types of marine-polysaccharide degrading enzymes, focusing on chitinase, chitosanase, alginate lyase, agarase, and carrageenase, and their product oligosaccharides. The state-of-the-art discussion of marine-polysaccharide degrading enzymes and their properties offers information that might enable more efficient production of marine oligosaccharides. We also highlight current problems in the field of marine-polysaccharide degrading enzymes and trends in their development. Understanding the properties, catalytic mechanisms, and modification of known enzymes will aid the identification of novel enzymes to degrade marine polysaccharides and facilitation of their use in various biotechnological processes. K E Y W O R D S agarase, alginate lyase, carrageenase, chitinase, chitosanase 1 INTRODUCTION Ocean occupies >70% of the global surface, and the vast oceanic water environment has spawned large and complex marine ecosystems with rich biodiversity. There is a wide variety of marine resources, however, they are largely unexploited. This situation makes the marine environment very attractive for bioprospecting to discover new functions and molecules (Arnaud-Haond, Arrieta, & Duarte, 2011). Research into marine compounds, especially polysaccharides from renewable biomass, is attracting increasing attention. Polysaccharides make up one of the largest

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“…The α-agarase AgaD from Thalassomonas sp. LD5 was shown to hydrolyze agarotetraose and generated agarotriose with a molecular weight of 486 g/mol and the G-A-G structural arrangement [24]. In the current study, rGaa16Bc produced neoagarotriose after hydrolyzing agarose, neoagarohexaose, and neoagarotetraose.…”

Section: Discussionmentioning

confidence: 50%

A Novel Neoagarotriose-Producing Agarase from the Marine Bacterium Gilvimarinus Agarilyticus JEA5

Lee¹,

Jo²,

Lee³

et al. 2020

Preprint

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Background The degradation of agar by bacterial agarases has many commercial and academic applications. We recently identified a novel neoagarotriose-producing β-agarase, Gaa16B, in the marine bacterium Gilvimarinus agarilyticus JEA5. This is the first report to describe neoagarotriose production from β-agarase.Results The Gaa16B agarase, which belongs to the glycoside hydrolase 16 (GH16) family of β-agarases, shows less than 70.9% amino acid similarity with previously characterized agarases. The coding region of Gaa16B is 1800 bp long, encoding 600 amino acids, and exhibits features typical of agarases belonging to the GH16 family. A recombinant Gaa16B lacking the carbohydrate binding region (rGaa16Bc) was overexpressed in Escherichia coli and purified as a maltose-binding protein (MBP) fusion protein. 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, neoagarotriose, and neoagarobiose, and the production of neoagarotriose by rGaa16Bc was successfully validated by high-resolution electrospray ionization mass spectrometry.Conclusion The biochemical properties of Gaa16B and the results of the hydrolytic pattern analysis suggest that Gaa16B could be useful to produce functional neoagaro-oligosaccharides for industrial applications.

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Characterization of an α-agarase from Thalassomonas sp. LD5 and its hydrolysate

Cited by 37 publications

References 27 publications

A Multifunctional Polysaccharide Utilization Gene Cluster in Colwellia echini Encodes Enzymes for the Complete Degradation of κ-Carrageenan, ι-Carrageenan, and Hybrid β/κ-Carrageenan

A Multifunctional Polysaccharide Utilization Gene Cluster in Colwellia echini Encodes Enzymes for the Complete Degradation of κ-Carrageenan, ι-Carrageenan, and Hybrid β/κ-Carrageenan

Marine‐polysaccharide degrading enzymes: Status and prospects

A Novel Neoagarotriose-Producing Agarase from the Marine Bacterium Gilvimarinus Agarilyticus JEA5

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