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Polymerized guluronates (polyG)-specific alginate lyase with lower polymerized mannuronates (polyM)-degrading activity, superior stability, and clear action mode is a powerful biotechnology tool for the preparation of AOSs rich in M blocks. In this study, we expressed and characterized a polyG-specific alginate lyase OUC-FaAly7 from Formosa agariphila KMM3901. OUC-FaAly7 belonging to polysaccharide lyase (PL) family 7 had highest activity (2743.7 ± 20.3 U/μmol) at 45 °C and pH 6.0. Surprisingly, its specific activity against polyG reached 8560.2 ± 76.7 U/μmol, whereas its polyM-degrading activity was nearly 0 within 10 min reaction. Suggesting that OUC-FaAly7 was a strict polyG-specific alginate lyase. Importantly, OUC-FaAly7 showed a wide range of temperature adaptations and remarkable temperature and pH stability. Its relative activity between 20 °C and 45 °C reached >90 % of the maximum activity. The minimum identifiable substrate of OUC-FaAly7 was guluronate tetrasaccharide (G4). Action process and mode showed that it was a novel alginate lyase digesting guluronate hexaose (G6), guluronate heptaose (G7), and polymerized guluronates, with the preferential generation of unsaturated guluronate pentasaccharide (UG5), although which could be further degraded into unsaturated guluronate disaccharide (UG3) and trisaccharide (UG2). This study contributes to illustrating the catalytic properties, substrate recognition, and action mode of novel polyG-specific alginate lyases., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Although chitooligosaccharide (COS) has attracted the attention of some researchers due to its good solubility and broad-spectrum antibacterial activity, our study found that Staphylococcus aureus treated with low concentration of COS actively entered the viable-but-nonculturable (VBNC) state to resist this environmental stress. In this study, the transcriptome of VBNC-state S. aureus after COS treatment was analyzed by RNA-sequencing. Compared with the control group, pathway enrichment analysis showed that COS-treated S. aureus adopted a series of adaptive adjustment strategies for survival, including significant up-regulation of the differential genes' expression of such as ABC transporters (metI, tagG), Sec dependent transport pathway (secDF), peptidoglycan synthesis pathway (murG) and alteration of their physiological metabolic patterns, where ATP depletion played a key role in the formation of the VBNC-state S. aureus. Further, by using oxidative phosphorylation uncoupling agent to adjust the initial level of ATP in S. aureus, it was found that the reduction of intracellular ATP level could accelerate the formation of VBNC state. Overall, our results preliminarily elucidated the molecular mechanism of COS inducing the VBNC-state S. aureus. It provided an important theoretical reference for further achieving effective bacterial inactivation by COS., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Ltd.)

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 His 654 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.

2'-Fucosyllactose (2'-FL) is one of the nutrient ingredients in human milk, which has various beneficial health effects. α-l-fucosidase is a biotechnological tool for 2'-FL preparation. Here, a novel and efficient α-l-fucosidase OUC-Jdch16 from the fucoidan-digesting strain Flavobacterium algicola 12076 was heterologously expressed and applied to produce 2'-FL in vitro. OUC-Jdch16 belongs to glycoside hydrolases (GH) family 29 and exhibits the highest 4-nitrophenyl-α-l-fucopyranoside-hydrolyzing activity at 25 °C and pH 6.0. OUC-Jdch16 could catalyze the synthesis of 2'-FL via transferring the fucosyl residue from pNP-α-fucose to lactose. Under the optimal transfucosylation conditions, the yield of the transfucosylation product reached 84.82% and 92.15% (mol/mol) from pNP-α-fucose within 48 h and 120 h, respectively. Moreover, OUC-Jdch16 was capable of transferring the fucosyl residue to other glycosyl receptors with the generation of novel fucosylated compounds. This study demonstrated that OUC-Jdch16 could be a promising tool to prepare 2'-FL and other novel glycosides., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Agaro-oligosaccharides originating from red algae have attracted increasing attention in both basic theoretical research and applied fields due to their excellent bioactivities, which indicates the wide prospects of agaro-oligosaccharides for application in the food, pharmaceutical and cosmetic industries. Thus, a considerable number of studies regarding functional agaro-oligosaccharides preparation as well as the bioactivities exploration have been carried out. Based on these studies, this review first introduced different methods that have been used in agar extraction from red algae, and further provided research progress on arylsulfatase. Then, different methods used for agaro-oligosaccharides production were summarized. Moreover, the abundant bioactivities of agaro-oligosaccharides were described in detail. Finally, this review has discussed current research problems and further provided critical aspects, which may be helpful for revealing the structure-function relationship of agaro-oligosaccharide., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Brown algae, as the main source of alginate, are a type of marine biomass with a very high output. Alginate, a polysaccharide composed of β-D-mannuronic acid (M) and α-L-guluronic acid (G), has great potential for applications in the food, cosmetic and pharmaceutical industries. Alginate lyases (Alys) can degrade alginate polymers into oligosaccharides or monosaccharides, resulting in a broad application field. Alys can be used for both the production of alginate oligosaccharides and the biorefinery of brown algae. In view of their important functions, an increasing number of Alys have been isolated and characterized. For better application, a comprehensive understanding of Alys is essential. Therefore, in this paper, we summarized recently discovered Alys, discussed their characteristics, and introduced their structural properties, degradation patterns and biological roles in alginate-degrading organisms. In addition, applications of Alys have been illustrated with examples. This paper provides a relatively comprehensive description of Alys, which is significant for Alys exploration and application., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Here we have explored the effect of neoagarotetraose (NAT) on liver injury caused by intense exercise. Our results showed that NAT treatment obviously decreased liver weight ( p < 0.01), improved the liver morphological structure, decreased ALT level ( p < 0.05) and endotoxin (LPS) ( p < 0.01). In addition, NAT could regulate bile acid profiles in feces and serum of mice, which indicated the potential of liver function, suggesting that NAT was effective to relieve intense exercise-induced liver injury. NAT could regulate the expression of colon genes. NAT tended to alter the microbial composition of mice under intense exercise. We uncovered the network interactions between liver traits and microbial communities in NAT treatment mice. Interestingly, our data indicated that intense exercise-induced liver injury may be related to Clostridiales . In summary, these results demonstrated that NAT relieved liver injury induced by intense exercise may be related to gut microbiota.

Scope: Exhaustive exercise stress has emerged as an important health issue, and gastrointestinal problems are a common concern during intense exercise. In this study, we investigated the potential antifatigue effects of neoagarotetraose (NAT) in mice under intense exercise stress., Materials and Methods: Exhaustive exercise stress significantly weakened several physiological and physical parameters of the mice, including decreased food intake, reduced body weight, and impaired integrity of the intestinal epithelial barrier. Our data showed that a 16-day NAT treatment resulted in a profound change in microbiome composition, which subsequently led to widespread shifts in the functional potential of the gut microbiome. Furthermore, NAT administration significantly increased the fecal concentration of total short-chain fatty acids (p < 0.01)., Conclusion: Together, our findings suggest that NAT may protect mice against intense exercise-induced fatigue and provide insights into the mechanisms of NAT as a potential prebiotic., (© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Neoagaro-oligosaccharides derived from agarose have been demonstrated to possess a variety of biological activities, such as anti-bacteria and anti-oxidative activities. In this study, we mainly explored the inhibitory effects and the mechanisms of neoagaro-oligosaccharide monomers against LPS-induced inflammatory responses in mouse macrophage RAW264.7 cells. The results indicated that neoagaro-oligosaccharide monomers especially neoagarotetraose could significantly reduce the production and release of NO in LPS-induced macrophages. Neoagarotetraose significantly suppressed the expression and secretion of inducible nitric oxide synthase (iNOS) and proinflammatory cytokines such as TNF-α and IL-6. The inhibition mechanisms may be associated with the inhibition of the activation of p38MAPK, Ras/MEK/ERK and NF-κB signaling pathways. Thus, neoagarotetraose may attenuate the inflammatory responses through downregulating the MAPK and NF-κB signaling pathways in LPS-stimulated macrophages. In summary, the marine-derived neoagaro-oligosaccharide monomers merit further investigation as novel anti-inflammation agents in the future.

AgWH50A, a novel β-agarase, was cloned from Agarivorans gilvus WH0801 by degenerate and nested PCR. It consists of 942 amino acids (105 kDa), including a 21-amino acid signal peptide. AgWH50A shares the highest amino acid sequence homology with AgaD02 from Agarivorans sp. QM38 (53%). The recombinant agarase gene was expressed in Escherichia coli and purified by affinity chromatography. Maximum enzymatic activity (Km 5.97 mg/mL and Vmax 0.781 U/mg) was observed at pH 6.0 and 30 °C. Using matrix-assisted laser desorption/ionisation-time-of-flight mass spectrometry, Fourier transform-nuclear magnetic resonance spectrometry and thin-layer chromatography, we analysed the hydrolysis products and concluded that AgWH50A is a neoagarotetraose-forming β-agarase, which can cleave agarose into neoagarotetraose. This novel agarase has potential applications in the industrial production of neoagarotetraose and provides a new agarose hydrolysis model for future research., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

λ-carrageenan oligosaccharides can be widely applied in the food, pharmaceutical, medicine and cosmetic industries due to their abundant bioactivities, and they are important products for the high-value utilization of λ-carrageenan. However, oligosaccharides with different degrees of polymerization have different properties, and the final products of λ-carrageenase reported so far are mainly λ-neocarrabiose, λ-neocarratetraose and λ-neocarrahexaose without longer-chain oligosaccharides. Further research is consequently required. Herein, a mutant λ-carrageenase was constructed by deleting the pyrroloquinoline quinone-like domain of OUC-CglA derived from Maribacter vaceletii. Interestingly, it was discovered that the majority of final products of the mutant OUC-CglA-DPQQ were long-chain oligosaccharides with a polymerization degree of 10–20, which underwent significant changes compared to that of OUC-CglA. Additionally, without the pyrroloquinoline quinone-like domain, fewer inclusion bodies were produced throughout the expression process, and the yield of the λ-carrageenase increased about five-fold. However, compared to its parental enzyme, significant changes were made to its enzymatic properties. Its optimal temperature and pH were 15 °C and pH 7.0, and its specific activity was 51.59 U/mg. The stability of the enzyme decreased. Thus, it was found that the deleting domain was related to the formation of inclusion bodies, the stability of the enzyme, the activity of the enzyme and the composition of the products. [ABSTRACT FROM AUTHOR]

Fucoidan is one of the main polysaccharides of brown algae and echinoderm, which has nutritional and pharmacological functions. Due to the low molecular weight and exposure of more sulfate groups, oligo-fucoidan or fucoidan oligosaccharides have potential for broader applications. In this research, a novel endo-α-1,4-L-fucoidanase OUC-FaFcn1 which can degrade fucoidan into oligo-fucoidan was discovered from the fucoidan-digesting strain Flavobacterium algicola 12,076. OUC-FaFcn1 belongs to glycoside hydrolases (GH) family 107 and shows highest activity at 40 °C and pH 9.0. It can degrade the α-1,4 glycosidic bond, instead of α-1,3 glycosidic bond, of the fucoidan with a random tangent way to generate the principal product of disaccharide, which accounts for 49.4% of the total products. Therefore, OUC-FaFcn1 is a promising bio-catalyst for the preparation of fucoidan-derived disaccharide. These results further enrich the resource library of fucoidanase and provide the basis for the directional preparation of fucoidan-derived oligosaccharide with specific polymerization. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

β-Galactosidases have a wide application in the food and pharmaceutical industries. Recently, β-galactosidase was also found to participate in agar degradation. In this study, the second reported agarolytic β-galactosidase was found in the marine bacterium Agarivorans gilvus WH0801 and characterized. The β-galactosidase named AgWH2A (83 kDa) exhibited good activities under optimal hydrolysis conditions of pH 8.0 and 40 °C. AgWH2A could cleave the first D-galactose of agarooligosaccharides from its nonreducing end to produce neoagarooligosaccharides, but could not act on the neoagarooligosaccharides. AgWH2A has great potential in the comprehensive utilization of marine red algae. [ABSTRACT FROM AUTHOR]

Carrageenan, the major carbohydrate component of some red algae, is an important renewable bioresource with very large annual outputs. Different types of carrageenolytic enzymes in the carrageenan metabolic pathway are potentially valuable for the production of carrageenan oligosaccharides, biofuel, and other chemicals obtained from carrageenan. However, these enzymes are not well-developed for oligosaccharide or biofuel production. For further application, comprehensive knowledge of carrageenolytic enzymes is essential. Therefore, in this review, we first summarize various carrageenolytic enzymes, including the recently discovered β-carrageenase, carrageenan-specific sulfatase, exo -α-3,6-anhydro-D-galactosidase (D-ADAGase), and exo-β-galactosidase (BGase), and describe their enzymatic characteristics. Subsequently, the carrageenan metabolic pathways are systematically presented and applications of carrageenases and carrageenan oligosaccharides are illustrated with examples. Finally, this paper discusses critical aspects that can aid researchers in constructing cascade catalytic systems and engineered microorganisms to efficiently produce carrageenan oligosaccharides or other value-added chemicals through the degradation of carrageenan. Overall, this paper offers a comprehensive overview of carrageenolytic enzymes, providing valuable insights for further exploration and application of these enzymes. [ABSTRACT FROM AUTHOR]

Red algae are important renewable bioresources with very large annual outputs. Agarose is the major carbohydrate component of many red algae and has potential to be of value in the production of agaro-oligosaccharides, biofuels and other chemicals. In this review, we summarize the degradation pathway of agarose, which includes an upstream part involving transformation of agarose into its two monomers, D-galactose (D-Gal) and 3,6-anhydro-α-L-galactose (L-AHG), and a downstream part involving monosaccharide degradation pathways. The upstream part involves agarolytic enzymes such as α-agarase, β-agarase, α-neoagarobiose hydrolase, and agarolytic β-galactosidase. The downstream part includes the degradation pathways of D-Gal and L-AHG. In addition, the production of functional agaro-oligosaccharides such as neoagarobiose and monosaccharides such as L-AHG with different agarolytic enzymes is reviewed. Third, techniques for the setup, regulation and optimization of agarose degradation to increase utilization efficiency of agarose are summarized. Although heterologous construction of the whole agarose degradation pathway in an engineered strain has not been reported, biotechnologies applied to improve D-Gal utilization efficiency and construct L-AHG catalytic routes are reviewed. Finally, critical aspects that may aid in the construction of engineered microorganisms that can fully utilize agarose to produce agaro-oligosaccharides or as carbon sources for production of biofuels or other value-adding chemicals are discussed. [ABSTRACT FROM AUTHOR]

• A novel PL-6 family alginate lyase was characterized. • This enzyme shows high stability under 40 °C as well as in pH 7.0–10.0 conditions. • The degradation pattern of PL-6 family alginate lyase was elucidated. • This study provides a novel insight into PL-6 family alginate lyases' action mode. Alginate is the main component of brown algae which contributes to a huge biomass. The alginate oligosaccharides (AOs) have been widely used in food, cosmetic and pharmaceutical industries due to their various physiological activities. In this study, we expressed and characterized a novel PL-6 alginate lyase, named OUC-ScCD6. The results indicated that OUC-ScCD6 showed highest activity at 50 °C and pH 9.0. OUC-ScCD6 prefers to degrade poly M blocks and could digest poly G blocks as well. Endolytic action mode towards polysaccharides contributes to the creation of AOs with the degrees of polymerization 2–6. Degradation towards saturated oligosaccharides showed that saturated trisaccharides (M3 and G3) were minimum identifiable substrates. Furthermore, OUC-ScCD6 shows an even-numbered glycosidic bonds preference from non-reducing end which provided clearer insights into the substrate recognition and action mode of PL-6 family alginate lyases. [ABSTRACT FROM AUTHOR]