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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]

A galactose oxidase gene, gao-5f, was cloned from Fusarium odoratissimum and successfully expressed in E. coli. The galactose oxidase GAO-5F belongs to the AA5 family and consists of 681 amino acids, with an estimated molecular weight of 72 kDa. GAO-5F exhibited maximum activity at 40 °C and pH 7.0 and showed no change in activity after 24 h incubation at 30 °C. Moreover, GAO-5F exhibited 40% of its maximum activity after 24 h incubation at 50 °C and 60% after 40 h incubation at pH 7.0. The measured thermostability of GAO-5F is superior to galactose oxidase's reported thermostability. The enzyme exhibited strict substrate specificity toward D-galactose and oligosaccharides/polysaccharides containing D-galactose. Further analysis demonstrated that GAO-5F specifically oxidized agarose to a polyaldehyde-based polymer, which could be used as a polyaldehyde to crosslink with gelatin to form edible packaging films. To our knowledge, this is the first report about the modification of agarose by galactose oxidase, and this result has laid a foundation for the further development of edible membranes using agarose. [ABSTRACT FROM AUTHOR]

Chitin deacetylase can be used in the green and efficient preparation of chitosan from chitin. Herein, a novel chitin deacetylase SbCDA from Streptomyces bacillaris was heterologously expressed and comprehensively characterized. SbDNA exhibits its highest deacetylation activity at 35 °C and pH 8.0. The enzyme activity is enhanced by Mn2+ and prominently inhibited by Zn2+, SDS, and EDTA. SbCDA showed better deacetylation activity on colloidal chitin, (GlcNAc)5, and (GlcNAc)6 than other forms of the substrate. Molecular modification of SbCDA was conducted based on sequence alignment and homology modeling. A mutant SbCDA63G with higher activity and better temperature stability was obtained. The deacetylation activity of SbCDA63G was increased by 133% compared with the original enzyme, and the optimal reaction temperature increased from 35 to 40 °C. The half-life of SbCDA63G at 40 °C is 15 h, which was 5 h longer than that of the original enzyme. The improved characteristics of the chitin deacetylase SbCDA63G make it a potential candidate to industrially produce chitosan from chitin. [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]

Neoagarotetraose (NT), a hydrolytic product of agar by β-agarase, is known to possess bioactive properties. However, the mechanisms via which NT alleviates intestinal inflammation remain unknown. In this study, a dextran sulfate sodium (DSS)-induced murine model was developed to evaluate the effect of NT on gut microbiome and microbial metabolism using 16S rRNA gene sequencing and untargeted metabolomics. Our data demonstrate that NT ingestion improved gut integrity and inflammation scores. NT reversed the abundance of Proteobacteria from an elevated level induced by DSS and significantly increased the abundance of Verrucomicrobia. Further, NT significantly increased the abundance of Akkermansia and Lactobacillus and concomitantly decreased that of Sutterella, which were among the important features identified by random forests analysis contributing to classification accuracy for NT supplementation. A microbial signature consisting of Adlercreutzia (denominator) and Turicibacter (numerator) predicted the NT supplementation status. Moreover, NT significantly modulated multiple gut metabolites, particularly those related to histidine, polyamine and tocopherol metabolism. Together, our findings provided novel insights into the mechanisms by which NT modulated the gut microbiome and metabolome and should facilitate the development of NT as a potent prebiotic for colitis management. [ABSTRACT FROM AUTHOR]

The world-famous Antarctic krill (Euphausia superba) plays a fundamental role in the Antarctic food chain. It resides in cold environments with the most abundant biomass to support the Antarctic ecology and fisheries. Here, we performed the first genome survey of the Antarctic krill, with genomic evidence for its estimated genome size of 42.1 gigabases (Gb). Such a large genome, however, is beyond our present capability to obtain a good assembly, although our sequencing data are a valuable genetic resource for subsequent polar biomedical research. We extracted 13 typical protein-coding gene sequences of the mitochondrial genome and analyzed simple sequence repeats (SSRs), which are useful for species identification and origin determination. Meanwhile, we conducted a high-throughput comparative identification of putative antimicrobial peptides (AMPs) and antihypertensive peptides (AHTPs) from whole-body transcriptomes of the Antarctic krill and its well-known counterpart, the whiteleg shrimp (Penaeus vannamei; resident in warm waters). Related data revealed that AMPs/AMP precursors and AHTPs were generally conserved, with interesting variations between the two crustacean species. In summary, as the first report of estimated genome size of the Antarctic krill, our present genome survey data provide a foundation for further biological research into this polar species. Our preliminary investigations on bioactive peptides will bring a new perspective for the in-depth development of novel marine drugs. [ABSTRACT FROM AUTHOR]

Enzymes are catalysts with high specificity. Different compounds could be produced by different enzymes. In case of agaro-oligosaccharides, agarooligosaccharide (AOS) can be produced by α-agarase through cleaving the α-1,3-glycosidic linkages of agarose, while neoagarooligosaccharide (NAOS) can be produced by β-agarase through cleaving the β-1,4-glycosidic linkages of agarose. However, in this study, we showed that β-agarase could also be used to produce AOSs with high purity and yield. The feasibility of our route was confirmed by agarotriose (A3) and agaropentaose (A5) formation from agaroheptaose (A7) and agarononoses (A9) catalyzed by β-agarase. Agarose was firstly liquesced by citric acid into a mixture of AOSs. The AOSs mixture was further catalyzed by β-agarase. When using the neoagarotetraose-forming β-agarase AgWH50B, agarotriose could be produced with the yield of 48%. When using neoagarotetraose, neoagarohexaose-forming β-agarase DagA, both agarotriose and agaropentaose could be produced with the yield of 14% and 13%, respectively. Our method can be used to produce other value-added agaro-oligosaccharides from agarose by different agarolytic enzymes. [ABSTRACT FROM AUTHOR]

A thermostable and alkaline-stable novel esterase (Est7) was identified through the whole genome sequencing of Stenotrophomonas maltophilia OUC_Est10. The open reading frame of this gene encoded 617 amino acid residues. After heterologous expression in Escherichia coli BL21 (DE3), the purified Est7 was separated as a single protein and presented a molecular mass of 70.6 kDa. Multiple sequence alignment indicated that Est7 had a typical catalytic triad (Ser-Asp-His) and the conserved sequence (GDSL) typical of the family II lipid hydrolase proteins. Est7 showed good stability in alkaline buffers, especially in Tris-HCl buffer at pH 9.0 (residual activity 93.8% after 96 h at 4 °C) and in the medium temperature conditions (residual activity 70.2% after 96 h at 45 °C and pH 8.0). The enzyme also retained higher stability toward several hydrophilic and hydrophobic organic solvents (e.g., after incubation in 100% acetonitrile or in n-hexane the enzyme retained about 97% and 84% of the activity in the absence of organic solvent, respectively). Furthermore, Est7 could catalyze the transesterification reaction of vinylacetate with 2-phenylethanol and cis-3-hexen-1-ol to their corresponding acetate esters in petroleum ether or tert-butyl methyl ether. These results indicate Est7 as a promising biocatalyst for applications of Est7 in non-aqueous media. [ABSTRACT FROM AUTHOR]

In this study, silica-coated magnetic nanoparticles (Fe3O4/SiO2) were synthesized and applied in the immobilization of phospholipase D (PLDa2) via physical adsorption and covalent attachment. The immobilized PLDa2 was applied in the synthesis of functional phosphatidylserine (PS) through a transphophatidylation reaction. The synthesis process and characterizations of the carriers were examined by scanning electron microscope (SEM), transmission electron microscope (TEM), and Fourier-transform infrared spectroscopy (FT-IR). The optimum immobilization conditions were evaluated, and the thermal and pH stability of immobilized and free PLDa2 were measured and compared. The tolerance to high temperature of immobilized PLDa2 increased remarkably by 10°C. Furthermore, the catalytic activity of the immobilized PLDa2 remained at 40% after eight recycles, which revealed that silica-coated magnetic nanoparticles have potential application for immobilization and catalytic reactions in a biphasic system. [ABSTRACT FROM AUTHOR]

In this research, a two-step extraction approach was developed for chitin preparation from shrimp shells by utilizing citric acids and deep eutectic solvents (DESs), which effectively removed minerals and proteins. In the first step, minerals of shrimp shells were removed by citric acid, and the demineralization efficiency reached more than 98%. In the second step, the removal of protein was carried out using deep eutectic solvents with the assistance of microwave, and the deproteinization efficiency was more than 88%. The results of scanning electron microscopy (SEM), Fourier transform infrared (FT-IR) spectroscopy, X-ray diffraction analysis (XRD), and thermogravimetric analysis (TGA) showed that the quality of DES-prepared chitin was comparable to that of traditional acid/alkali-prepared chitin. These results were realized without utilizing hazardous chemicals, which are detrimental to the environment. This research indicates that a DES-based preparation approach has the potential for application in the recovery of biopolymers from natural resources. [ABSTRACT FROM AUTHOR]

Chitin deacetylase can be used in the green and efficient preparation of chitosan from chitin. Herein, a novel chitin deacetylase Sb CDA from Streptomyces bacillaris was heterologously expressed and comprehensively characterized. Sb DNA exhibits its highest deacetylation activity at 35 °C and pH 8.0. The enzyme activity is enhanced by Mn 2+ and prominently inhibited by Zn 2+ , SDS, and EDTA. Sb CDA showed better deacetylation activity on colloidal chitin, (GlcNAc) 5 , and (GlcNAc) 6 than other forms of the substrate. Molecular modification of Sb CDA was conducted based on sequence alignment and homology modeling. A mutant Sb CDA63G with higher activity and better temperature stability was obtained. The deacetylation activity of Sb CDA63G was increased by 133% compared with the original enzyme, and the optimal reaction temperature increased from 35 to 40 °C. The half-life of Sb CDA63G at 40 °C is 15 h, which was 5 h longer than that of the original enzyme. The improved characteristics of the chitin deacetylase Sb CDA63G make it a potential candidate to industrially produce chitosan from chitin.