Tamlana laminarinivorans sp. nov. and Tamlana sargassicola sp. nov., two novel species isolated from Sargassum, show genomic and physiological adaptations for a Sargassum-associated lifestyle.

The genus Tamlana from the Bacteroidota currently includes six validated species. Two strains designated PT2-4T and 62-3T were isolated from Sargassum abundant at the Pingtan island coast in the Fujian Province of China. 16S rRNA gene sequence analysis showed that the closest described relative of strains PT2-4T and 62-3T is Tamlana sedimentorum JCM 19808T with 98.40 and 97.98% sequence similarity, respectively. The 16S rRNA gene sequence similarity between strain PT2-4T and strain 62-3T was 98.68 %. Furthermore, the highest average nucleotide identity values were 87.34 and 88.97 % for strains PT2-4T and 62-3T, respectively. The highest DNA-DNA hybridization (DDH) value of strain PT2-4T was 35.2 % with strain 62-3T, while the DDH value of strain 62-3T was 37.7 % with T. sedimentorum JCM 19808T. Growth of strains PT2-4T and 62-3T occurs at 15-40 °C (optimum, 30 °C) with 0-4 % (w/v) NaCl (optimum 0-1 %). Strains PT2-4T and 62-3T can grow from pH 5.0 to 10.0 (optimum, pH 7.0). The major fatty acids of strains PT2-4T and 62-3T are iso-C15 : 0 and iso G-C15 : 1. MK-6 is the sole respiratory quinone. Genomic and physiological analyses of strains PT2-4T and 62-3T showed corresponding adaptive features. Significant adaptation to the growth environment of macroalgae includes the degradation of brown algae-derived diverse polysaccharides (alginate, laminarin and fucoidan). Notably, strain PT2-4T can utilize laminarin, fucoidan and alginate via specific carbohydrate-active enzymes encoded in polysaccharide utilization loci, rarely described for the genus Tamlana to date. Based on their distinct physiological characteristics and the traits of utilizing polysaccharides from Sargassum, strains PT2-4T and 62-3T are suggested to be classified into two novel species, Tamlana laminarinivorans sp. nov. and Tamlana sargassicola sp. nov. (type strain PT2-4T=MCCC 1K04427T=KCTC 92183T and type strain 62-3T=MCCC 1K04421T=KCTC 92182T).

Patient-specific 3D printed models for enhanced learning of immediate implant procedures and provisionalization.

INTRODUCTION: This study aimed to describe the fabrication, implementation and evaluation of 3D-printed patient-specific models for unskilled students to enhance learning in immediate implant procedures and provisionalization. MATERIALS AND METHODS: The individualized simulation models were designed and processed based on CT and digital intraoral scanning of a patient. Thirty students performed simulation implant surgery and provisionalized the implant sites on the models and answered questionnaires to assess their perceptions before and after the training. The scores of the questionnaires were analysed using the Wilcoxon signed-rank test. RESULTS: Significant differences before and after training were found in the students' responses. Students reported better results in understanding of surgical procedures, knowledge in prosthetically driven implantology, understanding of minimally invasive tooth extraction, confirming the accuracy of surgical template, usage of the guide rings and usage of the surgical cassette after simulation training. The overall expenditure on the simulation training involving 30 students amounted to 342.5 USD. CONCLUSIONS: The patient-specific and cost-efficient 3D printed models are helpful for students to improve theoretical knowledge and practical skills. Such individualized simulation models have promising application prospects.

Bioconversion of bamboo shoot shells through the cultivation of the edible mushrooms Volvariella volvacea.

Bamboo shoot shell (BSS), as agricultural waste, is mostly burned or discarded, causing serious environment pollution. In this study, the degradation and utilization of BSS by the edible fungus Volvariella Volvacea was investigated. The composition of V. volvacea fruit body was determined by HPLC-MS, GC-MS and ICP-OES. The activities of CMCase and xylanase were monitored by DNS (3,5-dinitrosalicylic acid) method. Laccase activity was assayed by the oxidation reaction of ABTS [2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonate)]. The degraded bamboo shoot shell powder was characterized by FTIR and SEM. The results showed that the mycelium of V. volvacea could degrade and utilize BSS for growth. The activities of carboxymethyl cellulase and laccase were increased during the cultivation. At the same time, the physical structure of the shell fiber becames porous and rough. Most of the products of decayed fibers contain alkanes, ethyl or methyl groups. Moreover, the biological efficiency (fruiting body yield) of V. volvacea cultivated on BSS was 1.52-fold higher than that of straw cultivation. The contents of total lipid, elaidic acid (C18:1n-9), total essential amino acids, total amino acids and iron in V. volvacea fruit bodies grown on BSS were 1.11, 1.66, 1.52, 1.60 and 1.30-fold higher than those of straw treatment, respectively. This study provides an effective method to solve the environmental pollution caused by BSS, and provides a new way for the potential utilization of BSS in edible fungi cultivation.

Biochemical Characterization of a Novel Thermostable Ulvan Lyase from Tamlana fucoidanivorans CW2-9.

Ulvan is a complex sulfated polysaccharide extracted from Ulva, and ulvan lyases can degrade ulvan through a ß-elimination mechanism to obtain oligosaccharides. In this study, a new ulvan lyase, EPL15085, which belongs to the polysaccharide lyase (PL) 28 family from Tamlana fucoidanivorans CW2-9, was characterized in detail. The optimal pH and salinity are 9.0 and 0.4 M NaCl, respectively. The Km and Vmax of recombinant EPL15085 toward ulvan are 0.80 mg·mL-1 and 11.22 µmol·min -1 mg-1·mL-1, respectively. Unexpectedly, it is very resistant to high temperatures. After treatment at 100 °C, EPL15085 maintained its ability to degrade ulvan. Molecular dynamics simulation analysis and site-directed mutagenesis analysis indicated that the strong rigidity of the disulfide bond between Cys74-Cys102 in the N-terminus is related to its thermostability. In addition, oligosaccharides with disaccharides and tetrasaccharides were the end products of EPL15085. Based on molecular docking and site-directed mutagenesis analysis, Tyr177 and Leu134 are considered to be the crucial residues for enzyme activity. In conclusion, our study identified a new PL28 family of ulvan lyases, EPL15085, with excellent heat resistance that can expand the database of ulvan lyases and provide the possibility to make full use of ulvan.

Mutation of conserved residues in the laminarinase Lam1092 increases the antioxidant activity of the laminarin product hydrolysates.

Laminarinases from the glycoside hydrolase 16 (GH16) family are hydrolases that break ß-1,3-glycosidic bonds in laminarin, which is the major storage polysaccharide present in brown algae or microalgae. We explored a laminarinase from the marine Flavobacteriaceae species Tamlana sp. PT2-4 at the structural and functional levels. Based on a homology model of Lam1092-substrate interactions, the large active groove crossing Lam1092 was deemed a reasonable pathway for the bent substrates for hydrolysis. Eight residues (Gly361, Asn364, Arg400, His466, Asp449, Glu452, Ser477 and Thr538) were selected for mutagenesis based on the interactions of Lam1092 in complex with Lam4/Lam6. Ultimately, we generated eight mutants of Lam1092, and the antioxidant activities of the hydrolysates of two mutants (G361A and H466A) showed significant improvement. These results show that the antioxidant activity of laminarin can be improved by laminarinase mutation, which will be beneficial for developing efficient approaches to engineer the substrate specificity of laminarinases and improve the application of bioactive laminarioligosaccharides.

Insights into Algal Polysaccharides: A Review of Their Structure, Depolymerases, and Metabolic Pathways.

In recent years, marine macroalgae with extensive biomass have attracted the attention of researchers worldwide. Furthermore, algal polysaccharides have been widely studied in the food, pharmaceutical, and cosmetic fields because of their various kinds of bioactivities. However, there are immense barriers to their application as a result of their high molecular size, poor solubility, hydrocolloid nature, and low physiological activities. Unique polysaccharides, such as laminarin, alginate, fucoidan, agar, carrageenan, porphyran, ulvan, and other complex structural polysaccharides, can be digested by marine bacteria with many carbohydrate-active enzymes (CAZymes) by breaking down the limitation of glycosidic bonds. However, structural elucidation of algal polysaccharides, metabolic pathways, and identification of potential polysaccharide hydrolases that participate in different metabolic pathways remain major obstacles restricting the efficient utilization of algal oligosaccharides. This review focuses on the structure, hydrolase families, metabolic pathways, and potential applications of seven macroalgae polysaccharides. These results will contribute to progressing our understanding of the structure of algal polysaccharides and their metabolic pathways and will be valuable for clearing the way for the compelling utilization of bioactive oligosaccharides.

An electrostatically regulated organic self-assembly for rapid and sensitive detection of heparin in serum.

Heparin (Hep) is a highly negatively charged linear glycosaminoglycan involved in various physiological processes, especially blood coagulation. Hep is also a first-line drug for anticoagulation and prevention of thromboembolism, but its overdose will cause serious side effects. Herein, we designed a long-wavelength double-charged cationic fluorescent probe PYPN, and studied its aggregation state and detection performance for Hep. PYPN was readily synthesized through a one-step reaction without complicated purification. In aqueous medium, PYPN molecules with an amphiphilic structure spontaneously form nano-assemblies, which can be immediately decomposed by Hep due to the formation of a PYPN-Hep complex based on electrostatic attraction. The assembly shows a fast, sensitive and ratiometric fluorescence response to Hep, without being obviously interfered by other compounds. In various serum matrices, the fluorescence intensity ratio F610/F470 has a good linearity with Hep concentration (0-12 µg mL-1), and the detection limit (0.11-0.12 U mL-1) is lower than the minimum concentration (0.2 U mL-1) used in clinical treatment. Our study provides an easy-to-prepare and feasible tool for the selective and sensitive quantification of Hep in serum.