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Endoscopic submucosal dissection (ESD) is clinically used to remove early gastric cancer in stomach. The aim of the current study is to examine a therapeutic capacity of pulsed Ho:YAG laser for the development of laser-assisted ESD under various surgical parameters. Ex vivo porcine stomach tissue was ablated with 1-J Ho:YAG pulses at 10 Hz at different number of treatments (NT = 1, 2, and 3) and treatment speeds (TS = 0.5, 1, and 2 mm/s) without and with saline injection. Regardless of saline injection, straight tissue ablation showed that ablation depth increased with increasing NT and decreasing TS. At NT = 3 and TS = 0.5 mm/s, no saline injection yielded the maximum ablation depth (3.4 ± 0.3 mm), partially removing muscularis propria. However, saline injection confined the tissue ablation within a submucosal layer (2.1 ± 0.3 mm). Thermal injury was found to be 0.7~1.1 mm in the adjacent tissue with superficial carbonization. Circular tissue ablation (2 cm in diameter) at NT = 3 and TS = 0.5 mm/s presented that no saline injection yielded a reduction in the lesion area, whereas saline injection maintained the ablated lesion area. Histological analysis revealed that unlike no saline injection, saline injection ablated the entire mucosal layer without perforation in the muscular propria. The pulsed Ho:YAG laser can be a potential surgical tool for clinical ESD to incise a target lesion without adverse perforation. Further investigations will validate the efficacy and safety of the Ho:YAG laser-assisted ESD in in vivo porcine stomach models for clinical translation. [ABSTRACT FROM AUTHOR]

Two putative methylglyoxal synthases, which catalyze the conversion of dihydroxyacetone phosphate to methylglyoxal, from Oceanithermus profundus DSM 14,977 and Clostridium difficile 630 have been characterized for activity and thermal stability. The enzyme from O. profundus was found to be hyperthermophilic, with the optimum activity at 80 °C and the residual activity up to 59% after incubation of 15 min at 95 °C, whereas the enzyme from C. difficile was mesophilic with the optimum activity at 40 °C and the residual activity less than 50% after the incubation at 55 °C or higher temperatures for 15 min. The structural analysis of the enzymes with molecular dynamics simulation indicated that the hyperthermophilic methylglyoxal synthase has a rigid protein structure with a lower overall root-mean-square-deviation value compared with the mesophilic or thermophilic counterparts. In addition, the simulation results identified distinct regions with high fluctuations throughout those of the mesophilic or thermophilic counterparts via root-mean-square-fluctuation analysis. Specific molecular interactions focusing on the hydrogen bonds and salt bridges in the distinct regions were analyzed in terms of interatomic distances and positions of the individual residues with respect to the secondary structures of the enzyme. Key interactions including specific salt bridges and hydrogen bonds between a rigid beta-sheet core and surrounding alpha helices were found to contribute to the stabilisation of the hyperthermophilic enzyme by reducing the regional fluctuations in the protein structure. The structural information and analysis approach in this study can be further exploited for the engineering and industrial application of the enzyme. [ABSTRACT FROM AUTHOR]

Carbon dioxide (CO2) emissions are related to global warming. However, CO2 can be used as an abundant and cheap carbon source for production of valuable chemicals using carbon capture and storage technology. Here, the genes related to carbon flux toward pyruvate biosynthesis in E. coli were deleted to enhance the incorporation of CO2 for succinate production. The codonoptimized carbonic anhydrase gene (SP(-)HCCA) derived from Hahella chejuensis KCTC 2396 and the phosphoenolpyruvate carboxylase gene (ppc) of E. coli W3110 were co-overexpressed to enhance carbon flux toward oxaloacetate synthesis in E. coli. Finally, we constructed SGJS134, which shows the highest production of succinate derived from CO2 compared with other strains. SGJS134 produced approximately 6.5 mM succinate from CO2 and yielded approximately 13.0 mM succinate per dry cell weight. These results may be useful for enhancing the incorporation of CO2 for succinate production in E. coli. Additionally, the metabolic engineering method used in this study will propose the potential of E. coli to convert CO2 to valuable chemicals. [ABSTRACT FROM AUTHOR]

In this study, phosphoenolpyruvate carboxylase (PEPC) derived from Oceanimonas smirnovii (OS) was expressed as a soluble protein in Escherichia coli BL21(DE3). We isolated OS-PEPC (a recombinant PEPC protein) by his-tag purification. The purified protein showed a single band upon analysis with SDS-PAGE, and it had an apparent molecular mass of 98 kDa. Pufied OS-PEPC showed a specific activity value of 21.8 ± 0.495 U/mg protein. Especially, OS-PEPC showed the enzymatic activity between 40 and 50 °C. It maintained enzymatic activity in basic pH conditions (pH value, 9-10). We also measured OS-PEPC PEP and HCO saturation kinetics and confirmed the effect of divalent cation on OS-PEPC activity. [ABSTRACT FROM AUTHOR]

Objective: To produce 10-ketostearic acid from oleic acid. Results: Oleic acid was converted to 10-ketostearic acid by a recombinant Corynebacterium glutamicum ATCC 13032 expressing oleate hydratase from Stenotrophomonas maltophilia and a secondary alcohol dehydrogenase from Micrococcus luteus under the control of a synthetic constitutive promoter. Optimal conditions for 10-ketostearic acid production were pH 7.5 and 30 °C with 5 g cells l and 2.5 g oleic acid l. Under these conditions, the cells produced 1.96 g 10-ketostearic acid l from oleic acid in 6 h, with a conversion yield of 78 % (w) and a maximum volumetric productivity of 1.67 g l h. Conclusion: This is the first report of 10-ketostearic acid production using a recombinant C. glutamicum. [ABSTRACT FROM AUTHOR]