• Journal of Microbiology and Biotechnology
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• v.29 no.4
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• pp.625-632
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• 2019

The unified saccharification and fermentation (USF) system was developed for direct production of ethanol from agarose. This system contains an enzymatic saccharification process that uses three types of agarases and a fermentation process by recombinant yeast. The $pGMF{\alpha}-HGN$ plasmid harboring AGAH71 and AGAG1 genes encoding ${\beta}-agarase$ and the NABH558 gene encoding neoagarobiose hydrolase was constructed and transformed into the Saccharomyces cerevisiae 2805 strain. Three secretory agarases were produced by introducing an S. cerevisiae signal sequence, and they efficiently degraded agarose to galactose, 3,6-anhydro-L-galactose (AHG), neoagarobiose, and neoagarohexose. To directly produce ethanol from agarose, the S. cerevisiae $2805/pGMF{\alpha}-HGN$ strain was cultivated into YP-containing agarose medium at $40^{\circ}C$ for 48 h (for saccharification) and then $30^{\circ}C$ for 72 h (for fermentation). During the united cultivation process for 120 h, a maximum of 1.97 g/l ethanol from 10 g/l agarose was produced. This is the first report on a single process containing enzymatic saccharification and fermentation for direct production of ethanol without chemical liquefaction (pretreatment) of agarose.

• Journal of Life Science
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• v.34 no.7
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• pp.443-452
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• 2024

This study aimed to examine the influence of 3,6-anhydroxygalactose (L-AHG) on the pro-inflammatory M1 polarization and pro-inflammatory responses observed in the RAW264.7 mouse macrophage cell line following stimulation with lipopolysaccharides (LPS). L-AHG exhibited a significant and dose-dependent inhibition of inducible nitric oxide synthase (iNOS) expression, a hallmark of M1 polarization, and subsequent NO production in LPS-stimulated RAW264.7 cells. Furthermore, the LPS-induced upregulation of cyclooxygenase-2 (COX-2), which drives the production of prostaglandin E2, an inflammatory mediator, was also inhibited by L-AHG. L-AHG did not affect the LPS-triggered Toll-like receptor 4 (TLR4)-mediated pro-inflammatory signaling pathway, which culminated in the activation of transforming growth factor-β-activated kinase 1 (TAK1). However, it was observed to inhibit the generation of reactive oxugen species (ROS) in a dose-dependent manner, as well as the TAK1-driven activation of JNK and p38 MAPK. Given that the active p38 MAPK is known to contribute to the assembly of active nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, which catalyzes the intracellular generation of pro-inflammatory ROS in LPS-stimulated macrophages, the dose-dependent reduction in the LPS-induced ROS generation by L-AHG may be mainly due to the prevention of TAK1-driven activation of p38 MAPK. Together, these results demonstrate that the L-AHG-mediated inhibition of the TAK1-JNK/p38 MAPK activation phase of the pro-inflammatory signaling pathway in LPS-stimulated RAW264.7 cells by L-AHG represents a promising mechanism for suppressing M1 polarization and pro-inflammatory responses in macrophages.

• Journal of Fisheries and Marine Sciences Education
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• v.29 no.1
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• pp.108-117
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• 2017

Laver, Porphyra, is distinctive for its high content of proteins and polysaccharides such as porphyran and hemicellulose. The chemical properties of the polysaccharides extracted with different extraction methods such as hot water, dilute acid(pH 4.0) or alkali solution(2N NaOH) were examined to investigate the suitable extraction conditions for porphyran and hemicellulose from laver. For porphyran extraction, dilute acid solution was more preferable to hot water and alkali solution because of its higher 3,6-anhydrogalactose content and lower protein content. However, alkali solution was more suitable to extract the hemicellulose because of higher mannose content indicating the extraction of mannan. To decrease contamination of the polysaccharides with protein, the dried lavers were pretreated with enzymes (Protamex, Flavourzyme, Alcalase, Viscozyme) before hot water extraction. All enzyme pretreatments increased the yield of polysaccharides by compared with control (enzyme unpretreated) and Flavourzyme pretreatment was most effective to decrease protein contamination in the polysaccharide. All viscosities of porphyran solutions pretreated by enzymes were lower compared to the control porphyran solution and showed pseudoplastic behavior with yield stress. In case of alkali extraction of residues obtained after enzyme hydrolysis and hot water extraction, protease pretreatment increased the mannose contents in the polysaccharide while the xylose content was increased by Viscozyme pretreatment.