• Preventive Nutrition and Food Science
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• v.22 no.3
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• pp.184-190
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• 2017

Matrix metalloproteinases (MMPs) are endopeptidases that take significant roles in extracellular matrix degradation and therefore linked to several complications such as metastasis of cancer progression, oxidative stress, and hepatic fibrosis. Hizikia fusiformis, a brown algae, was reported to possess bioactivities, including but not limited to, antiviral, antimicrobial, and anti-inflammatory partly due to bioactive polysaccharide contents. In this study, the potential of H. fusiformis against cancer cell invasion was evaluated through the MMP inhibitory effect in HT1080 fibrosarcoma cells in vitro. H. fusiformis crude extract was fractionated with organic solvents, $H_2O$, n-BuOH, 85% aqueous MeOH, and n-hexane (n-Hex). The non-toxicity of the fractions was confirmed by MTT assay. All fractions inhibited the enzymatic activities of MMP-2 and MMP-9 according to the gelatin zymography assay. Cell migration was also significantly inhibited by the n-Hex fraction. In addition, both gene and protein expressions of MMP-2 and -9, and tissue inhibitor of MMPs (TIMPs) were evaluated by reverse transcription-polymerase chain reaction and Western blotting, respectively. The fractions suppressed the mRNA and protein levels of MMP-2, MMP-9 while elevating the TIMP-1 and TIMP-2, with the $H_2O$ fraction being the least effective while n-Hex fraction the most. Collectively, the n-Hex fraction from brown algae H. fusiformis could be a potential inhibitor of MMPs, suggesting the presence of various derivatives of polysaccharides in high amounts.

• Journal of Microbiology and Biotechnology
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• v.15 no.6
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• pp.1250-1257
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• 2005

Two groups of exopolysaccharides (designated as Fr-I EPS and Fr-II EPS) were isolated from the culture filtrate of new fungal strain Trichoderma erinaceum DG-312 by Sepharose CL-6B chromatography. The structures of the exopolysaccharides were investigated using gas chromatography (GC), Fourier transform-infrared (FT-IR) spectroscopy, GCMS analysis, and NMR. GC analysis indicated that Fr-I EPS was composed of mainly mannose ($78.9\%$) and galactose ($21.1\%$), whereas Fr-II EPS contained mannose ($68.4\%$), galactose ($26.2\%$), and glucose ($5.4\%$). In the anomeric region ($950-700cm_{-1}$) of the FT-IR spectrum, both EPSs exhibited obvious characteristic absorption of $810\;cm_{-1}$, indicating the existence of mannose. The spectra of $\alpha-and\;\beta$-configurations were assigned at 880 and $914\;cm_{-1}$, respectively. The results of GC-MS analyses confirmed that both EPSs were complex heteropolysaccharides with a ($1{\rightarrow}3$)-linked mannan backbone. The C-1 region that appeared in the $^{13}C-NMR$ spectra of these EPSs indicated a typical anomeric carbon signal. The Fr-I EPS showed two anomeric carbon signals at 102.6 and 99.6 ppm, whereas the Fr-II EPS displayed four anomeric carbon signals at 102.5, 99.6, 98.5, and 94.3 ppm. The molecular characteristics of the EPSs were further investigated using a size exclusion chromatography/multi-angle laser light scattering (SEC/MALLS) system. The SEC/MALLS system revealed that the average molar masses of the EPSs were $6.592{\times}10^{4}$ (Fr-I EPS) and $1.920{\times}10^{4}$ (Fr-II EPS) g/mol, and the molecular conformation of both EPSs in aqueous solution was random coils.

• KSBB Journal
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• v.28 no.6
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• pp.408-414
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• 2013

In order to obtain functional materials from aloe callus, we cultured Aloe vera L. leaf on MS medium added 0.2 mg/L IAA, 0.3 mg/L kinetin and 100 mg/L grape or/and apple juice for 30 days. While a callus differentiation during callus culture did not show, the cultured leaves were uniquely released extracellular material into the agar plate. After cultivation for 18 days, the cultured leaf and agar were harvested for extraction a functional material. The materials extracted were measured on the amount of total phenols, flavonoids and polysaccharides and determined on the antioxidant and antimicrobial activity. In result, callus extracts of additive free (CT) and added apple juice (2T) had more amount of phenol compound ($659{\mu}g/mL$, $533{\mu}g/mL$) and flavonoid ($580{\mu}g/mL$, $501{\mu}g/mL$) than natural leaf (p: $525{\mu}g/mL$, f: $301{\mu}g/mL$). However, the extract of natural leaf had the better effect of lipid peroxidation and polysaccharide content than the culture extract. All samples extracted had same effect on the nitrite scavenging activity. On the other hand, only 2T extract showed excellent 72% nitric oxide scavenging activity. The agar extract was also confirmed to contain polyphenol compound and polysaccharide content that had antioxidant and antimicrobial activity partly.

• KSBB Journal
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• v.8 no.2
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• pp.164-171
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• 1993

The production of extracellular polysaccharide by Methylomonas mucosa (NRRL B-5696) was investigated. The microorganism uses methanol as the carbon source for their growth and produces exopolysaccharides. The productivity of exopolysaccharides was investigated under various culture modes: batch, fed-batch and continuous culture. In flask culture the growth of cell mass and the production of polysaccharide were inhibited at above 1% (v/v) methanol. At 1%(v/v) methanol maximum specific growth rate was obtained. As C/N ratio (g methanol/g ammonium sulfate) increased, polysaccharide production increased and cells mass decreased. Magnesium ion was also found to be essential for the polysaccharide production. In batch culture the production of polysaccharides was more affected by the specific growth rate than the cell concentration. In fed-batch culture the concentration of polysaccharide was 4 times higher than that of batch culture, but the yield was lower. The productivity of fed-batch with continuous feeding was higher than that of batch or fed-batch with intermittent feeding. This is due to no methanol limitation or inhibition that used to occur in fed-batch culture with intermittent feeding. In continuous culture pure oxygen was supplied to avoid the oxygen limitation. As the dilution rate in- creased up to 0.21 h-1, the yield and productivity increased. The solution viscosity of the produced polysaccharide obtained from above increased exponentially with the concentration of polysaccharide.

• Microbiology and Biotechnology Letters
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• v.13 no.3
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• pp.173-178
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• 1985

Extracellular amylase from tile filtrate of the submerged culture of Ganoderma lucidum was partially purified by ammonium sulfate precipitation and its properties were studied. The optimum pH and temperature of the enzyme activity were 5.5 and 5$0^{\circ}C$. respectively. This enzyme was most stable at pH 5.0 and stable up to 3$0^{\circ}C$, but it lost completely the activity when it was treated at 6$0^{\circ}C$ for 10 min. The enzyme was activated by the addition of M $n^{++}$, $C^{++}$ and C $u^{++}$, but inhibited by H $g^{++}$, A $g^{++}$ And various enzyme inhibitors and chemical reagents did not affect the enzyme activity. The enzyme hydrolyzed the boiled amylaceous polysaccharides, but it hydrolyzed raw starches very slowly. The activation energy of the enzyme for soluble starch was calculated and found to be 7.06 Kcal per mole. The Km values of the enzyme for soluble starch, amylose, amylopectin and glycogen were 0.16, 0.37, 0.19, and 0.16mg/$m\ell$, respectively. Maltose was found to inhibit the enzyme activity and kinetic analysis revealed a competitive type of inhibition.n.n.n.n.n.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.5
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• pp.680-692
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• 2010

This paper gives an overview of the availability, nutritive quality, and possible strategies to improve the utilization of rice straw as a feed ingredient for ruminants. Approximately 80% of the rice in the world is grown by small-scale farmers in developing countries, including South East Asia. The large amount of rice straw as a by-product of the rice production is mainly used as a source of feed for ruminant livestock. Rice straw is rich in polysaccharides and has a high lignin and silica content, limiting voluntary intake and reducing degradability by ruminal microorganisms. Several methods to improve the utilization of rice straw by ruminants have been investigated in the past. However, some physical treatments are not practical because of the requirement for machinery or treatments are not economical feasible for the farmers. Chemical treatments, such as NaOH, $NH_3$ or urea, currently seem to be more practical for onfarm use. Alternative treatments to improve the nutritive value of rice straw are the use of ligninolytic fungi (white-rot fungi), with their extracellular ligninolytic enzymes, or specific enzymes degrading cellulose and/or hemicellulose. The use of fungi or enzyme treatments is expected to be a more practical and environmental-friendly approach for enhancing the nutritive value of rice straw and can be costeffective in the future. Using fungi and enzymes might be combined with the more classical chemical or physical treatments. However, available data on using fungi and enzymes for improving the quality of rice straw are relatively scarce.

• Research in Plant Disease
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• v.21 no.3
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• pp.161-179
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• 2015

Plant have developed sophisticated defence mechanisms against microbial pathogens. The recent accumulated information allow us to understand the nature of plant immune responses followed by recognition of microbial factors/determinants through cutting-edge genomics and multi-omics techniques. However, the practical approaches to sustain plant health using enhancement of plant immunity is yet to be fully appreciated. Here, we overviewed the general concept and representative examples on the plant immunity. The fungal, bacterial, and viral determinants that was previously reported as the triggers of plant immune responses are introduced and described as the potential protocol of biological control. Specifically, the role of chitin, glucan, lipopolysaccharides/extracellular polysaccharides, microbe/pathogen-associated molecular pattern, antibiotics, mimic-phytohormones, N-acyl homoserine lactone, harpin, vitamins, and volatile organic compounds are considered. We hope that this review stimulates scientific community and farmers to broaden their knowledge on the microbial determinant-based biological control and to apply the technology on the integrated pest management program.

• Microbiology and Biotechnology Letters
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• v.22 no.3
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• pp.240-245
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• 1994

Relation the phage defense mechanism of phage resistant Lactococcus lactis subsp. cremoris ATCC 11602-A1 to its cell wall components was investigated. To determine whether teichoic acid which is known to be one of the phage receptor site present on the cell wall, phage adsorption was examined after treatment 5% TCA(60%$\CIRC $C) and concanavalin A to the cell wall of A1 and parent strain. However, the adsorption rate of two strains did not change. Total amount of phosphate after TCA treatment did not change in both strains, but a difference between the two strains was observed. Ribitol and glycerol, components of teichoic acid, could not be detected in the cell walls of two strains by GC analysis. These results suggest that although teichoic acid was not present in the cell walls of both strains, the composition of cell wall of two strains was not identical. Measurement of amount of protein and SDS-polyacryamide gel electrophoresis were carried out to examine the involvement of cell wall protein in phage resistance, showing that protein is nothing to do with phage adsorption of parent strain, but phage resistance of A1 is related to protein. Cell wall carbohydrates of A1 contained rhamnose, glucose, and galactose. Total amount of carbohydrate of 1% SDS-treated A1 cell wall was reduced to the level of parent strain. The results suggest that phage resistance of A1 was due to the presence of a higher level of carbohydrates then parent strain, and to interaction of carbohydrate and protein.

• Journal of Microbiology and Biotechnology
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• v.21 no.11
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• pp.1174-1178
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• 2011

Exopolysaccharides (EPSs) are microbial polysaccharides that are released outside of the bacterial cell wall. There have been few studies on EPS-producing lactic acid bacteria that can enhance macrophage activity and the underlying signaling mechanism for cytokine expression. In the current study, EPS-overproducing Lactobacillus (L.) paracasei KB28 was isolated from kimchi and cultivated in conditioned media containing glucose, sucrose, and lactose. The whole bacterial cells were obtained with their EPS being attached, and the cytokine-inducing activities of these cells were investigated. Gas chromatography analysis showed the presence of glucose, galactose, mannose, xylose, arabinose, and rhamnose in EPS composition. EPS-producing L. paracasei KB28 induced the expression of tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-6, and IL-12 in mouse macrophages. This strain also caused the degradation of $I{\kappa}B{\alpha}$ and phosphorylation of the major MAPKs: Jun N-terminal kinase (JNK), p38, and extracellular signal-regulated kinase (ERK)1/2. The use of pharmacological inhibitors showed that different signaling pathways were involved in the induction of TNF-${\alpha}$, IL-6 and IL-12 by L. paracasei KB28. Our results provide information for a better understanding of the molecular mechanisms of the immunomodulatory effect of food-derived EPS-producing lactic acid bacteria.

• The Plant Pathology Journal
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• v.27 no.1
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• pp.89-92
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• 2011

Cyclic AMP receptor-like protein (Clp), is known to be a global transcriptional regulator for the expression of virulence factors in Xanthomonas campestris pv. campestris (Xcc). Sequence analysis showed that Xanthomonas oryzae pv. oryzae (Xoo) contains a gene that is strongly homologous to the Xcc clp. In order to determine the role of the Clp homolog in Xoo, a marker exchange mutant of $clp_{xoo4158}$ was generated. Virulence and virulence factors, such as the production of cellulase, xylanase, and extracellular polysaccharides (EPS) and swarming motility were significantly decreased in the $clp_{xoo4158}$ mutant. Moreover, the mutation caused the strain to be more sensitive to hydrogen peroxide and to over-produce siderophores. Complementation of the mutant restored the mutation-related phenotypes. Expression of $clp_{xoo4158}$, assessed by reverse-transcription realtime PCR and clp promoter activity, was significantly reduced in the rpfB, rpfF, rpfC, and rpfG mutants. These results suggest that the clp homolog, $clp_{xoo4158}$, is involved in the control of virulence and resistance against oxidative stress, and that expression of the gene is controlled by RpfC and RpfG through a diffusible signal factor (DSF) signal in Xanthomonas oryzae pv. oryzae KACC10859.