• Microbiology and Biotechnology Letters
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• v.46 no.1
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• pp.9-17
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• 2018

The aims of this work were to characterize and determine bioactivities of crude exopolysaccharide (EPS) extract from Bacillus tequilensis PS21 isolated from milk kefir from Kampaeng Petch, Thailand. B. tequilensis PS21 produced 112.1 mg dried EPS/l from initial 80 g/l lactose in modified TSB media at 52 h, with EPS product yield of 8.9 mg EPS/g lactose and specific product yield of 0.3 mg EPS/mg biomass. The FTIR result confirmed EPS to be a protein-bound polysaccharide and SEM analysis showed the morphology to be a grainy appearance with an uneven surface, covered with pores. HPLC analysis determined EPS as a heteropolysaccharide consisting of five sugar units with the following molar ratios; xylose (17.65), glucose (2.54), ribose (1.83), rhamnose (1.23), and galactose (1). Chemical components of this EPS were predominantly carbohydrate at 697.8 mg/g EPS (65%), protein 361.4 mg/g EPS (34%), and nucleic acid 12.5 mg/g EPS (1%). The EPS demonstrated antioxidant activities at 57.5% DPPH scavenging activity, $37.2{\mu}M\;Fe(II)/mg$ EPS and $34.9{\mu}M\;TEAC\;{\mu}M/mg$ EPS using DPPH, FRAP and ABTS assays, respectively. EPS also exhibited anti-tyrosinase activity at 34.9% inhibition. This work represents the first finding of EPS produced by Bacillus sp. from Thai milk kefir which shows potential applications in the production of antioxidant functional foods and whitening cosmetics. However, optimization of EPS production for industrial exploitation requires further study to ascertain the economic potential.

• Journal of Microbiology and Biotechnology
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• v.30 no.2
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• pp.155-162
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• 2020

Acetyl xylan esterase (AXE; E.C. 3.1.1.72) is one of the accessory enzymes for xylan degradation, which can remove the terminal acetate residues from xylan polymers. In this study, two genes encoding putative AXEs (LaAXE and BhAXE) were cloned from Lactobacillus antri DSM 16041 and Bacillus halodurans C-125, and constitutively expressed in Escherichia coli. They possess considerable activities towards various substrates such as p-nitrophenyl acetate, 4-methylumbelliferyl acetate, glucose pentaacetate, and 7-amino cephalosporanic acid. LaAXE and BhAXE showed the highest activities at pH 7.0 and 8.0 at 50℃, respectively. These enzymes are AXE members of carbohydrate esterase (CE) family 7 with the cephalosporine-C deacetylase activity for the production of antibiotics precursors. The simultaneous treatment of LaAXE with Thermotoga neapolitana β-xylanase showed 1.44-fold higher synergistic degradation of beechwood xylan than the single treatment of xylanase, whereas BhAXE showed no significant synergism. It was suggested that LaAXE can deacetylate beechwood xylan and enhance the successive accessibility of xylanase towards the resulting substrates. The novel LaAXE originated from a lactic acid bacterium will be utilized for the enzymatic production of D-xylose and xylooligosaccharides.

• Korean Journal of Environmental Agriculture
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• v.32 no.2
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• pp.108-116
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• 2013

BACKGROUND: Certain crop-based waste materials have been recognized as cost-effective and highly efficient adsorbents for removal and recovery of different kind of heavy metals from aqueous solutions. The ability is strongly attributed to the carboxyl functional group of some pectin substances such as galacturonic acid often found in fruit peels. The present manuscript was aimed at assessing the potential applicability of banana peel for metal removal from contaminated waters. METHODS AND RESULTS: As revealed by laboratory investigations, banana peel contains pectin (10-21%), lignin (6-12%), cellulose (7.6-9.6%), and hemicelluloses (6.4-9.4%). The pectin extraction is reported to have glucose, galactose, arabinose, rhamnose, xylose, and galactouroninc acid. Several studies conducted under different conditions proved that banana peel is capable of adsorbing 5.71, 2.55, 28.00, 6.88, 7.97, and 5.80 mg/g of $Cd^{2+}$, $Co^{2+}$, $Cu^{2+}$, $Ni^{2+}$, $Pb^{2+}$, and $Zn^{2+}$, respectively, from aqueous solutions. Adsorption capacity is, however, dependent upon several factors including solution pH, dose of adsorbent and metal concentration, contact time and shaking speed. CONCLUSION(S): Since the annual world production of banana exceeds 100 million tons, about 40 million tons of banana peel (40% of total weight of the fresh fruit) remains vastly unused. Exploring a sound technology with banana peel would therefore, not only address the much needed sustainable tool for cleaning contaminated waters, but of course bring an additional value to the banana industry worldwide.

• Microbiology and Biotechnology Letters
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• v.29 no.4
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• pp.221-226
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• 2001

Streptomyces themocyaneovio-laceus producing the thermostable xylanase was used for the production of xylooligosaccharides from xylan. The optimal conditions for the xylanase production were investigated in jar fermentor, which operated at 2 vvm aera-tion and 400 rpm agitation speed at $50^{\circ}C$ for 24 h. The optimal reaction condtion for the production of xylooli-gosaccharides with xylanases which were prepared by the percipitation with ammonium sulfate were obtained by the reaction at $60^{\circ}C$ for 12 h in the mixture composed of 10% birchwood xylan in 50 mM sodium phosphate buffer (pH 6.0)and 10 unit/ml of xylanase. In this optimal condition for the xylooligosaccharides production the mixture of xylooligosaccharides (58.8 g/I) which were composed of 20.1 g/I of xyobiose, 8.9 g/I of xylotriose 4.5 g/I of xylotetraose 16.2g/I of xylopentaose and 9.1 g/I xylohexaose and 5.0 g/I of xylose was produced from 100 g/I of birchwood xylan by the xylanases of S thermocyaneoviolaceus .

• Journal of Microbiology and Biotechnology
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• v.13 no.1
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• pp.125-133
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• 2003

Chemical properties and physiological activities of the freeze-dried synnemata of Beauveria bassiana were examined. A proximate analysis showed that the synnemata consisted mainly of carbohydrate (49.86%), protein 11.36%), and a moisture content of 30.64%. It contained a low amount of crude ash (4.76%) and fat (3.38%). The carbohydrate was composed mainly of mannose (52.3%), galactose (31.5%), glucose (13.2%), and rhamnose (3%). Trace amounts of arabinose, xylose, and fructose were present. Major amino acids In the synnemata were glutamic acid, glycine, aspartic acid, arginine, threonine, alanine, valine, leucine, lysine, and aspartic acid with the amounts of 30.42, 25.22, 17.17, 15.12, 12.65, 15.23, 12.47, 11.47, 14.24, and 17.17 mg/g, respectively. Among extracts from the synnemata, the hot-water extract showed 67% of anticomplementary activity compared to that of the positive control, followed by ethyl acetate extract (17%) and methanol extract (15%). The hot-water extract also had anticoagulant activity with 55 sec of coagulating time and this fraction exhibited the most potent Intestinal immune system modulating activity. The methanol extract showed the highest inhibitory activity (25%) on the 12-O-tetradecanoyl phorbol-13-acetate-induced superoxide ($O_2^-$) generation, followed by hot-water extract (18%) and ethyl acetate extract (10%). The data in the present study indicate that the extract of Beauveria bassiana synnemata contains some healthful chemical ingredients and it could provide beneficial physiological activities. These features of the synnemata should be of interest to the food industry as well as other industrial fields.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.11
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• pp.1555-1565
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• 2009

Anaerobic rumen fungi have been regarded as good genetic resources for enzyme production which might be useful for feed supplements, bio-energy production, bio-remediation and other industrial purposes. In this study, an expressed sequence tag (EST) library of the rumen anaerobic fungus Neocallimastix frontalis was constructed and functional genes from the EST library were analyzed to elucidate carbohydrate metabolism of anaerobic fungi. From 10,080 acquired clones, 9,569 clones with average size of 628 bp were selected for analysis. After the assembling process, 1,410 contigs were assembled and 1,369 sequences remained as singletons. 1,192 sequences were matched with proteins in the public data base with known function and 693 of them were matched with proteins isolated from fungi. One hundred and fifty four sequences were classified as genes related with biological process and 328 sequences were classified as genes related with cellular components. Most of the enzymes in the pathway of glucose metabolism were successfully isolated via construction of 10,080 ESTs. Four kinds of hemi-cellulase were isolated such as mannanase, xylose isomerase, xylan esterase, and xylanase. Five $\beta$-glucosidases with at least three different conserved domain structures were isolated. Ten cellulases with at least five different conserved domain structures were isolated. This is the first solid data supporting the expression of a multiple enzyme system in the fungus N. frontalis for polysaccharide hydrolysis.

• Journal of Microbiology and Biotechnology
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• v.14 no.3
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• pp.474-482
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• 2004

The $\alpha$-L-arabinofuranosidase (Arfase) gene of Bacillus stearothermophilus No. 236 was cloned and sequenced. The ORF of the gene, designated arfA, encoded a 507 -residue polypeptide with calculated molecular mass of 57 kDa. The Arfase produced by a recombinant Escherichia coli strain containing the arfA gene was purified to apparent homogeneity and characterized. The molecular mass of the Arfase determined by SDS-PAGE was 60 kDa. However, according to gel filtration, it was estimated to be approximately 190 kDa. These results indicated that the functional form of the Arfase is trimeric. The optimal pH and temperature for the enzyme activity were pH 6.5 and $55^{\circ}C$, respectively. The half-life of the enzyme at $60^{\circ}C$ was about 6 h. Kinetic experiments at $45^{\circ}C$ with pNPM (p-nitrophenyl $\alpha$-L-arabinofuranoside) as a substrate gave the $K_m and V_{max}$ values of 1.19 mM and 26.1 U/ mg, respectively. When the enzyme was combined with Bacillus stearothermophilus No. 236 endoxylanase and $\beta$-xylosidase, it hydrolyzed arabinoxylan into L-arabinose and xylose more efficiently than Arfase alone. This synergistic effect suggested that the complete hydrolysis of xylan with large amounts of arabinose side chains required Arfase as well as endoxylanase and $\beta$-xylosidase.

• Journal of Microbiology and Biotechnology
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• v.20 no.1
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• pp.45-53
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• 2010

We have developed an efficient and precise method for genome manipulations in Bacillus subtilis that allows rapid alteration of a gene sequence or multiple gene sequences without altering the chromosome in any other way. In our approach, the Escherichia coli toxin gene mazF, which was used as a counter-selectable marker, was placed under the control of a xylose-inducible expression system and associated with an antibiotic resistance gene to create a "mazF-cassette". A polymerase chain reaction (PCR)-generated fragment, consisting of two homology regions joined to the mazF-cassette, was integrated into the chromosome at the target locus by homologous recombination, using positive selection for antibiotic resistance. Then, the excision of the mazF-cassette from the chromosome by a single-crossover event between two short directly repeated (DR) sequences, included in the design of the PCR products, was achieved by counter-selection of mazF. We used this method efficiently and precisely to deliver a point mutation, to inactivate a specific gene, to delete a large genomic region, and to generate the in-frame deletion with minimal polar effects in the same background.

• Microbiology and Biotechnology Letters
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• v.22 no.2
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• pp.175-181
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• 1994

A bacterial strain producing highly viscous polysaccharide(A29 POL) was isolated from soil and identified sa Bacillus sp. A29. The cultural conditions of the Bacillus sp. A29 for the polysaccharide prouction were dextrin 12%, soytone 0.2%, SnCl$_{2}$ $\cdot $2H$_{2}$O 0.02%, Na$_{2}$HPO$_{4}$ $\CDOT $12H$_{2}$O 0.36%, L-alanine 0.01%, initial pH6.8, and 30$\circ $C at pH 3 FOR 4 days. Final viscosity of the culture broth was 65, 000 cp and then the amount of produced polysaccharide was 8.3 g/l. A29 POL was composed of glucose and xylose. A29 POL showed high viscosity at low concentration(0.1%) and in the presence of the salts such as NaCl or CaCl$_{2}$. A29 POL showed high viscosity acid condition and at alkali condition and high pseudoplasticity in the presence of a NaCl or CaCl$_{2}$. It was shown that the viscosity at high temperature(80$\circ $C) was decreased but it was recovered at low temperature (20$\circ $C. A29 POL was able to from film and gel in the presence of MgSO$_{4}$ $\CDOT $7H$_{2}$O, Na$_{2}$CO$_{3}$ \CDOT $H$_{2}$O, MnSO$_{4}$ $\CDOT $ 7H$_{2}$O. A29 POL had anionic charge.

• Nutrition Research and Practice
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• v.11 no.5
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• pp.381-387
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• 2017

BACKGROUND/OBJECTIVES: Vascular dementia (VaD) caused by reduced blood supply to the brain manifests as white matter lesions accompanying demyelination and glial activation. We previously showed that arabinoxylan consisting of arabinose and xylose, and arabinose itself attenuated white matter injury in a rat model of VaD. Here, we investigated whether larch arabinogalactan (LAG) consisting of arabinose and galactose could also reduce white matter injury. MATERIALS/METHODS: We used a rat model of bilateral common carotid artery occlusion (BCCAO), in which the bilateral common carotid arteries were exposed and ligated permanently with silk sutures. The rats were fed a modified AIN-93G diet supplemented with LAG (100 mg/kg/day) for 5 days before and 4 weeks after being subjected to BCCAO. Four weeks after BCCAO, the pupillary light reflex (PLR) was measured to assess functional consequences of injury in the corpus callosum (cc). Additionally, Luxol fast blue staining and immunohistochemical staining were conducted to assess white matter injury, and astrocytic and microglial activation, respectively. RESULTS: We showed that white matter injury in the the cc and optic tract (opt) was attenuated in rats fed diet supplemented with LAG. Functional consequences of injury reduction in the opt manifested as improved PLR. Overall, these findings indicate that LAG intake protects against white matter injury through inhibition of glial activation. CONCLUSIONS: The results of this study support our hypothesis that cell wall polysaccharides consisting of arabinose are effective at protecting white matter injury, regardless of their origin. Moreover, LAG has the potential for development as a functional food to prevent vascular dementia.