Browsing by Author "Junjarus Sermsathanaswadi"

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    Characterization and high-quality draft genome sequence of Herbivorax saccincola A7, an anaerobic, alkaliphilic, thermophilic, cellulolytic, and xylanolytic bacterium
    (Elsevier GmbH, 2018) Shimpei Aikawa; Sirilak Baramee; Junjarus Sermsathanaswadi; Phakhinee Thianheng; Chakrit Tachaapaikoon; Ayumi Shikata; Rattiya Waeonukul; Patthra Pason; Khanok Ratanakhanokchai; Akihiko Kosugi; A. Kosugi; Biological Resources and Post-Harvest Division, Japan International Research Center for Agricultural Sciences (JIRCAS), Tsukuba, 1-1 Ohwashi, 305-8686, Japan; email: akosugi@affrc.go.jp
    An anaerobic, cellulolytic-xylanolytic bacterium, designated strain A7, was isolated from a cellulose-degrading bacterial community inhabiting bovine manure compost on Ishigaki Island, Japan, by enrichment culture using unpretreated corn stover as the sole carbon source. The strain was Gram-positive, non-endospore forming, non-motile, and formed orange colonies on solid medium. Strain A7 was identified as Herbivorax saccincola by DNA-DNA hybridization, and phylogenetic analysis based on 16S rRNA gene sequences showed that it was closely related to H. saccincola GGR1 (= DSM 101079T). H. saccincola A7 (= JCM 31827 = DSM 104321) had quite similar phenotypic characteristics to those of strain GGR1. However, the optimum growth of A7 was at alkaline pH (9.0) and 55 �C, compared to pH 7.0 at 60 �C for GGR1, and the fatty acid profile of A7 contained 1.7-times more C17:0 iso than GGR1. The draft genome sequence revealed that H. saccincola A7 possessed a cellulosome-like extracellular macromolecular complex, which has also been found for Clostridium thermocellum and C. clariflavum. H. saccincola A7 contained more glycoside hydrolases (GHs) belonging to GH families-11 and -2, and more diversity of xylanolytic enzymes, than C. thermocellum and C. clariflavum. H. saccincola A7 could grow on xylan because it encoded essential genes for xylose metabolism, such as a xylose transporter, xylose isomerase, xylulokinase, and ribulose-phosphate 3-epimerase, which are absent from C. thermocellum. These results indicated that H. saccincola A7 has great potential as a microorganism that can effectively degrade lignocellulosic biomass. � 2018 Elsevier GmbH
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    Characterization of an Anaerobic, Thermophilic, Alkaliphilic, High Lignocellulosic Biomass-Degrading Bacterial Community, ISHI-3, Isolated from Biocompost
    (Elsevier Inc., 2018) Ayumi Shikata; Junjarus Sermsathanaswadi; Phakhinee Thianheng; Sirilak Baramee; Chakrit Tachaapaikoon; Rattiya Waeonukul; Patthra Pason; Khanok Ratanakhanokchai; Akihiko Kosugi; A. Kosugi; Biological Resources and Post-harvest Division, Japan International Research Center for Agricultural Sciences (JIRCAS), Tsukuba, 1-1 Ohwashi, 305-8686, Japan; email: akosugi@affrc.go.jp
    The generation of a complex microbial consortium is a promising approach for efficient biomass decomposition. An anaerobic thermophilic alkaliphilic microbial consortium with efficient degradation ability was screened from bovine manure compost using non-pretreated milling corn stover (CS) and rice straw (RS). A stable microbial consortium ISHI-3 with high degradation ability for CS and RS was isolated by the roll tube technique. ISHI-3 comprised Herbivorax saccincola and bacteria belonging to the classes Pelotomaculum, Tepidanaerobacter, and Tepidimicrobium, as determined by DGGE of the PCR-generated 16S rRNA genes. Furthermore, metagenomics analysis using a 16S rRNA library was carried out to determine the bacterial distribution during degradation of CS and RS. H. saccincola and bacteria belonging to Pelotomaculum were relatively abundant in the beginning to middle periods of culture with CS and RS whereas bacteria belonging to Tepidanaerobacter and Tepidimicrobium gradually increased in the population during the later stages. To understand the role of non-cellulolytic bacteria in the consortium, novel strains ET1 and GL4, which were most closely related to Tepidimicrobium ferriphilum and Tepidanaerobacter acetatoxydans, were isolated from ISHI-3. Based on their carbon source usage, morphology, and phylogenetic analysis, we propose that strains ET1 and GL4 should be classified as a novel genus or species. Bacteria ET1 and GL4 can utilize different organic compounds as carbon and energy sources such as organic acids, alcohols, sugars, and amino acids, showing a preference for organic acids and alcohols rather than sugars such as glucose and cellobiose. These results indicated that ET1 and GL4 help to accelerate efficient lignocellulose degradation of H. saccincola. � 2018 Elsevier Inc.
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    Formation of Rice Bran Glycosphingolipids Microemulsion Powders with Vitamin B1, B2, B12, and Folate as Additives for Elderly Food
    (Research and Development Institute Suan Dusit University, 2023) Junjarus Sermsathanaswadi; Siwawit Buasuwan; Dusit Angthararuk; Rittipun Rungruang; J. Sermsathanaswadi; Faculty of Science and Technology, Suan Dusit University, Bangkok, 10300, Thailand; email: janjaras_ser@dusit.ac.th
    This research investigated the extraction of glycosphingolipids from rice bran as an emulsifier to form microemulsions with vitamins B1, B2, B12 and folate being added to prepare the emulsion powder as the dietary supplement materials. Rice bran, a low-value agricultural material, was used as raw material for glycosphingolipids extraction using a solvent mixed between dichloromethane and methanol at a ratio of 2:1. The crude extract was observed in physical characteristics. The sphingosine in crude extract was analyzed by spectrophotometer technique. The emulsion was prepared using a mixture of crude extract, Polysorbate 80 and coconut oil. The average particle size of the emulsion was determined using Particle Size Analyzer. The emulsion was made into powder using various drying methods including hot air drying, spray drying and freeze-drying method. The mannan-oligosaccharides were used as an additive to replace the expensive mannitol or the energy-producing sucrose. The distribution property in distilled water of the emulsion powder was determined. The morphology and the surface of the powder emulsion were measured using SEM. The resistance to the imitation of the digestive system and encapsulation efficiency were determined. After extraction, the yield of crude glycosphingolipids extract from rice bran was 20.65%. The crude extract was a clear liquid, slightly yellow, insoluble in water and looks like oil. We found that the crude extract contained sphingosine 22.75 _g/g of crude extract from rice bran. When the emulsion was prepared, the characteristic of the emulsion showed the colloidal solution with a milky white color. The size of the emulsion without vitamins and with vitamins B1, B2, B12, and folate (total 100 ppm) were 70-75 nm and 74.4-78.1 nm, respectively. This result illustrated that the emulsion was classified as glycosphingolipid microemulsion. The emulsion powder was prepared and we found that the hot air drying and spray drying methods showed a viscous liquid with an oily smell. Whereas the freeze-drying method created the form of a light yellow, odorless and fine emulsion powder. The average particle size of emulsion was 80-100 nm. The solubility test showed that the emulsion powder was able to dissolve up to 300 g/L and 450 g/L at 25¡C and 70¡C, respectively. The morphological analysis showed that the powder emulsion was quite spherical with a diameter of less than 100 micrometers. In a simulated digestive system test, it was found that glycosphingolipid emulsion releases all vitamins in gastric simulated fluids and simulated small intestinal fluid conditions with values greater than 80%. © 2023, Research and Development Institute Suan Dusit University. All rights reserved.
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    Formation of Rice Bran Glycosphingolipids Microemulsion Powders with Vitamin B1, B2, B12, and Folate as Additives for Elderly Food
    (Graphicsite, 2024-12-18) Junjarus Sermsathanaswadi; Siwawit Buasuwan; Dusit Angthararuk; Rittipun Rungruang
    This research investigated the extraction of glycosphingolipids from rice bran as an emulsifier to form microemulsions with vitamins B1, B2, B12 and folate being added to prepare the emulsion powder as the dietary supplement materials. Rice bran, a low-value agricultural material, was used as raw material for glycosphingolipids extraction using a solvent mixed between dichloromethane and methanol at a ratio of 2:1. The crude extract was observed in physical characteristics. The sphingosine in crude extract was analyzed by spectrophotometer technique. The emulsion was prepared using a mixture of crude extract, Polysorbate 80 and coconut oil. The average particle size of the emulsion was determined using Particle Size Analyzer. The emulsion was made into powder using various drying methods including hot air drying, spray drying and freeze-drying method. The mannan-oligosaccharides were used as an additive to replace the expensive mannitol or the energy-producing sucrose. The distribution property in distilled water of the emulsion powder was determined. The morphology and the surface of the powder emulsion were measured using SEM. The resistance to the imitation of the digestive system and encapsulation efficiency were determined. After extraction, the yield of crude glycosphingolipids extract from rice bran was 20.65%. The crude extract was a clear liquid, slightly yellow, insoluble in water and looks like oil. We found that the crude extract contained sphingosine 22.75 micrograms/gram of crude extract from rice bran. When the emulsion was prepared, the characteristic of the emulsion showed the colloidal solution with a milky white color. The size of the emulsion without vitamins and with vitamins B1, B2, B12, and folate (total 100 ppm) were 70-75 nm and 74.4 -78.1 nm, respectively. This result illustrated that the emulsion was classified as glycosphingolipid microemulsion. The emulsion powder was prepared and we found that the hot air drying and spray drying methods showed a viscous liquid with an oily smell. Whereas the freeze-drying method created the form of a light yellow, odorless and fine emulsion powder. The average particle size of emulsion was 80-100 nm. The solubility test showed that the emulsion powder was able to dissolve up to 300 g/L and 450 g/L at 25°C and 70°C, respectively. The morphological analysis showed that the powder emulsion was quite spherical with a diameter of less than 100 micrometers. In a simulated digestive system test, it was found that glycosphingolipid emulsion releases all vitamins in gastric simulated fluids and simulated small intestinal fluid conditions with values greater than 80%.
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    Molecular characterization of hypothetical scaffolding-like protein S1 in multienzyme complex produced by Paenibacillus curdlanolyticus B-6
    (Springer Verlag, 2019) Patthra Pason; Junjarus Sermsathanaswadi; Rattiya Waeonukul; Chakrit Tachaapaikoon; Sirilak Baramee; Khanok Ratanakhanokchai; Akihiko Kosugi; A. Kosugi; Biological Resources and Post-harvest Division, Japan International Research Center for Agricultural Sciences (JIRCAS), Tsukuba, 1-1 Ohwashi, 305-8686, Japan; email: akosugi@affrc.go.jp
    Paenibacillus curdlanolyticus B-6 produces an extracellular multienzyme complex containing a hypothetical scaffolding-like protein and several xylanases and cellulases. The largest (280-kDa) component protein, called S1, has cellulose-binding ability and xylanase activity, thus was considered to function like the scaffolding proteins found in cellulosomes. S1 consists of 863 amino acid residues with predicted molecular mass 91,029�Da and includes two N-terminal surface layer homology (SLH) domains, but most of its sequence shows no homology with proteins of known function. Native S1 (nS1) was highly glycosylated. Purified nS1 and recombinant Xyn11A (rXyn11A) as a major xylanase subunit could assemble in a complex, but recombinant S1 (rS1) could not interact with rXyn11A, indicating that S1 glycosylation is necessary for assembly of the multienzyme complex. nS1 and rS1 showed weak, typical endo-xylanase activity, even though they have no homology with known glycosyl hydrolase family enzymes. S1 and its SLH domains bound tightly to the peptide-glycan layer of P. curdlanolyticus B-6, microcrystalline cellulose, and insoluble xylan, indicating that the SLHs of S1 bind to carbohydrate polymers and the cell surface. When nS1 and rXyn11A were co-incubated with birchwood xylan, the degradation ability was synergistically increased compared with that for each protein; however synergy was not observed for rS1 and rXynA. These results indicate that S1 may have a scaffolding protein-like function by interaction with enzyme subunits and polysaccharides through its glycosylated sites and SLH domains. � 2019, The Author(s).
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    Nitrogen-fixing Bacteria and Trends in Agricultural Applications
    (Graphicsite, 2023-09-26) Junjarus Sermsathanaswadi
    Bacteria that can increase the number of nutrients in the soil are important to plants, especially nitrogen-fixing bacteria that fix atmospheric nitrogen and change into the form that plants can use. In recent years, the use of nitrogen-fixing bacteria in agriculture has received a lot of attention because it offers an economically attractive and environmentally friendly method. Many species of nitrogen-fixing bacteria, symbiotic and non-symbiotic, that promote plant growth are used on a regular basis in order to improve crop yields. In addition to agricultural benefits, there are also potential benefits for environmental applications. Many nitrogenfixing bacteria which grow and multiply within plant tissues are called endophytes. They illustrate the tight association with the plant tissues without causing damage. Therefore, different types of endophytes which produce plant growth hormone provide benefit for many plants.
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    The C-terminal region of xylanase domain in Xyn11A from Paenibacillus curdlanolyticus B-6 plays an important role in structural stability
    (Springer Verlag, 2014) Junjarus Sermsathanaswadi; Somsak Pianwanit; Patthra Pason; Rattiya Waeonukul; Chakrit Tachaapaikoon; Khanok Ratanakhanokchai; Krisna Septiningrum; Akihiko Kosugi
    Paenibacillus curdlanolyticus B-6 produces an extracellular multienzyme complex containing a major xylanase subunit, designated Xyn11A, which includes two functional domains belonging to glycosyl hydrolase family-11 (GH11) and carbohydrate binding module family-36 (CBM36) and possesses a glycine and asparagine-rich linker (linker). To clarify the roles of each functional domain, recombinant proteins XynXL and XynX (CBM36 deleted and CBM36 and linker deleted, respectively) were constructed. Their xylanase activities were similar toward soluble xylan, whereas XynXL showed decreased hydrolysis activity toward insoluble xylan while XynX had no xylanase activity. To determine the significance of the linker and its neighbor region, XynX was subjected to secondary structural alignments using circular dichroism (CD) spectroscopy and three-dimensional (3D) structural analysis. A seven amino acid (NTITIGG) neighbor linker sequence was highly conserved among GH11 xylanases of Paenibacillus species. Although XynX exhibited a typical GH11 xylanase structure, conformational gaps were observed in the _6- and _12-sheets and in CD spectra. Flipping of the Arg163 side chains in the subsite was also observed upon analysis of superimposed models. Docking analysis using xylohexaose indicated that flipping of the Arg163 side chains markedly affected substrate binding in the subsite. To identify the amino acids related to stabilizing the substrate binding site, XynX with an extended C-terminal region was designed. At least seven amino acids were necessary to recover substrate binding and xylanase activity. These results indicated that the seven amino acid neighbor Xyn11A linker plays an important role in the activity and conformational stability of the xylanase domain. © 2014, Springer-Verlag Berlin Heidelberg.
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    The family 22 carbohydrate-binding module of bifunctional xylanase/_-glucanase Xyn10E from Paenibacillus curdlanolyticus B-6 has an important role in lignocellulose degradation
    (Elsevier Inc., 2017) Junjarus Sermsathanaswadi; Sirilak Baramee; Chakrit Tachaapaikoon; Patthra Pason; Khanok Ratanakhanokchai; Akihiko Kosugi; A. Kosugi; Biological Resources and Post-harvest Division, Japan International Research Center for Agricultural Sciences (JIRCAS), Tsukuba, 1-1 Ohwashi, 305-8686, Japan; email: akosugi@affrc.go.jp
    A newly isolated endo-_-1,4-xylanase (Xyn10E) from Paenibacillus curdlanolyticus B-6 has a modular structure consisting of a family 22 carbohydrate-binding module (CBM), a glycoside hydrolase (GH) family 10 catalytic domain, two fibronectin type III (Fn3) domains, and a family 3 CBM at the C-terminus. Intact Xyn10E (rXyn10E), CBM22-deleted Xyn10E (X-CBM3), CBM3-deleted Xyn10E (X-CBM22), and GH10 catalytic domain only (X-GH10) were expressed in Escherichia coli. rXyn10E showed bifunctional degradation activity toward xylan and _-glucan and also degraded microcrystalline cellulose. Although X-CBM3 and X-GH10 had drastically reduced xylanase and _-glucanase activities, X-CBM22 mostly retained these activities. Similar Km values were obtained for rXyn10E and X-CBM3, but kcat and kcat/Km values for X-CBM3 and X-GH10 were lower than those for rXyn10E, suggesting that CBM22 of Xyn10E may contribute to catalytic efficiency. In binding assays, X-CBM3 was still able to bind to _-glucan, soluble xylan, insoluble xylan, and cellulose through GH10 and CBM3. These results indicate that CBM22 has an important role not only in binding to xylan and _-glucan but also in feeding both polysaccharides into the neighboring GH10 catalytic domain. rXyn10E showed remarkable synergism with rXyn11A, a major xylanase subunit of P. curdlanolyticus B-6, in the degradation of untreated corn stover and sugarcane bagasse; however, the combination of X-CBM3 and rXyn11A was not synergistic. These results indicate that Xyn10E and Xyn11A act synergistically on lignocellulosic biomass, and CBM22 is essential for efficient degradation of lignocellulosic materials. � 2016 Elsevier Inc.
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    The GH67 _-glucuronidase of Paenibacillus curdlanolyticus B-6 removes hexenuronic acid groups and facilitates biodegradation of the model xylooligosaccharide hexenuronosyl xylotriose
    (Elsevier Inc., 2015) Krisna Septiningrum; Hiroshi Ohi; Rattiya Waeonukul; Patthra Pason; Chakrit Tachaapaikoon; Khanok Ratanakhanokchai; Junjarus Sermsathanaswadi; Lan Deng; Panida Prawitwong; Akihiko Kosugi
    4-O-Methylglucuronic acid (MeGlcA) side groups attached to the xylan backbone through _-1,2 linkages are converted to hexenuronic acid (HexA) during alkaline pulping. _-Glucuronidase (EC 3.2.1.139) hydrolyzes 1,2-linked MeGlcA from xylooligosaccharides. To determine whether _-glucuronidase can also hydrolyze HexA-decorated xylooligosaccharides, a gene encoding _-glucuronidase (AguA) was cloned from Paenibacillus curdlanolyticus B-6. The purified protein degraded hexenuronosyl xylotriose (_X3), a model substrate prepared from kraft pulp. AguA released xylotriose and HexA from _X3, but the Vmax and kcat values for _X3 were lower than those for MeGlcA, indicating that HexA side groups may affect the hydrolytic activity. To explore the potential for biological bleaching, _X3 degradation was performed using intracellular extract from P. curdlanolyticus B-6. The intracellular extract, with synergistic _-glucuronidase and _-xylosidase activities, degraded _X3 to xylose and HexA. These results indicate that _-glucuronidase can be used to remove HexA from _X3 derived from pulp, reducing the need for chemical treatments in the pulping process. � 2015 Elsevier Inc.