• Microbiology and Biotechnology Letters
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• v.20 no.1
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• pp.14-19
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• 1992

The xylanase producing microorganisms occurring on rotten woods were selectively isolated on the modified Czapek-Dox medium supplemented with 0.5% xylan as a sole carbon source. Among more than three-hundred isolates of xylanase producing microorganisms, only two bacterial isolates were turned out to be more potent xylanase producer than the reference strain of xylanase producer, Aureobaszdium pullulans NRRL Y-2311. The exo-xylanase producer, bacterial isolate No. 33 was identified as a strain of Pseudomonas sp. on the basis of morphological and biochemical characterizations as well as cellular fatty acid composition. Optima of pH and of temperature for enzyme reactions of xylanase were 5.5 and $50^{\circ}C$ respectively. The enzyme was stable in a range of pH 5.0~7.0 and below $45^{\circ}C$. Among the number of carbohydrate substrates, xylose was turned out to be a potent inducer of Pseudomonas sp. No.33 exo-xylanase. Among the raw materials tested, rice straw was the best material for xylanase production by Pseudomonas sp. strain No. 33.

• Microbiology and Biotechnology Letters
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• v.20 no.5
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• pp.574-582
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• 1992

Exo-xylanase encoded by the xylA gene of Bacillus stearothermoPhillus was produced from Escherichia coli ]M109 carrying a recombinant plasmid pMGL Synthesis of the enzyme was observed to be cell-associated, and about 94% of the enzyme synthesized was located in the cytoplasmic region. The maximum production was attained when the E. coli strain was grown at $37^{\circ}C$ for 8 hours on the medium containing 0.5% fructose, 1.0% tryptone, 1.0% sodium chloride, and 0.5% yeast extract. The exo-xylanase was purified to homogeneity using a combination of salting out with ammonium sulfate, DEAE-Sepharose CL-6B ion exchange chromatography, Sephadex G-IOO gel filtration, and Sephadex G-150 gel filtration. The' purified enzyme was most active at pH 6.0 and $45^{\circ}C$. $Ca^{2+}$ and $Co^{2+}$ activated the exo-xylanase activity by about 20% while $Ag^{2+}$, $Fe^{2+}$, $Mg^{2+}$ and $Zn^{2+}$ inhibited the enzyme activity by up to 60%. The $K_m$, value on p-nitrophenyl-$\beta$-D-xylanopyranoside was 2.75 mM. The enzyme had a pI value of 4.7. The estimated molecular weight of the native protein was 200,000 daL SDS-polyacrylamide gel electrophoresis analysis suggested that the native enzyme was a trimer composed of three identical 66,000 da!. polypeptides. The purified enzyme efficiently converted all the xylo-oligosaccharides tested to xylose. It was also confirmed that the enzyme split xylans in an exo-manner even though the degree of hydrolysis was fairly low. The xylanolytic enzyme was, therefore, classified to be one of the few bacterial exo-xylanases lacking transferase activity.

• Microbiology and Biotechnology Letters
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• v.17 no.4
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• pp.289-294
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• 1989

A bacterial strain capable of producing high level of extracellular xylanase was isolated from soil. The characteristics of the isolated strain No.236 were identified to be Bacillus stearothermophilus. The maximal xylanase production was observed in the medium containing 0.75% xylan, 0.35% yeast extract, 1.06% $K_2$HPO$_4$and 0.05% CaCO$_3$with initial pH of 6.5 when the strain was cultured at 5$0^{\circ}C$ for 28 hrs with reciprocal shaking. Hydrolysis of xylan by the xylanase revealed that xylose was the only product of the reaction. This suggested that the enzyme produced by Bacillus stearothermophilus No. 236 was an exe-acting xylanase.

• The Plant Pathology Journal
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• v.24 no.2
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• pp.143-151
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• 2008

To define the functions of the rpf genes in Xanthomonas oryzae pv. oryzae (Xoo), which regulates pathogenicity factors in Xanthomonas campestris pv. campestris (Xcc), marker-exchange mutants of each rpf gene were generated. When the mutants were inoculated on a susceptible cultivar, the lesion lengths caused by the rpfB, rpfC, rpfF, and rpfG mutants were significantly smaller than those caused by the wild type, whereas those caused by the rpfA, rpfD, and rpfI mutants were not. Several virulence determinants, including extracellular polysaccharide (EPS) production, xylanase production, and motility, were significantly decreased in the four mutants. However, the cellulase activity in the mutants was unchanged. Complementation of the rpfB and rpfC mutations restored the virulence and the expression of the virulence determinants. Expression analysis of 14 virulence genes revealed that the expression of genes related to EPS production (gumG and gumM), LPS (xanA, xanB, wxoD, and wxoC), phytase (phyA), xylanase (xynB), lipase (lipA), and motility (pitA) were reduced significantly in the mutants rpfB, rpfC, rpfF, and rpfG. In contrast, the expression of genes related to cellulase (eglxob, clsA), cellobiosidase (cbsA), and iron metabolism (fur) was unchanged. The results of this study clearly show that rpfB, rpfC, rpfF, and rpfG are important for the virulence of Xoo KACC10859, and that virulence genes are regulated differently by the Rpfs.

• Proceedings of the Korea Technical Association of the Pulp and Paper Industry Conference
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• 2011.04a
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• pp.81-91
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• 2011

Current fuel ethanol research and development deals with process engineering trends for improving biotechnological production of ethanol. Recently, a large amount of studies regarding the utilization of lignocellulosic biomass as a good feedstock for producing fuel ethanol is being carried out worldwide. The plant biomass is mainly composed of cellulose, hemicellulose and lignin. The main challenge in the conversion of biomass into ethanol is the complex, rigid and harsh structures which require efficient process and cost effective to break down. The isolation of microorganisms is one of the means for obtaining enzymes with properties suitable for industrial applications. For these reasons, crude cultures containing cellulosic biomass degrading microorganisms were isolated from rice field soil, cow farm soil and rotten rice straw from cow farm. Carboxymethyl cellulose (CMC), xylan and Avicel (microcrystalline cellulose) degradation zone of clearance on agar platefrom rice field soil resulted approximately at 25 mm, 24 mm and 22 mm respectively. As for cow farm soil, CMC, xylan and Avicel degradation clearancezone on agar plate resulted around at 24mm, 23mm and 21 mm respectively. Rotten rice straw from cow farm also resulted for CMC, xylan and Avicel degradation zone almost at 24 mm, 23 mm and 22 mm respectively. The objective of this study is to isolatebiomass degrading microbial strains having good efficiency in cellulose hydrolysis and observed the effects of different substrates (CMC, xylan and Avicel) on the production of cellulase enzymes (endo-glucanase, exo-glucanase, cellobiase, xylanase and avicelase) for producing low cost biofuel from cellulosic materials.