• Journal of Microbiology and Biotechnology
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• v.24 no.11
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• pp.1559-1565
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• 2014

Cellulase and xylanase are main hydrolysis enzymes for the degradation of cellulosic and hemicellulosic biomass, respectively. In this study, our aim was to develop and test the efficacy of a rapid, high-throughput method to screen hydrolytic-enzyme-producing microbes. To accomplish this, we modified the 3,5-dinitrosalicylic acid (DNS) method for microwell plate-based screening. Targeted microbial samples were initially cultured on agar plates with both cellulose and xylan as substrates. Then, isolated colonies were subcultured in broth media containing yeast extract and either cellulose or xylan. The supernatants of the culture broth were tested with our modified DNS screening method in a 96-microwell plate, with a $200{\mu}l$ total reaction volume. In addition, the stability and reliability of glucose and xylose standards, which were used to determine the enzymatic activity, were studied at $100^{\circ}C$ for different time intervals in a dry oven. It was concluded that the minimum incubation time required for stable color development of the standard solution is 20 min. With this technique, we successfully screened 21 and 31 cellulase- and xylanase-producing strains, respectively, in a single experimental trial. Among the identified strains, 19 showed both cellulose and xylan hydrolyzing activities. These microbes can be applied to bioethanol production from cellulosic and hemicellulosic biomass.

• Journal of Microbiology and Biotechnology
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• v.27 no.2
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• pp.342-349
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• 2017

Polylactic acid (PLA) has been highlighted as an alternative renewable polymer for the replacement of petroleum-based plastic materials, and is considered to be biodegradable. On the other hand, the biodegradation of PLA by terminal degraders, such as microorganisms, requires a lengthy period in the natural environment, and its mechanism is not completely understood. PLA biodegradation studies have been conducted using mainly undefined mixed cultures, but only a few bacterial strains have been isolated and examined. For further characterization of PLA biodegradation, in this study, the PLA-degrading bacteria from digester sludge were isolated and identified using a polymer film-based screening method. The enrichment of sludge on PLA granules was conducted with the serial transference of a subculture into fresh media for 40 days, and the attached biofilm was inoculated on a PLA film on an agar plate. 3D optical microscopy showed that the isolates physically degraded the PLA film due to bacterial degradation. 16S rRNA gene sequencing identified the microbial colonies to be Pseudomonas sp. MYK1 and Bacillus sp. MYK2. The two isolates exhibited significantly higher specific gas production rates from PLA biodegradation compared with that of the initial sludge inoculum.

• Korean Journal of Agricultural Science
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• v.28 no.2
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• pp.125-131
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• 2001

Trametes versicolor(CV5) selected as a white-rot fungus with strong lignin degrading activity, in the previous paper, was investigated on a properties of degradation of wood lignin. Lignins of hardwoods, especially oak(Querous acutissima carruth) an paulownia (Paulownia coreana Uyeki) were considerably delignified by the CV5, however, softwoods used in this experiment were not delignified. Bavendamm's reaction was positive with several phenols on agar plates for the confirmation of a phenoloxidase secreted Through the morphologies of decayed wood chip observed with the aid of scanning electron microscopy, it was found that the hypha of CV5 penetrated the ray cells and vessels caused separation of the wood cellulose.

• Journal of Microbiology and Biotechnology
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• v.23 no.1
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• pp.15-21
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• 2013

We previously observed that Bacillus subtilis spores from sspE mutants presented a lower germination capacity in media containing high salt concentrations (0.9M NaCl). This deficiency was attributed to the absence of SASP-E (gamma-type small-acid-soluble protein), rich in osmocompatible amino acids released by degradation. Herein we observed that, in addition, this mutant spore presented a reduced capacity to use L-alanine as germinant (L-ala pathway), required longer times to germinate in calcium dipicolinate ($Ca^{2+}$-DPA), but germinated well in asparagine, glucose, fructose, and potassium chloride (AGFK pathway). Moreover, mild sonic treatment of mutant spores partially recovered their germination capacity in L-ala. Spore qualities were also altered, since sporulating colonies from the sspE mutant showed a pale brownish color, a higher adherence to agar plates, and lower autofluorescence, properties related to their spore coat content. Furthermore, biochemical analysis showed a reduced partition in hexadecane and a higher content of $Ca^{2+}$-DPA when compared with its isogenic wild-type control. Coat protein preparations showed a different electrophoretic pattern, in particular when detected with antibodies against CotG and CotE. The complementation with a wild-type sspE gene in a plasmid allowed for recovering the wild-type coat phenotype. This is the first report of a direct involvement of SASP-E in the spore coat assembly during the differentiation program of sporulation.

• Journal of the Korean Wood Science and Technology
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• v.48 no.1
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• pp.107-116
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• 2020

Termiticidal and fungicidal activities of wood vinegar from Cinnamomum parthenoxylon (CP) stem wood have been evaluated against Coptotermes curvignathus and wood rotting fungi (Schizophyllum commune and Fomitopsis palustris). The utilized CP wood vinegar was produced in the operating temperature range 250-300℃ pyrolysis. A no-choice test was applied for evaluating termiticidal activity with 33 active termites and antifungal activity using the agar media assay. The result showed that an increase in the concentrations of CP wood vinegar significantly raised the mortality of termite. CP wood vinegar showed high termiticidal activity, organic acids (acetic acid 42.91%, 3-butenoic acid 6.89%, butanoic acid, 2-propenyl ester 2.26%), and ketones (1-hydroxy-2-propanone 5.14%, 3-methylcyclopentane-1,2-dione 2.34%) might be largely contributed to termiticidal activity in addition to other minor components. Furthermore, CP wood vinegar exhibited significant inhibition of fungal growth. These data showed that CP wood vinegar was more toxic to white-rot fungi (S. commune) than brown-rot (F. palustris). The results suggested that phenolic compounds from lignin degradation were responsible for good antifungal activity.

• Journal of the Korea Organic Resources Recycling Association
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• v.19 no.4
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• pp.84-89
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• 2011

Cellulose-degrading bacteria were isolated and identified from cocopeat which has a good quality as a bulking agent in composting. Various bacteria from different sourecs of cocopeat were detected on CMC agar media, and these were found to be Burkholderi2a sp., Bacillu subtilis, Sphingomonas sp., Rhodotorula sp. & Pseudomonas sp. etc. Among these, four bacteria were further selected and analyzed for their biochemical characteristics and CMCase activities. CMCase activities of four bacteria, P. aeruginosa, P. stutzeri, B. subtilis, and P. luteola were found to be 83%, 40%, 8%, 6%, respectively, compared with that of the standard strain Cellulomonas sp.

• Journal of Applied Biological Chemistry
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• v.60 no.1
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• pp.79-86
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• 2017

Microbes in forest are very important due to not only to enhance soil fertility but also maintain a healthy ecosystem by supplying the energy available to living organisms by producing various kinds of enzymes related to degradation of lignocellulosic biomass. In order to isolate a lignocellulosic biomass degrading bacterial strain from the Jurassic park located in Gyeongnam National University of Science and Technology, We used the Luria-Bertani-Carboxymethyl cellulose (CMC) agar trypan blue method containing 0.4 % carboxymethyl cellulose and 0.01 % trypan blue. As a result, we isolated a bacterial strain showing both activity on the CMC and xylan. To identify the isolated strain, 16S rRNA sequencing and API kit analysis were used. The isolated strain turned out to belong to Bacillus species and then named Bacillus sp. GJY. In the CMC zymogram analysis, it showed that one active band of about 28kDa in size is present. Xylan zymogram analysis also showed to have one active band of about 25kDa in size. The optimal growth temperature of Bacillus sp. GJY was $37^{\circ}C$. The maximal activities of CMCase and xylanase were 12 hour after incubation. The optimal pH and temperature for CMCase were 5.0 and $40^{\circ}C$, respectively, whereas the optimal pH and temperature for xylanase was 4.0 and $40^{\circ}C$. Both activities for CMCase and xylanase showed to be thermally stable at 40and $50^{\circ}C$, while both activities rapidly decreased at over $60^{\circ}C$.

• Korean Journal of Environmental Agriculture
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• v.19 no.4
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• pp.284-293
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• 2000

For the artificial removal of air pollutants such as pesticides, environmental toxicants, and pathogenic microorganisms in the greenhouse or the living environment, the accelerated photodegradation and the biocidal effects of some photosensitizers (PS)/photocatalysts (PC) were tested under the sunlight and/or artificial light. The selected photosensitizers/photocatalysts included the semiconductors (PC-1 and PC-2), the oxidizers (PC-3, PC-4, PC-5 and PC-6), the aromatic ketone (PS-7) and the aromatic amine (PS-8). In the case of dichlorvos, all the photocatalysts selected showed more accelerated photodegradation than the control without photocatalysts under both the sunlight and artificial light. Whereas, only the photocatalyst PC-1 accelerated the degradation of methyl tert-butyl ether about 17 times more than the control under both the sunlight and artificial light. Procymidone was much more degraded by the photosensitizer PS-8 and the two photocatalysts (PC-1, PC-6) than by PS-7. In the preliminary experiments to diminish the population of the microorganisms in the air, the photocatalyst PC-1 added to the suspensions of Pseudomonas putida, Phytophthora capsici, and Salmonella typhimurium obviously inhibited the microbial growth under the artificial light. The photocatalyst PC-1 showed a bactericidal activity against Salmonella typhimurium spread on the nutrient broth agar medium. These results suggest that the photosensitizers/photocatalysis under the light can remove some air pollutants and hence they can be used to reduce the exposure of the workers in the horticultural facilities and/or the public in the environment to the harmful pollutants.

• Journal of Microbiology and Biotechnology
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• v.27 no.1
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• pp.179-188
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• 2017

White-rot basidiomycetes are the organisms that decompose lignin most efficiently, and Trametes villosa is a promising species for ligninolytic enzyme production. There are several publications on T. villosa applications for lignin degradation regarding the expression and secretion of laccase and manganese peroxidase (MnP) but no reports on the identification and characterization of lignin peroxidase (LiP), a relevant enzyme for the efficient breakdown of lignin. The object of this study was to identify and partially characterize, for the first time, gDNA, mRNA, and the corresponding lignin peroxidase (TvLiP) protein from T. villosa strain CCMB561 from the Brazilian semiarid region. The presence of ligninolytic enzymes produced by this strain grown in inducer media was qualitatively and quantitatively analyzed by spectrophotometry, qPCR, and dye fading using Remazol Brilliant Blue R. The spectrophotometric analysis showed that LiP activity was higher than that of MnP. The greatest LiP expression as measured by qPCR occurred on the $7^{th}$ day, and the ABSA medium (agar, sugarcane bagasse, and ammonium sulfate) was the best that favored LiP expression. The amplification of the TvLiP gene median region covering approximately 50% of the T. versicolor LPGIV gene (87% identity); the presence of Trp199, Leu115, Asp193, Trp199, and Ala203 in the translated amplicon of the T. villosa mRNA; and the close phylogenetic relationship between TvLiP and T. versicolor LiP all indicate that the target enzyme is a lignin peroxidase. Therefore, T. villosa CCMB561 has great potential for use as a LiP, MnP, and Lac producer for industrial applications.

• Journal of Microbiology and Biotechnology
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• v.23 no.11
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• pp.1519-1528
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• 2013

Bacillus pumilus SG2, a halotolerant strain, expresses two major chitinases designated ChiS and ChiL that were induced by chitin and secreted into the supernatant. The present work aimed to obtain a mutant with higher chitinolytic activity through mutagenesis of Bacillus pumilus SG2 using a combination of UV irradiation and nitrous acid treatment. Following mutagenesis and screening on chitin agar and subsequent formation of halos, the mutated strains were examined for degradation of chitin under different conditions. A mutant designated AV2-9 was selected owing to its higher chitinase activity. To search for possible mutations in the whole operon including ChiS and ChiL, the entire chitinase operon, including the intergenic region, promoter, and two areas corresponding to the ChiS and ChiL ORF, was suquenced. Nucleotide sequence analysis of the complete chitinase operon from the SG2 and AV2-9 strains showed the presence of a mutation in the catalytic domain (GH18) of chitinase (ChiL). The results demonstrated that a single base change had occurred in the ChiL sequence in AV2-9. The wild-type chitinase, ChiL, and the mutant (designated ChiLm) were cloned, expressed, and purified in E. coli. Both enzymes showed similar profiles of activity at different ranges of pH, NaCl concentration, and temperature, but the mutant enzyme showed approximately 30% higher catalytic activity under all the conditions tested. The results obtained in this study showed that the thermal stability of chitinase increased in the mutant strain. Bioinformatics analysis was performed to predict changes in the stability of proteins caused by mutation.