• Journal of Mushroom
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• v.15 no.1
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• pp.8-13
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• 2017

An auxin-producing bacterial strain, designated 4-3, was isolated from waste button mushroom compost in Boryeong-si, Chungnam. The strain 4-3 was classified as a novel strain of Leucobacter tardus, based on chemotaxonomic and phylogenetic analyses. TLC and HPLC the isolated L. tardus strain 4-3 produced indole-3-acetic acid (IAA), the auxin. Maximum IAA productionof $94.3mg\;L^{-1}$ was detected for bacteria cultured in R2A medium with 0.1% l-tryptophan, incubated for 24 h at $35^{\circ}C$. Negative correlationwas observed between IAA production and pH of the culture medium, indicating that the increase inIAA caused acidification ofthe medium. The effect of supplementation with varying concentrations of l-tryptophan, a known precursor of IAA, was also assessed. production was maximal at 0.1% l, but decreased at lconcentrations above 0.2%. To investigate the plant growth-promoting effects of the bacterium, L. tardus strain 4-3 culture broth was used to inoculate water cultures and seed pots of mung bean. We found thatadventitious root induction and root growth were 2.2-times higher in thethan in the non-inoculated plants.

• Journal of Applied Biological Chemistry
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• v.53 no.1
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• pp.1-7
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• 2010

xylA promoter is a major promoter in xylose operon of Escherichia coli. xylA promoter is sufficient as the promoter for the construction of new expression vector because this promoter was tightly controlled and induced by the addition of xylose. For the construction of xylose-inducible expression vector, 600 bp of xylA promoter was ligated between AatII and HindIII of pUC18, named pXA600. In order to investigate the effect of XylR protein encoded by xylR gene on the xylA promoter, 1,988 bp of xylR gene including its promoter was ligated into downstream of multiple cloning site to the opposite direction of xylA promoter in pXA600, named pXAR600. For the measurement of expression level, 3,048 bp of lacZ structural gene was fused into xylA promoter in both plasmids pXA600 and pXAR600 as a reporter gene, named pXA600-lacZ and pXAR600-lacZ, respectively. The $\beta$-galactosidase activity of pXA600-lacZ and pXAR600-lacZ in E. coli JM109 was determined to be 1,641 and 2,304 unit by the induction with xylose in LB medium, respectively. The $\beta$-galactosidase activity of pXAR600-lacZ/JM109 was about 1.4 times higher by the induction with xylose than that of pXA600-lacZ/JM109. The $\beta$-galactosidase activity of pXA600-lacZ and pXAR600-lacZ in E.coli JM109 showed 6,282 and 9,320 unit by the induction with xylose in DM minimal medium, respectively. A regulator, xylR protein works as an activator for the gene expression by the addition of xylose in the xylose-inducible vectors because the level of gene expression in pXA600 is increased by the insertion of xylR gene into the same vector. The xynA gene of Streptomyces thermocyaneoviolaceus cloned in pXA600 and pXAR600 was successfully expressed in E. coli BLR(DE3). As a result, plasmids pXA600 and pXAR600 using xylA promoter are sufficient as new expression system to produce a foreign protein in E. coli.

• Current Research on Agriculture and Life Sciences
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• v.10
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• pp.125-135
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• 1992

Nine strains of xyl mutants that could not utilize xylose as a carbon source were isolated from E. coli JM109 by the treatment of NTG in order to investigate the regulation of xylose operon and to use recipient cells for the cloning of xylR gene. For the characterization of all isolated mutants, colony colors of all mutants on MacConkey-xylose and MacConkey-xylulose agar plate were observed for the utilization of xylose and xylulose, and the growth level and the activity of xylose isomerase and xylulokinase were determined in need. The isolated xylR/T mutants formed the white colony on MacConkey-xy-lose and MacConkey-xylulose agar plate. They did not detect the activity of xylose isomerase, and the activity of xylose isomerase was not restored in transformants of xylR/T mutant with pEX13 which contained xylA gene. xylR and xylT mutants were classified from xylR/T mutants depending upon the growth level in minimal medium. xylT mutants; DH13, DH121 and DH125 could grow a little in that medium, but xylR mutants; DH10, DH53, and DH60 could not grow that medium.

• Journal of Mushroom
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• v.16 no.1
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• pp.45-50
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• 2018

This study was performed to compare medium characteristics during the composting stage for medium turning performed using an excavator agitator and a prototype medium turner in button mushroom cultivation. The changes in temperature in the medium did not significantly differ between the treatments until the 3rd turn performed using the excavator agitator. However, during the 4th and 5th turns, the temperature increased during turning with the prototype medium turner. During outdoor composting, various types of microorganisms such as thermophilic bacteria (Bacillus spp.), Actinomycetes, fluorescent Pseudomonas spp., and filamentous fungi were found to be distributed in the medium. The counts of aerobic bacteria and fluorescent Pseudomonas spp. did not significantly differ between treatments, and the counts of thermophilic bacteria and thermophilic actinomycetes were slightly higher during turning with the prototype medium turner. The rice straw was slightly shorter and water content lower for the prototype medium turner. There was no significant difference between pH and EC treatments. The L, a, and b values tended to increase on turning with the prototype medium turner.

• Journal of Mushroom
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• v.9 no.2
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• pp.53-58
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• 2011

In order to confirm the presence of putative glucoamylase gene in Tricholoma matsutake genome, the genomic DNA was prepared from T. matsutake NBRC30773 strain and was used as template to clone the glucoamylases gene (TmGlu1). We obtained the nucleotide sequence of TmGlu1 and its franking region. The coding region (from ATG to stop codon) is 2,186 bp. The locations of exons and introns were determined from the nucleotide sequences of 3'- and 5'-RACE PCR and RT-PCR products. On the other hand, to investigate the relationship between composition of medium and glucoamylase expression, we checked the expression level of glucoamylase gene by realtime reverse transcription PCR and measurement of glucoamylase enzyme activity. It was found that enzyme activity of glucoamylase was very low in different medium. Expression of glucoamylases gene appeared to not be affected by different carbon source.

• Journal of Mushroom
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• v.21 no.3
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• pp.110-117
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• 2023

Cordycepin (3'-deoxyadenosine) is a nucleoside analog known for its diverse range of biological activities. This study investigated the effect of different types of sawdust on the production of the bioactive compound cordycepin. The results of the study showed that different types of wood sawdust affected the biosynthesis of cordycepin and a significant increase was observed when the conventional SDB medium was replaced with 1% NaOH treated pine sawdust. To optimize cordycepin production from Paecilomyces tenuipes in a medium containing 1% NaOH-pretreated pine sawdust, we employed Response Surface Methodology (RSM) in its Box-Behnken design (BBD) canonical form. The optimal conditions were determined as follows: a particle size of 109.5111-mesh (140 ㎛) for 1% NaOH-pretreated pine sawdust, an input weight of 21.1679 g/L, and an incubation time of 73.8423 hours. According to our model, this combination is expected to yield a maximum cordycepin content of 896.1428 ㎍/mL. Experimental validation of this prediction was performed using the suggested optimal conditions, resulting in an average cordycepin content of 922.6771 ㎍/mL across three replicates, thus confirming the model's accuracy.

• Journal of Mushroom
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• v.20 no.3
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• pp.102-106
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• 2022

This study was conducted to investigate the growth and antioxidant activity of Pleurotus ostreatus mycelia grown in medium supplemented with Aronia. The diameter and dry weight of the mycelia were increased in the medium supplemented with Aronia compared with the untreated medium. The total polyphenol content of mycelia grown in medium supplemented with Aronia and untreated medium was 6.54 mg GAE/g and 5.77 mg GAE/g, respectively. The 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity of mycelia grown in medium supplemented with Aronia and untreated medium was 61.81% and 49.65%, respectively. Moreover, the 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging activity of mycelia grown in medium supplemented with Aronia and untreated medium was 59.83% and 52.66%, respectively. These results confirmed that P. ostreatus mycelial growth and antioxidant activity were increased when Aronia was added to the culture medium.

• Proceedings of the Korean Society of Life Science Conference
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• 2001.11a
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• pp.3-8
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• 2001

Isopentenyl diphosphate (IPP) is the common, five-carbon building block in the biosynthesis of all carotenoids, IPP in Escherichia coli is synthesized through the non-mevalonate pathway. The first reaction of IPP biosynthesis in E. coli is the formation of 1-deoxy-D-xylulose-5-phosphate (DXP), catalyzed by DXP synthase and encoded by dxs. The second reaction in the pathway is the reduction of DXP to 2-C-methyl-D-erythritol-4-phosphate, catalyzed by DXP reductoisomerase and encoded by dxr. To determine if one or more of the reactions in the non-mevalonate pathway controlled flux to IPP, dxs and dxr were placed on several expression vectors under the control of three different promoters and transformed into three E. coli strains (DH5(, XL1-Blue, and JM101) that had been engineered to produce lycopene. Lycopene production was improved significantly in strains transformed with the dxs expression vectors. When the dxs gene was expressed from the arabinose-inducible araBAD promoter (PBAD) on a medium-copy plasmid, lycopene production was 2-fold higher than when dxs was expressed from the IPTG-inducible trc and lac promoters (Ptrc and Plac, respectively) on medium-copy and high-copy plasmids, Given the low final densities of cells expressing dxs from IPTG-inducible promoters, the low lycopene production was probably due to the metabolic burden of plasmid maintenance and an excessive drain of central metabolic intermediates. At arabinose concentrations between 0 and 1.33 mM, cells expressing both dxs and dxr from PBAD on a medium-copy plasmid produced 1.4 - 2.0 times more lycopene than cells expressing dxs only. However, at higher arabinose concentrations lycopene production in cells expressing both dxs and dxr was lower than in cells expressing dxs only. A comparison of the three E. coli strains transformed with the arabinose-inducible dxs on a medium-copy plamid revealed that lycopene production was highest in XL1-Blue.

• Journal of Mushroom
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• v.20 no.4
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• pp.235-240
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• 2022

This study aimed to replace the imported Chinese complete medium for Lentinula edodes with a newly-developed complete medium that is suitable for export as well. Three media compositions that supported fast mycelium growth and higher density, compared to that in the control, were chosen. Culture in the T13 medium reduced the incubation period for 'Nongjingo' by 12 days and for 'Cham-aram' by 10 days, compared to that required for the control; in addition, the number of days required for browning was greatly reduced for both varieties. The quantity of each mixed medium was increased according to the composition from the 1^st to the 5^th cycle by 5.9% for 'Nongjingo' and 12.6% for 'Cham-aram' in T13, compared to that in the control. A mixed medium comprising oak sawdust + Douglas fir sawdust + corn flour (40:40:20, v/v) was selected as the most suitable complete medium.

• Journal of Mushroom
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• v.22 no.2
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• pp.67-72
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• 2024

In this study, we investigated the microbial community of oyster mushrooms at different growth stages at the species level. Gram-positive bacteria were predominant in the presterilized medium. On the other hand, Gram-negative bacteria were predominant in the culture-completed medium, post-harvest medium, and fruiting bodies. In addition, Pseudomonas tolaasii, which is known to cause disease in mushrooms, was confirmed in the cultured medium, post-harvest medium, and fruiting bodies, and it was determined that the mycelium culture stage was contaminated, and the reason why no disease occurred was Sphingobacterium psychroaquaticum. It was confirmed that this was because the growth of Pseudomonas tolaasii was suppressed by producing a component called tolacin. As a result of confirming the diversity of microorganisms, it was confirmed that the presterilization medium contains a variety of microorganisms compared to other growth stages, and the diversity decreases in the order of culture completion medium, fruiting body, and post-harvest medium. showed a trend. As a result of microbial similarity analysis, it was confirmed that the cultured medium and the post-harvest medium showed similar microbial communities, and in the case of fruiting bodies, there were some similarities but overall differences.