• Korean Journal of Environmental Agriculture
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• v.32 no.3
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• pp.240-243
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• 2013

BACKGROUND: Several yeast species have potential applications in biotechnology and the identification of such yeast species is of great interest. The first step in the identification of yeasts is the establishment of an effective isolation method. Thus, we compared the efficacy of different yeast media in the isolation of yeast associated with Aloe vera and Aloe saponaria. METHODS AND RESULTS: In this study, we spread homogenized A. vera and A. saponaria leaves onto 4 different yeast selective media containing chloramphenicol, streptomycin, Triton X-100 and L-sorbose. We observed high selectivity for yeast and many colonies on media. We isolated 67 yeast strains from A. vera and 42 yeast strains from A. saponaria. We used phylogenetic analysis to identify the yeast isolates based on ITS region sequencing and performed sequence analysis on representative isolates from each agar plate. Further, we compared the sequences obtained with reference sequences. The yeast species isolated from A. vera were as follows: 56 isolates of Meyerozyma, 9 isolates of Cryptococcus, and 1 isolate each of Rhodotorula and Sporobolomyces. Those isolated from A. saponaria were as follows: 41 isolates of Rhodosporidium and 1 isolate of Sporobolomyces. CONCLUSION(S): All the isolates obtained using large agar plate containing chloramphenicol, streptomycin, Triton X-100 and L-sorbose were identified as yeast. Therefore, we concluded that this method is useful for selective screening of yeast species.

• Journal of the Korean Society of Food Science and Nutrition
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• v.18 no.4
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• pp.435-443
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• 1989

The regulation of the synthesis of alcohol-oxidase(E. C. 1. 1. 3. 13) was investigated in the methanol-utilizing yeasts during growth on different carbon sources. For this experiment, Pichia pastoris CBS 2612 and Pichia pastoris CBM 10 were cultured in mineral salt medium by changing its carbon sources. The production of alcohol-oxidase was varied by the carbon sources. For example, alcohol-oxidase was undetectable in all strains submitted to the test in the medium with glucose, but its production was rapidely increased when the carbon source was changed from glucose to methanol after 48hrs of incubation. Moreover, this enzyme was not synthesized during growth on the primary aliphatic alcohols alone(ethanol, propanol, butanol or pentanol) or on the mixed substrates(0.5% methanol+0.5% primary aliphatic alcohols). When cells were grown on the various carbon sources(glucose, xylose, lactose, glycerol, galactose, saccharose, sorbose, lactic acid or acetic acid), The alcohol-oxidase activity was detected a very little amounts. These carbon sources together with methnol yieled far better synthesis of alcohol-oxidase than in case of carbon sources alone. Especially, the alcohol-oxidase activity of the cells grown on sorbose, lactose or lactic acid together with methanol was far better or similar than that of cells grown on methanol alone. The apparent Km values for the methanol of Pichia pastoris CBS 2612 and Pichia pastoris CBM 10 enzymes were 1.92 and 210 mM, respectively. It is also active towards alcohols of shorter alkyl-chain length than $C_7$, insaturated alcohols(allylalcohol, crotyl-alcohol) and secondary alcohols (iso-amylacohol, iso-butylalcohol). The affinity of alcohol-oxidase for this alcohols decreased with the increasing length of the alkyl-chain.

• Microbiology and Biotechnology Letters
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• v.20 no.5
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• pp.543-550
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• 1992

The production of xylanase and $\beta$-xylosidase was investigated in submerged culture of Aspergillus niger NRC 107. The maximum production occurred when the pH was controlled at 6.0 during the fermentation. Among the various carbon sources investigated, corn-cob xylan (1.5%, w/v) yielded maximal production of the enzymes. The $NaNO_{3}$ was the most favorable nitrogen source for enzyme production and $KH_2P0_4$ concentration at 0.3%(w/v) was found to be optimum. Incorporation of wheat bran to the culture medium improved xylanase production. Addition of L( -) sorbose to the culture medium promoted the secretion of $\beta$-xylosidase. It was possible to increase the production of xylanase (39.43 units/ml) and that of $\beta$-xylosidase (4.2 unitslml) by submerged culturing the A. niger NRC 107 in the modified medium.

• Journal of Microbiology and Biotechnology
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• v.20 no.2
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• pp.415-424
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• 2010

The purpose of this study was to investigate the relationship between the global regulatory mechanism known as quorum sensing and expression of virulence factors in Escherichia coli O157:87. A nonpolar luxS deletion was introduced into the chromosome of strain CI03J, a human clinical isolate from South Korea, to create the ${\Delta}luxS$ mutant strain ML03J. Phenotypic characterization of wild-type and mutant strains demonstrated that ML03J had no obvious growth or metabolic defects on 0.2% glucose LB medium, produced a functionally defective flagellum, and could not utilize sorbose; the biological significance of sorbose utilization is unknown. Omics-based analysis revealed the involvement of LuxS in the transcriptional activation of several flagella/chemotaxisrelated genes (flhD; fliA, C, D, S, Z; and cheA, Y, Z), repression of glutamate-dependent acid resistance genes (gadAB), and expression of virulence factors including Shiga toxin, hemolysin, and SepD within the LEE pathogenicity island.

• Journal of Microbiology and Biotechnology
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• v.7 no.2
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• pp.127-131
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• 1997

Alkaline phosphatase (APase) was found to be inducible in Anabaena sp. strain CA Growth was less than control in presence of most amino acids except glycine and serine, but most amino acids enhanced APase activity. Highest APase activity was recorded in tyrosine supplemented culture followed by hydroxyproline, cystein, valine and glutamic acid. Threonine supplemented material showed lowest APase level (1.8 nmol/mg protein/min). Lactose, glucose, sodium pyruvate and succinate stimulated growth but not APase activity. APase activity was high in the presence of sucrose, mellibiose, mannitol, arabinose, maltose and sorbose, even though the growth in these supplements was less than in control. Organic phosphate sources supported good growth of the organism. Best growth occurred in presence of inorganic phosphate, adenosine diphosphate, fructose 1,6-diphosphate or ribulose 1,5-diphosphate, followed by other phosphorus sources tested. APase activity in presence of any of the organic phosphate sources was 3 to 5 fold low as compared to phosphate limited culture. Also, there was no APase activity in cultures grown on inorganic phosphate. These data indicate that most amino acids and a few carbohydrates (sucrose, mellibiose, arabinose and sorbose) are suitable for APase production. Lactose, glucose, pyruvate or succinate may be used as a carbon source during photoheterotrophic growth of the cyanobacterium. Glycine and serine are preferred nitrogen sources for its growth. Phosphate repressible APase activity has been found in Anabaena sp. strain CA.

• Journal of Life Science
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• v.14 no.4
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• pp.582-588
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• 2004

Eighty one bacterial strains of parathion degrading bacteria were isolated from soil samples that were contaminated with pesticide in Daejeon area. Among them, one bacterial strain was finally selected in media containing parathion as the sole source of carbon and energy, and this strain was identified as Pseudomonas rhodesiae H5 through physiological and biochemical tests, and analysis of its 16S rRNA sequence. Pseudomonas rhodesiae H5 was able to utilize various carbohydrates but did not utilize sorbose as sole carbon source. Pseudomonas rhodesiae H5 was resistance to ampicillin, spectinomycin, and mitomycin C but sensitive to kanamycin and chloramphenicol. And this strain showed high resistance up to several milligrams of heavy metals such as $BaCl_2$, LiCl, and $MnSO_4$. Optimal growth condition for temperature and pH of P. rhodesiae H5 was 3$0^{\circ}C$, and pH 7.0, respectively. It can be presumed that P. rhodesiae H5 hydrolyzed an organophosphate bond of parathion, forming p-nitrophenol, and then metabolized via ortho-ring cleavage mechanism.

• KSBB Journal
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• v.7 no.1
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• pp.15-20
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• 1992

The use of Jerusalem artichoke containing $\beta$-1, 2-fructose oligomer in the production of sorbitol that is used as food additives and precursor for the L-sorbose has been studied. Coimmobilization of both inulinase and oxidoreductase was considered for the simultaneous reaction for hydrolysis of inulin and conversion of glucose and fructose liberated from inulin to sorbitol. Both inulinase and oxidoreductase were immobilized in chitin(5%, w/v) and K-carrageenan(4%, w/v), The activity of oxidoreductase was specified by permeabilization of Zymomonas mobilis cell with 0.2% CTAB(Cetyltrimethylammonlumbromide). The use of inulinase for hydrolysis of inulin resulted in 36.65g/l of glucose and 85.32g/1 of fructose respectively. These are valuable substrates for sorbitol production. Using these hydrolyzates, accumulation of 35.64g/l for sorbitol occurred at $38^{\circ}C$ and pH6.2. When permeabilized cells and inulinase were coimmobilized, sorbitol produced at 30.15g/l although it is low compared with 35.64g/l in separated reactor system.

• Journal of Microbiology and Biotechnology
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• v.31 no.8
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• pp.1154-1162
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• 2021

The transcriptional capacities of target genes are strongly influenced by promoters, whereas few studies have focused on the development of robust, high-performance and cross-species promoters for wide application in different bacteria. In this work, four novel promoters (P_k.rtufB, P_k.r1, P_k.r2, and P_k.r3) were predicted from Ketogulonicigenium robustum and their inconsistency in the -10 and -35 region nucleotide sequences indicated they were different promoters. Their activities were evaluated by using green fluorescent protein (gfp) as a reporter in different species of bacteria, including K. vulgare SPU B805, Pseudomonas putida KT2440, Paracoccus denitrificans PD1222, Bacillus licheniformis and Raoultella ornithinolytica, due to their importance in metabolic engineering. Our results showed that the four promoters had different activities, with P_k.r1 showing the strongest activity in almost all of the experimental bacteria. By comparison with the commonly used promoters of E. coli (tufB, lac, lacUV5), K. vulgare (Psdh, Psndh) and P. putida KT2440 (JE111411), the four promoters showed significant differences due to only 12.62% nucleotide similarities, and relatively higher ability in regulating target gene expression. Further validation experiments confirmed their ability in initiating the target minCD cassette because of the shape changes under the promoter regulation. The overexpression of sorbose dehydrogenase and cytochrome c551 by P_k.r1 and P_k.r2 resulted in a 22.75% enhancement of 2-KGA yield, indicating their potential for practical application in metabolic engineering. This study demonstrates an example of applying bioinformatics to find new biological components for gene operation and provides four novel promoters with broad suitability, which enriches the usable range of promoters to realize accurate regulation in different genetic backgrounds.

• Journal of the Korean Applied Science and Technology
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• v.34 no.1
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• pp.33-40
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• 2017

To isolate and identify the yeast strains associated with D. morbifera, homogenized D. morbifera root samples were spread onto GPY, DG18, SCG and DOB agar media containing antibiotics, Triton X-100, and l-sorbose. Total 81 yeast isolates were analyzed by sequencing of internal transcribed spacer (ITS) region of the ribosomal DNA. The results showed that the root-associated yeast species were composed of the genera Vanderwaltozyma (40 isolates), Cryptococcus (40 isolates), and Kluyveromyces (one isolate). Moreover, the Kluyveromyces isolate exhibited high bioethanol productivity. In addition, the Vanderwaltozyma and Cryptococcus were dominant in D. morbifera roots. The specific yeast community associated with D. morbifera roots was identified by phylogenetic sequence analyses. These yeast isolates may have industrial applications as biosurfactant and bioethanol.

• Korean Journal of Environmental Agriculture
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• v.35 no.2
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• pp.137-142
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• 2016

BACKGROUND: Identification of endophytic yeasts inhabiting the internal roots of the Mankyua chejuense tree requires techniques involving biotechnology. There is a need for a culture-based method to isolate and identify yeast strains associated with M. chejuense.METHODS AND RESULTS: We spread homogenized M. chejuense root samples onto glucose-peptone- yeast agar containing antibiotics, Triton X-100, and L-sorbose. A total of 152 yeast isolates were obtained and identified via phylogenetic analysis based on ITS gene sequencing. The results revealed that the root-associated yeast species included the genera Cyberlindnera (140 isolates), Candida (11 isolates), and Kluyveromyces (one isolate). Additionally, three yeast isolates showed high bioethanol production.CONCLUSION: We identified the specific yeast community associated with M. chejuense roots. These yeast isolates may have industrial applications as bioethanol producers. Our findings revealed that Cyberlindnera isolates included C. suaverolens and C. satumus, while Kluyveromyces isolates showed high bioethanol production.