• Journal of the Korean Society of Food Science and Nutrition
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• v.36 no.10
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• pp.1341-1350
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• 2007

This study was designed to ultimately develop a highly efficient mass production technology of bacterial cellulose isolated from the citrus gel fermented by G. hansenii TL-2C. Pilot equipment made with FRP vessel length (665 mm) ${\times}$ width (375 mm) ${\times}$ height (210 mm) was developed for mass production of the citrus gel. To develop the optimal conditions for mass production of citrus gel, comprised of citrus juice (6,000 mL) diluted 100 times, containing 5% seed bacteria, 10% sucrose, and 1% ethanol, citrus juice was fermented at $30^{\circ}C$ for 14 days, and gel productivity in pilot system was examined. BC was isolated and purified from the citrus gel, and their chemical composition and physicochemical properties were investigated.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.1
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• pp.170-175
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• 2004

This study was conducted to determine the minimum concentration of citrus juice for basal medium and also to search for an additional carbon source for the best production of the gel. A concentrate of citrus fruit juice of 65$^{\circ}$Brix, it was diluted to be used as a basal medium. Static cultivation of Gluconacetobacter hansenii TL-2C for 14 days at 3$0^{\circ}C$ produced the best gel with 7.5$\pm$0.4 mm thickness in the 6-fold diluted citrus Juice concentrate without any additional nutrient. However, the same thickness could be obtained with 60 to 100-fold diluted juice concentrate when refined white sugar was added at appropriate concentrations. Glucose was the most effective sugar for the both of gel and acid production, and optimal concentration of the sugar was 10$^{\circ}$Brix. Ethyl alcohol at 1.0% had synergistic effects in combination with refined sugar and increased the gel thickness up to 15.1 mm which was 1.85 times thicker than that of refined sugar alone. However, acetic acid was not effective. Gel productivity with supplement of ethanol was 172.6$\pm$8.4 g wet/L, and it was approximately equal to 4.7 g of dry gel/L.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.1
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• pp.176-181
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• 2004

Gluconacetobacter hansenii TF-2, an isolate from black tea fungus, was statically cultivated to ferment cellulose gel from citrus juice. The juice prepared by press filtering of peeled citrus fruit contained 135.5 mg of total sugar/mL, 1.23% of total acid, and average pH of the juice was 3.98. The bacterium produced cellulose gel optimally on the surface of culture broth containing 17% of citrus juice and 10$^{\circ}$Brix of total sugar. The optimum temperature was 3$0^{\circ}C$ for producing acetic acid and gel formation. The bacterium could not produce acetic acid on gel formation at 4$0^{\circ}C$. The optimum pH was 3.0∼4.0 but was not significantly different between pH 3.0∼4.0. The cultivation for 18 days under optimal conditions produced gel as 14.2$\pm$0.6 mm of thickness and acids equivalent to 1.90$\pm$0.22% of acetic acid. The pH of culture broth was stabilized at 2.6∼2.8 during the cultivation. Remaining sugar content was 27.1$\pm$4.2 mg/mL of total sugar and 6.9 mg/mL of reducing sugar. The gel productivity was 137.8$\pm$9.7 g/L.

• The Plant Pathology Journal
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• v.34 no.6
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• pp.499-505
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• 2018

Huanglongbing (HLB, Citrus greening disease) is one of the most devastating diseases that threaten citrus production worldwide. Although HLB presents systemically, low titer and uneven distribution of these bacteria within infected plants can make reliable detection difficult. It was known loop-mediated isothermal amplification (LAMP) method has the advantages of being highly specific, rapid, efficient, and laborsaving for detection of plant pathogens. We developed a new LAMP method targeting gene contained tandem repeat for more rapid and sensitive detection of Candidatus Liberibacter asiaticus (CLas), putative causal agent of the citrus huanglongbing. This new LAMP method was 10 folds more sensitive than conventional PCR in detecting the HLB pathogen and similar to that of real-time PCR in visual detection assay by adding SYBR Green I to mixture and 1% agarose gel electrophoresis. Positive reactions were achieved in reaction temperature 57, 60 and $62^{\circ}C$ but not $65^{\circ}C$. Although this LAMP method was not more sensitive than real-time PCR, it does not require a thermocycler for amplification or agarose gel electrophoresis for resolution. Thus, we expect that this LAMP method shows strong promise as a reliable, rapid, and cost-effective method of detecting the CLas in citrus and can be applied for rapid diagnosis is needed.

• Journal of Microbiology
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• v.41 no.3
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• pp.212-218
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• 2003

Citrus phytoene synthase (CitPsy) and ${\beta}$-carotene hydroxylase (CitChx), which are involved in caroteinoid biosynthesis, are distantly related to the corresponding bacterial enzymes from the point of view of amino acid sequence similarity. We investigated these enzyme activities using Pantoea ananatis carotenoid biosynthetic genes and Escherichia coli as a host cell. The genes were cloned into two vector systems controlled by the T7 promoter. SDS-polyacrylamide gel electrophoresis showed that CitPsy and CitChx proteins are normally expressed in E. coli in both soluble and insoluble forms. In vivo complementation using the Pantoea ananatis enzymes and HPLC analysis showed that ${\beta}$-carotene and zeaxanthin were produced in recombinant E. coli, which indicated that the citrus enzymes were functionally expressed in E. coli and assembled into a functional multi-enzyme complex with Pantoea ananatis enzymes. These observed activities well matched the results of other researchers on tomato phytoene synthase and Arabidopsis and pepper ${\beta}$-carotene hydroxylases. Thus, our results suggest that plant carotenoid biosynthetic enzymes can generally complement the bacterial enzymes and could be a means of carotenoid production by molecular breeding and fermentation in bacterial and plant systems.

• Journal of Microbiology
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• v.35 no.2
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• pp.134-140
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• 1997

Botrytis cinerea T91-1 has been shown to produce at least four different polygalacturonases into a liquid medium containing citrus pectin, a carbon sousrce. One of the enzymes, which had an apparent molecular weight of 66 kDa estimated by denatured polyacrylamide gel electrophoresis, was purified to electrophoretic homogeneity by a series of procedures including a cetone precipitation, ion exchange, heparin affinity, and reverse phase column chromatographies. The molecular weight of native enzyme was determined to be 64 kDa by gel permeation chromatography indicating the enzyme to be a single polypeptide chain. By viscometric analysis, the enzyme was revealed as exo-polygalacturonase. The enzyme activity was inhibited by divalent cations such as $Ca^{2+}$, $Mg^{2+}$, and Cu$^{2+}$. Km and Vmax for polygalacturonic acid hydrolysis were 0.33 mg/ml and 28.6 nM/min, respectively. The optimum temperature for enzymatic activity was 5$0^{\circ}C$. And the enzyme showed optimal pH values between 4.0 and 5.0. The enzyme was stable upto 12 hours in the range of pH 3 to 8 and at temperature below 3$0^{\circ}C$.

• The Korean Journal of Mycology
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• v.25 no.4 s.83
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• pp.330-339
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• 1997

Botrytis cinerea T91-1 has shown to produce at least four different polygalacturonases in a liquid medium containing citrus pectin as a carbon source. One of the enzymes, its molecular weight was estimated as 37 kDa by denatured polyacrylamide gel electrophoresis, was purified by a series of procedures including acetone precipitation, ion exchange, heparin affinity, and reverse phase column chromatographies. By viscometric analysis, the enzyme was revealed as an endo-polygalacturonase. The enzyme activity was inhibited by divalent cations such as $Ca^{2+}$, $Co^{2+}$, and $Cu^{2+}$. Km and Vmax for polygalacturonic acid hydrolysis were 0.33 mg/ml and 28.6 nM/min, respectively. The optimum temperature for enzymatic activity was $55^{\circ}C$ and the enzyme showed optimal pH values between 4.0 and 4.5. The enzyme was stable up to 12 hours in the range of pH 4 to 7 and at the temperature below $30^{\circ}C$. Amino acid sequence from N-terminal up to 6 amino acids determined by Edman degradation showed little homology with polygalacturonases from fungi and plants.