• Journal of Microbiology and Biotechnology
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• v.20 no.2
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• pp.340-345
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• 2010

The culture variables were optimized to increase 1,3-dihydroxyacetone (DHA) production by Gluconohacter oxydans ZJB09112 in shake flasks and bubble column bioreactors. After fermentation in the optimized medium (g/l: yeast extract 5, glycerol 2.5, mannitol 22.5, $K_2HPO_4$ 0.5, $KH_2PO_4$ 0.5, $MgSO_4{\cdot}7H_2O$ 0.1, $CaCO_3$ 2.0, pH 5.0), when five times of glycerol feeding were applied, $161.9{\pm}5.9\;g/l$ of DHA was attained at a $88.7{\pm}3.2%$ conversion rate of glycerol to DHA.

• Journal of Microbiology and Biotechnology
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• v.26 no.11
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• pp.1908-1917
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• 2016

Wild strain L-6 was subjected to combined mutagenesis, including UV irradiation, atmospheric and room temperature plasma, and ion beam implantation, to increase the yield of 1,3-dihydroxyacetone (DHA). With application of a high-throughput screening method, mutant Gluconobacter oxydans I-2-239 with a DHA productivity of 103.5 g/l in flask-shake fermentation was finally obtained with the starting glycerol concentration of 120 g/l, which was 115.7% higher than the wild strain. The cultivation time also decreased from 54 h to 36 h. Compared with the wild strain, a dramatic increase in enzyme activity was observed for the mutant strain, although the increase in biomass was limited. DNA and amino acid sequence alignment revealed 11 nucleotide substitutions and 10 amino acid substitutions between the sldAB of strains L-6 and I-2-239. Simulation of the 3-D structure and prediction of active site residues and PQQ binding site residues suggested that these mutations were mainly related to PQQ binding, which was speculated to be favorable for the catalyzing capacity of glycerol dehydrogenase. RT-qPCR assay indicated that the transcription levels of sldA and sldB in the mutant strain were respectively 4.8-fold and 5.4-fold higher than that in the wild strain, suggesting another possible reason for the increased DHA productivity of the mutant strain.

• Microbiology and Biotechnology Letters
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• v.28 no.3
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• pp.134-138
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• 2000

Extensive screening for cellulose-producing bacteria was done using differential media. Fifty seven strains were isolated totally from the fruits and the vinegar, respectively; the isolate A9 strain from apples was selected and examined to determine its taxonomical characteristics. The bacterium was identified as the genus Acetobacter sp_ based on morphological, cultural and biochemical properties. A9 strain produced acetic acid from ethanol and decomposed acetic acid to $CO_2$ and $H_2O$. They produced dihydroxyacetone from glycerol but did not produce y-pyrone from glucose and fructose. When A9 strain was cultivated statically in Hestrin and Schramm liquid medium(HS medium). thick cellulose pellicle was formed_ Higher cellulose production was obtained in the shaken culture using HS medium at 100 rpm.

• Journal of Microbiology and Biotechnology
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• v.30 no.2
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• pp.271-278
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• 2020

Glycerol dehydrogenase (GlyDH) catalyzes the oxidation of glycerol to dihydroxyacetone (DHA), which is the first step in the glycerol metabolism pathway. GlyDH has attracted great interest for its potential industrial applications, since DHA is a precursor for the synthesis of many commercially valuable chemicals and various drugs. In this study, GlyDH from Klebsiella pneumoniae (KpGlyDH) was overexpressed in E. coli and purified to homogeneity for biochemical and molecular characterization. KpGlyDH exhibits an exclusive preference for NAD⁺ over NADP⁺. The enzymatic activity of KpGlyDH is maximal at pH 8.6 and pH 10.0. Of the three common polyol substrates, KpGlyDH showed the highest k_cat/K_m value for glycerol, which is three times higher than for racemic 2,3-butanediol and 32 times higher than for ethylene glycol. The k_cat value for glycerol oxidation is notably high at 87.1 ± 11.3 sec^-1. KpGlyDH was shown to exist in an equilibrium between two different oligomeric states, octamer and hexadecamer, by size-exclusion chromatography analysis. KpGlyDH is structurally thermostable, with a T_m of 83.4℃, in thermal denaturation experiment using circular dichroism spectroscopy. The biochemical and biophysical characteristics of KpGlyDH revealed in this study should provide the basis for future research on its glycerol metabolism and possible use in industrial applications.

• Journal of the Korean Society of Food Science and Nutrition
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• v.32 no.5
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• pp.684-688
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• 2003

The objective of this study was to search for the best strain as a source of L- $\alpha$-glycerophosphate oxidase (GPO) production and to establish the process technology for the purification of GPO on an industrial scale. The GPO was produced by culturing Streptococcus faecium, and purified by ammonium sulfate, DEAE-cellulose and hydroxyapatite chromatography. The relative activity was 60 units/L for 5. faecim ATCC 12755, 65 units/L for 5. faecium ATCC 19634, and 67 units/L for 5. faecium $M_{74}$.LC, respectively. The optimum condition for fermentation was $37^{\circ}C$ for temperature, 300 rpm for stir rate, 0.5 L/min for aeration rate and 17 hours. The main culture medium prepared by the modified AC medium. AC medium consists of 0.1% glucose, 0.2% glycerol, 1.0% tryptone and 1.0% yeast extract, 0.5% $K_2HP0_4$, pH 7.0. The GPO was purified by ammonium sulfate fractionation and ion exchange column chromatography, The yield and purity were 17.2% and 5.3 fold, respectively.