Biomedical subjects

Jung-Hoe Kim

Publications and source records attributed to Jung-Hoe Kim.

7 recordsLinked to original sources

PubMed2006

Over-production of beta-carotene from metabolically engineered Escherichia coli.

To produce recombinant beta-carotene in vitro, synthetic operons encoding genes governing its biosynthesis were engineered into Escherichia coli. Constructs harboring these operons were introduced into either a high-copy or low-copy cloning vector. beta-Carotene production from these recombinant E. coli cells was either constitutive or inducible depending upon plasmid copy number. The most efficient beta-carotene production was with the low-copy based vector. The process was increased incrementally from a 5 l to a 50 l fermentor and finally into a 300 l fermentor. The maximal beta-carotene yields achieved using the 50 l and 300 l fermentor were 390 mg l(-1) and 240 mg l(-1), respectively, with overall productivities of 7.8 mg l(-1) h(-1) and 4.8 mg l(-1) h(-1).

Drug Industry↗

PubMed2006

Sequential and simultaneous statistical optimization by dynamic design of experiment for peptide overexpression in recombinant Escherichia coli.

The production of recombinant anti-HIV peptide, T-20, in Escherichia coli was optimized by statistical experimental designs (successive designs with multifactors) such as 2(4-1) fractional factorial, 2(3) full factorial, and 2(2) rotational central composite design in order. The effects of media compositions (glucose, NPK sources, MgSO4, and trace elements), induction level, induction timing (optical density at induction process), and induction duration (culture time after induction) on T-20 production were studied by using a statistical response surface method. A series of iterative experimental designs was employed to determine optimal fermentation conditions (media and process factors). Optimal ranges characterized by %T-20 (proportion of peptide to the total cell protein) were observed, narrowed down, and further investigated to determine the optimal combination of culture conditions, which was as follows: 9, 6, 10, and 1 mL of glucose, NPK sources, MgSO4, and trace elements, respectively, in a total of 100 mL of medium inducted at an OD of 0.55-0.75 with 0.7 mM isopropyl-beta-D-thiogalactopyranoside in an induction duration of 4 h. Under these conditions, up to 14% of T-20 was obtained. This statistical optimization allowed the production of T-20 to be increased more than twofold (from 6 to 14%) within a shorter induction duration (from 6 to 4 h) at the shake-flask scale.

Anti-HIV Agents↗

PubMed2006

Statistical medium formulation and process modeling by mixture design of experiment for peptide overexpression in recombinant Escherichia coli.

The medium formulation and robust process modeling for anti-HIV peptide (T-20) production by recombinant Escherichia coli overexpression were studied by employing a crossed experimental design. The crossed design, a mixture design combined with process factor (induction duration), was used to find the optimal medium formulation and process time. The optimal settings for three major components (7.75 mL of NPK sources, 5.5 mL of glucose, and 11.75 mL of MgSO4) characterized by %T-20 (14.45%), the proportion of peptide to the total protein, were observed in a total of 100 mL of medium inducted at an optical density of 0.67 with 0.7 mM isopropyl-beta-D-thiogalactopyranoside) for a 3-h induction duration at shake-flask scale. These conditions were further investigated to find robust process conditions (8.2 mL of NPK sources, 5.6 mL of glucose, and 11.3 mL of MgSO4, and a 3.5-h induction duration time) for T-20 production (13.9%) by applying propagation of error.

Anti-HIV Agents↗

PubMed2005

Correlation between serum CEA level and metabolic volume as determined by FDG PET in postoperative patients with recurrent colorectal cancer.

To determine the correlation between serum CEA level and the metabolic volume by FDG PET in postoperative patients with recurrent colorectal cancer, FDG PET was performed in 29 consecutive patients with recurrent or metastatic colorectal cancer whose CEA levels were higher than 5 ng/ml. A whole body emission scan was performed 60 minutes after injecting 370-555 MBq of F-18 FDG. "PET volume" and "PET metabolic volume" of tumors were measured on FDG PET images. Based on an isocontour plot of tumor mass at 2.5 SUV (standardized uptake value), the metabolically active tumor "PET volume" was calculated. "PET metabolic volume" was obtained by multiplying the "PET volume" by the mean SUV of the tumor. All recurrent or metastatic lesions were single or multiple lesions of measurable size (axial diameter > 1 cm, minimum "PET volume" 3.5 cm3), and were verified by operation or by other imaging modalities (CT or MRI). There was a linear associations between "PET volume" and serum CEA level. Further regression analysis by least squares showed a highly significant model with an equation of volume = 41.2 + 0.471 x CEA (r2 = 0.629). However, no such association was found between "PET metabolic volume" and serum CEA level according to the residual normality test. In conclusion, "PET volume" measured by FDG PET and serum CEA level in colorectal cancer are significantly correlated. Tumor volume determined by FDG PET can be used as an effective marker of tumor burden in postoperative patients with colorectal carcinoma.

Biomarkers, Tumor↗

PubMed2003

Enhanced conversion of sucrose to isomaltulose by a mutant of Erwinia rhapontici.

Mutagenesis of Erwinia rhapontici was performed to enhance the production of isomaltulose from sucrose. A mutant strain, BN 68089, was obtained through a screening process involving automated and miniaturized cultivation in Bioscreen C. This high-throughput, miniaturized screening system was optimized to identify the mutant strain, which had a conversion yield (90%) and productivity (194 g l(-1) h(-1)). The BN 68089 mutant cells were immobilized in sodium alginate and when operated in a packed bed reactor gave a yield of 89% and a productivity of 144 g l(-1) h(-1) of at 30 degrees C, the optimal temperature. Immobilized BN 68089 cells exhibited 8% and 15% higher yield and productivity, respectively, than those of the wild-type strain.

Bioreactors↗

PubMed2003

Purification and characterization of a novel erythrose reductase from Candida magnoliae.

Erythritol biosynthesis is catalyzed by erythrose reductase, which converts erythrose to erythritol. Erythrose reductase, however, has never been characterized in terms of amino acid sequence and kinetics. In this study, NAD(P)H-dependent erythrose reductase was purified to homogeneity from Candida magnoliae KFCC 11023 by ion exchange, gel filtration, affinity chromatography, and preparative electrophoresis. The molecular weights of erythrose reductase determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration chromatography were 38,800 and 79,000, respectively, suggesting that the enzyme is homodimeric. Partial amino acid sequence analysis indicates that the enzyme is closely related to other yeast aldose reductases. C. magnoliae erythrose reductase catalyzes the reduction of various aldehydes. Among aldoses, erythrose was the preferred substrate (K(m) = 7.9 mM; k(cat)/K(m) = 0.73 mM(-1) s(-1)). This enzyme had a dual coenzyme specificity with greater catalytic efficiency with NADH (k(cat)/K(m) = 450 mM(-1) s(-1)) than with NADPH (k(cat)/K(m) = 5.5 mM(-1) s(-1)), unlike previously characterized aldose reductases, and is specific for transferring the 4-pro-R hydrogen of NADH, which is typical of members of the aldo/keto reductase superfamily. Initial velocity and product inhibition studies are consistent with the hypothesis that the reduction proceeds via a sequential ordered mechanism. The enzyme required sulfhydryl compounds for optimal activity and was strongly inhibited by Cu(2+) and quercetin, a strong aldose reductase inhibitor, but was not inhibited by aldehyde reductase inhibitors and did not catalyze the reduction of the substrates for carbonyl reductase. These data indicate that the C. magnoliae erythrose reductase is an NAD(P)H-dependent homodimeric aldose reductase with an unusual dual coenzyme specificity.

Aldehyde Reductase↗

PubMed2002

Cytotoxic activity of aminoderivatized cationic chitosan derivatives.

Chitosan derivatives were prepared by dialkylaminoalkylation and reductive amination followed by quaternization. In this study, the cytotoxic activity of the chitosan derivatives was investigated and a relationship between structure and activity is suggested. The cationic chitosan derivatives elicited dose-dependent inhibitory effects on the proliferation of tumor cell lines.

Antineoplastic Agents↗