Hyun Joong Kim

Optimization of Direct Lysine Decarboxylase Biotransformation for Cadaverine Production with Whole-Cell Biocatalysts at High Lysine Concentration.

Cadaverine (1,5-diaminopentane) is an important industrial chemical with a wide range of applications. Although there have been many efforts to produce cadaverine through fermentation, there are not many reports of the direct cadaverine production from lysine using biotransformation. Whole-cell reactions were examined using a recombinant Escherichia coli strain overexpressing the E. coli MG1655 cadA gene, and various parameters were investigated for the whole-cell bioconversion of lysine to cadaverine. A high concentration of lysine resulted in the synthesis of pyridoxal-5-phosphate (PLP) and it was found to be a critical control factor for the biotransformation of lysine to cadaverine. When 0.025 mM PLP and 1.75 M lysine in 500 mM sodium acetate buffer (pH6) were used, consumption of 91% lysine and conversion of about 80% lysine to cadaverine were successfully achieved.

Takotsubo Cardiomyopathy Following Cerebral Infarction Involving the Insular Cortex

Background Takotsubo cardiomyopathy is characterized by clinical features similar to those of acute myocardial ischemia, but without angiographic evidence of obstructive coronary artery disease. We present a patient with takotsubo cardiomyopathy following acute infarction involving the left insular cortex. Case Report A 52-year-old man was admitted with acute infarction of the left middle cerebral artery territory and acute chest pain. Acute myocardial infarction was suspected because of elevated serum troponin levels and hypokinesia of the left ventricle on echocardiography. However, a subsequent coronary angiography revealed no stenosis within the coronary arteries or ballooning of the apical left ventricle. Conclusions We postulated that catecholamine imbalance due to the insular lesion could be responsible for these interesting features.

Biotransformation of lysine into cadaverine using barium alginate-immobilized Escherichia coli overexpressing CadA

In this study, Escherichia coli cells overexpressing lysine decarboxylase (CadA) were used for cadaverine production. Barium alginate was selected as a matrix for immobilization of E. coli YH91. Free cells and immobilized cells (IC) were characterized for their physiochemical properties, and the optimum pH and temperature were determined as 6 and 37xa0°C, respectively. Immobilized cells were three times more thermally stable compared to free cells at the optimum temperature and had a half-life (t1/2) of 131xa0h. The free cells lost most of lysine decarboxylase activity after nine cycles, but in contrast immobilized cells retained 56xa0% of their residual activity even after the 18th cycle. The immobilized cells gave a maximum production of cadaverine (75.8xa0g/L) with 84xa0% conversion.

Microbial biodiesel production from oil palm biomass hydrolysate using marine Rhodococcus sp. YHY01

The effect of various biomass derived inhibitors (i.e. furfural, hydroxymethylfurfural (HMF), vanillin, 4-hydroxy benzaldehyde (4-HB) and acetate) was investigated for fatty acid accumulation in Rhodococcus sp. YHY 01. Rhodococcus sp. YHY01 was able to utilize acetate, vanillin, and 4-HB for biomass production and fatty acid accumulation. The IC50 value for furfural (3.1mM), HMF (3.2mM), vanillin (2.0mM), 4-HB (2.7mM) and acetate (3.7mM) was calculated. HMF and vanillin affect fatty acid composition and increase saturated fatty acid content. Rhodococcus sp. YHY 01 cultured with empty fruit bunch hydrolysate (EFBH) as the main carbon source resulted in enhanced biomass (20%) and fatty acid productivity (37%), in compression to glucose as a carbon source. Overall, this study showed the beneficial effects of inhibitory molecules on growth and fatty acid production, and support the idea of biomass hydrolysate utilization for biodiesel production by avoiding complex efforts to remove inhibitory compounds.

A Fractured Sirolimus-Eluting Stent With a Coronary Aneurysm

A 55-year-old man had undergone successful percutaneous intervention with a sirolimus-eluting stent, placed in the right coronary artery (2.5 x 33 mm) and distal left circumflex artery (3.0 x 28 mm) without high pressure ballooning. Twelve months later he presented with unstable angina. Angiography revealed two fracture sites on the right coronary artery-deployed stent, with a large aneurysm and an aneurysmal dilatation of the left circumflex artery without stent fracture. Due to the potential risk of aneurysmal rupture, he underwent coronary artery bypass grafting and ligation of the aneurysm.

Identification and Functional Characterization of an α-Amylase with Broad Temperature and pH Stability from Paenibacillus sp.

Amylases are important industrial enzymes that have been applied widely in the food, detergent, and pulp industries and fermentation processes. In the present study, a gene encoding an alpha-amylase from the genomic DNA library of Paenibacillus sp. was identified and characterized. The amylase gene designated amy1 was shown to consist of 1,980xa0bp and shared sequence identity towards α-amylase genes from other Bacillus sp. The deduced amino acid sequence for Amy1 indicated 80xa0% sequence identity with other Bacillus strains. Heterologous expression of recombinant Amy1 in Escherichia coli BL21(DE3) facilitated the recovery of this protein in soluble form. Enzyme kinetic data revealed Amy1 to have a Km of 23.83xa0mg/mL and Kcat of 48.74xa0min−1 and Kcat/Km of 2xa0min−1xa0mg−1xa0mL−1 for starch. The activity of this protein was found to be enhanced by Mn2+, and furthermore, Amy1 remained active at a broad pH range (4–10) and temperature (30–90xa0°C). The ability of Amy1 to act on food waste under broad temperature and pH conditions, together with its ability to produce simple sugars, shows many advantages for further application in the food industry.

Metal removal and reduction potential of an exopolysaccharide produced by Arctic psychrotrophic bacterium Pseudomonas sp. PAMC 28620

An exopolysaccharide (EPS) was produced from psychrotrophic Arctic glacier fore-field soil bacterium Pseudomonas sp. PAMC 28620 using glycerol enriched medium and the maximum productivity 7.24 ± 0.31 g L−1 of EPS was obtained after 168 h of fermentation. The EPS was purified and analysed by HPLC, GC-MS, FT-IR, 1H and 13C NMR. The EPS obtained from Arctic strain PAMC 28620 exhibits a distinctive structural composition and the constituent sugar monomers are rhamnose, galactose, glucose, fucose, mannose and ribose. The purified EPS has shown excellent flocculating and emulsification capacities with promising biotechnological and ecological implications. From the metal removal experiments, the EPS exhibited remarkable metal adsorption (99%) potential adopting the order Fe2+ > Cu2+ > Mg2+ > Zn2+ > Mn2+ > Ca2+. FE-SEM combined with EDX analysis has shown that the metal ions were complexed or immobilized onto the EPS matrix and further reduced to nanoparticles (150–950 nm). This study is significant in terms of metal removal and reduction potential of Arctic bacterial EPS and the possible ecological roles of the EPS in Arctic environment.

The serum protein fetuin-B is involved in the development of acute myocardial infarction.

The rupture of an atherosclerotic plaque is one of the main causes of coronary artery thrombotic occlusion, leading to myocardial infarction. However, the exact mechanism and causal risk factors for plaque rupture remain unclear. To identify a potential molecule that can influence atherosclerotic plaque rupture, we investigated protein expression in serum from patients with acute myocardial infarction (AMI) and stable angina (SA), using proteomic analysis. The expression of six proteins, including fibrinogen, fetuin-B, keratin 9, proapolipoprotein and fibrinogen, were altered in serum from patients with AMI compared with serum from those with SA. Of these, fetuin-B, proapolipoprotein, fibrinogen γ-B-chain precursors and fibrinogen expression were greater in serum from patients with AMI than from patients with SA. Increased fetuin-B expression in serum from AMI patients was also confirmed by Western blot analysis. Treatment with recombinant human fetuin-B increased the migration in monocytes and macrophages in a concentration-dependent manner. Fetuin-B also affected vascular plaque-stabilizing factors, including lipid deposition and cytokine production in macrophages, the activation of matrix metalloproteinase (MMP)-2 in monocytes, and the activation of apoptosis and MMP-2 in vascular smooth muscle cells. Moreover, in vivo administration of fetuin-B decreased the collagen accumulation and smooth muscle cell content and showed an increased number of macrophages in the vascular plaque. From these results, we suggest that fetuin-B may act as a modulator in the development of AMI. This study may provide a therapeutic advantage for patients at high risk of AMI.

A Liquid-Based Colorimetric Assay of Lysine Decarboxylase and Its Application to Enzymatic Assay.

A liquid-based colorimetric assay using a pH indicator was introduced for high-throughput monitoring of lysine decarboxylase activity. The assay is based on the color change of bromocresol purple, measured at 595 nm in liquid reaction mixture, due to an increase of pH by the production of cadaverine. Bromocresol purple was selected as the indicator because it has higher sensitivity than bromothymol blue and pheonol red within a broad range and shows good linearity within the applied pH. We applied this for simple determination of lysine decarboxylase reusability using 96-well plates, and optimization of conditions for enzyme overexpression with different concentrations of IPTG on lysine decarboxylase. This assay is expected to be applied for monitoring and quantifying the liquid-based enzyme reaction in biotransformation of decarboxylase in a high-throughput way.

Phosphorylation of chloramphenicol by a recombinant protein Yhr2 from Streptomyces avermitilis MA4680.

Although phosphorylation of chloramphenicol has been shown to occur in the chloramphenicol producer, Streptomyces venezuelae, there are no reports on the existence of chloramphenicol phosphorylase in other Streptomyces species. In the present study, we report the modification of chloramphenicol by a recombinant protein, designated as Yhr2 (encoded by SAV_877), from Streptomyces avermitilis MA4680. Recombinant Yhr2 was expressed in Escherichia coli BL21 (DE3) and the cells expressing this recombinant protein were shown to phosphorylate chloramphenicol to a 3-O-phosphoryl ester derivative, resulting in an inactivated form of the antibiotic. Expression of yhr2 conferred chloramphenicol resistance to E. coli cells up to 25 μg/mL and in an in vitro reaction, adenosine triphosphate (ATP), guanosine triphosphate (GTP), adenosine diphosphate (ADP) and guanosine diphosphate (GDP) were shown to be the phosphate donors for phosphorylation of chloramphenicol. This study highlights that antibiotic resistance conferring genes could be easily expressed and functionalized in other organisms that do not produce the respective antibiotic.