Young Tae Kim

Keunho Ji, Yeeum Kim and Young Tae Kim. Microbiol. Biotechnol. Lett. 2021;49:167-73. https://doi.org/10.48022/mbl.2012.12001

Overproduction and accumulation of melanin cause a number of skin diseases. The inhibitors of tyrosinase are important for the treatment of skin diseases associated with hyper-pigmentation after UV exposure and application in cosmetics for whitening and depigmentation. Reactive oxygen species (ROS) including hydrogen peroxide are generated by chemical substances and metabolic intermediates and cause various diseases including cancer and heart diseases. We have isolated four different lactic acid bacteria (LAB) strains from dairy cow feces and investigated the tyrosinase inhibition and anti-oxidative effects of culture filtrates prepared from the isolated bacteria, which are designated as EA3, EB2, PC2, and PD3. To investigate optimal culture conditions isolated LAB strains, the measurements of tyrosinase inhibitory and anti-oxidative activities were performed. The results of tyrosinase inhibitory activities revealed that Enterococcus sp. EA3 showed about 65% at culture conditions (14 h, 30 °C, pH 8, and 0% NaCl), Enterococcus sp. EB2 about 65% at culture conditions (12 h, 30 °C, pH 9, and 0% NaCl), Pediococcus sp. PC2 about 80% at culture conditions (20 h, 30 °C, pH 6, and 0% NaCl), and Pediococcus sp. PD3 about 80% at culture conditions (20 h, 30 °C, pH 8, and 0% NaCl), respectively. In addition, anti-oxidative activities against four different LAB strains showed approximately more than 30% at optimal conditions for the measurements of tyrosinase inhibitory activities. From the results, we have suggested that the isolated four LAB strains could be useful for a potential agent for developing anti-oxidants and tyrosinase inhibitors.

The astaxanthin biosynthesis gene cluster in Paracoccus haeundaensis consists of six genes: crtW, crtZ, crtY, crtI, crtB, and crtE contain 726, 486, 1158, 1503, 912, and 879 base pairs, respectively. Individual carotenoid biosynthesis genes of P. haeundaensis have now been expressed in E. coli and each gene product has been purified to homogeneity. Their molecular characteristics, including enzymatic activities, are reported here.

Acetohydroxy acid synthase (AHAS, EC 2.2.1.6; also known as acetolactate synthase, ALS) catalyzes the first common step in the biosynthesis of valine, leucine, and isoleucine in plants and microorganisms. AHAS is the target of several classes of herbicides. In the present study, the role of three well-conserved arginine residues (R141, R372, and R376) in tobacco AHAS was determined by site-directed mutagenesis. The mutated enzymes, referred to as R141A, R141F, and R376F, were inactive and unable to bind to the cofactor, FAD. The inactive mutants had the same secondary structure as that of the wild type. The mutants R141K, R372F, and R376K exhibited much lower specific activities than the wild type, and moderate resistance to herbicides such as Londax, Cadre, and/or TP. The mutation R141K showed a strong reduction in activation efficiency by ThDP, while the mutations R372K and R376K showed a strong reductions in activation efficiency by FAD in comparison to the wild type enzyme. Taking into account the data presented here and the homology model constructed previously [Le et al. (2004) Biochem. Biophys. Res. Commun. 317, 930–938], it is suggested that the three amino acid residues studied (R141, R372, and R376) are located essentially at the enzyme active site, and, furthermore, that residues R372 and R376 are possibly responsible for the binding of the enzyme to FAD.

Ornithine decarboxylase (ODC) is a homodimeric enzyme dependent on pyridoxal 5’-phosphate. We identified a complementary DNA clone corresponding to ODC from the brain of adult flounder (Paralichthys olivaceus). The flounder ODC cDNA consisted of 2939 bp encoding 272 amino acid residues. The flounder ODC showed 80.3% sequence identity to zebrafish and 70.8% to rat at the amino acid level. Comparison of the structure and nucleotide sequence of the ODC genes revealed that the gene is highly conserved in the flounder, zebrafish, and rat. The presence of ODC mRNA species in brain, kidney, liver, and embryo was confirmed using the reverse transcriptase polymerase chain reaction. The recombinant protein of flounder ODC containing a short histidine tag at the carboxyl terminus was overexpressed in Escherichia coli BL21 (DE3) codon plus using an inducible T7 expression system, and was purified by Ni-NTA affinity chromatography.