• Journal of Food Hygiene and Safety
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• v.35 no.3
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• pp.252-260
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• 2020

This study was conducted to improve the standard method for vitamin B₁₂ and biotin contained in foods for special dietary uses to ensure the specificity of the complex matrix properties of foods. For the food code, the test method was improved to determine vitamin B₁₂ and biotin by high-performance liquid chromatography (HPLC)-UV using column-switching after concentration using immunoaffinity column. The immunoaffinity columns contain a gel suspension of monoclonal antibody specific to the vitamin of interest so that it can be used to concentrate the vitamin B₁₂ and biotin and remove interferences from the food extracts. Moreover, validation of advanced new methods was carried out to support the suitability of the proposed analytical procedure (specificity, linearity, detection limits (LOD), quantitative limits (LOQ), accuracy, and precision). The improved analytical method is being used to monitor relevant food items on sale. The results of this study showed that the new analytical method is suitable and appropriate for managing food intended for special dietary uses.

• Journal of the Korean Magnetic Resonance Society
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• v.12 no.1
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• pp.1-13
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• 2008

Acetyl-CoA carboxylase (ACC) catalyzes the first step in fatty acid biosynthesis: the synthesis of malonyl-CoA from acetyl-CoA. As essential regulators of fatty acid biosynthesis and metabolism, ACCs are regarded as therapeutic targets for the treatment of metabolic diseases such as obesity, In ACC, the biotinoyl domain performs a critical function by transferring an activated carboxyl group from the biotin carboxylase domain to the carboxyl transferase domain, followed by carboxyl transfer to malonyl-CoA. Despite the intensive research on this enzyme, only the bacterial and yeast ACC structures are currently available, To explore the mechanism of ACC holoenzyme function, we determined the structure of the biotinoyl domain of human ACC2 and analyze its characteristics using NMR spectroscopy. The 3D structure of the hACC2 biotinoyl domain has a similar folding topology to the previously determined domains from E. coli and P. Shermanii, however, the 'thumb' structure is absent in the hACC2 biotinoyl domain. Observations of the NMR signals upon the biotinylation indicate that the biotin group of hACC2 does not affect the structure of the biotinoyl domain, while the biotin group for E. coli ACC interacts directly with the thumb residues that are not present in the hACC2 structure. These results imply that, in the E. coli ACC reaction, the biotin moiety carrying the carboxyl group from BC to CT can pause at the thumb of the BCCP domain. The human biotinoyl domain, however, lacks the thumb structure and does not have additional non-covalent interactions with the biotin moiety; thus, the flexible motion of the biotinylated lysine residue must underlie the "swinging arm" motion. This study provides insight into the mechanism of ACC holoenzyme function and supports the "swinging arm" model in human ACCs.

• Journal of Life Science
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• v.28 no.12
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• pp.1416-1423
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• 2018

With strong biotin binding affinity ($K_D=10^{-14}M$), the tetrameric feature of streptavidin could be used to increase the antigen binding activity of a camel heavy chain (VHH) antibody through their fusion, here stained with biotinylated horseradish peroxidase and subsequent immunoassays ELISA and Western blot analysis. For this application, we cloned the streptavidin gene amplified from the Streptomyces avidinii chromosome by PCR, and this was fused to the gene of the 8B9 VHH antibody which is specific to green fluorescent protein (GFP) antigens. To express a soluble fusion protein in Escherichia coli, we used the pUC119 plasmid-based expression system which uses the lacZ promoter for induction by IPTG, the pelB leader sequence at the N-terminus for secretion into the periplasmic space, and six polyhistidine tags at the C-terminus for purification of the expressed proteins using an $Ni^+$-NTA-agarose column. Although streptavidin is toxic to E. coli because of its strong biotin binding property, this soluble fusion protein was expressed successfully. In SDS-PAGE, the size of the purified fusion protein was 122.4 kDa in its native condition and 30.6 kDa once denatured by boiling, suggesting the tetramerization of the monomeric subunit by non-covalent association through the streptavidin moiety fusing to the 8B9 VHH antibody. In addition, this fusion protein showed biotin binding activity similar to streptavidin as well as GFP antigen binding activity through both ELISA and Western blot analysis. In conclusion, the protein resulting from the fusion of an 8B9 VHH antibody with streptavidin was successfully expressed and purified as a soluble tetramer in E. coli; it showed both biotin and GFP antigen binding activity suggesting the possible production of a tetrameric and bifunctional VHH antibody.

• Microbiology and Biotechnology Letters
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• v.1 no.2
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• pp.89-92
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• 1973

In the previous paper the biological characteristics and some reaction mechanisms of Astradix-P, yeaststatic substance, were reported. Especially yeaststatic activity of Astradix-P was anta-gonistically inhibited by alkaline amino acids, arginine, lysine and histidine, which were added as a nitrogen source in the yeast growing medium. In this paper the effects of alkaline nitrogen containing substance and several vitamins on the yeaststatic activity were investigated. The antagonistic action of alkaline nitrogen containing substance; adenine and vitamins; thiamine, riboflavin, pyridoxin, cobalamin, nicotinic acid, folic acid, biotin, p-aminobenzoic acid, inositol, and pantothenic acid to Astradix-P were not observed, thus evidencing that the yeaststatic activity of Astradix-P was not inhibited by a alkaline nitrogen containing substance and several vitamins.

• The Korean Journal of Food And Nutrition
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• v.8 no.4
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• pp.275-279
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• 1995

A bacterium L-10 which produce mush of glutamic acid was Isolated from soil and identified as the genus Pserdomonas. The maximal glutamic acid production was obtained when the strain was cultured at 3$0^{\circ}C$ for 30 hrs in the optimal medium containing 5% glucose, 0.5% each of urea and yeast extract, 0.1% K2HP04, 0.02% MgSO4.7H20, 0.3% (NH, )rHP04, 0.5ug/l biotin and Initial pH 7.0, and then final glutamic acid production under the above conditions was 1.2mg/ml of cell cultures.

• Korean Journal of Veterinary Research
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• v.33 no.2
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• pp.255-261
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• 1993

For a immunohistochemical diagnosis of the frozen and paraffin-embedded tissues against rabies virus, mice were intracerebrally inoculated with challege virus standard(CVS) rabies virus and then were used to detect the rabies viral antigen by the immunoperoxidase(IP) and the avidin-biotin complex(ABC) method. In this study, the results confirmed that ABC and IP methods, although the former showed more specific and sensitive than the latter, were reliable and effective for the demonstration of rabies virus in both frozen and paraffin-embedded brain tissues prepared from rabies-infected mice. Additionally, IP technique using the monoclonal antibody against rabies virus could be recommended as a standard diagnostic tool instead of the present immunofluorescent method for the local veterinary services in Korea.

• Korean Journal of Veterinary Research
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• v.33 no.1
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• pp.119-123
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• 1993

The present study was intended to use the avidin-biotin-peroxidase-antiperoxidase complex (ABPAP) method for the identification of Mycobacterium bovis in the tissue sections of infected cattle. Antibodies and linksera for ABPAP procedure used in incubated order were rabbit anti-Mycobacterium polyvalent antibodies, goat anti-rabbit IgG, rabbit peroxidase-antiperoxidase complex, biotinyl-horse anti-rabbit IgG, and avidin-biotin-peroxidase complex. Where the bacterial antigen was localized by ABPAP, a dark brown deposit occurred in the cytoplasms of macrophages and Langerhans' giant cells of the granulomatous lesions. The method approved to be highly specific for the identification of the bacteria and allowed a precise localization of the bacterial antigen in infected cells.

• Korean Journal of Microbiology
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• v.25 no.2
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• pp.129-136
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• 1987

Nitrogen fixing activity associated with 40 varieties of rice was assayed at heading stage by an in situ acetylene reduction method. The in situ acetylene reduction activity and population of nitrogen fixing bacteria obtained on nitrogen-free malate medium for Azospirillum spp. enrichment showed positive correlation. Six Azospirillum spp. with high nitrogenase activity were isolated from the rice roots, from which five spp. were identified as A. lipoferum and one was A. brasilense. The physiological characteristics of the six Azospirillum isolates, that is, carbon source utilization, biotin requirement, antibiotic resistance, indole acetic acid excretion, plasmid profile and protein patterns were compared.

• Bulletin of the Korean Chemical Society
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• v.34 no.2
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• pp.565-568
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• 2013

Acetyl-CoA carboxylases (ACCs) play critical roles in fatty acid synthesis and oxidation by the catalytic activity of the carboxylation of acetyl-CoA to malonyl-CoA. It is known that ACCs are inactivated through reversible phosphorylation by AMP-activated protein kinase (AMPK) and allosterically activated by citrate. Here, we determined the crystal structures of biotin carboxylase (BC) domain of human ACC2 phosphorylated by AMPK in the presence of citrate in order to elucidate the activation mechanism by citrate. This structure shows that phosphorylated Ser222 is released from the dimer interface, and thereby facilitating the dimerization or oligomerization of the BC domain allosterically. This structural explanation is coincident with the experimental result that the phosphorylated Ser222 was dephosphorylated more easily by protein phosphatase 2A (PP2A) as the citrate concentration increases.

• Journal of Microbiology
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• v.33 no.4
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• pp.322-327
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• 1995

Tolerance of Mycobacterium sp. against organic solvents has been induced for the cholesterol side chain degradation by adding chemicals associated with synthesis of fatty acids or alcohols. Biotin of 300 .mu.g/1 and 0.5% aqueous ethanol solution were optima for the enhancement of ethanol tolerance of the microorganism. The induction of ethanol tolerance by biotin was found to be due to increase of degree of unsaturation of the fatty acids in membranous phospholipid of the cell, especially due to increase of oleic acid content. However when 0.5% of ethanol was added for the ethanol tolerance induction, there was an ambiguous correlation between ethanol tolerance and degree of unsaturation of the fatty acids, in spite of the fact that the induction increased the content of unsaturated fatty acids. Addition of 0.5% of ethanol induced several ethanol shock proteins having molecular weight similar to that of heat shock proteins.