• Microbiology and Biotechnology Letters
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• v.32 no.3
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• pp.286-289
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• 2004

Transglycosylation from sucrose to phenolic compounds by the recombinant sucrose phosphorylase from Bifidobacterium longum was studied. HPLC analysis revealed that the enzyme transferred glucosyl residue of sucrose to 1,2-dihydroxybenzene, 1,4-dihydroxybenzene, 1,2,3-trihydroxybenzene, and 2-hydroxybenzyl alcohol. The enzyme could transfer the glucosyl moiety of sucrose to phenolic compounds which have phenolic OH or alcoholic (hydroxymethyl) OH group.

• Microbiology and Biotechnology Letters
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• v.21 no.6
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• pp.570-576
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• 1993

For the production of glucose-1-phosphate from sucrose, bacteria having sucrose phosphorylase were isolated from Kimchi. Among them, JS-05, newly isolated strain having high activity of sucrose phosphorylase was selected and identified as Leuconostoc sp. The specific activity of sucrose phosphorylase of Leuconostoc sp. JS-05 was the highest when the strain was cultured at 25C for 20 hrs in the medium (pH 7.5) containing 10 g sucrose, 5g corn steep liquor, and 2.5g yeast extract per liter.

• Microbiology and Biotechnology Letters
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• v.32 no.3
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• pp.199-205
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• 2004

The nucleotide sequence of 8.6-kb EcoRI fragment containing sucrose phosphorylase gene isolated from Bifidobacterium longum SJ32 was determined. It was found that the fragment contained five open reading frames including the gene cluster for sucrose utilization such as a sucrose phosphorylase (ScrP), a sucrose transporter (ScrT), and a GalR-LacI-type transcriptional regulator (ScrR) identified by amino acid homology. Each gene showed over 94% amino acid homology among various B. longum strains. Genomic organization of the gene cluster is the same as those of other strains of B. longum but different from that of B. lactis. In spite of high homology of each gene among B. longum strains, the difference of flanking sequences of the gene cluster between strains SJ32 and NCC2705 insinuates the horizontal transfer of scrPTR between B. longum strains. The increase of sucrose phosphorylase activity in heterologous E. coli system by the co-expression of scrT with scrP against the single expression of scrP was measured. It seems to be the result of sucrose uptake increment by scrT in the host and is an indirect evidence that scrT is the gene for sucrose transport. The existence of multiple sucrose uptake systems in B. longum is supposed from the findings of several genes besides scrPTR involved in sucrose uptake in the genome of B. longum NCC2705.

• Korean Journal of Microbiology
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• v.54 no.4
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• pp.445-447
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• 2018

Leuconostoc lactis CCK940, which was isolated from kimchi obtained from a Korean traditional market, produced an oligosaccharide with a degree of polymerization of more than 4. In this study, the draft genome sequence of L. lactis CCK940 was reported by using PacBio 20 kb platform. The genome of this strain was sequenced and the genome assembly revealed 2 contigs. The genome was 1,741,511 base pairs in size with a G + C content of 43.33%, containing 1,698 coding sequences, 12 rRNA genes, and 68 tRNA genes. L. lactis CCK940 contained genes encoding glycosyltransferase, sucrose phosphorylase, maltose phosphorylase, and ${\beta}$-galactosidase which could synthesize oligosaccharide.

• Korean Journal of Pharmacognosy
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• v.17 no.2
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• pp.134-138
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• 1986

Based on a tracer study, starch phosphorylase was implicated as an agent in the starch synthesis in sweet potato roots. The enzyme was purified from the tissue as a cluster of isozymes with an average mw of 205K (fresh roots) or 159K (roots stored for 3 mon.). On SDS polyacrylamide gel electrophoresis, one large subunit of 98K mw and several small ones of 47${\sim}57K mw were observed. From the mw data and the results of peptide mapping and immunoelectrophoretic blotting using mono- and polyclonal antibodies, it was deduced that a large part of the large subunit was cleaved at the middle part of the peptide chain to give rise to the small subunits, and on storage, the enzyme molecules were further modified by proteolysis. During the course of phosphorylase purification, a proteinaceous inhibitor of the enzyme was isolated. It had a mw of 250K and was composed of 5 identical subunits of 51K mw. In the direction of starch synthesis, the inhibitor showed a noncompetitive kinetics with a Ki of $1.3{\times}10^{-6}\;M$. By immunohistochemical methods, both the enzyme and the inhibitor were located on the cell wall and amyloplast. Crossreacting materials of the inhibitor were present in spinach leaf, potato tuber and rice grain. These findings indicate the wide occurrence of the inhibitor and also imply its possible participation in regulating starch phosphorylase activity in vivo.

• KSBB Journal
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• v.19 no.5
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• pp.363-367
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• 2004

Leuconostoc mesenteroides NRRL B-1149 produces various glucoseyltransferases for the synthesis of dextran, levan and glucose-1-phosphate using sucrose as a substrate. A sucrose phosphorylase (1149SPase) was purified from L. mesenteroides NRRL B-1149 culture by using hollow fiber filtration (30 kDa cut off), Toyopearl DEAE 650 M column chromatography and following two times of DEAE-Sepharose column chromatographies. The specific activity of the purified 1149SPase was 25.7 (U/mg) with $16\%$ yield. The 1149SPase showed a molecular size of 56 kDa on denatured $10\%$ SDS-PAGE. The N-terminal amino acid sequence of the enzyme was MEIQNKAM. The optimum pH and temperature of this enzyme were 6.2~6.5 and 37^{circ}C, respectively. It had an apparent K_{m} of 6.0 mM and K_{cat} of 1.62/s for sucrose. 1149SPase crystal was formed by hanging drop diffusion technique using 20 mM calcium chloride dihydrate, 100 mM sodium acetate trihydrate pH 4.6 and $30\%$ 2-methyl-2,4-pentanediol as vaporizing and reservation solution. The 1149SPase catalyzes transferring of glucose from isomaltose or sucrose to salicin and salicyl alcohol by disproportionation reaction or acceptor reaction and synthesized two acceptor products, respectively.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2018.04a
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• pp.5-5
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• 2018

Ginseng (Panax ginseng C.A. Meyer), one of the most widely used medicinal plants in traditional oriental medicine, is used for the treatment of various diseases. It has been classified according to its cultivation environment, such as field cultivated ginseng (FCG) and mountain cultivated ginseng (MCG). However, little is known about differences in gene expression in ginseng roots between field cultivated and mountain cultivated ginseng. In order to investigate the whole transcriptome landscape of ginseng, we employed High-Throughput sequencing technologies using the Illumina HiSeqTM2500 system, and generated a large amount of sequenced transcriptome from ginseng roots. Approximately 77 million and 87 million high-quality reads were produced in the FCG and MCG roots transcriptome analyses, respectively, and we obtained 256,032 assembled unigenes with an average length of 1,171 bp by de novo assembly methods. Functional annotations of the unigenes were performed using sequence similarity comparisons against the following databases: the non-redundant nucleotide database, the InterPro domains database, the Gene Ontology Consortium database, and the Kyoto Encyclopedia of Genes and Genomes pathway database. A total of 4,207 unigenes were assigned to specific metabolic pathways, and all of the known enzymes involved in starch and sucrose metabolism pathways were also identified in the KEGG library. This study indicated that alpha-glucan phosphorylase 1, putative pectinesterase/pectinesterase inhibitor 17, beta-amylase, and alpha-glucan phosphorylase isozyme H might be important factors involved in starch and sucrose metabolism between FCG and MCG in different environments.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2001.06a
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• pp.179-182
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• 2001

Two intracellular enzymes, cellobiose phosphorylase (CBP) and cellodextrin phosphorylase (CDP) are involved in the phosphorolytic pathway in cellulose degradation. Those enzymes are considered to be useful in syntheses of oligosaccharides because the reactions are reversible. CBP from Cellvibrio gilvus and CDP from Clostridium thermocellum YM-4 were cloned and over-expressed in Escharichia coli. Both the enzyme reactions showed ordered bi bi mechanism. Acceptor specificity of CBP in the reverse reaction was determined. Several $\beta$-l,4-glucosyl disaccharides were synthesized by using the reaction. A new substrate inhibition pattern, competitive substrate inhibition, was also found in the reverse reaction of CBP Cellobiose was produced from sucrose at a high yield by a combined action of three enzymes including CBP

• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.407-407
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• 2005

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2002.10a
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• pp.71-71
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• 2002