• Title/Summary/Keyword: glucansucrase

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Oligosaccharide Production by Leuconostoc lactis CCK940 Which Has Glucansucrase Activity (Leuconostoc lactis CCK940의 Glucansucrase 활성에 의한 올리고당 생산 최적화)

  • Lee, Sulhee;Park, Young-Seo
    • Food Engineering Progress
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    • v.21 no.4
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    • pp.383-390
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    • 2017
  • Glucansucrase is an enzyme classified as a glycoside hydrolase (GH) 70 family, which catalyzes the synthesis of glucooligosaccharides with a low molecular weight using sucrose as a donor of D-glucopyranose and maltose as a carbohydrate acceptor. In this study, glucansucrase-producing lactic acid bacteria strain was isolated from the fermented foods collected in traditional markets, and the optimum conditions for the oligosaccharide production were investigated. The strain CCK940 isolated from Chinese cabbage kimchi was selected as an oligosaccharide-producing strain due to its high glucansucrase activity, with 918.2 mU/mL, and identified as Leuconostoc lactis. The optimum conditions for the production of oligosaccharides using Leu. lactis CCK940 were to adjust the initial pH to 6.0, add 5% (w/v) sucrose and 10% (w/v) maltose as a donor and acceptor molecules, respectively, and feed 5% (w/v) sucrose at 4 and 8 h of cultivation. When Leu. lactis CCK940 was cultured for 12 h at optimum conditions, at least four oligosaccharides with a polymerization degree of 2-4 were produced.

Modification of Acetobacter xylinum Bacterial Cellulose Using Dextransucrase and Alternansucrase

  • Kim, Do-Man;Kim, Young-Min;Park, Mi-Ran;Park, Don-Hee
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.704-708
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    • 1999
  • In addition to catalyzing the synthesis of glucan from sucrose as a primary reaction, glucansucrase also catalyzes the transfer of glucose from sucrose to other carbohydrates that are present or are added to the reaction digest. Using dextransucrase and altemansucrase, prepared from Leuconostoc mesenteroides B-742CBM and B-1355C, respectively, we modified the bacterial cellulose in Acetobacter xylinum ATCC10821 culture, and then produced a characteristic cellulose that is soluble and has a new structure. There were also some partially modified insoluble cellulose and oligosaccharides in the modification culture. After methylation and following acid hydrolysis of both the soluble and insoluble glucans, there were ($1{\rightarrow}4$) as well as ($1{\rightarrow}6$) and ($1{\rightarrow}3$) glycosidic linkages in the soluble glucan.

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Characterization of novel EGCG (Epigallocatechin gallate) analogues synthesized by glucansucrase from Leuconostoc mesenteroides B-1299CB

  • Moon, Young-Hwan;Lee, Jin-Ha;Nam, Seung-Hee;Kim, Do-Won;Oh, Deok-Kun;Kim, Do-Man
    • 한국생물공학회:학술대회논문집
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    • 2005.10a
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    • pp.502-506
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    • 2005
  • Three EGCG analogues were synthesized by the acceptor reaction of a glucansucrase from Leuconostoc mesenteroides B-1299CB with EGCG and sucrose. The transfer products was purified using Sephadex LG-20 column chromatography and high performance liquid chromatography (HPLC). EGCG-G1 and EGCG-G2 were novel compounds for the first time reported in this paper. EGCG glycosides showed similar or slower antioxidative effects according to their structures $(EGCG{\geq}EGCG-G1>EGCG-G1'>EGCG-G2)$. However, the water solubilities of the EGCG-G1, EGCG-G1' and EGCG-G2 were 52, 76 and 140 times higher than that of EGCG. Furthermore, they showed more browning resistance against UV irradiation than EGCG.

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Simultaneous Biocatalytic Synthesis of Panose During Lactate Fermentation in Kimchi

  • Han, Nam-Soo;Jung, Yoon-Seung;Eom, Hyun-Ju;Koh, Young-Ho;Robyt, John F.;Seo, Jin-Ho
    • Journal of Microbiology and Biotechnology
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    • v.12 no.1
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    • pp.46-52
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    • 2002
  • As a functional additive for intestinal microflora, panose ($6^2-{\alpha}$-D-glucopyranosylmaltose) was synthesized during kimchi fermentation using the glucose transferring reaction of glucansucrase from Leuconostoc mesenteroides. For the glucose transferring reaction, sucrose and maltose were added ($2\%$ each, w/v) to dongchimi-kimchi as the glucosyl donor and acceptor molecule, respectively. After five days of incubation at $10^{\circ}C$, referring to the initial phase for the production of lactic acid in kimchi, over $60\%$ (w/v) of the total sugars were converted into panose and other branched oligosaccharides. Thereafter, the kimchi was stored at $4^{\circ}C$ and the amount of panose remained at a constant level for three weeks, thereby indicating the stability of panose to microbial degradation during the period of kimchi consumption. The use of maltose as the acceptor molecule in the kimchi also facilitated a lower viscosity in the kimchi-juice by preventing the synthesis of a dextran-like polymer which caused an unfavorable taste. Accordingly, the application of this new method of simultaneous biocatalytic synthesis of oligosaccharides during lactate fermentation should facilitate the extensive development of new function-added lactate foods.

Functional, Genetic, and Bioinformatic Characterization of Dextransucrase (DSRBCB4) Gene in Leuconostoc mesenteroides B-1299CB4

  • Kang, Hee-Kyoung;Kim, Young-Min;Kim, Do-Man
    • Journal of Microbiology and Biotechnology
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    • v.18 no.6
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    • pp.1050-1058
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    • 2008
  • A gene encoding a dextransucrase (dsrBCB4) that synthesizes only ${\alpha}$-1,6-linked dextran was cloned from Leuconostoc mesenteroides B-1299CB4. The coding region consisted of an open reading frame (ORF) of 4,395 bp that coded a 1,465-amino-acids protein with a molecular mass of 163,581 Da. The expressed recombinant DSRBCB4 (rDSRBCB4) synthesized oligosaccharides in the presence of maltose or isomaltose as an acceptor, plus the products included ${\alpha}$-1,6-linked glucosyl residues in addition to the maltosyl or isomaltosyl residue. Alignments of the amino acid sequence of DSRBCB4 with glucansucrases from Streptococcus and Leuconostoc identified conserved amino acid residues in the catalytic core that are critical for enzyme activity. The mutants D530N, E568Q, and D641N displayed a 98- to 10,000-fold reduction of total enzyme activity.

Potential Probiotic Properties of Exopolysaccharide Producing Lactic Acid Bacteria Isolated from Fermented Soybean Product (장류유래 Exopolysaccharide 생성 유산균의 잠재적 Probiotic 특성)

  • Ahn, Yu-Jin;Choi, Hye-Sun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.5
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    • pp.749-755
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    • 2014
  • Exopolysaccharides (EPSs) have been widely used in the food industry as viscofying, stabilizing, and emulsifying agents as well as in the pharmaceutical industry for their immunomodulatory, anti-tumor, and anti-inflammatory effects. A total of 458 lactic acid bacteria (LAB) strains isolated from several kinds of soybean pastes were screened for the production of homo-EPS (HoPS). LAB isolates were primarily screened using thin layer chromatography (TLC) and further screened polymerase chain reaction (PCR) targeting genes involved in HoPS production. Six LAB isolates producing high amounts of HoPS were identified by TLC. Among these isolates, glucansucrase gene was amplified in two strains (JSA57, JSB22), whereas the fructansucrase gene was detected in three strains (JSA57, JSB22, JSB66). After isolating the strains, their morphological characteristics and 16S rDNA sequences were determined. Six species were identified as L. alimentarius HSB15, L. plantarum JSA22, L. pentosus JSA57, L. brevis JSB22, L. alimentarius JSB66, and L. parabrevis JSB89. To evaluate the potential probiotic properties of these LAB, their survival rates against a simulated intestinal environment were determined. After 2 hr of incubation in artificial gastric juice, survival rates of JSA57, JSB90, JSB22, and JSB66 were all greater than 50%. After 2 hr of incubation in bile juice, viable cell count of JSB22 was similar with initial vial cell counts. Growth of the six LAB was screened in arabino-oligosaccharide (AOS)-containing MRS broth. Results showed that growth of the isolates selectively increased after culture in AOS-containing media. Strain JSB22 (6 hr), JSB66 (6 hr), HSB15 (20 hr), and JSA22 (29 hr) showed maximum growth rate. Especially, JSB22 showed the highest growth rate. These results suggest that EPS-producing LAB isolated from Deonjang could be applied as synbiotics.

Enzymatic Synthesis of New Oligosaccharides Using Glucansucrases. (Glucansucrases를 이용한 새로운 올리고당의 합성)

  • ;;;;;John F. Robyt
    • Microbiology and Biotechnology Letters
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    • v.26 no.2
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    • pp.179-186
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    • 1998
  • Dextransucrase hyper-producing Leuconostoc mesenteroides B-512FMCM and dextransucrase constitutive mutants B-742CB and B-1355C catalyzed the transfer of glucose from sucrose to other carbohydrates which were present or were added to the reaction digests. When the acceptor was a maltose, gentiobiose, lactose or raffinose, there was produced a series of oligosaccharide acceptor products or single product based on the kinds of enzymes and reaction conditions. To obtain the quantitative information about the yield and the distribution of acceptor products and dextran two experimental parameters were studied: a) the ratio of acceptor to sucrose and b) the amount of enzyme at constant carbohydrate concentration (100 mM). As the amount of enzyme increased, the synthesis of acceptor products (of maltose or gentiobiose) increased, and the formation of dextran decreased. As the ratio of acceptor to sucrose increased, the amount of dextran and the number of acceptor-products decreased and the amount of acceptor-products increased. When maltose or gentiobiose was an acceptor, the glucose from sucrose was transferred to the C-6 hydroxyl group of the nonreducing-end glucose residue of accepters to give a homologous series of isomaltosyl dextrins. In case of lactose or raffinose, there was produced only one acceptor product from B-512FMCM dextransucrase reaction. In the lactose acceptor reaction, the glucose from sucrose was transferred to the C-2 hydroxyl of the reducing end glucose residue of lactose. To get a series of oligosaccharides from lactose or raffinose acceptor reaction we used B-742CB dextransucrase or B-1355C alternansucrase with 500 mM sucrose in reaction digest.

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