• KSBB Journal
• /
• v.29 no.3
• /
• pp.131-138
• /
• 2014

In this study, we isolated two novel chitinase producing bacterial strains from yellow loess samples collected from Jullanamdo province. The chitinase producing bacteria were isolated based on the zone size of clearance in the chitin agar plates. Both of them were gram positive, rod ($2{\sim}3{\times}0.3{\sim}0.4{\mu}m$), spore-forming, and motility positive. They were facultative anaerobic, catalase positive and hydrolyzed starch, gelatin, and casein. From the 16s rRNA gene sequence analysis, the isolates were labeled as Bacillus licheniformis KYLS-CU01 and B. licheniformis KYLS-CU02. The isolates showed higher extracellular chitinase activities than B. licheniformis ATCC 14580 as a control. The optimum temperature and pH for chitinase production were $40^{\circ}C$ and pH 7.0, respectively. Response Surface Methodology (RSM) was used to optimize the culture medium for efficient production of the chitinase. Under this optimal condition, 1.5 times higher chitinase activity of B. licheniformis KYLS-CU02 was obtained. Extracellular chitinases of the two isolates were purified through ammonium sulfate precipitation and anion-exchange DEAE-cellulose column chromatography. The specific activities of purified chitinase from B. licheniformis KYLS-CU01 and B. licheniformis KYLS-CU02 were 7.65 and 5.21 U/mg protein, respectively. The molecular weights of the two purified chitinases were 59 kDa. Further, the purified chitinase of B. licheniformis KYLS-CU01 showed high antifungal activity against Fusarium sp.. In conclusion, these two bacterial isolates can be used as a biopesticide to control pathogenic fungi.

• Journal of Life Science
• /
• v.8 no.5
• /
• pp.497-503
• /
• 1998

Cyclodextrinase from B. stearothemophilus KJ16 that can produce both cyclodextrin(CD) glucanotransferase and cyclodextrinase was purified 87.6-fold with 7% yield by ammonium sulfate precipitation, DEAE-cellulose chromatog-raphy, Sephadex G-100 chromatography, and FPLC. The molecular weight of the purified enzyme was about 68,000 dalton by SDS-PAGE. The optimal pH and temperature were 6.0 and 55$^{\circ}C$, respectively. The enzyme was stable at 5$0^{\circ}C$ for 2 hr in the pH range of 5.5 and 8.5. The enzyme activity was inhibited strongly by mercaptoethanol, di-thiothreitol, p-chloromercuribenzoate, N-bromosuccinimide, $Cu^{+2}$and $Hg^{+2}$. The purified enzyme hydrolyzed CDs with$\gamma$-CD>$\beta$-CD>$\alpha$-CD. The enzyme also hydrolyzed linear maltodextrins and polysaccharides, but the rates of hyd-rolysis for such substrates were slow as compared to that for $\gamma$-CD. The final degradation products with all substrates were maltose and glucose. Maltose was not further hydrolyzed.

• Korean Journal of Food Science and Technology
• /
• v.49 no.3
• /
• pp.229-234
• /
• 2017

For elderly individuals with swallowing difficulties, texture-modified food containing thickeners is needed. The plant materials freeze-dried yam, lotus root, and artichoke were evaluated for their potential use as thickeners and compared to a commercial thickener (corresponding to 0.5, 1, or 2 bag viscosity, respectively). Yam powder showed a high peak viscosity, corresponding to that of rice starch. Although artichoke showed no pasting viscosity, it may be useful as a thickener when combined with xanthan gum. The certain amount of yam, lotus root, and artichoke powders added to commercial soup showed similar viscosities as the addition of 0.5 or 1 bag commercial thickener at $60^{\circ}C$. However, sufficient viscosity corresponding to that of 2 bags of commercial thickener was not achieved by using the plant material itself. Sensory tests by elderly subjects revealed that the soup with yam powder was preferred compared to soup made with other materials.

• Journal of Microbiology and Biotechnology
• /
• v.9 no.6
• /
• pp.811-819
• /
• 1999

The ${\beta}-CGTase$ gene of alkalophilic Bacillus firmus var. alkalophilus was cloned into E. coli using $pZErO^{TM}-2$ as a vector. The cloned gene encoded a total of 710 amino acid residues consisting of 674 amino acids of the matured protein and 36 amino acids of the signal peptide, including 20 amino acids from the lacZ gene in the vector. Although the cloned ${\beta}-CGTase$ gene did not contain the promoter and start codons, it was expressed by the lac promoter and lacZ start codon in the $pZErO^{TM}$ vector. A comparison was made with the amino acid sequence and ten other CGTases from Bacillus sp. Also, ten highly conserved regions, which are important amino acid residues in catalysis of CGTase, were identified. The lac promoter used for expression of the ${\beta}-CGTase$ gene was induced constitutively in recombinant E. coli even without IPTG possibly because of a lack of the lacI gene in both host and vector, repressing the lacZ gene in the lac operon. Its expression was catabolically repressed by glucose, however, its repression was reduced by soluble starch, mainly because of the extremely high increase of the cAMP level. ${\beta}-CGTase$ can be overproduced in the recombinant E. coli by maintaining intracellular cAMP levels mostly through the intermittent feeding of glucose during cultivation.

• Preventive Nutrition and Food Science
• /
• v.9 no.2
• /
• pp.113-120
• /
• 2004

Selected microbiological and biochemical characteristics of the steeping process for the production of yukwa, a traditional Korean oil-puffed snack made of waxy rice, were investigated during steeping of waxy rice in water for 15 days. The lengthy steeping process was largely predominated by lactic acid bacteria (LAB), particularly, Lactobacillus and Leuconostoc. The predominat type of bacterium isolated was the Y26 strain tentatively identified as Lactobacillus plantarum. The titratable acidity of the steeping medium increased from 0.01 to 1.13%, in parallel with the decrease in pH ranging from 6.3 to 4.2 as the steeping period increased from 0 to 15 days. A high amount of lactic acid and to a much lesser extent, butyric acid, acetic acid, propionic acid, and succinic acid were detected during the steeping process. The amount of reducing sugars in the steeping medium increased from 0.61 to 10.43 mg/mL, whereas sucrose decreased from 0.46 mg% to an undetectable level. Starch degradation products including glucose, maltose and oligosaccharides ranging G3-G7 were not initially noticed, but their content increased during the steeping process until completion. However, no oligosaccharides larger than G8 were detected in the steeping medium. The activities of $\alpha$-amylase, $\beta$-amylase and protease in the steeping medium of waxy rice tended to rise increase with time during the steeping process. From these results, the lengthy steeping process in yukwa production can be characterized as the spontaneous fermentation, dominated by lactic acid bacteria, which is a necessary process for inducing biochemical modification of waxy rice.

• Food Science and Biotechnology
• /
• v.14 no.6
• /
• pp.860-865
• /
• 2005

A gene for a putative glucoamylase, stg, of a hyperthermophilic archae on Sulfolobus tokodaii was cloned and expressed in Escherichia coli. The recombinant glucoamylase (STGA) had an optimal temperature of $80^{\circ}C$ and was extremely thermostable with a D-value of 17 hr. The pH optimum of the enzyme was 4.5. Being different from fungal glucoamylases, STGA hydrolyzed maltotriose (G3) most efficiently. Gel permeation chromatography and sedimentation equilibrium analytical ultracentrifugation analysis showed that the enzyme existed as a dimer. STGA was stable enough to hydrolyze liquefied com starch to glucose in 4 hr at $90^{\circ}C$ with a yield of95%. Comparison of the $k_{cat}$ values for the hydrolysis and the reverse reaction at $75^{\circ}C$ and $90^{\circ}C$ indicated that glucose production by STGA was more efficient at $90^{\circ}C$ than $75^{\circ}C$. Therefore, STGA showed great potential for application to the industrial glucose production process due to its high thermostability.

• Applied Biological Chemistry
• /
• v.40 no.6
• /
• pp.465-471
• /
• 1997

The cyclodextrin glycosyltransferase (CGTase, EC 3.2.1.19) from Bacillus brevis CD162 was purified by precipitating with ammonium sulfate, DEAE-Sepharose CL-6B column chromatography and Sephadex G-150 column chromatography. The molecular mass and pI of the purified enzyme were estimated to be 74,000 and 6.3 by SDS-PAGE and isoelectric focusing, respectively. The purified enzyme was clearly identified as the CGTase by zymogram after SDS-PAGE. The optimum pH and temperature for the enzyme activity were 8.0 and $55^{\circ}C$, respectively. The enzyme was stable at the range of pH $5.5{\sim}9.0$, and up to $50^{\circ}C$. The amino acid sequence from the $NH_2-terminal$ of the purified CGTase was Ser-Val-Thr-Asn-Lys-Val-Asn-Tyr-Ser-Lys-Asp-Val-Ile-Tyr-Gln. The yields of the products from starch as the substrate were 1.3% for ${\alpha}-$, 33.9% for ${\beta}-$, and 9.7% for ${\gamma}-cyclodextrin$.

• Journal of Ginseng Research
• /
• v.40 no.4
• /
• pp.453-461
• /
• 2016

Background: This study aims to describe the characterization of Paenibacillus polymyxa GBR-1 (GBR-1) with respect to its positive and negative effects on plants. Methods: The morphological characteristics of GBR-1 were identified with microscopy, and subjected to Biolog analysis for identification. Bacterial population and media optimization were determined by a growth curve. The potential for GBR-1 as a growth promoting agent, to have antagonistic activity, and to have hydrolytic activity at different temperatures was assessed. The coinoculation of GBR-1 with other microorganisms and its pathogenicity on various stored plants, including ginseng, were assessed. Results: Colony morphology, endospore-bearing cells, and cell division of GBR-1 were identified by microscopy; identification was performed by utilizing the Biolog system, gas chromatography of fatty acid methyl esters (GC-FAME). GBR-1 showed the strongest antagonistic activity against fungal and bacterial pathogens. GBR-1 cell numbers were relatively higher when the cells were cultured in brain heart infusion (BHI) medium when compared with other media. Furthermore, the starch-hydrolytic activity was influenced by GBR-1 at higher temperature compared to low temperatures. GBR-1 was pathogenic to some of the storage plants. Coinoculation of GBR-1 with other pathogens causes differences in rotting on ginseng roots. A significant growth promotion was observed in tobacco seedlings treated with GBR-1 suspensions under in vitro conditions, suggesting that its volatile organic compounds (VOCs) might play a role in growth promotion. Conclusion: The results of this study indicate that GBR-1 has both positive and negative effects on ginseng root and other stored plants as a potential biocontrol agent and eliciting in vitro growth promotion.

• Korean Journal of Food Science and Technology
• /
• v.26 no.5
• /
• pp.633-637
• /
• 1994

A bacterial strain KSM-35 producing maltotetraose forming amylase was isolated from compost and identified as Streptomyces based on its morphological, cultural, and physiological characteristics. The amylase from Streptomyces sp. KSM-35 culture filtrate was purified by ammonium sulfate precipitation, followed by the liquid chromatographic procedures using DEAE-Toyo pearl and sephadex G-100 with 27.1% activity recovery. The molecular weight of the enzyme was estimated to be 50,000 and the isoelectric point 4.3. The main product by the amylase from soluble starch was maltotetraose which accounted for 56% of all the oligosaccharides detected after 26 hrs of reaction. Maltose (20%o) and maltotriose (16%) were the next important byproducts while glucose and maltopentaose were detected as traces.

• Journal of Microbiology and Biotechnology
• /
• v.20 no.10
• /
• pp.1378-1385
• /
• 2010

A novel agarolytic bacterium, KY-YJ-3, producing extracellular agarase, was isolated from the freshwater sediment of the Sincheon River in Daegu, Korea. On the basis of Gram-staining data, morphology, and phylogenetic analysis of the 16S rDNA sequence, the isolate was identified as Cellvibrio sp. By ammonium sulfate precipitation followed by Toyopearl QAE-550C, Toyopearl HW-55F, and MonoQ column chromatographies, the extracellular agarase in the culture fluid could be purified 120.2-fold with a yield of 8.1%. The specific activity of the purified agarase was 84.2 U/mg. The molecular mass of the purified agarase was 70 kDa as determined by dodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The optimal temperature and pH of the purified agarase were $35^{\circ}C$ and pH 7.0, respectively. The purified agarase failed to hydrolyze the other polysaccharide substrates, including carboxymethyl-cellulose, dextran, soluble starch, pectin, and polygalacturonic acid. Kinetic analysis of the agarose hydrolysis catalyzed by the purified agarase using thin-layer chromatography showed that the main products were neoagarobiose, neoagarotetraose, and neoagarohexaose. These results demonstrated that the newly isolated freshwater agarolytic bacterium KY-YJ-3 was a Cellvibrio sp., and could produce an extracellular ${\beta}$-agarase, which hydrolyzed agarose to yield neoagarobiose, neoagarotetraose, and neoagarohexaose as the main products.