• Journal of Animal Science and Technology
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• v.51 no.6
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• pp.527-536
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• 2009

A potent protein degrading bacterium was isolated from soil samples of different environments. Polyphasic taxonomic studies and phylogenetic 16S rRNA sequence analyses led to identify the isolate IB No. 11 as a strain of Bacillus subtilis. The isolated strain was recognized to produce protease constitutively, and the maximum production (1.64 units/ml) was attained in a shake flask culture when the isolate was grown at $40^{\circ}C$, for 32 h in basal medium supplemented with starch (0.25%) and gelatin (1.25%) as sole carbon and nitrogen source, respectively. The optimum pH and temperature for the protease activity were determined to be pH 7.0 and $50^{\circ}C$, respectively. $Ca^{2+}$ and $Mn^{2+}$ enhanced remarkably the protease activity but neither showed positive effect on the protease's thermal stability. In addition, it was observed that the protease was fairly stable in the pH range of 6.5-8.0 and at temperatures below $50^{\circ}C$, and it could be a good candidate for an animal feed additive. The inhibition profile of the protease by various inhibitors indicated that the enzyme is a member of serine-proteases. A combination of UV irradiation and NTG mutagenesis allowed to develop a protease hyper-producing mutant strain coded as IB No. 11-4. This mutant strain produced approximately 3.23-fold higher protease activity (6.74 units/mg) than the parent strain IB No. 11 when grown at $40^{\circ}C$ for 32h in the production medium. The protease production profile of the selected mutants was also confirmed by the zymography analysis.

• Food Science and Biotechnology
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• v.18 no.3
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• pp.776-781
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• 2009

A putative cyclomaltodextrinase (BHCD) gene was found from the genome of Bacillus halodurans C-125, which encodes 578 amino acids with a predicted molecular mass of 67,279 Da. It shares 42-59% of amino acid sequence identity with common cyclomaltodextrinase (CDase)-family enzymes. The corresponding gene was cloned by polymerase chain reaction (PCR) and the dimeric enzyme with C-terminal 6-histidines was successfully overproduced and purified from recombinant Escherichia coli. BHCD showed the highest activity against ${\beta}-CD$ at pH 7.0 and $50^{\circ}C$. Due to its versatile hydrolysis and transglycosylation activities, BHCD has been confirmed as a member of CDases. However, BHCD can be distinguished from other typical CDases on the basis of its novel multisubstrate specificity. While typical CDases have over 10 times higher activity on ${\beta}-CD$ than starch or pullulan, the CD-hydrolyzing activity of BHCD is only 2.3 times higher than pullulan. In particular, it showed significantly higher activity ratio of maltotriose to acarbose than other common CDase-family enzymes.

• Journal of Fisheries and Marine Sciences Education
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• v.23 no.4
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• pp.662-672
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• 2011

This study was investigated to obtain basic data which can be applied to processing of canned seasoned kwamaegi. Commercial Kwamaegi was cut into $2{\times}3cm$ lengths, filled 90 g into can (301-3) and added with 60 g water before precooking for 10 min. at $100^{\circ}C$. The precooked Kwamaegi was packed into the can, and added with 60 g seasoning sauces, which was prepared by mixing soy sauce 23%, monosodium glutamate (MSG) 2%, sorbitol 2%, sesame oil 1%, vinegar 2%, starch syrup 17%, sake 5%, water 48%. The cans were sealed using a vacuum seamer and then sterilized for various Fo values (Fo 8~12 min.) in a steam system retort at $121^{\circ}C$. The factors such as pH, VBN, amino-N, total amino acid, free amino acid, color value (L, a, b), texture profile, TBA value, mineral, sensory evaluation and viable bacterial count of the canned seasoned Kwamaegi produced with various sterilization condition(Fo 8~12 min.) were measured. There was no remarkable difference between sterilization conditions and sensual characteristics. The results showed that the product sterilized at Fo 8 min. was the most desirable because this condition is most economical.

• Journal of Microbiology
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• v.44 no.6
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• pp.600-606
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• 2006

Spore forming actinobacteria (sporoactinobacteria) isolated from soils with an acidic pH in Pinus thunbergii forests and coal mine waste were subjected to taxonomic characterization. For the isolation of acidophilic actinobacteria, acidified starch casein agar (pH adjusted to 4-5) was used. The numbers of actinobacteria growing in acidic media were between $3.2{\times}10^4$ and $8.0{\times}10^6$ CFU/g soil. Forty three acidophilic actinobacterial strains were isolated and their 16S rDNA sequences were determined. The isolates were divided into eight distinctive phylogenetic clusters within the variation encompassed by the family Streptomycetaceae. Four clusters among them were assigned to the genus Streptacidiphilus, whereas the remaining four were assigned to Streptomyces. The clusters belonging to either Streptomyces or Streptacidiphilus did not form a monophyletic clade. The growth pH profiles indicated that the representative isolates grew best between pH 5 and 6. It is evident from this study that acidity has played a critical role in the differentiation of the family Streptomycetaceae, and also that different mechanisms might have resulted in the evolution of two groups, Streptacidiphilus (strict acidophiles) and neutrotolerant acidophilic Streptomyces. The effect of geographic separation was clearly seen among the Streptacidiphilus isolates, which may be a key factor in speciation of the genus.

• Preventive Nutrition and Food Science
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• v.7 no.3
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• pp.310-316
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• 2002

Extracellular cyclodextrin glucanotransferase (CGTase) from Paenibacillus sp. JK-12 was purified through sev-eral purification steps consisting of ammonium sulfate precipitation and chromatographies on DEAE-sephadex A-50 and Mono QIM HR5/5. The purified CGTase exhibited a single band on SDS-PAGE and was estimated to be approximately 82 kDa. The isoelectric point of the enzyme was 7.2 as determined by isoelectric focusing. The CGTase from Paenibacillus sp. JK-12 had a transglucosylation activity at the C-2 position of L-ascorbic acid. The optimum pH and temperature for the CGTase activity were 8.0 and 5$0^{\circ}C$, respectively. The enzyme activity was stable from pH 6.0 to 9.() and at temperatures up to 55$^{\circ}C$ at pB 8.0, having 80% residual activity. The activity of the CGTase was strongly resistant to metals such as A $g^{+}$ and $Ba^{2+}$ but slightly inhibited by H $g^{+}$, N $i^{2+}$ and $Mg^{2+}$. The enzymeproduced $\alpha$ -cyclodextrin ($\alpha$-CD) and $\beta$-CD as the main products from starch, but not ${\gamma}$-CD.X>-CD.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.42 no.2
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• pp.109-115
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• 2009

The properties of surimi gel from Olive flounder Paralichthys olivaceus were evaluated with addition of various food additives and by heating under different conditions. The optimum heating conditions for get good textured surimi gel from Olive flounder was 40 minutes at $85^{\circ}C$. Optimum texture characteristics such as hardness, cohesiveness, chewiness, gumminess and gel strength of olive flounder fish paste can be acquired by washing the paste 2 times and by adding sodium chloride (2.5%), sodium polyphosphate (0.3%) and starch (4%), respectively. Furthermore, we compared gel texture characteristics and strength of manufactured Olive flounder fish surimi with commercial Alaska pollack to determine their commercial applicability. Texture characteristics and gel strength of Olive flounder surimi were higher than those of the commercial Alaska pollack surimi. In addition, both lightness and whiteness were higher in surimi gel from Olive flounder than from the commercial Alaska pollack.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.89-92
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• 2001

Aspergillus niger has been used as a host system to express many heterologous proteins. It has various advantages over other expression systems in that it is a small eukaryotic GRAS (Generally Recognized aS Safe) organism with a capacity of secreting large amount of foreign proteins. However, it has been known that the presence of an abundant protease is a limiting factor to express a heterologous protein. The proteases deficient mutants of A. niger were obtained using UV -mutagenesis. A total of 1 ${\times}$ $10^5$ spores were irradiated with 10-20% survival dose of UV, 600J/M2 at 280nm, and the resulting spores were screened on the casein -gelatin plates. Ten putative protease deficient mutants were further analyzed on the starch plates to differentiate the pro from the secretory mutant. An endogenous extracellular enzyme, glucose oxidase, was also examined to confirm that the mutant phenotype was due to the proteases deficiency rather than the mutation in the secretory pathway. The reduced proteolytic activity was measured using SDS-fibrin zymography gel, casein degradation assay, and bio-activity of a supplemented hGM -CSF (human Granulocyte-Macrophage Colony Stimulating Factor). Comparing with the wild type strain, less than 30 % of proteolytic activity was observed in the culture filtrate of the protease deficient mutant (pro -20) without any notable changes in cell growth and secretion.

• Journal of Physiology & Pathology in Korean Medicine
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• v.27 no.1
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• pp.107-111
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• 2013

To investigate the possible usage as probiotics, we isolated and identified one strain of bacillus from Korean traditional food, Jeotgal. The isolate was identified as Bacillus megaterium KS12 by examining its biochemical properties and 16S rRNA sequence analysis (99% similarity). B. megaterium KS12 exhibited high viability in artificial gastric juice for 3 h and in artificial bile salts for 24 h, about 14.4 and 14%, respectively. The herbal digestive medicine containing the isolate showed high starch hydrolytic activity and proteolytic activities (about 6-fold) compared to traditional herbal digestive medicine. The viable cells in the herbal digestive medicine containing the isolate were higher than those of traditional herbal digestive medicine.

• Microbiology and Biotechnology Letters
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• v.14 no.2
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• pp.205-208
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• 1986

The 23 serovars of Bacillus thuringiensis strain were commonly gram-positive and motile, formed endotoxin crystals, produced acid and alkali in the KIA media, and acid from glucose, hydrolyzed starch, and reduced nitrate but did not produce H$_2$S, oxidase and indole, did not decompose lysine, ornithine, phenylalanine, malonate, lactose, dulcitol, adonitol, inositol, sorbitol, arabinose, raffinose, rhamnose, maltose, and xylose. Eighteen serovars were positive in the MR tests and 15 in the VP tests. Four serovars used citrate. Five serovars produced urease, 5 $CO_2$ from glucose, 2 DNase, and 15 lecithinase. Twelve serovars decomposed arginine, 11 did sucrose, 2 manitol, and 9 salicin Serovar tohokuensis did not hemolyze, but the others did.

• Journal of Microbiology and Biotechnology
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• v.4 no.1
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• pp.41-48
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• 1994

The xylanase from alkali-tolerant Bacillus sp. YA-14 was purified to homogeneity by CM-cellulose, Sephadex G-50, and hydroxyapatite column chromatographies. The molecular weight of the purified enzyme was estimated to be 20, 000 Da by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The purified enzyme slightly hydrolyzed carboxymethyl cellulose and Avicel, but did not hydrolyze soluble starch, dextran, pullulan, and ${\rho}-nitrophenyl-{\beta}$-D-xylopyranoside. The maximum degree of hydrolysis by enzyme for birchwood xylan and oat spelts xylan were 47 and 40%, respectively. The Michaelis constants for birchwood xylan and oat spelts xylan were calculated to be 3.03 mg/ml and 5.0 mg/ml, respectively. The activity of the xylanase was inhibited reversibly by $HgCl_2$, and showed competitive inhibition by N-bromosuccinimide, which probably indicates the involvement of tryptophan residue in the active center of the enzyme. The Xylanase was identified to be xylose-producing endo-type xylanase and did not show the enzymatic activities which cleave the branch point of the xylan structure.