• Applied Microscopy
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• v.24 no.3
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• pp.67-77
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• 1994

Storage material of endosperm cells digested by various enzymes should be transported to the embryo. At this time, the cellulose of the endosperm cell wall is guessed to be hydrolyzed by the cellulase enabling to transfer the storage material from the endosperm cells to the embryo. Therefore, this study has been carried out to investigate the ultrastructure of endosperm and the localization of the cellase activity on Cannabis sativa L. during germination. Endosperm cells contain a large number of lipid bodies and protein bodies with globoids as the storage material. During gemination they are gradually degenerated, however, the former almost remain until the cells are completely digested. Electron-microscopical reaction products of cellulase on endosperm cells are present. The closer the embryo, the more amount of reaction products on the endosperm cell wall are appeared.

• Korean journal of food and cookery science
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• v.16 no.2
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• pp.151-159
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• 2000

The effects of bromelain and $\alpha$-chymotrypsin treatments on the functional properties(foaming capacity, foaming stability, emulsifying capacity, and emulsifying stability) of soy protein isolate(SPI) and the addition of various sweeteners(sucrose, sorbitol, xylitol) on the quality attributes(viscosity, overrun ratio, melt-down property, and sensory characteristic) of soy ice cream were studied. SPI was more effectively hydrolyzed with $\alpha$-chymotrypsin than bromelain, resulting in a better foaming and emulsifying capacity. Adding xylitol could significantly improve the viscosity, overrun and melt-down property of soy ice creams while the effect was the lowest in the sucrose addition. Bromelain treatment caused a lower apparent viscosity of SPI suspension compared with $\alpha$-chymotrypsin treatment and untreated. The overrun ratios of the soy ice cream prepared with bromelain and $\alpha$-chymotrypsin treated SPI were 18.9∼25.9% and 24.9∼40.3%, respectively as a result of freezing with agitation for 20 min in an ice cream maker. Comparatively, untreated SPI could bring only 15.8∼21.4% overrun ratios after operating for 15 min. The bromelain treatment caused high melt-down tendency of the product while soy ice cream with untreated SPI showed an opposite trend. In sensory characteristics, no significant differences in the strength of beany flavor were noted among the samples. Sweetness, bitter taste, icy feel, and mouthfeel of the product were greatly affected by the enzyme-treatment of SPI. Soy ice cream added with xylitol after $\alpha$-chymotrypsin treatment was the most acceptable among all samples.

• Korean Journal of Food Science and Technology
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• v.23 no.1
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• pp.25-30
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• 1991

Chicken liver was enzymatically hydrolyzed with an alkaline protease and determined the optimal conditions of reaction temperature and time, pH and enzyme to substrate ratio(E/S ratio) for possible utilization as a protein supplementary ingredient. The functional properties of hydolysate measured were water and oil absorption capacity, emulsifying activity and viscosity and sensory properties were also evaluated. It was found that hydrolysis at $60^{\circ}C$ and pH 8.0 were most effective and the degree of hydrolysis increased with increasing E/S ratio. A decrease in water and oil absorption capacity and an increase in viscosity were found during hydrolysis. The lowest emulsifying activity and highest water absorption were measured for 1/2 hour-hydrolysate and little difference was found for those treated more than 1 hour. The sensory characteristics of odor showed no significant difference among the chicken liver hydrolysates while the brightness increased and red decreased significantly(p<0.01) as the hydrolysis proceeded.

• Journal of Microbiology and Biotechnology
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• v.16 no.4
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• pp.613-618
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• 2006

Two endoxylanases from an alkaliphilic bacterium, Bacillus halodurans C-1, were purified 3.8- and 7.9- fold with specific activities of 9.4 and 19.8U/mg protein, respectively. The molecular masses of both purified enzymes were 23 and 47 kDa, respectively, and 23 kDa xylanase I (Xyl I) exhibited an optimum pH at 7.0, whereas 47 kDa xylanase II (Xyl II) showed a broad pH range of 5.0 to 9.0. The temperature optima of both xylanases were $60^{\circ}C\;and\;70^{\circ}C$, respectively. Both were stable in the pH range of 6.0 to 9.0 and 5.0 to 10.0, respectively, and they were stable up to $60^{\circ}C\;and\;70^{\circ}C$, respectively. The $K_m\;and\;V_{max}$ of Xyl I were 4.33mg/ml and $63.5{\mu}mol/min/mg$, respectively, whereas Xyl II had a $K_m$ value of 0.30 mg/ml and $V_{max}$ of $210{\mu}mol/min/mg$. Both xylanases hydrolyzed xylans from birchwood, oat spelt, and larchwood. However, they showed different modes of action; a series of xylooligosaccharides larger than xylotriose were released as the major products by Xyl I, whereas xylobiose and xylotriose were the main products by Xyl II. The maximum synergistic action of the two enzymes on hydrolysis of xylan was 2.16 with the ratio of Xyl I to Xyl II at 1:9.

• Journal of Microbiology and Biotechnology
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• v.14 no.3
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• pp.474-482
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• 2004

The $\alpha$-L-arabinofuranosidase (Arfase) gene of Bacillus stearothermophilus No. 236 was cloned and sequenced. The ORF of the gene, designated arfA, encoded a 507 -residue polypeptide with calculated molecular mass of 57 kDa. The Arfase produced by a recombinant Escherichia coli strain containing the arfA gene was purified to apparent homogeneity and characterized. The molecular mass of the Arfase determined by SDS-PAGE was 60 kDa. However, according to gel filtration, it was estimated to be approximately 190 kDa. These results indicated that the functional form of the Arfase is trimeric. The optimal pH and temperature for the enzyme activity were pH 6.5 and $55^{\circ}C$, respectively. The half-life of the enzyme at $60^{\circ}C$ was about 6 h. Kinetic experiments at $45^{\circ}C$ with pNPM (p-nitrophenyl $\alpha$-L-arabinofuranoside) as a substrate gave the $K_m and V_{max}$ values of 1.19 mM and 26.1 U/ mg, respectively. When the enzyme was combined with Bacillus stearothermophilus No. 236 endoxylanase and $\beta$-xylosidase, it hydrolyzed arabinoxylan into L-arabinose and xylose more efficiently than Arfase alone. This synergistic effect suggested that the complete hydrolysis of xylan with large amounts of arabinose side chains required Arfase as well as endoxylanase and $\beta$-xylosidase.

• Journal of Microbiology and Biotechnology
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• v.17 no.6
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• pp.905-912
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• 2007

A novel ${\beta}-glucosidase$ gene, bglA, was isolated from uncultured soil bacteria and characterized. Using genomic libraries constructed from soil DNA, a gene encoding a protein that hydrolyzes a fluorogenic analog of cellulose, 4-methylumbelliferyl ${\beta}-D-cellobioside$ (MUC), was isolated using a microtiter plate assay. The gene, bglA, was sequenced using a shotgun approach, and expressed in E. coli. The deduced 55-kDa amino acid sequence for bglA showed a 56% identity with the family 1 glycosyl hydrolase Chloroflexus aurantiacus. BglA included two conserved family 1 glycosyl hydrolase regions. When using $p-nitrophenyl-{\beta}-D-glucoside$ (pNPG) as the substrate, the maximum activity of the purified ${\beta}-glucosidase$ exhibited at pH 6.5 and $55^{\circ}C$, and was enhanced in the presence of $Mn^{2+}$. The $K_m\;and\;V_{max}$ values for the purified enzyme with pNPG were 0.16 mM and $19.10{\mu}mol/min$, respectively. The purified BglA enzyme hydrolyzed both pNPG and $p-nitrophenyl-{\beta}-D-fucoside$. The enzyme also exhibited substantial glycosyl hydrolase activities with natural glycosyl substrates, such as sophorose, cellobiose, cellotriose, cellotetraose, and cellopentaose, yet low hydrolytic activities with gentiobiose, salicin, and arbutin. Moreover, BglA was able to convert the major ginsenoside $Rb_1$ into the pharmaceutically active minor ginsenoside Rd within 24 h.

• Journal of Microbiology and Biotechnology
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• v.10 no.2
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• pp.238-243
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• 2000

An exoinulinase (${\beta}-D-fructofuranosidase$) gene was cloned by chromosome walking along the upstream region of the endoinulinase gene of Pseudomonas mucidolens isolated from soil. the exoinulinase gene consisted of an ORF of 0,506 bp encoding a polypeptide of 501 amino acids with a deduced molecular weight of 55,000. The exoinulinase produced by the recombinant Escherichia coli $DH5{\alpha}$ strain was also purified to homogeneity as determined by SDS-PAGE and a zymogram. The molecular weight of the purified exoinulinase according to both SDS-PAGE and gel filtration matched the deduced molecular weight of the protein described above, thereby indicating that the native form of the exoinulinase was a monomer. The purified enzyme hydrolyzed activity value of 2.0. Furthermore, no inulo-oligomers were liberated from the inulin substrate in the enzymatic reaction mixtures incubated for 90 min at $55^{\circ}C$. Taken together, these results indicate that the purified ${\beta}-D-fructofuranosidase$ was an exoinulinase. The pH and temperature optima of the exoinulinase were pH 6.0 and $55^{\circ}C$, respectively. the enzymehad no apparent requirement for a cofactor, and its activity was completely inactivated by $Ag^{+},{\;}Hg^{2+},{\;}and{\;}Zn^{2+}$. Kinetic experiments gave $K_m,{\;}V_{max},{\;}and{\;}K_{cat}$ values for inulin of 11.5 mM, 18 nM/s, and $72{\;}s^{-1}$, respectively. the exoinulinase was fairly stable in broad pH conditions (pH 5-9), and at pH 6.0 it showed a residual activity of about 70% after 4 h incubation at $55^{\circ}C$.

• Microbiology and Biotechnology Letters
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• v.25 no.2
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• pp.166-172
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• 1997

Aspergillus niger was selected as a strain producing the potent raw starch hydorlyzing enzyme. These experiments were conducted to investigate the conditions of the glucoa- mylase production, the purification of the enzyme, some characteristics of the purified enzyme and hydrolysis rate on various raw starches such as com, rice, potato, glutinous rice, sweet potato, wheat and barley. The optimum cultural temperature and time for the enzyme production on wheat bran medium were $30^{\circ}C$ and 96hrs, respectively. The respective addition of yeast extract and nutrient broth on wheat bran medium increased slightly the enzyme production. The enzyme was purified by ammonium sulfate fractionation and DEAE-cellulose column chromatography. The specific activity of the purified enzyme was 30.7u/mg-protein and the yield of enzyme activity was 25.8%. The purified enzyme showed a single band on polyacrylamide disc gel electrophoresis and its molecular weight was estimated to be 56,000 by SDS-polyacrylamide disc gel electrophoresis. The isoelectric point for the purified enzyme was pH3.7. The optimum temperature and pH were $65^{\circ}C$ and pH 4.0, respectively. The purified enzyme was stable in the pH range of pH 3.0-9.5 and below $45^{\circ}C$, and its thermal stability was slightly increased by the addition of $Ca^{2+}$. The purified enzyme was activated by $Co^{2+},\;Sr^{2+},\;Mn^{2+},\;Fe^{2+},\;Cu^{2+}$. Raw rice starch, raw corn starch, raw glutinous rice starch, raw sweet potato starch, raw wheat starch and raw barley starch showed more than 90% hydrolysis rate in 48hrs incubation. Even raw potato starch, most difficult to be hydrolyzed, showed 80% hydrolysis rate. The purified enzyme was identified as glucoamylase.

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

• Preventive Nutrition and Food Science
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• v.7 no.4
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• pp.451-453
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• 2002

Alkali treatment following acidic hydrolysis (ATAH) of defatted soybean is currently used to reduce the level of 3-chloropropane-1,2-diol (3-MCPD), a known carcinogen, in commercial Korean soy sauce. 3-MCPD concentrations in commercial soy sauces made by ATAH were compared with those made only by acidic hydrolysis, and products that combine soy sauce made by acid hydrolysis (followed by alkalinization) and enzymatic methods. The four soy sauces made by ATAH had lower 3-MCPD concentrations (below 0.078 ppm) than 4 commercial products (0.147∼0.481 ppm) made only by acidic hydrolysis. On the other hand, 3-MCPD concentrations in 4 commercial products made by combining soy sauces made enzymatically with that made from acid (with alkali treatment)-hydrolyzed soybean protein in varying ratios were in a range of 0.016∼0.053 ppm. The 3-MCPD concentrations in commercial Korean soy sauces, with the exception of 2 of the soy sauces made only by acidic hydrolysis, were lower than allowable limit of 0.3 ppm in Korea. These results demonstrated that currently produced commercial soy sauces on the Korean market hate toxicologically save 3-MCPD concentrations. It is also provides evidence that ATAH is an effective process for reducing 3-MCPD concentrations in commercial soy sauce.