• Preventive Nutrition and Food Science
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• v.11 no.2
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• pp.120-126
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• 2006

Fish backbone, a major by-product in the fish processing industry, accounts for about 15% of whole fish weight. In this study, recovery of bioavailable calcium from Alaska pollack (Theragra chalcogramma) backbone by-products using enzymatic hydrolysis was investigated. Finely ground fish backbones were hydrolyzed with two proteolytic enzymes (pepsin and protease) to obtain soluble calcium from the by-products. The pepsin digest had a higher degradation efficiency (88%) than protease. Four different concentrations of the fish backbone calcium (100, 250, 500 and 1000 mg/L) prepared by the pepsin digest were treated with $Na_2HPO_4$ at a concentration gradient (0, 1, 2, 4, 8, 10, 15 and 20 mM) to evaluate their solubility, revealing that solubilities of the fish backbone calcium were superior to those of $CaCl_2$ at all the calcium and $Na_2HPO_4$ concentrations. Among the tested concentrations the highest solubility was found in the pepsin digest containing a calcium concentration of 1000 mg/L. Thus, hydrolyzing with pepsin is an effective mode of recovering bioavailable calcium from Alaska pollack fish backbones.

• Journal of Applied Biological Chemistry
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• v.50 no.3
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• pp.132-135
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• 2007

An efficient production method of chlorella extract was developed by enzymatic treatment using cell lytic and proteolytic enzymes. The suitable dosage of Tunicase, a cell lytic enzyme, was found to be 1.0% (w/w). Proteolytic enzymes were screened to obtain high chlorella growth factor (CGF) index, which indicates crude CGF content and solid recovery. Among the seven tested proteases, Esperase, whose optimal dosage was 1.0% (w/w), was selected. By co-treatment using optimal dosages of Tunicase and Esperase, the highest CGF index and solid recovery were obtained. The CGF index and solid recovery of co-treatment were remarkably enhanced by 250 ($4.36{\rightarrow}15.21$) and 220% ($12.65%{\rightarrow}40.15%$), respectively, than those of the non-treated extracts.

• Journal of Sericultural and Entomological Science
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• v.18 no.2
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• pp.101-105
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• 1976

The study was carried out to investigate the variations in the fibroin hydrolysis of leading silkworm varieties in Korea, and the relationship between the fibroin hydrolysis of the silkworm varieties with some characteristics and silk morphological property. The results obtained are summarized as follows: 1) The fibroin hydrolyzing ratio of both parent silkworm cocoons was reduced in the F$_1$hybrids. 2) A positive corelation (r=＋0.86) was approved between the mid-parent value and the F$_1$hybrid in the fibroin hydrolysis. 3) It was disclosed that the fibroin hydrolyzing ratio of Jam 103${\times}$Jam 104 was the lowest among the leading silkworm F$_1$hybrids. 4) The fibroin hydrolysis was affected by the silk morphological property, showing a positive corelation between the size of cocoon filament and the fibroin hydrolyzing ratio. 5) It was thought that the determination of the fibroin hydrolyzing ratio was useful as a marker for the selection of its related economical characters in silkworm breeding, because the fibroin hydrolysis of parents silkworm cocoons was heritable in their F$_1$hybrids.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2001.06a
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• pp.40-45
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• 2001

Raw starch-digesting amylase (BF-2A, M.W. 93, 000 Da) from Bacillus circulans F-2 was converted to two components during digestion with subtilisin. Two components were separated and designated as BF-2A' (63, 000 Da) and BF-2B (30, 000 Da), respectively. BF-2A' exhibited the same hydrolysis curve for soluble starch as the original amylase (BF-2A). Moreover, the catalytic activities of original and modified enzymes were indistinguishable in $K_{m}$, Vmax for, and in their specific activity for soluble starch hydrolysis. However, its adsorbability and digestibility on raw starch was greatly decreased. Furthermore, the enzymatic action pattern on soluble starch was greatly different from that of the BF-2A. A smaller peptide (BF-2B) showed adsorb ability onto raw starch. By these results, it is suggested that the larger peptide (BF-2A') has a region responsible for the expression of the enzyme activity to hydrolyze soluble substrate, and the smaller peptide (BF-2B) plays a role on raw starch adsorption. A similar phenomenon is observed during limited proteinase K, thermolysin, and endopeptidase Glu-C proteolysis of the enzyme. Fragments resulting from proteolysis were characterized by immunoblotting with anti-RSDA. The proteolytic patterns resulting from proteinase K and subtilisin were the same, producing 63- and 30-kDa fragments. Similar patterns were obtained with endopeptidase Glu-C or thermolysin. All proteolytic digests contained a common, major 63-kDa fragment. Inactivation of RSDA activity results from splitting off the C-terminal domain. Hence, it seems probable that the protease sensitive locus is in a hinge region susceptible to cleavage. Extracellular enzymes immunoreactive toward anti-RSDA were detected through whole bacterial cultivation. Proteins of sizes 93-, 75-, 63-, 55-, 38-, and 31-kDa were immunologically identical to RSDA. Of these, the 75-kDa and 63-kDa proteins correspond to the major products of proteolysis with Glu-C and thermolysin. These results postulated that enzyme heterogeneity of the raw starch-hydrolysis system might arise from the endogeneous proteolytic activity of the bacterium. Truncated forms of rsda, in which the gene sequence encoding the conserved domain had been deleted, directed the synthesis of a functional amylase that did not bind to raw starch. This indicates that the conserved region of RSDA constitutes a raw starch-binding domain, which is distinct from the active centre. The possible role of this substrate-binding region is discussed.d.

• Korean Journal of Food Science and Technology
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• v.16 no.2
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• pp.211-217
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• 1984

To study affinity of proteolytic enzymes to soy proteins, the physicochemical and functional properties of enzymatically modified protein products, kinetic parameters and degree of hydrolysis were measured using trypsin, alcalase (serine type protease) and pronase. Bacterial alcalase and pronase showed much greater affinity to soy protein than animal intestinal trypsin. This effect was very significant when unheated soy isolate was used as a substrate. Specific activities of these enzymes decreased with the increment of substrate concentration (over 2.0%, w/v) when heat denatured soy protein was used as a substrate. However, the decrease in specific activity was negligible at substrate concentrations lower than 2.0%. Polyacrylamide gel electrophoretic results showed that the pattern of 2S protein band changed distinctly in alcalase hydrolysis as compared with those of trypsin and pronase. Protein solubilities of alcalase and pronase hydrolyzates increased by 25-30%, at their pI (pH 5.0) over the control. Virtually no change was observed in solubility by trypsin hydrolysis. Heat coagulability and calcium-tolerance of the protein increased by enzymatic hydrolysis. No clear tendency, however, was observed for emulsion properties, foam expansion and the amount of free -SH groups. The enzyme treatment considerably decreased foam stability.

• Journal of Life Science
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• v.11 no.2
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• pp.138-143
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• 2001

Marsh calm(Corbicular elatior)with a short-term storage in raw and a law-rate of utilization has been increasing the needs to develop new marsh calm processing products for a temporary mass treatment and long-term distribution, Therefore the processing conditions of marsh calm beverage using proteolytic enzyme hydrolysis were investigated. A partial hydrolysis at 6$0^{\circ}C$ for 1 hour after adding 3% Alcalase as more effective than a hot water extraction to develop taste compounds from the marsh calm. The result of ommission test showed that nucleotides and their related compounds were contributed in the taste of the marsh calm hydrolysates rather than free amino acids. The taste of the hydrolysates was produced by association with these compounds rather than only one compound s the hydrolystes taste differently for the control when one of these compound was omitted. The hydrolysates were fractionated to molecular weight below 500 dalton to eliminate bitter taste and to improve it flavor from the hydrolysates, 0.05% bay leaf was more effective to improve the odor than other herbs.

• Proceedings of the Korean Society of Sericultural Science Conference
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• 2002.10a
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• pp.94-94
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• 2002

No Abstract, See Full Text

• KSBB Journal
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• v.10 no.4
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• pp.374-385
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• 1995

Nutritional requirements and cultural conditions for the production of extracellular gelatin-degrading proteolytic enzyme by Bacillus subtilis B0021 were investigated. Optimum carbon source for proteolytic enzyme production was salicin, but it was substituted by glucose for economical reason. The fermentation medium giving a maximum proteolytic enzyme activity was consisted of 1.5%(w/v) glucose, 2.5%(w/v) yeast extract, and 0.001%(w/v) manganese sulfate and 0.002%(w/v) ferrous sulfate. Proteolytic enzyme activity of B. subtilis B0021 was completely inhibited by 0.5%(w/v) tannic acid. Initial pH was optimal at 7.0 and the enzyme activity in the flask culture usually reached a maximal level after 36 hours of fermentation at $30^{\circ}C$. In the $5\ell$ fermentor fermentation at $30^{\circ}C$, enzyme activity was maximum at 36 hour of cultivation but after this enzyme activity was decreased rapidly. Initial viscosity of 45%(w/v) gelatin(2,800mPas) was decreased rapidly to 96%(mPas) after hydrolysis for 4hr at $40^{\circ}C$ by crude enzyme of B. subtilis B0021.

• Food Engineering Progress
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• v.21 no.1
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• pp.88-92
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• 2017

We attempted to investigate antibacterial and proteolytic activities of bacteria isolated from three ethnic fermented seafoods in the east coast of South Korea, gajami sikhae, squid jeotgal, and fermented jinuari (Grateloupia filicina). Bacillus cereus ATCC 14579, Listeria monocytogenes ATCC 15313, Staphylococcus aureus KCTC 1916, Escherichia coli O157:H7 ATCC 43895, and Salmonella enterica serovar Typhimurium ATCC 4931 were selected to determine the antibacterial activity of the bacterial isolates. Among 233 isolates from the three foods, 36 isolates (15.5%) showed antibacterial activity against B. cereus ATCC 14579, the highest incidence of inhibition, followed by S. aureus KCTC 1916 (7.7%) and L. monocytogenes ATCC 15313 (6.0%). However, only five and three strains among the isolates exhibited inhibitory activity against Gram-negative indicators, E. coli ATCC 43895 and Sal. enterica ATCC 4931, respectively. The proteolytic activity of the isolates was determined via hydrolysis of skim milk after 24, 48, and 72 h incubation. After 72 h incubation, 72 out of 233 isolates (30.9%) showed proteolytic activity, and the isolates of fermented jinuari exhibited the highest incidence of proteolytic activity (60%, 36 isolates). These results suggest that ethnic fermented seafoods in the east coast of South Korea might be a promising source of bacterial strains producing antibacterial and proteolytic compounds.

• Mass Spectrometry Letters
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• v.8 no.3
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• pp.53-58
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• 2017

Glycated hemoglobin ($HbA_{1c}$) has been commonly used to screen and diagnose for patients with diabetes mellitus. Here the accelerated procedure of microwave-assisted sample treatment from acid hydrolysis to enzyme digestion followed by isotope dilution liquid chromatography-tandem mass spectrometry (ID-LC-MS/MS) was optimized and applied to measure $HbA_{1c}$ in an effort to speed up analysis time. First, two signature peptides of $HbA_{1c}$ and hemoglobin $A_0$ were certified with amino acid analysis by setting optimized acid hydrolysis conditions to $150^{\circ}C$, 1.5 h and $10{\mu}M$ sample concentration in 8 M hydrochloric acid. Consequently, the accurate certified peptides above were used as calibration standards to implement the proteolytic procedure with endoproteinase Glu-C at $37^{\circ}C$, 700 W for 6 h. Compared to the traditional method, the microwave heating not only shortened dramatically sample preparation time, but also afforded comparable recovery yields. The optimized protocol and analytical conditions in this study are suitable for a primary reference method of $HbA_{1c}$ quantification with full SI-traceability and other similar proteins in complex biological samples.