• Korean Journal of Food Science and Technology
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• v.16 no.2
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• pp.254-256
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• 1984

The amounts of husk materials from barley kernel were determined by an enzymatic method and compared with the values determined by conventional methods involving acid or alkaline treatments. The enzymatic method consists of boiling in distilled water and pressing to help squeeze out the gelatinized starch from the husk matrix, and enzymatic removal of starch by ${\alpha}-amylase$ and weighing the residual husk materials after washing 3 times with hot water and then drying at $95^{\circ}C$. Husk materials amounted about 15 of the covered barley (Gangbori and Olbori) and 10-12% of naked variety (Backdong and Sedohadaga) and the values were always somewhat higher than those obtained by the conventional methods. The husk materials prepared by the enzymatic procedure contained protein 4-8%, lipid 5-10%, ash 0.2-0.6% and crude fiber 20-40%. Although it took longer time, the enzymatic procedures can provide more intack husk materials for further characterization of the materials.

• Journal of Life Science
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• v.20 no.5
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• pp.683-690
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• 2010

The yeast strains of Saccharomyces diastaticus produce one of three isozymes of an extracellular glucoamylase I, II or III, a type of exo-enzyme which can hydrolyse starch to generate glucose molecules from non-reducing ends. These enzymes are encoded by the STA1, STA2 and STA3 genes. Another gene, sporulation-specific glucoamylase (SGA), also exists in the genus Saccharomyces which is very homologous to the STA genes. The SGA has been known to be produced in the cytosol during sporulation. However, we hypothesized that the SGA is capable of being secreted to the extracellular region because of about 20 hydrophobic amino acid residues at the N-terminus which can function as a signal peptide. We expressed the cloned SGA gene in S. diastaticus YIY345. In order to compare the biochemical properties of the extracellular glucoamylase and the SGA, the SGA was purified from the culture supernatant through ammonium sulfate precipitation, DEAE-Sephadex A-50, CM-Sephadex C-50 and Sephadex G-200 chromatography. The molecular weight of the intact SGA was estimated to be about 130 kDa by gel filtration chromatography with high performance liquid chromatography (HPLC) column. Sodium dedecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed it was composed of two heterogeneous subunits, 63 kDa and 68 kDa. The deglycosylation of the SGA generated a new 59 kDa band on the SDS-PAGE analysis, indicating that two subunits are glycosylated but the extent of glycosylation is different between them. The optimum pH and temperature of the SGA were 5.5 and $45^{\circ}C$, respectively, whereas those for the extracellular glucoamylase were 5.0 and $50^{\circ}C$. The SGA were more sensitive to heat and SDS than the extracellular glucoamylase.

• The Korean Journal of Food And Nutrition
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• v.18 no.1
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• pp.11-18
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• 2005

Angiotensin Ⅰ converting enzyme(ACE) inhibitory activities of laver(Porphyra tenera) protein hydrolysates were investigated by enzymes used for hydrolysis, molecular fractions and drying methods. For the enzymatic hydrolysis, crude laver protein, separated by filtration of water extract of dried laver extracted with 20 times(w/v) water for 3 hours at boiling temperature, were hydrolyzed with three commercial protease, Pepsin, alcalase and maxazyme NNP at optimal conditions. The yield of hydrolysis and ACE inhibitory activities of which were high in order of pepsin, alcalase and maxazyme NNP. ACE inhibitory activities of laver hydrolysates by molecular levels were high in order of 3 kDa > 10 kDa > 3∼10 kDa, and the IC/sub 50/ ACE inhibitory activities by molecular lebels were 4 mg/mL(3 kDa), 5 mg/mL(total hydrolysate), and 20 mg/mL(10 kDa), respectively. The storage stability of dried laver hydrolysates at 20℃ were strongly affected by drying methods, hot air dried of which were much stabler than freeze-dried one.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.20 no.5
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• pp.431-440
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• 1987

In order to develop a new type of food source for the effective utilization of fish protein, plastein reaction was applied to improve the functional properties of filefish protein. Plasteins were synthesized from a peptic filefish protein hydrolysate by papain, pepsin, $\alpha-chymotrypsin$ and protease(from Streptomyces griceus) under the optimum conditions of previous paper). Also, L-glutamic acid diethylester and L-leucine ethylester were incorporated into plastein during the plastein reaction by papain. And, General composition, yield, molecular weight, amino acid composition, color and IR spectrum of plasteins were measured. The protein, ash and lipid content of the plasteins were $72\~78\%,\;7.4\~11.8\%\;and\;0.3\~0.9\%$ respectively. The yield of plasteins were papain $55.0\%,\;pepsin\;47.6\%,\;\alpha-chymotrypsin\;38.3\%,\;protease\;23.6\%$, glutamic acid-incorporated plastein (Glu-Plastein) $35.0\%$, and leucine-incorporated plastein (Leu-plastein) $45.7\%$. The glutamic acid and leucine content in Glu-plastein and Leu-plastein were $38.7\%,\;41,7\%$, respectively, while the contents in the peptic filefish protein hydrolysate were $16.01\%\;and\;8.16\%$, respectively. The amino acid compositions were similar to that of the original filefish muscle protein. The major molecular weights of the peptic hydrolysate estimated by gel filteration were 2,000 and 310, and those of plasteihs were 21,000 and 4,900 for papain, 24,000 for pepsin, 18,500 for $\alpha-chymotrypsin$ 6,700 for protease, 24,000 for Glu-plastein and 17,000 for Leu-plastein. The structural changes in freeze-dried filefish meat, the FPC and hydrolysate were not observed on the IR spectrum. But plasteins showed amide I band in $1,600\~l,700cm^{-1}$ range and resulted in a strong band in $800\~850\;cm^{-1},\;700\~750\;cm^{-1}\;and\;650\~700\;cm^{-1}$. The amide I band of Glu-plastein was wider than those of other plasteins and had also a small band at $1,440\;cm^{-1}$.

• Korean journal of food and cookery science
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• v.6 no.4 s.13
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• pp.1-8
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• 1990

For the purpose of supplying the imformation to improve the acceptability of soy paste as the condiment, the changes of enzyme activity, general component and flavor compounds (Free amino acid, Nucleic acid related compounds, and peptide) during improved soy paste fermentation were determined. The results were as follows; 1. The protease activity during fermentation were increased continuously, but amylase activity were decreased in 45 day fermentation. Cellulase activity were slowly increased until 45 day, and then slowly decreased. 2. Total nitrogen contents were almost constant during fermentation, but amino nitrogem were increased rapidly. Reducing sugar were not constant, but increased in the end of fermentation. PH were decreased to pH 4.97. 3. Total contents of free amino acid as flavor compound were rapidly increased in 10 day fermentation, but were constant in $30{\sim}60$ day. Aspartic acid contents were increased continuously, but glutamic acid were increased slowly until 30 day fermentation and were almost constant. IMP and GMP contents showed increasing pattern during fermentation.

• Journal of the Korean Wood Science and Technology
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• v.32 no.5
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• pp.39-44
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• 2004

This study was carried out to investigate the tenderizing effect of Neungi mushroom (Sarcodon aspratus) powder and its protease. The addition of Neungi mushroom powder and its protease enhanced water retention values (WRY) of meat. The WRY of meat was increased 26.8% by protease addition, compared to 13.8% WRV by sugar addition. This increase in WRY derived to the increase of water soluble fraction in the meat texture by hydrolysis of meat protein, and had the meat tenderized. Concerned to the meat tenderizing effect, the addition of Neungi mushroom powder and its protease have decreased of meat hardness and gave similar tenderizing effect, as compared to commercial tenderizer, papain. The decreasing rates of meat hardness were 51.6% of Neungi mushroom powder, 58.5% of its protease, and 563% of commercial tenderizer, papain. This tenderizing effect of protease attributed to the degradation of muscle fiber protein in meat, such as actin, myosin and connectin etc. The addition of Neungi mushroom to foods gives significant changes in food color, mainly decreasing lightness.

• Korean Journal of Agricultural Science
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• v.11 no.1
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• pp.120-132
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• 1984

In order to clarify the effect of the fish meat hydrolysate on the growth of lactic acid bacteria(Str. lactis, Str. thermophibus, L. bulgaricus, L. acidophilus, and L. helveticus), the optimum conditions for hydrolyzing the fish meat were examined, and changes of the acid production, viable cell count of lactic acid bacteria and the charge of pH of the culture medium by addition of the fish meat hydrolysate were tested. The results were as follows: 1. When the hydrolysis of back muscle of mackerel was proceeded at $50^{\circ}C$ and at pH 8, for 48 hours adding 6% pancreatin of the protein content in the substrate, the best result was obtained. 2. The composition of the fish meat hydrolysate were 53.6% moisture, 32.4% protein, 1.0% fat, 10.7% carbohydrate, and 3.2% ash. 3. Above 0.1% of the fish meat hydrolysate in the culture medium, the acidity of the culture medium by Sir. lactis and Str. thermophilus were increased remarkably. The acidity of the culture medium by L. acidophilus and L. helveticus were increased in above 0.2% fish meat hydrolysate in the culture medium. but L. bulgaricus was not effected by the fish meat hydrolysate. 4. The pH of the culture medium during incubating Str. laclis and Sir. thermophilus failed obviously by adding the fish meat hydrolysate. But in the cases of L. bulgaricus, L. acidophilus, and L. helveticus, the pH were not changed clearly. 5. The viable cell count in all bacterial strains tested here were elevated by increasing the concentration of the fish meat hydrolysate.

• Microbiology and Biotechnology Letters
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• v.15 no.6
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• pp.375-380
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• 1987

D-$\alpha$-Amino-$\varepsilon$-carpolactam racemase (EC 5.1.1) and L-$\alpha$-amino-$\varepsilon$-caprolactam hydrolase (EC 3.5.2) were fractionated from cell-free extracts of Alcaligenes eutrophus A52 using ammonium sulfate precipitation and DEAE-cellulose ion exchange chromatography. It was made sure that D-$\alpha$-amino-$\varepsilon$-caprolactam was converted to L-$\alpha$-amino-$\varepsilon$-caprolactam by racemase, and then hydrolyzed into L-lysine by hydrolase in Alcaligenes eutrophus A52. For the conversion of D-$\alpha$-amino-$\varepsilon$-caprolactam into L-lysine by cell-free extracts of Alcaligenes eutrophus A52, the optimum temperature and pH were 6$0^{\circ}C$ and 8.5 respectively. The results showed that 0.5% D-$\alpha$-amino-$\varepsilon$-caprolactam was converted to L-lysine at 55$^{\circ}C$ for 10 hr with a conversion rate of 98% by cell-free extracts containing 3.1mg of protein.

• Korean Journal of Food Science and Technology
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• v.32 no.1
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• pp.161-167
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• 2000

The protease produced by a newly isolated Aspergillus wentti from Korean traditional Meju was purified and characterized. The optimal medium composition and culture conditions for maximum protease production were ; bran :1% glucose solution =1 : 1, pH 9.0, $30^{\circ}C$, and 4 days of fermentation. Protease was purified by QAE-Sephadex, SP-Sephadex ion exchange chromatography and Sephadex G-100 chromatography. The specific activity and the purification fold of the purified enzyme were 213 unit/mg protein and 27.3, respectively. The molecular weight of purified protease was found to be 32 kDa by SDS-PAGE. Km and Vmax value's for hammastein milk casein were $3.049{\times}10^{-4}\;M\;and\;151.1\;{\mu}g/min$, respectively. Kinetic parameters showed that the enzyme has higher affinity to casein than isolated soybean protein, hemoglobin and bovine serum albumin. Optimal pH and temperature for reaction of the purified enzyme were 9.0 and $50^{\circ}C$, respectively. The enzyme was stable at pH 4.0-11.0, below $40^{\circ}C$, and the activity was not stimulated by metal ions. 1mM phenylmethylsulfonyl fluoride inhibited the enzyme activity by 98.5%. It means that the enzyme is one of serine protease.

• Microbiology and Biotechnology Letters
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• v.40 no.1
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• pp.1-9
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• 2012

This study's aim was to isolate microorganisms producing agarase with a high activity, with possible applications in improving the performance of the pretreatment processes for bioethanol production. Marine algaes were collected from the south coast of Korea, from which three kinds of microorganisms were isolated. After a 4-day culture of these strains at $25^{\circ}C$, crude enzymes were obtained from culture supernatant or cell-free extract by ammonium sulfate precipitation and membrane dialysis. Agarase activity was observed in these crude enzymes. Notably higher specific activity was observed in the crude enzyme obtained from the culture supernatant rather than that from the cell-free extract. This indicates that a secreted enzyme has a much greater activity than a cellular enzyme. Crude enzymes from the GNUM08122 strain were inferred to have ${\alpha}$-agarase activity because release of p-nitrophenol was observed, possibly due to the cleavage of p-nitrophenyl-${\alpha}$-D-galactopyranoside. The 16S rRNA sequence of GNUM08122 showed a close relationship to Pseudoalteromonas issachenkonii KMM 3549 (99.8%) and Pseudoalteromonas tetraodonis IMA 14160 (99.7%), which led us to assign it to the genus Pseudoalteromonas. Biochemical and physiological study revealed that this strain can grow well at $40^{\circ}C$ under a wide range of pH (pH 4~8) in high-salt conditions (10% NaCl).