• The Korean Journal of Mycology
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• v.26 no.4 s.87
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• pp.496-501
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• 1998

The mycelia of Tricholoma matsutake DGUM 26001, 26101, 26210 and FRI 91024 were used to determine the extracellular enzyme activity in mycelia. When the filtrate of culture broth after 30-day cultivation at $24^{\circ}C$ was used as a crude solution of extracellular enzyme, the average specific activity of ${\alpha}-amylase$ was 6142.3 unit/mg protein. The specific activity of xylanase was comparatively high. However, little or no enzyme activities were found for ${\beta}-glucosidase$, ligninase, CMCase, chitinase, protease, and lipase.

• Microbiology and Biotechnology Letters
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• v.35 no.4
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• pp.304-309
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• 2007

Soybean is useful source of protein, especially in Asia. But soybean needs heat inactivation or fermentation process before consumption, since it contains the toxic lectin and various protease inhibitors. Therefore, production of soybean boiling-waste liquor (SBWL) as a byproduct is inevitable. In this study, the chemical composition of SBWL and the optimization of culture conditions for Bacillus pumilus JB-1, a selected strain for functional chungkuk-jang fermentation, using SBWL were investigated. The SBWL contains 88% water, 9.5% free sugar, 1.6% crude protein, 0.3% crude fat, 0.1% crude fiber and 2.1% ash, respectively. The contents of total polyphenol, total flavonoids and free amino acid in SBWL were 55%, 76%, and 30% of those of raw soybean, respectively. Culture conditions for B. pumilus JB-1 in SBWL were optimized. The 1/10-diluted, 0.1 % of $(NH_4)_2SO_4$ added SBWL without pH adjustment and carbon source addition was cultured at $37^{\circ}C$ for 48 h with agitation (120 rpm). The 0.5% of inoculation was enough. The large scale fermentation in 5-L jar fermentor showed that the SBWL is a good resource for production of chungkuk-jang starter and functional ingredients.

• Microbiology and Biotechnology Letters
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• v.7 no.3
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• pp.157-164
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• 1979

These experiments were performed to investigate the culture condition, characteristic of crude enzyme and the heat resistance of the acid protease by Aspergillus awamori U-3. The results obtained were as follows: 1. The optimum culture temperature and time on wheat bran medium and defatted rice bran medium were 3$0^{\circ}C$ and 72 hrs, respectively. The optimum amount of added water was 100~120 ％ on wheat bran medium and 100~130 ％ on de-fatted rice bran medium. 2. Of the these various ingredients, addition of KN $O_3$, glutamic acid and glucose on wheat bran medium and addition Of KN $O_3$, (N $H_4$)$_2$S $O_4$, glucose, lactose, K $H_2$P $O_4$ and MgC $l_2$ on defatted rice bran medium were very effective. On wheat bran medium, concentration of addition of glucose, KN $O_3$ and glutamic acid were 3.0~4.0％, 0.2~0.4 ％ and 1.0％, respectively. 3. The optimum pH for the enzyme action was 2.4 ％, the optimum temperature about 45$^{\circ}C$ and the stable pH range 2.0~5.0, The enzyme was stable below 5$0^{\circ}C$ and was inactivated rapidly above 5$0^{\circ}C$. 4. The addition of CaC $l_2$ and CaS $O_4$ as the heat resistance agents showed the slight resistance. 5. When the enzyme solution added with the heat resistance agents (CaC1$_{2}$ and CaS $O_2$) was heated for 10-30 minutes at 6$0^{\circ}C$, their remaining activities were decreased largely above 20 minutes and The heat resistance effects of CaC $l_2$ and CaS $O_4$ were not observed almost at 8$0^{\circ}C$.

• Journal of Life Science
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• v.27 no.7
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• pp.805-811
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• 2017

In the previous paper, we isolated a bacterium that can hydrolyze various organic materials from soybean paste, including cellulose, lipids, starch, and protein. The activity and chemical properties of the crude enzymes produced by the isolate Bacillus subtilis CK-2 were further investigated. Cellulase showed the highest activity at pH 5.0 and $55^{\circ}C$. The stability of cellulase was maintained within the ranges of pH 5.0~10.0 and $20{\sim}50^{\circ}C$. Cellulolytic enzymes were activated by a $Co^{2+}$ ion, demonstrating the highest activity at a 0.45%(w/v) concentration of $Co^{2+}$. The optimal conditions for amylase were pH 5.0 and $50^{\circ}C$. The activity of amylase was stable within the ranges of pH 4.0~5.0 and $20{\sim}50^{\circ}C$. The $Co^{2+}$ ion was also necessary for amylase activity, which was the highest at a 0.2%(w/v) concentration of $Co^{2+}$. The optimal pH and temperature conditions of protease were pH 8.0 and $50^{\circ}C$. The activity of protease was stable within the ranges of pH 7.0~8.5 and $20{\sim}50^{\circ}C$. Protease activity was catalyzed by $Mn^{2+}$, which was the highest at a 0.125%(w/v) concentration of $Mn^{2+}$. The isolate B. subtilis CK-2 demonstrated a high activity of autolysin. Based on these results, we identified and suggested the optimal pH, temperature, and metal ion concentration in the use of the hydrolytic enzymes of B. subtilis CK-2 for industrial purposes.

• Applied Biological Chemistry
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• v.22 no.3
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• pp.135-141
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• 1979

These experiments were conducted to investigate the conditions of the acid protease production by Rhizopus oryzae and the characteristics of crude enzyme. The results obtained were as follows: 1. The optimum culture time and the optimum amount of added water to the wheat bran medium were about 48 hrs and $80{\sim}120%$, respectively. 2. The addition of $(NH_4)_6Mo_7O_{24},\;(NH_4)_2SO_4,\;NH_4NO_3$, casein, and albumin, respectively, as nitrogen sources to the wheat bran medium was effective. Of these, the optimum concentrations of addition of $(NH_4)_6Mo_7O_{24}$ and casein which were the most effective were 0.1% and 1.0%, respectively. 3. The addition of glucose, galactose, maltose, lactose, and soluble starch, respectively, as carbon sources to the wheat bran medium was effective. Of these, the optimum concentration of addition of glucose which was the most effective was 3.0%. 4. The addition of $KH_2PO_4$ as a phosphate salt to the wheat bran medium was effective. The optimum concentration of addition of $KH_2PO_4$ was 0.3%. 5. The optimum pH for the enzyme action was 2.4, the optimum temperature about $40^{\circ}C$, and the stable pH range $2.0{\sim}5.0$. The enzyme was stab1e below $40^{\circ}C$. 6. The enzyme activity increased rapidly for 10 minutes after addition, thereafter it increased slowly. 7. The enzyme activity increased rapidly to 2 ml of addition, but nearly did not increase at the amounts greater than 2ml.

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.1
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• pp.108-114
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• 2017

This study was performed to evaluate the characterization of fermented mulberry leaf using Bacillus subtilis by focusing on protein degradation and antioxidant activity. The crude protein and total amino acid compositions of mulberry leaf were 21.40% (w/w) and 105.06 mg/g, respectively. The pH level decreased sharply in mulberry leaf extracts fermented using B. subtilis, in accordance with an increase in bacteria cell populations (9.49 log CFU/mL) during 36 h of fermentation. The protease activity of mulberry leaf increased to 97.45 units/mL after 5 days of fermentation. After fermentation, free amino acid contents in fermented mulberry leaf increased from $486.91{\mu}g/g$ to $644.35{\mu}g/g$ due to considerable elevation of isoleucine (6-fold), alanine (4.67-fold), leucine (4.52-fold), and valine (4.21-fold). The DPPH radical scavenging activity of fermented mulberry leaf also increased from 25.93% to 73.22% after 5 days of fermentation. These results suggest that mulberry leaf fermentation using B. subtilis can improve nutritional quality and antioxidant activity.

• Microbiology and Biotechnology Letters
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• v.41 no.1
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• pp.33-43
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• 2013

A bacterium producing a fibrinolytic enzyme was isolated from Cheonggukjang. The bacterium was identified as a strain of Bacillus amyloliquefaciens by 16S rDNA analysis and designated as B. amyloliquefaciens HC188. The optimum culture medium appeared to be one containing 0.5% (w/v) maltose and 0.5% (w/v) soytone. Bacterial growth in the optimal medium at $37^{\circ}C$ reached the stationary phase after 27 h of incubation and the fibrinolytic enzyme showed optimum activity at 24 h. The enzyme was purified by 20-80% ammonium sulfate precipitation, CM Sepharose fast flow ion exchange chromatography, and Sephacryl S-200HR column chromatography. Its specific activity was 38359.3 units/mg protein and the yield was 5.5% of the total activity of the crude extracts. The molecular weight was 24.7 kDa and the amino acids of the N-terminal sequence were AQSVPYGVSQIKAPA. The fibrinolytic enzyme activity had an optimum temperature of $40^{\circ}C$ and an optimum pH of 8.0, and the enzyme was stable in the ranges $20-40^{\circ}C$ and pH 6.0-8.0. Enzyme activity was increased by $Ca^{2+}$ and $Co^{2+}$ but inhibited by $Cu^{2+}$, EDTA, and PMSF. It is suggested that the purified enzyme is a metallo-serine protease.

• Korean Journal of Food Science and Technology
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• v.30 no.5
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• pp.1236-1242
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• 1998

Korean Lentinus edodes SR-1 was cultured to multiply the mycelia in the complete broth medium (C/N=13.1) for mushroom, and protein-bound polysaccharides were extracted from the cultured broth containing mycelia (The whole cultured broth was used to increase the yields: 80% of protein-bound polysaccharides were existed at the cell wall of mycelia and 20% of those were secreted extracellularily in this culture). Protein-bound polysaccharides in the cultured broth containing mycelia were extracted by using three different methods: 1) Extraction with hot water, 2) Disintegration of cell wall by glass bead mill treatment before extraction with hot water, and 3) Cellulase treatment before extraction with hot water. The highest yield was obtained (930 mg polysaccharides/100 mL culture broth) when protein-bound polysaccharides were extracted with 2) method. The extracted crude protein-bound polysaccharides were purified using protease, DEAE-cellulose and Sephadex G-100. The growth inhibition activity for $P_{388}$, mouse leukemic cell, increased (53.7, 62.2, 93.7% and 97.4%) as the purification level increased. Protein-bound polysaccharides contained 46.1% of polysaccharides, 7.3% of protein, and trace amounts of minerals. Polysaccharides contained glucose, galactose, xylose and mannose. The content of proline and glycine were high, however, methionine and leucine were not found. The major minerals were Na, K, Zn, and Ca.

• Korean Journal of Food Science and Technology
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• v.15 no.4
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• pp.385-391
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• 1983

The change of free amino acid contents and nitrogen compounds in the course of the Chungkookjang fermentation that occurred by utilizing Bacillus natto and Bacillus subtilis are to the following effects. pH, during the growth period, that is 6.35 in pH at the first stage of fermentation, were turned into 8.2 after 72 hours. Crude protein content increased irregularly from 16.82%-18% and total sugar decreased. Increasing with the progress of fermentation time, protease activity showed the maximum value between 48-60 hours, but Bacillus natto activated a little than Bacillus subtilis. Amino nitrogen and water soluble nitrogen content increased but difference was found that is, Bacillus natto increased more than Bacillus subtilis. Glutamic acid content was the highest among the contents of free amino acid between both Bacillus sp. and the order of the next contents showed as leucine, phenylalanine, histidine alanine. arginine, but difference was found between Bacillus sp., that is, Bacillus natto was higher than Bacillus subtilis. In view of the results as above, Bacillus natto was excellent than Bacillus subtillus as Bacillus strains of Chungkookjang koji production.

• Journal of Animal Science and Technology
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• v.45 no.5
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• pp.805-812
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• 2003

This study was conducted to examine effects of feeding dry TMR(DTMR), wet TMR(WTMR) and fermented TMR(FTMR) on rumen fermentation, enzyme activity and digestibility in the total tract of sheep. Three rumen cannulated sheep were used in a 3${\times}$3 latin square design. The present results showed that pH, NH3-N, total and individual VFA, A/P ratio and enzymes (CMCase, Xylanase and Protease) activity in the rumen were higher in WTMR and FTMR compared with DTMR. In addition, dry matter, organic matter, crude protein, ether extract, NDF and ADF digestibility in the total tract were also higher in WTMR and FTMR compared with DTMR. Therefore, the present results showed that WTMR and FTMR are better than DTMR for rumen fermentation and nutrients digestibility.