• Korean Journal of Microbiology
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• v.14 no.2
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• pp.47-56
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• 1976

Enzyme activities, such as glucoamylase dextrinogenic amylase, cellulase, acid protase and neutral protease, of Rhizopus isolated from various substrates collected throughout South Korea are measured, and their enzyme activities are surveyed from taxonomical, ecological and physiological viewpoint. Effect of carbon sources and phytohormones on the amylalse production of Rhizopus are also measured. Among the 735 strains of Phizopus isolated, strain number 587 exhibiting most prominent dextrinogenic amylase and netral protease activity is selected as the best strain, and the strain number 673, 108, 329, 165 and 728 are seleted for their predominant cellulase, acid protease, glucoamylase, dextrinogenic amylase and neutral protease activities, respectively. R.acidus and R.nigricans which exhibited relatively higher callulalse activity, showed lower activities for both amylase. R.tritici exhibited higher protease activity. The relations between activities and various substrates of wild strains are not outstnading difference, although the strains isolated from inland region exhibited more or less higher amylase and cellulase activities, than those of coast region, generally. Lactose and dextrin are most effective carbon sources for glucoamylase and dextrinogenic amylase production of the Rhizopus niveus, respectively. Although all phytohormones tested are effective for production of amylase by the Rhizopus strains, except nicotinamide for glucoamylase production, biotin and ascorbate are most effective for dextrinogenic amylase and glucoamylase production, respectively.

• Microbiology and Biotechnology Letters
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• v.20 no.3
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• pp.295-301
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• 1992

An alkaline protease producing microorganism was isolated from soil and identified as Streptomyces griseus HC-1141. The optimum pH and temperature for the purified enzyme activity were 8.0 and $60^{\circ}C$, respectively. The enzyme was relatively stable in the pH range of 7.0-9.0 and at the temperature below $60^{\circ}C$. The activity of purified enzyme was inhibited by $Hg^{2+}$, $Cu^{2+}$, $Zn^{2+}$, $Ba^{2+}$ and $Fe^{2+}$, whereas activated by $Mn^{2+}$ and $Ca^{2+}$. $\varepsilon$-Amino caproic acid, 2,4-dinitrophenol and iodine did not show inhibitory effect on the activity of alkaline protease, but p-chloromercuribenzoic acid, ethylendiaminetetraacetic acid showed inhibitory effect on the enzyme activity. These result suggested that the protease was metalloenzyme, and require a reactive SH group for the activity. The reaction of this enzyme follows typical Michaelis-Menten kinetics with the $K_m$ value of $2.229{\times}10^{-4}$M and the $V_{max}$ of $46.08 {\mu}$g/min for casein. The activation energy for the alkaline protease calculated by Arrhenius equation was 3.643 kcal/mol. This enzyme hydrolyzed casein more rapidly than the hemoglobin and egg albumin.

• Korean Journal of Microbiology
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• v.18 no.2
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• pp.51-58
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• 1980

Mutational experiments were performed to improved to improve the protease productivity of Aspergillus flavus KU 153, which is selected among the wild strains. A UV-induced mutant strain having high protease productivity was obtained by the use of the clear zone method as a simple criterion for a primary screening test. Neutral and alkaline protease activities of hte mutant strain were higher than 1.8 times, comopared with those of the parental strain, respectively, while in the case of acid protease, it was 2.7 times. The mutant strain selected was more powerful in the production of cellulase and amylase, as well s protease in wheat bran, compared with those of the parental strain. protease production of the parental strain has reached maximum level at 3 days culture, while alkaline nad neutral protease production of the mutantstrain has reached at 2 days culture. On the other hand, the mutant strain formed the spore slowly, compared with the parental strain. Column chromatography of the neutral protease on DEAE-Sephadex A-50 showed that the mutant strain was not induced the formation of another neutral protease isozyme, but induced the variation in the function of regulatory gene.

• Journal of Microbiology and Biotechnology
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• v.8 no.6
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• pp.639-644
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• 1998

Extracellular serine proteases were isolated from a soil bacterium, alkalophilic coryneform bacterium TU-19, which have been grown in a liquid medium optimized at 3$0^{\circ}C$ and pH 10.0. Three different sizes, 120 kDa (protease I), 80 kDa (protease II), and 45 kDa (protease III), of serine pro teases were purified using Sephadex G-150 and QAE-Sephadex chromatography (Kang et al. 1995. Agric. Chem Biotech. 38: 534-540). SDS-PAGE showed that the 120 kDa protease was degraded into the 80 kDa protease in 20 mM Tris-HCI (pH 8.0) buffer solution. This degradation was enhanced in the presence of 0.5 M NaCl and 5 mM EDTA, but was inhibited in the presence of 5 mM $CaCl_2$. These results indicated that the $Ca^{2＋}$ ion seems to stabilize the 120 kDa protease like other proteases derived from Bacillus species. The $NH_2$-terminal amino acid sequences of the 10 residues of both proteases were completely identical: Met-Asn-Thr-Gln-Asn-Ser-Phe-Leu-Ile-Lys. In contrast to this, the 80 kDa protease has 1.5 times higher specific activity than the 120 kDa protease does (Kang et al. 1995. Agric. Chern. Biotech. 38: 534-540). Therefore the C-terminal of the 120 kDa protease seems to be autolyzed to the 80 kDa protease but this autolysis did not decrease the protease activity. Optimum pH and temperature of both 80 kDa and 120 kDa proteases were pH 10.5 and $45^{\circ}C$, respectively, and pH and thermal stability were almost identical. Several divalent ions except the $Fe^{2＋}$ ion showed similar effects on activities of both proteases, which are similarly resistant to three different detergents.

• Journal of Animal Science and Technology
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• v.48 no.1
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• pp.49-58
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• 2006

This study was conducted to evaluate effects of mud flat bacteria origin protease supplementation on growth performance, amino acid digestibility, blood characteristics, meat quality, fecal VFA (volatile fatty acids) and NH3-N (ammonia nitrogen) concentration in finishing pigs. Eighty pigs (Landrace×Yorkshire×Duroc, 60.08±2.69 kg average initial body weight) were used during experimental period. Dietary treatments included 1) high nutrient density diet, 2) high nutrient density diet+0.1% protease, 3) low nutrient density diet and 4) low nutrient density diet+0.1% protease. For overall period, ADG was improved in treatment of high nutrient density diet added protease compared with treatment of low nutrient density diet without protease (P<0.05). DM and N digestibilities were improved in treatments of high nutrient density diet and low nutrient density diet added protease compared with treatment of low nutrient density diet without protease (P<0.05). Essential amino acid digestibility was improved in treatment of low nutrient density diet added protease compared with other treatments (P<0.05). Nonessential amino acid digestibility was improved in treatment of high nutrient density diet added protease compared with treatments of high and low nutrient density diet (P<0.05). BUN (blood urea nitrogen)　concentration in blood was increased in treatment of high nutrient density diet added protease compared with treatment of low nutrient density diet without protease (P<0.05). L*value of M. longissimus dorsi muscle was increased in treatments of low nutrient density diet compared with treatments of high nutrient density diet (P<0.05). In conclusion, mud flat bacteria origin protease was effective for improving growth performance, amino acid digestibility and influencing BUN concentration and meat color in finishing pigs.

• Food Science and Biotechnology
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• v.15 no.1
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• pp.28-32
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• 2006

Protease inhibitor of 72.6 kDa was successively purified from chum salmon (Oncorhynchus keta) eggs by ion exchange, gel permeation, and affinity chromatographies. Protease inhibitor was purified with yield and purification fold of 1.50% and 58.11, respectively. SDS-PAGE results showed purified protease inhibitor consisted of two protein subunits of 54.0 and 18.6 kDa. Chum salmon inhibitor exhibited stability between 20 and $40^{\circ}C$ in weak acid environment (PH 6), and inhibited papain and cathepsin, members of cysteine protease, but not chymotrypsin. The protein inhibited cathepsin more effectively than did egg white protease inhibitor, whereas the reverse was true for papain. These results indicate chum salmon egg inhibitor is heterodimer, thus the inhibitor was classified as cysteine protease inhibitor.

• Korean Journal of Food Science and Technology
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• v.30 no.6
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• pp.1333-1338
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• 1998

Several enzymes of kimchi ingredients were assayed to improve the product quality using these quality related enzyme information. Among various hydrolases, amylase and protease were selected with respect to lactic acid fermentation. Peroxidase and ascorbic acid oxidase were studied for off flavor production and ascorbic acid destruction. The amount of protein in kimchi ingredients, specific and total enzyme activity of sample were compared. Regarding total enzyme activity of sample, ${\alpha}-amylase$ activity of salted and fermented anchovy, dried red pepper and salted and fermented shrimp were higher than other ingredients. Activity of salted and fermented anchovy was 2,790.0 units/g sample. Salted and fermented anchovy, oyster and Chinese radish showed the highest ${\beta}-amylase$ activity (4.4, 2.1, 1.0 units/g sample, respectively). Salted and fermented anchovy showed the highest protease activity of 13.4 PU/g sample, followed by salted and fermented shrimp and dried red pepper. For peroxidase, Chinese radish, cucumber, green onion showed the highest activity of 7.2, 6.8 and 5.6 units/g sample, respectively. In case of ascorbic acid oxidase, salted and fermented anchovy showed the strongest enzyme activity (331.4 units/g sample), followed by dried red pepper and salted and fermented shrimp.

• Journal of the Korean Society of Food Science and Nutrition
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• v.18 no.3
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• pp.348-355
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• 1989

This experiment was conducted to investigate the characteristics of alkaline protease from Aspergillus fumigatus which was isolated from soil as a superior strain for the production of the alkaline protease. The optimum temperature for enzyme activity was $50^{\circ}C$ and optimum pH was 9.0. The enzyme was stable at pH 8.0 to 10.0 and thermal inactivation was shown $30^{\circ}C$. The activity of the enzyme was increased by the addition of $Mn^{++},\;Cu^{++},\;Ba^{++},\;Mg^{++},\;$wheras it was inhibitied by $K^+,\;Fe^{+++},\;Ag^+,\;Pb^{++},\;Na^+,\;Ca^{++},\;Hg^+,\;Zn^{++}$. EDTA. 2, 4-DNP, ${\varepsilon}-amino$ caproic acid did not show inhibitory effect on the proteolytic activity of alkaline protease but P-chloromercuribenzoic acid inhibited the enzyme activity, indicating that reactive sulfhydryl group is required for the enzymatic activity. The reaction of this enzyme followed typical Michael-Menten Kinetics with the Km value of $8.33{\times}10^{-4}mole/{\ell}$ with the Vmax of $47.62{\mu}g/min$. This enzyme had stronger proteolytic activity than trypsin on substrate such as casin and hemoglibin.

• Food Engineering Progress
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• v.15 no.1
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• pp.41-47
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• 2011

The defatted rice bran (DRB) was enzymatically hydrolyzed using eight commercial proteases for 4hr at optimum pH and temperature. Proteolytic hydrolysates were examined in supernatant and precipitate using lowry, semimicro kjeldahl and gravimetric method using weight difference before and after enzymatic hydrolysis. In lowry and kjeldahl protein assay method, two proteases (Alcalase and Protease N) were found to be the most effective enzymes. In gravimetric method, 60.6~118.3 mg protein/g DRB was hydrolyzed after eight commercial proteases treatments. Similar to lowry and kjeldahl method, 118.3 and 107.1 mg protein/g DRB were hydrolyzed after Alcalase and Protease N treatments, respectively. When two or three effective proteases (Protamex, Alcalase and Protease N) were applied at one time to obtain synergistic effect, significant increase (P<0.05) was observed when three proteases were applied at one time (63.4 mg protein/g DRB in lowry method and 204.5 mg protein/g DRB in gravimetric method). This result suggests that Alcalase and Protease N were the most effective enzymes for proteolysis of DRB and three commercial enzymes (Protamex, Alcalase and Protease N) showed the synergistic effect on the hydrolysis of DRB.

• The Korea Journal of Herbology
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• v.22 no.4
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• pp.21-28
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• 2007

Objectives : Kyenegum has been popularly used long as the digestive. The purpose of this study was to investigate the purification and characteristics of protease obtained from Kyenegum crude enzyme. Methods : Kyenegum protease was purified by precipitation with ammonium sulfate followed by SP-Spharose ion exchange chromatography. The molecular weight of Kyenegum protease was estimated by SDS-PAGE electrophoresis. Results : Kyenegum protease was 3,087 units/mg protein specific activity, 14.5 purification fold and 9.8 % recovery. The molecular weight of protease was estimated to be 18 kDa. The isoelectric point was pI 8.97 and values of Km and Vmax of its were 48 mg/mL and 2 units/min, respectively. Conclusion : The result suggests that the protease obtained from Kyenegum has excellent stability of temperature, acid and collagen substrate specificity.