• Microbiology and Biotechnology Letters
• /
• v.26 no.5
• /
• pp.427-434
• /
• 1998

An alkaline protease was 4-fold purified, yielding 2.3% of recovery by ammonium sulfate precipitation, CM-cellulose column chromatography and Sephadex G-100 column chromatography. The purified enzyme was estimated to be monomeric with molecular weight of about 62,000 from polyacrylamide gel eletrophoresis (PAGE) and sodiumdodecylsulfate polyacrylamide gel electrophoresis (SDS-FAGE). The optimal pH and temperature of the alkaline pretense activity were 11.0 and 50$^{\circ}C$, respectively, exhibiting high stability at pH value from 6.0 to 11.0 at 50$^{\circ}C$ for 30 minute. The alkaline pretense was activated by MnSO$_4$, CaCl$_2$, and was inhibited by CuSO$_4$, ZnSO$_4$, HgCl$_2$, EDTA and EGTA. Also, the enzyme was found to be a metaloenzyme requiring Mn$\^$2+/ as cofactor. The NH$_2$-terminal amino acid of alkaline protease was alanine. The Km and Vmax values of this enzyme for casein was 4.0 mg/$m\ell$ and 5,500 unit/$m\ell$, respectively.

• Applied Biological Chemistry
• /
• v.49 no.1
• /
• pp.7-14
• /
• 2006

This study was carried out to examine the quality characteristics of soybean pastes made of Meju with mold-producing pretense isolated from traditional Meju. The changes in moisture content, enzyme activity (amylase, protease, lipase), reducing sugar, amino-type nitrogen contents and anti-oxidant activity were investigated during the aging period. The moisture contents decreased gradually with time. Amylase activity decreased during the aging period while pretense and lipase activities increased until 30-45 days of aging, but decreased thereafter, Especially pretense activity in soybean paste with mold was greater than that in the control. Amino-type nitrogen and ammoniacal nitrogen contents increased, but reducing sugar contents decreased with time. Amino-type nitrogen contents were greater in soybean paste made of Meju with isolated mold than those in the control. Antioxidative activity was also confirmed in soybean paste.

• Preventive Nutrition and Food Science
• /
• v.7 no.4
• /
• pp.397-404
• /
• 2002

Soybean curd residue (SCR) was fermented by lactic acid bacteria, Lactobacillus rhamnosus LS and Entercoccus faecium LL, isolated from SCR. The pH, titratable acidify and viable cell counts were determined from the fermented SCR to evaluate the lactic acid production and growth of lactic acid bacteria. Optimal amounts of pretense enzyme and glucose, and ideal fermentation time for SCR fermentation were estimated by response surface methodology (RSM). Raw SCR fermented by indigenous microorganisms had 0.78 % titratable acidity, The acid production in SCR fermented by L. rhamnosus LS was greatly enhanced by the addition of glucose and lactose. However only glucose increased acid production by Ent. faecium LL. The proof test of SCR fermentation demonstrated that similar results for titratable acidity, tyrosine content and viable cell counts to that predicted could be obtained by the at optimized fermentation conditions. In the presence of 0.029 % (w/w) pretense enzyme and 0.9% (w/w) glucose, the SCR fermented by Ent. faecium LL showed 1.07% (w/v) of titratable acidity, 1.02 mg% tyrosine content and 2$\times$10$^{9}$ (cfu/g) of viable cell counts. With the SCR fortified with 0.033% pretense enzyme and 1.7% glucose, L. rhamnosus LS showed 1.8% (w/v) of titratable acidity, 0.92 mg% of tyrosine content and 2$\times$10$^{9}$ (cfu/g) of viable cell counts.

• Microbiology and Biotechnology Letters
• /
• v.16 no.6
• /
• pp.526-531
• /
• 1988

Procedure for the purification of pretense from culture broth of Streptomyces sp. SMF301 was developed. It was evident that the strain produced two different proteases of which molecular weights were estimated to be 23, 500 and 38, 900 dalton. It was found that the optimum pH of the smaller was 9.0 and that of the larger was 1.0. The optimal temperature of the alkaline pretense was 5$0^{\circ}C$ and that of the neutral pretense was much more stable than neutral protease at extreme condition viz. high temperature, and pH.

• Applied Biological Chemistry
• /
• v.49 no.4
• /
• pp.304-308
• /
• 2006

To extract insoluble proteins from silkworm pupa meal, the meal was treated with pretense produced by Bacillus sp. JH-209. The extraction of insoluble silkworm pupa protein was enhanced at alkaline pHs ranged from 7 to 11 by treatment with the protease. The optimum extraction temperature was $40^{\circ}C$ for in soluble protein treated with pretense. The optimum protease treatment time for extraction of protein was 11 hrs and optimum amount of enzyme treated for extraction of protein was 60 Unit, respectively. The treatment of enzyme extracted more protein than ordinary extraction method without pretense. The foaming capacity, foaming stability, emulsion capacity, and emulsion stability of silkworm pupa meal protein extracted by the treatment of the enzymes increased at all pH ranges. Further more oil absorption as well as water absorption capacities of the protein extracted by the treatment of the enzymes were also increased.

• Journal of Aquaculture
• /
• v.18 no.4
• /
• pp.245-251
• /
• 2005

This study was investigated the condition of their maximum activity to assay the enzymes of rotifer, Brachionus rotundiformis의 $\alpha$-amylase, total alkaline Protease, trypsin and TG-lipase activities of rotifer were higher and more sensitive in phosphate-NaOH buffer than Tris-HCl buffer. $\alpha$-amylase, trypsin and TG-lipase activities were appeared the maximum at pH 8.0, and total alkaline protease activity showed the maximum activity at pH 7.0. $\alpha$-amylase activity showed the highest activity at $40^{\circ}C$, and total alkaline protease and trypsin activities were assayed the highest at $55{\~}60^{\circ}C$. However, TG-lipase activity was appeared the highest at $25{\~}30^{\circ}C$. The optimum substrate concentration of enzyme activity of a-amylase, total alkaline protease, rypsin and TG-lipase were $3.5\%$ starch, $\0.6%$ azo-casein, $87.5{\mu}M$ BApNA and 81.2 mM olive oil, respectively. The optimum reaction time of enzyme activity of $\alpha$-amylase, total alkaline protease, trypsin and TG-lipase were increased up to 40, 60, 30 and 25 min., respectively. The data obtained in this study could be used for the digestive enzyme research of rotifer, B. rotundiformis.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.22 no.3
• /
• pp.138-146
• /
• 1989

This experiment was conducted to utilize the water soluble protein from the fish meal in wastewater as nitrogen source by alkaline protease producting bacteria and to investigate the culture condition of the production. G-12 and G-14 strains having the strong activity of the alkaline pretense were isolated from sea water. These strains were identified as Pseudomonas chlororaphis and Pseudomonas alcaligenes according to physiologycal characteristics, respectively. In enzyme production, galactose and casein for G-12 strain, and raffinose and the water soluble protein of the fish meal wastewater for G-14 strain was favorable as carbon and nitrogen source. An action of inhibition appeared in all of the metal salts used. The optimal temperature of enzyme production was $30^{\circ}C$ for all strains. Optimal initial pH for the enzyme formation in G-12 and G-14 strains was pH 10.0 and 8.0. When these two strains were incubated for $30\~35$ hours in the optimal production medium, the enzyme production reached at maximum.

• Microbiology and Biotechnology Letters
• /
• v.34 no.4
• /
• pp.323-328
• /
• 2006

A bacterium producing the extracellular protease was isolated from insect-eating plant and has been identified as a member of the genus Bacillus based on partial 165 rRNA sequences. In order to develop the medium composition, effects of ingredients including nitrogen sources, carbon source, metal ions and phosphate were examined for protease production of the isolate, SH-8. Soluble starch increased the protease productivity, while glucose repressed it. Yeast extract was effective nitrogen source for enzyme production, but the pretense production of Bacillus sp. SH-8 was reduced by large amount of yeast extract. The calcium was found to induce pretense activity as well as protease productivity. However, cell growth and enzyme production was completely inhibited by divalent ions such as $Zn^{2+}$, $Cu^{2+}$, $Co^{2+}$ and $Mn^{2+}$. The maximum protease productivity was reached 435 unit/ml in the optimized medium consisting of soluble starch (2%), yeast extract (0.3%), $CaCl_2$ (0.3%), $K_2HPO_4$ (0.01%) and $KH_2PO_4$ (0.01%). The pretense activity of culture filtrate was dramatically decreased after incubation for 26 h.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.17 no.4
• /
• pp.312-319
• /
• 1988

The functional properties of plasteins have been compared with those of sardine protein concentrate and egg albumin. The solubility of plasteins was higher than that of FPG and the glu-plastein had 84% solubility in the range of pH 3-10. The dispersibility of plasteins was lower than that of egg albumin, however those of plasteins was higher than that of sardine protein concentrate. The water holding capacity of plasteins was higher than that of egg albumin. Lipid absorption of leu-papain plastein was the highest, holding 2.2m119, and that of the other plastein was higher than that of egg albumin. The emulsifying activity of leu-papain plastein was the highest, holding 66.4%, and that of glu-papain plastein was the lowest, holding 51.2%, The emulsifying stability of plasteins was similar to that of the emulsifying activity. The foaming capacitt of leu-papain plastein was the highest, holding 460%, and those of the other plasteins was higher than that of egg albumin. The foaming stability of plasteins was superior to that of egg albumin. The viscosity of plasteins was lower than that of see albumin. The in vitro digestibility of plasteins was 67.6-78.0% range. The digestibility by four pretense were somewhat lower in the glu-papain plastein than in the FPG. The digest of plasteins treated with the microbiol pretense such as molsin and pretense(from Streptomyces griceus), which had a storage broth taste.

• Applied Biological Chemistry
• /
• v.39 no.6
• /
• pp.460-465
• /
• 1996

The production of bacterial protease and its characteristics were investigated with Bacillus subtilis globigii CCKS-118 which was isolated from Korean traditional soy sauce. The optimum culture condition of the strain for the production of alkaline protease was as follow : 2% soluble starch, 0.2% yeast extract, 0.1% $(NH_4)_2SO_4$, 0.2% $MgSO_4$, pH 7.5, $35^{\circ}C$ and 20h rs. The optimum pH and temperature for the enzyme action of alkaline protease producing Bacillus subtilis globigii CCKS-118 were pH 9.0 and $50^{\circ}C$, respectively. The enzyme was relatively stable at $pH\;6.0{\sim}9.0$ and at temperature below $40^{\circ}C$. The activity of the enzyme was inhibited by $Hg^{2+}$ whereas $Cu^{2+}$ gave rather activating effects on the enzyme activity. The enzyme was inhibited by phenylmethane-sulfonyl fluoride indicating serine pretense metal ion group are required for the enzyme activity. Km value was $1.242{\times}10^{-4}M$, $V_{max}$ value was $25.99\;{\mu}g/min$. This enzyme hydrolyzed casein more rapidly than the hemoglobin.