• Food Science and Preservation
• /
• v.10 no.1
• /
• pp.80-88
• /
• 2003

In order to establish the enzymatic hydrolysis system improving of taste and flavor in the preparation of soy protein hydrolysates using the enzymes with excellent hydrolytic ability and different hydrolysis pattern of soy protein, Degree of hydrolysis(DH) and surface hydrophobicity under the optimal conditions of enzyme reaction, hydrolysis patterns by the SDS electrophoresis and sensory evaluation of soy protein hydrolysates by enzyme reactions were investigated. Four enzyme reactions were highly activated at pH 7.0, 45$^{\circ}C$ under the optimal conditions. As result of changes on the pattern of soy-protein hydrolysates by SDS-electrophoresis, high molecular peptides of hydrolysates by No. 5(Mucor circinelloides M5) and No. 16(Bacillus megaterium B16) enzymes were slowly decrease and 66KD band of these were remained after 3hours reaction. Production of low molecular peptides of hydrolysates by No. 4(Aspergillus oryzae M4) and No. 95(Bacillus subtilis YG 95) enzymes were remarkably detected during the proceeding reactions. As results of HPLC analysis, low molecular peptides of 15∼70KD were mainly appeared during the proceeding enzyme reactions. And, the more DH was increased, the more SDS-surface hydrophobicity was decreased. Hydrolysates by No. 4 enzyme was not only the highest DH of all hydrolysates, but the strongest bitter taste in a sensory evaluation. Sweat taste among the hydrolysates showed little difference. But, when combinative enzymes were treated, combinative enzyme of No. 4(Aspergillus oryzae M4)and No. 16(Bacillus megaterium B16) showed the strongest sweat taste. In conclusion, we assumed that it will be possible to prepare the hydrolysates having functionality when soy-protein were hydrolyzed by these specific enzymes.

• Microbiology and Biotechnology Letters
• /
• v.20 no.5
• /
• pp.519-526
• /
• 1992

The gene coding for urease of alkalophilic Bacillus pasteurii had been cloned in Escherichia coli previously. The urease protein was purified 63.1-fold by TEAE-cellulose, DEAE-Sephadex A-50, Sephadex G-150 and Sephadex G-200 chromatographies with a 7.3% yield from the sonicated fluid of the E. coli HB1Ol(pBUll) encoding B. pasteurii urease gene. The ureases of E. coli (pBUll) and B. pasteurii possessed as a $K_m$ for urea, 42.1 mM and 40.4 mM, respectively. They hydrolyzed urea with $V_{max}$ of 86.9$\mu$mol/min and 160$\mu$mol/min, respectively. Both ureases were composed with four subunits (Mrs 67,000) and a subunit (Mr 20,000). The molecular weight of both native enzymes was Mr 280,OOO$pm$10,000 determined by gel filtration chromatography and Coomassie blue staining of the subunits. The optimal reaction pH of both ureases were pH 7.5. The ureases were stabled in pH 5.5-10.5. The optimal reaction temperature of both ureases were $60^{\circ}C$, and the ureases were stable for an hour at $50^{\circ}C$, 40min at $60^{\circ}C$ and 10 min at $70^{\circ}C$ The activity of both enzymes were inhibited completely by $Ag^{2+}$, $Hg^{2+}$, $Zn^{2+}$, $Cu^{2+}$, and were inhibited 60% by CoH, 30% by $Fe^{2+}$ and 10% by $Pb^{2+}$. However it was increased by the addition of $Sn^{2+}$, $Mn^{2+}$, $Mg^{2+}$ at concentration of $1{\times}10^{-3}$M. Both ureases were inhibited completely by p-CMB and acetohydroxamic acid. The urease expressed in E. coli (pBU11) was inhibited 70% by SDS. The urease of B. pasteurii was inhibited 40% by hydroxyurea, whereas the recombinant urease of E. coli strain was inhibited 17%. Both enzymes were not inhibited by cyclohexanediaminetetraacetic acid (CDTA) and ethylendiaminetetraacetic acid (EDTA).

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.17 no.2
• /
• pp.117-124
• /
• 1984

Sarine scrap usually comprises about $40\%$ of the raw fish in processing. The purpose of this study is to establish the desirable methods for proteinaceous materials from the sardine scrap through autolysis or enzymatic digestion and to convert them into useful by-products such as sardine sauce. Sardine scrap was chopped and mixed with equal weight of water, and be hydrolyzed them by autolysis and/or by addition of commercial proteolytic enzyme and various concentrations of sodium chloride. The optimal conditions for hydrolysis of sardine scrap were revealed in temperature at $55^{\circ}C$ and 4 hours digestion with bromelain($0.4\%$) and commercial complex enzyme ($6.0\%$), and those conditions were also applicated in autolysis. The maximum hydrolyzing rate of protein and released amino nitrogen were $82.5\%,\;5.2\%$ through autolysis, $84.3\%,\;5.8\%$ by bromelain digestion and $92.5\%,\;5.9\%$ by complex enzyme, respectively. In the products prepared from sardine scrap through autolysis or bromelain digestion, hypoxanthine was dominant, as $17.4 {\mu}mole/g$, dry matter for autolysis and $16.0 {\mu}mole/g$, dry matter, for bromelain digestion among the nucleotidcs and their related compounds, respectively. The abundant free amino acids were leucine, glutamic acid, lysnie, valine and alanine. The contents of those amino acids were $51.3\%,\;48.3\%$ of the total free amino acids, respectively. And the contents of 5'-IMP and TMAO were negligible but total creatinine was developed in value from $9.2\%\;to\;10\%$ of total extracted nitrogen. The flavor of sardine sauce prepared from sardine scrap by autolysis or enzyme digestion were not inferior to that of traditional Korean soy sauce by sensory evaluation.