• Microbiology and Biotechnology Letters
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• v.16 no.2
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• pp.92-97
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• 1988

Cysteinylglycinase was partially purified from Proteus mirabilis by consecutive procedure. The specific activity was increased about 16-fold to that of cell-free extract. The enzyme was found rather unstable on ammonium sulfate precipitation ann the precipitated enzyme protein became partially insoluble during dialysis. The precipitated enzyme was found to be solubilized by treatment of 4% Triton X-100 effectiviely, The optimum temperature and pH of the enzyme activity were 35$^{\circ}C$ and 7.3, respectively. After heat treatment of the enzyme at 5$0^{\circ}C$ for 30 min, it lost the activity to 70%. The enzyme was stable at pH 7.0-8.0. The molecular weight of the cysteinylglycinase was found to be about 190,000 by Sephadex G-150 gel filtration. The enzyme was activated by the addition of Mn$^{2+}$ and $Mg^{2+}$ ions. The maximal activation was obtained in preincubation with $Mg^{2+}$ ion for 30 min. The enzyme catalyzed the hydrolysis of various dipeptides and tripeptides. The Km and Vmax values for cysteinylglycine were 1.60 mM and 0.24 m unit/ mg, respectively.

• Microbiology and Biotechnology Letters
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• v.35 no.3
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• pp.226-230
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• 2007

In order to achieve high-level expression of interferon-${\alpha}1$ (IFN-${\alpha}1$) during fed-batch fermentation of recombinant E. coli, effects of oxygen supply and induction temperature on the expression of recombinant proteins were evaluated. Supplementation of oxygen and its transfer into cells is one of the most important parameters involved in the design and operation of mixing-sparging equipment for bioreactors. Generally, higher oxygen supply stimulates cell growth of aerobic microorganism and consequently the amount of products is increased. In this study, the optimum aeration strategy for the higher production of IFN-${\alpha}1$ during fed-batch fermentation of recombinant E. coli was surveyed. The growth of the cells was also monitored with four different concentrations of dissolved oxygen (DO; limiting, 20%, 35%, 50%) conditions. The DO was controlled by varying aeration rates of air and pure oxygen. Oxygen uptake rate (OUR) and specific oxygen uptake rate (SOUR) were evaluated and compared for the enhanced growth and induction of the cells and IFN-${\alpha}1$, respectively. We confirmed that increased DO by additional oxygen supply, up to 35%, can improve the production of IFN-${\alpha}1$ during the fermentation.

• Microbiology and Biotechnology Letters
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• v.42 no.4
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• pp.324-330
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• 2014

A marine bacterial strain producing agar-degrading enzymes was isolated from a mud flat in Jeboo-do (Korea) using a selective artificial sea water (ASW) agar plate containing agar as the sole carbon source. The isolate, designated as SH-1, was gram-negative, aerobic, and motile with single polar flagellum. 16S rRNA gene sequence similarity analysis showed the isolate SH-1 had the highest homology (96.5%) to marine bacterium Neiella marina J221. Cells could grow at $28-37^{\circ}C$ but not at $42^{\circ}C$, and the agarase activity of the cell culture supernatant was higher when grown at $28^{\circ}C$ than when grown at $37^{\circ}C$. Cells could grow when concentrations of 1-5% (w/v) NaCl were added to the growth media with the best growth observed at 3% NaCl, and the agardegrading enzyme activity of the cell culture supernatant was best when grown at 3% NaCl-containing growth media under the conditions we examined. The crude enzyme prepared from 48-h culture broth of strain SH-1 exhibited an optimum pH and temperature for agar-degrading activity at 7.0 and $40^{\circ}C$, respectively. Zymogram analysis of the crude supernatant and cell extract showed that strain SH-1 produced at least 3 agar-degrading enzymes with molecular weights of 15, 35, and 52 KD. Thinlayer chromatography (TLC) analysis also suggested that HS-1 produces ${\beta}$-agarase to degrade agarose to neoagarooligosaccharides.

• Journal of Microbiology and Biotechnology
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• v.30 no.11
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• pp.1720-1728
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• 2020

We have previously characterized AprESJ4, the major fibrinolytic enzyme from Bacillus subtilis SJ4 (Yao et al., 2019). During that study, we observed a 68 kDa protein with fibrinolytic activity. In this study, we cloned the gene (vprSJ4) encoding the 68 kDa protein, a mature Vpr and minor protease secreted by Bacillus species. vprSJ4 encodes a preproenzyme consisting of 810 amino acids (aa) including signal sequence (28 aa) and prosequence (132 aa). The mature enzyme (650 aa) has a predicted molecular weight of 68,467.35. Unlike Vprs from other B. subtilis strains, VprSJ4 has 4 additional amino acids (DEFA) at the C-terminus. vprSJ4 was overexpressed in Escherichia coli. PreproVprSJ4 was localized in inclusion bodies, and subjected to in vitro renaturation and purification by an affinity column. SDS-PAGE and western blot showed that autoprocessing of preproVprSJ4 occurred and 68 kDa and smaller proteins were produced. The optimum pH and temperature of the recombinant VprSJ4 were pH 7.0 and 40℃, respectively. Kinetic parameters of recombinant VprSJ4 were measured by using an artificial substrate, N-succinyl-ala-ala-pro-phe-p-nitroanilide. Coexpression of vprSJ4 and aprESJ4 using pHY300PLK increased the fibrinolytic activity a further 117% when compared with aprESJ4 single expression using the same vector in B. subtilis WB600.

• Journal of Microbiology and Biotechnology
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• v.30 no.11
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• pp.1659-1669
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• 2020

1,3-α-3,6-anhydro-L-galactosidase (α-neoagarooligosaccharide hydrolase) catalyzes the last step of agar degradation by hydrolyzing neoagarobiose into monomers, D-galactose, and 3,6-anhydro-L-galactose, which is important for the bioindustrial application of algal biomass. Ahg943, from the agarolytic marine bacterium Gayadomonas joobiniege G7, is composed of 423 amino acids (47.96 kDa), including a 22-amino acid signal peptide. It was found to have 67% identity with the α-neoagarooligosaccharide hydrolase ZgAhgA, from Zobellia galactanivorans, but low identity (< 40%) with the other α-neoagarooligosaccharide hydrolases reported. The recombinant Ahg943 (rAhg943, 47.89 kDa), purified from Escherichia coli, was estimated to be a monomer upon gel filtration chromatography, making it quite distinct from other α-neoagarooligosaccharide hydrolases. The rAhg943 hydrolyzed neoagarobiose, neoagarotetraose, and neoagarohexaose into D-galactose, neoagarotriose, and neoagaropentaose, respectively, with a common product, 3,6-anhydro-L-galactose, indicating that it is an exo-acting α-neoagarooligosaccharide hydrolase that releases 3,6-anhydro-L-galactose by hydrolyzing α-1,3 glycosidic bonds from the nonreducing ends of neoagarooligosaccharides. The optimum pH and temperature of Ahg943 activity were 6.0 and 20℃, respectively. In particular, rAhg943 could maintain enzyme activity at 10℃ (71% of the maximum). Complete inhibition of rAhg943 activity by 0.5 mM EDTA was restored and even, remarkably, enhanced by Ca²⁺ ions. rAhg943 activity was at maximum at 0.5 M NaCl and maintained above 73% of the maximum at 3M NaCl. K_m and V_max of rAhg943 toward neoagarobiose were 9.7 mg/ml and 250 μM/min (3 U/mg), respectively. Therefore, Ahg943 is a unique α-neoagarooligosaccharide hydrolase that has cold- and high-salt-adapted features, and possibly exists as a monomer.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.4
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• pp.342-347
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• 2002

Ciliates have the possibility of a new live food in marine finfish culture because of their wide range of body size, thin tell wall, show motility, and fast reproduction rate. In this research, six species of ciliates were isolated from south coast and salt pond in Korea. The fitness of these species as a live food was evaluated in terms of size, motility, suspensibility and cell density. As the result, Euplotes sp. (K-1) was found suitable to be a new live food which might substitute rotifers, Brachionus plintilis and B. rotundiformis in fish larvae culture. The modified $F{\emptyset}yn's$ Erdschreiberd media, MErds-2 with the addition of glycine, glucose and yeast extract increased six times higher growth rate of Euplotes sp. (K-1) than the basic F$\emptyset$yn's Erdschreiberd media. The optimum water temperature, pH and light intensity for this ciliates were $22.5^{\circ}C$, 8 and 2,000 lux, respectively, and its culture environmental range was relatively wide, On the other hand, this ciliate fed baker's yeast, Saccharomyces cererisiae grew up to 1,240 inds./mL with the inocula of 100 inds./mL within 7 days. The results of the study showed that Euplotes sp. (K-1) has a potential to be utilized as a new live food in fish larvae culture.

• Korean Journal of Food Science and Technology
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• v.27 no.4
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• pp.506-515
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• 1995

The cultural characteristics of 60 strains of psychrotrophic lactic acid bacteria which were isolated from kimchi, a Korean traditional fermented vegetable food, and identified as Leuconostoc mesenteroides subsp. mesenteroides, Leu. mesenteroides subsp. dextranicum, Leu. paramesenteroides, Leu. lactis, Lactobacillus bavaricus and Lac. homohiochii were tested. All strains grew at $5^{\circ}C,\;10^{\circ}C\;and\;37^{\circ}C$ in tomato glucose broth, but not at $45^{\circ}C$. The optimum growth temperature of Leu. mesenteroides and Lactobacillus sp. were $33{\sim}34^{\circ}C\;and\;34{\sim}36^{\circ}C$, respectively. All strains of Leu. mesenteroides and Lactobacillus sp. grew at 4.8 and 4.2 of initial pH, but not at 4.0. The final pH of Leu. mesenteroides and Lactobacillus sp. in glucose broth were $3.84{\sim}4.10\;and\;3.82{\sim}3.99$, respectively. None of the 60 strains clotted milk nor reduced litmus in litmus milk. All strains of Leu. mesenteroides and Lactobacillus sp. grew in tomato glucose broth containing 7% ethanol or 6.5% NaCl, but not in the broth containing 15% ethanol or 10% NaCl. All strains grew in tomato glucose broth containing 40% bile juice and survived in the artificial gastric juice of pH 3.5. Furthermore, all strains of Leu. mesenteroides survived in the artificial gastric juice of pH 3.0. Since many strains of lactic acid bacteria tested in this study showed differences in several physiological characteristics from those described in Bergey's Manual of Systematic Bacteriology, it was considered that further tests would be necessary to clarify their positions in taxonomic system.

• Korean Journal of Food Science and Technology
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• v.34 no.5
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• pp.780-786
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• 2002

Optimum condition of the drying process and the changes in aroma components during dehydration were determined for Sarcodon aspratus. The drying curve of mushrooms consisted of short constant rate period followed by long falling rate period. The drying rate increased with increasing drying temperature and air velocity. Results showed that mushrooms dried at $50^{\circ}C$ and air velocity of 1.5 m/sec had the greatest peak area of aroma compound. The aromatic components of the dried mushrooms were 1-octen-3-one, 1-octen-3-ol, 3-octanone, 1-octanol, 2-octen-1-ol, 3-octanol, 3-octanone, 1-octanol, 2-octen-1-ol, and 3-octanol. Peak areas of mushroom alcohol and aromatic compounds of mushrooms including 1-octen-3-ol, 1-octanol, 2-octen-1-ol, 3-octanol decreased significantly, whereas those of 1-octen-3-one and 3-octane increased during the drying period. New unfavorable compounds including butyric acid, propanoic acid, and 3-methyl thiopropanol were formed during the drying period.

• Korean Journal of Food Science and Technology
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• v.22 no.1
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• pp.13-18
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• 1990

Crude lipase activator (L. Activator) was extracted with 0.85M NaCl solution from green pepper, Capsicum annuun Lin and then fractionated by 0.2 saturation with ammonium sulfate. The activity of crude L. Activator preparation $(OD_{280}=1.0)$ had proportional relation with its added amounts below 1.0ml. The L.Activator showed optimum temperature at $35^{\circ}C$. The L.Activator was very stable at the temperatures below $50^{\circ}C$ and at pH range of $7{\sim}9$, and its activities also remained 60% even at $100^{\circ}C$, 72% at pH 3, and 85% at pH 10, respectively. The activities of L.Activator decreased by most metal ions besides $Na^+,\;Mg^{++},\;and\;Ca^{++}$. The decreasing effects of heavy metal ions such as $Ag^+\;and\;Hg^{++}$ on L.Activator activity were not, however, so great as compared with the commonly known great effects of them on most enzyme activity. Crude L.Activator was separated into 4 peaks by the cellulofine column chromatography and the main active peak of L.Activator seemed to be contained in the same components as those of the activatory peak from crude L.Inhibitor.

• Journal of the Korean Crystal Growth and Crystal Technology
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• v.24 no.1
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• pp.15-20
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• 2014

In this research, the geopolymer was fabricated using municipal solid waste incineration ash (denoted as MSWIA) slag and alkali activator, NaOH and its properties were analyzed. Particularly, the effects of NaOH molarity, particle size of MSWIA, and liquid/solids ratio on the compressive strength of geopolymers were investigated. The compressive strength of geopolymers fabricated increased with finer grain size of MSWIA, and optimum value of the liquid/solids ratio was identified as 0.13. As the molarity of the NaOH increased, the compressive strength of geopolymers was increased. Even more the 20 M of NaOH, but the strength was not increased. The calcium aluminum silicate and calcium aluminum silicate hydrate zeolites were generated in the geopolymer fabricated with more than 20 M of NaOH, with some unreacted silica and unknown crystals remained. The highest compressive strength, 163 MPa, of geopolymer was appeared at conditions of curing temperature $70^{\circ}C$, and 20 M of NaOH, indicating that the high concentration of NaOH accelerates the geopolymer reaction and dense microstructure. The high-strength geopolymer fabricated in the present study is expected to contribute significantly to develop the field of cement alternative substances and to improve the recycling rate of MSWIA slag.