• Journal of Microbiology and Biotechnology
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• v.28 no.4
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• pp.588-596
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• 2018

An endo-${\beta}$-1,4-glucanase gene, cel9K, was cloned using the shot-gun method from Paenibacillus sp. X4, which was isolated from alpine soil. The gene was 2,994 bp in length, encoding a protein of 997 amino acid residues with a predicted signal peptide composed of 32 amino acid residues. Cel9K was a multimodular enzyme, and the molecular mass and theoretical pI of the mature Cel9K were 103.5 kDa and 4.81, respectively. Cel9K contains the GGxxDAGD, PHHR, GAxxGG, YxDDI, and EVxxDYN motifs found in most glycoside hydrolase family 9 (GH9) members. The protein sequence showed the highest similarity (88%) with the cellulase of Bacillus sp. BP23 in comparison with the enzymes with reported properties. The enzyme was purified by chromatography using HiTrap Q, CHT-II, and HiTrap Butyl HP. Using SDS-PAGE/activity staining, the molecular mass of Cel9K was estimated to be 93 kDa, which is a truncated form produced by the proteolytic cleavage of its C-terminus. Cel9K was optimally active at pH 5.5 and $50^{\circ}C$ and showed a half-life of 59.2 min at $50^{\circ}C$. The CMCase activity was increased to more than 150% in the presence of 2 mM $Na^+$, $K^+$, and $Ba^{2+}$, but decreased significantly to less than 50% by $Mn^{2+}$ and $Co^{2+}$. The addition of Cel9K to a commercial enzyme set (Celluclast 1.5L + Novozym 188) increased the saccharification of the pretreated reed and rice straw powders by 30.4% and 15.9%, respectively. The results suggest that Cel9K can be used to enhance the enzymatic conversion of lignocellulosic biomass to reducing sugars as an additive.

• Journal of Mushroom
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• v.13 no.3
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• pp.270-273
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• 2015

Hypsizigus marmoreus is commercially the most important edible mushroom in Japan. This mushroom is usually cultivated for a longer period (about 85~120 days) than other mushroom. In order to develop a new cultivar that has a shortened cultivation period, the genome analysis of this strain has been considered. This study aims to obtain parental monokaryotic strains reproducing 'Haemi' cultivar in Hypsizigus marmoreus for reference genome sequencing. The mycelia were cultured in MCM and MYG media for various incubation periods. Homogenized mycelia were treated with commercial cell wall degrading enzymes to maximize protoplasts production yield from Hypsizigus marmoreus. The greatest number of protoplasts was obtained from mycelia cultured in MCM media for 3 days using Novozyme enzyme. The isolated protoplasts were grown in regeneration agar media after two weeks. Regenerated colonies were picked and moved on separated dishes for microscopic observation. Neohaplonts regenerated from dikayotic strains were identified by the absence of clamp connections. We confirmed that one of monokaryotic strains is a parental strain by crossing with an original compatible strain of 'Haemi' cultivar. This parental strain will be used for reference genome sequence analysis.

• Food Science of Animal Resources
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• v.29 no.1
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• pp.15-23
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• 2009

In order to develop a new starter for fermented milk, 2082 bacteria were isolated from raw milk. The strain that showed excellent acid forming and ${\gamma}$-aminobutyric acid (GABA) production ($711.40{\mu}g/g$ D.W) characteristics after incubation at $37^{\circ}C$ for 18 hr was selected and identified as Lactobacillus acidophilus by the result of API carbohydrate fermentation pattern and 16S rDNA sequence. L. acidophilus RMK567 was investigated for its physiological characteristics. RMK67 strain showed good GABA production compared with commercial lactic acid bacteria. The optimum growth temperature of L. acidophilus RMK567 was $40^{\circ}C$ and cultures took 15 hr to reach pH 4.3. L. acidophilus RMK567 showed higher sensitivity to penicillin-G, novobiocin, as compared to other 14 different antibiotics. However, it showed more resistance to kanamycin, neomycin, streptomycin. It showed higher leucine arylamidase and ${\beta}$-galactosidase activities compared to 16 other enzymes. It was comparatively tolerant to bile juice and able to survive at pH 2 for 3 hr. It showed resistence to Escherichia coli, Salmonella typhimurium and Staphylococcus aureus with rates of 29.2%, 39.1% and 51.4%, respectively. Based on these and previous results, L. acidophilus RMK567 could be an excellent starter culture for fermented milk with excellent GABA contents.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.9
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• pp.1273-1278
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• 2016

In this study, we optimized conditions for extraction of eleutherosides B and E from stem powder of Acanthopanax. To enhance eleutheroside B and E yields, Acanthopanax sessiliflorus Seeman and Eleutherococcus senticosus Maxim were pre-incubated with the following enzymes: Celluclast, Viscozyme L, Lactozyme, Lecitase, and Novozyme 33095. Treatment with Novozyme 33095, a commercial pectinase, for 3 h resulted in the highest yields of eleutherosides B and E from A. sessiliflorus and E. senticosus. Compared with extraction at $121^{\circ}C$ for 120 min, at $121^{\circ}C$ for 15 min after Novozyme 33095 pre-incubation increased eleutheroside B and E yields from A. sessiliflorus by 7% and 17%, respectively. In the case of E. senticosus, eleutheroside B and E yields increased by 25% and 29%, respectively.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.44 no.3
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• pp.191-196
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• 2011

In this study, we investigated the chemical and enzymatic properties of unmarketable cultured bastard halibut (UCBH) Paralichthys olivaceus harvested at different times (March, May, July, September, November, and January), and we examined the physical properties of surimi gel from UCBH as a potential source of surimi and surimi gel. The moisture and crude protein contents of UCBH harvested in July and January were >78% and <19%, respectively, which is greater than the moisture content in UCBH harvested in May, March, and September, but lower than the crude protein content. Regardless of the month of harvest, the UCBH had a higher crude protein content than Alaska pollock, which is the largest fishery biomass used for surimi and surimi gel, but a lower moisture content. Regardless of the month of harvest, the enzymatic activity in crude extracts of UCBH muscle ranged from 0.31-0.59 U/mg for casein (pH 6.0 and 9.0) and 11.7-12.7 U/mg for LeuPNA. These findings suggest that autolytic enzymes were unaffected by gel formation. Gel strength was highest in the surimi gel prepared from UCBH harvested in September, November, and January; second highest in that prepared from UCBH harvested in March and May; and lowest in that prepared from UCBH harvested in July. Compared to the gel strength of surimi gel from grade SA commercial Alaska pollock surimi, the strength of the surimi gels prepared from UCBH harvested in March, May, September, November, and January were superior, whereas that of the surimi gel prepared from UCBH harvested in July was similar.

• Journal of the Korean Society of Food Science and Nutrition
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• v.14 no.4
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• pp.401-408
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• 1985

The yeast cell wall lytic action of the alkaline AL-protease, which was found out of the crude Zymolyase that a kind of yeast cell wall lytic $endo-{\beta}-1$, 3-glucanase produced from Arthrobacter luteus, was investigated with the viable cells of S. sake and it's cell wall preparation. AL-protease on the lysis of the viable yeast cells showed very low activities with the alone, but the lytic activities were highly increased with the combination of AL-protease and Zymolyase. On the stepwise treatment of the viable yeast cells with AL-protease and Zymolyase, the cells were lysed highly only by the course having a treatment with Zymolyase after pretreatment with AL-protease. Thus synergistic action of AL-protease was not observed with any some commercial enzymes, known as a type of alkaline and serine protease such as AL-protease, and was also found to be affected greatly by the culture conditions and species of the yeast tested. AL-protease caused the release of some peptide and a lot of sugar from the cell wall preparation, but could not lysed the cell wall more than 66%. Whereas Zymolyase could lysed the cell walls almost completely with alone. On the basis of these results, the synergistic action of AL-protease on the lysis of S. sake cells is hypothesized that at first AL-protease bind to the yeast cell surface layer consisting of mannan and protein, and then changes their conformation to facilitate the penetration of Zymolyase from the outside to the inside framework layer constituted of alkali insoluble ${\beta}-1,\;3-glucan$.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.7
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• pp.998-1003
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• 2016

Although exogenous protease enzymes have been used in poultry diets quite extensively, this has not been the case for pig diets. In general, due to their better gut fermentative capacity and longer transit time, pigs have greater capacity to digest dietary proteins than poultry. However, in early-weaned piglets, the stress brought about by weaning adversely affects the digestion of dietary proteins. Therefore, a study was conducted to determine the effects of a commercial protease enzyme in weanling pigs. Indices of growth, nutrient digestibility, blood profiles, fecal microflora, fecal gas emission and fecal scores were measured during the study. A total of 50 weanling pigs ($6.42{\pm}0.12kg$) at 28 d of age were randomly assigned to receive 1 of 2 dietary treatments: i) control diet (corn-soy based) with no supplemental protease (CON), and ii) control diet+200 g/ton protease (PROT) for 42 d. A completely randomized design consisting of 2 treatments, 5 replicates, and 5 pigs in each replicate was used. Growth performance in terms of body weight ($27.04{\pm}0.38kg$ vs $25.75{\pm}0.39kg$; p<0.05) and average daily gain ($491{\pm}7.40g$ vs $460{\pm}7.46g$; p<0.05) in PROT fed pigs were increased significantly, but gain per feed ($0.700{\pm}0.01$ vs $0.678{\pm}0.01$; p>0.05) was similar between treatments at d 42. Relative to CON pigs, PROT fed pigs had increased (p<0.05) apparent total tract digestibility ($84.66%{\pm}0.65%$ vs $81.21%{\pm}1.13%$ dry matter and $84.02%{\pm}0.52%$ vs $80.47%{\pm}1.22%$ nitrogen) and decreased (p<0.05) $NH_3$ emission ($2.0{\pm}0.16ppm$ vs $1.2{\pm}0.12ppm$) in the feces at d 42. Except for a decreased (p<0.05) in blood creatinine level, no differences were observed in red blood cell, white blood cell, lymphocyte, urea nitrogen, and IgG concentrations between treatments. Fecal score and fecal microflora (Lactobacillus and E. coli) were also similar between CON and PROT groups. Overall, the supplementation of protease enzyme in weanling pigs resulted in improved growth rate and nutrient digestibility. Exogenous protease enzyme reduced fecal $NH_3$ emission, thus, potentially serving as a tool in lowering noxious gas contribution of livestock production in the environment.

• Journal of the Korean Wood Science and Technology
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• v.39 no.3
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• pp.221-229
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• 2011

Due to the increase of oil price and the environmental issue such as the emission of volatile organic compounds, the necessity for developing alternative resins of petroleum-based adhesive resins, which have extensively been used for the manufacture of wood-based products, has been speculation since the early 1990. In our study, rapeseed flour (RSF), which is the by-product of bio-diesel produced from rapeseed, were hydrolyzed by enzymes. As a crosslinking agents of the RSF hydrolyzates, phenol-formaldehyde prepolymers (PF) were prepared. The RSF hydrolyzates and PF were mixed to complete the formulation of RSF-based adhesive resins, and the resins were applied to make the laminated veneer lumber (LVL). The physical and mechanical properties of the LVL were measured to examine whether RSF can be used as raw materials of adhesive resins for the fabrication of LVL or not. The average moisture content and soaking delamination rate of the LVL bonded with RSF-based adhesive resins exceeded the minimum requirement of KS standard. Moreover, thermal analysis of the RSF-based resins showed similar tendencies except for the RSF-based adhesive resins formulated with pectinase-hydrolyzed RSF. The bending strengths of the LVL were higher than that of the LVL made with commercial PF resins. These results showed the potential of RSF as a raw material of alternative adhesives for the production of LVL. Further works on the optimal conditions of RSF hydrolysis and spreading characteristics for RSF-based adhesive resins is required to improve the adhesive performance of RSF-based resins.

• Korean Journal of Environmental Agriculture
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• v.31 no.4
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• pp.293-299
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• 2012

BACKGROUND: Soil utilization pattern can be the main factor affecting soil physico-chemical properties, especially in soil phosphorus (P). Understanding the distribution and bioavailability of P is important for developing management to minimize P release from arable soils to environment. This study was conducted to evaluate the potential bioavailability of soil organic P by using phosphatase hydrolysis method. METHODS AND RESULTS: Twenty-four soils from onion-rice double cropping and 30 soils from plastic film house were selected from Changyeong and Daegok in Gyeongnam province, respectively. The P accumulation pattern (total P, inorganic P, organic P, residual P) and water soluble P were characterized. Commercial phosphatase enzymes were used to classify water-extractable molybdate unreactive P from arable soils into compounds that could be hydrolysed by (i) alkaline phosphomonoesterase (comprising labile orthophosphate monoesters), (ii) a combination of alkaline phosphomonoesterase and phosphodiesterase (comprising labile orthophosphate monoesters and diesters), and (iii) phytase (including inositol hexakisphosphate). Available P was highly accumulated with 616 and 1,208 mg/kg in double cropping system and plastic film house, respectively. Dissolved reactive P (DRP) and dissolved unreactive P (DUP) had similar trends with available P, showing 24 and 109 mg/kg in double cropping and 37 and 159 mg/kg in plastic film house, respectively, indicating that important role of dissolved organic P in the environments had been underestimated. From the result of phosphatase hydrolysis, about 39% and 66% of DUP was evaluated as bioavailable in double cropping and plastic film house, respectively. CONCLUSION(S): Orthophosphate monoester and orthophosphate diester accounted for high portion of dissolved organic P in arable soils, indicating that these organic P forms give important impacts on bioavailability of P released from P accumulated soils.

• Journal of Food Hygiene and Safety
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• v.35 no.1
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• pp.83-92
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• 2020

This study investigated the optimal conditions of enzymatic reaction for production of isomaltooligosaccharides (IMO) using rice flour. To manufacture IMO, commercial enzymes (Termamyl 2X, Maltogenase L, Promozyme D2, Fungamyl 800L and Transglucosidase L) were used. The sugar composition and amount of IMO were examined by HPLC with charged aerosol detector (HPLC-CAD) in each manufacturing process. Liquefaction reaction was performed according to different Termamyl 2X concentrations (0.025%, 0.05%, 0.075%, 0.1%) and reaction times (1 h, 2 h). As a result, the reducing sugar content was the highest at 138.26 g/L when 0.075% Termamyl 2X was added for 2 hours. In order to optimize simultaneous saccharification and transglucosylation, experiments on enzyme selection, enzyme concentration and enzyme reaction time were conducted. Reaction with 0.0015% Maltogenase L, 0.05-0.1% Promozyme D2 and 0.1% Tansglucosidase L was effective in decreasing glucose content and increasing content of IMO with a high degree of polymerization. A change in sugar content was observed every 6 hours to determine the optimal reaction time, and the highest IMO was produced after 36 hours of reaction (75.36 g/L). The IMO prepared under optimal conditions showed isomaltose, 35.11 g/L; panose, 11.97 g/L; isomaltotriose, 19.95 g/L; isomaltotetraose, 7.46 g/L; isomaltopentaose, 1.05 g/L at 18 brix and the ratio of IMO in the total sugar was 56.37%.