• Journal of The Korean Society of Grassland and Forage Science
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• v.13 no.2
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• pp.86-92
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• 1993

The variation of the esterase isozyme, germination rate, chemical composition and digestibility of field bean(G1ycine soja S. and Z.) were estimated. The results are as follows; 1. The banding patterns of the esterase isozyme in field bean were varied with the tissue and habitat. 2. The enzyme activity of the Est-I, Est-2, Est-3 and Est-4 in field bean showed a high value compared with the other enzyme. 3. The range of germination temperature in field bean was 10-40C and the optimum germination temperature was 25- 38^{\circ}C.$. 4. The crude protein(CP) contents was 19.9% in the whole plant, 27.8% in the leaf and 45.9% in the seed, the cellulose contents was 29.5% in the whole plant, 23.0% in the leaf and 13.8% in the seed, the neutral detergent fiber(NDF) was 62.6% in the whole plant, 47.9% in the leaf and 47.9% in the seed and the acid detergent fiber(ADF) was 44.5% in the whole plant, 28.4% in the leaf and 28.4% in the seed, respectively. 5. The digestibility of the field bean was 44.1% in the whole plant, 49.6% in the leaf and 75.1% in the seed, NDF was 26.2% in the whole plant 46.2% in the leaf, ADF was 29.0% in the whole plant, 47.7% in the leaf and 58.0% in the seed and Cellulose was 48.7% in the whole plant, 58.0% in the leaf and 70.2% in the seed, respectively. 6. Total digestible nutrients(TDN) of the field bean was 47.4% in the whole plant, 51.5% in the leaf and 70.2% in the seed, respectively. The digestible energy(DE) value was 2.1 kcal/g in the whole plant, 2.27 kcal/g in the leaf and 3.10 kcal/g in the seed and the metabolizable energy(ME) value was 1.72 kcal/g in the whole plant, 1.86 kcal/g in the leaf and 3.23 kcal/g in the seed, respectively.

• Microbiology and Biotechnology Letters
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• v.17 no.4
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• pp.358-364
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• 1989

A crystalline alkaline pretense- producing Streptomyce sp. YSA-130 was isolated from soil in alkaline medium(pH 10.5). The optimum culture condition of Streptomyce sp. YSA-130 for the production of alkaline protease was as follows; 2.0% soluble starch, 1.0% soytone, 0.3% $K_2$HPO$_4$, 0.02% MgSO$_4$.7$H_2O$, 0.8% Na$_2$CO$_3$, pH 10.5, 3$0^{\circ}C$, and 12 hr. The alkaline pretense from the culture broth of Streptomyce sp. YSA-130 was purified about 24 folds by ammonium sulfate precipitation , dialysis, DEAE-cellulose ion exchange chromatography, gel filtration on Sephadex G-15 and crystallization. Optimum temperature and pH of purified enzyme were 6$0^{\circ}C$, and 11.5. Temperature and pH stability of purified enzyme were 5$0^{\circ}C$, and 5.5-12.0. Calcium ion was effective to stabilize the enzyme at higher temperature. The molecular weight of the purified enzyme was approximately 30,000. The purified enzyme was inactivated by diisopropyl flurophosphate(DFP) but not affected by metal ion, EDTA, sulfhydryl reagent and stable detergent.

• Journal of Microbiology and Biotechnology
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• v.22 no.1
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• pp.84-91
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• 2012

Two staphylococcal lipases were obtained from Staphylococcus epidermidis S2 and Staphylococcus aureus S11 isolated from sebaceous areas on the skin of the human face. The molecular mass of both enzymes was estimated to be 45 kDa by SDS-PAGE. S2 lipase displayed its highest activity in the hydrolysis of olive oil at $32^{\circ}C$ and pH 8, whereas S11 lipase showed optimal activity at $31^{\circ}C$ and pH 8.5. The S2 lipase showed the property of cold-adaptation, with activation energy of 6.52 kcal/mol. In contrast, S11 lipase's activation energy, at 21 kcal/mol, was more characteristic of mesophilic lipases. S2 lipase was stable up to $45^{\circ}C$ and within the pH range from 5 to 9, whereas S11 lipase was stable up to $50^{\circ}C$ and from pH 6 to 10. Both enzymes had high activity against tributyrin, waste soybean oil, and fish oil. Sequence analysis of the S2 lipase gene showed an open reading frame of 2,067 bp encoding a signal peptide (35 aa), a pro-peptide (267 aa), and a mature enzyme (386 aa); the S11 lipase gene, at 2,076 bp, also encoded a signal peptide (37 aa), pro-peptide (255 aa), and mature enzyme (399 aa). The two enzymes maintained amino acid sequence identity of 98-99% with other similar staphylococcal lipases. Their microbial origins and biochemical properties may make these staphylococcal lipases isolated from facial sebaceous skin suitable for use as catalysts in the cosmetic, medicinal, food, or detergent industries.

• Applied Biological Chemistry
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• v.48 no.4
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• pp.322-325
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• 2005

Pseudomonas sp. JH1014 was isolated from stream water as a detergent-compatible alkaline protease producing microorganism. The strain produced no detectable cellulolytic activity in LB medium. The addition of carboxymethyl cellulose induced the production of carboxymethyl cellulase (CMCase) without causing any significant change in the growth pattern of the strain. The strain reached its maximum growth after 9 to 12 h at $37^{\circ}C$, and the production of CMCase in the presence of the substrate reached its maximum after 21 h of growth at $37^{\circ}C$. The optimum pH of the crude enzyme preparation was pH 6.0. The enzyme had an optimal temperature at $55^{\circ}C$, and retained 70% of its original activity when preincubated at $70^{\circ}C$ for 10 min. Activity staining of the crude enzyme preparation separated on an SDS-PAGE gel showed two active bands with molecular masses of 54 and 30 kDa, indicating that Pseudomonas sp. JH1014 produced at least 2 kinds of CMCase.

• BMB Reports
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• v.35 no.3
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• pp.330-336
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• 2002

The lgtB genes that encode $\beta$-1,4-galactosyltransferases from Neisseria meningitidis ATCC 13102 and gonorrhoeae ATCC 31151 were isolated by a polymerase chain reaction using the pfu DNA polymerase. They were expressed under the control of lac and T7 promoters in Escherichia coli M15 and BL21 (DE3). Although the genes were efficiently expressed in E. coli M15 at $37^{\circ}C$ (33 kDa), most of the $\beta$-1,4-galactosyltransferases that were produced were insoluble and proteolysed into enzymatically inactive polypeptides that lacked C-terminal residues (29.5 kDa and 28 kDa) during the purification steps. When the temperature of the cell growth was lowered to $25^{\circ}C$, however, the solubility of the $\beta$-1,4-galactosyltransferases increased substantially. A stable N-terminal his-tagged recombinant enzyme preparation could be achieved with E. coli BL21 (DE3) that expressed lgtB. Therefore, the cloned $\beta$-1,4-galactosyltransferases were expressed under the control of the T7 promoter in E. coli BL21 (DE3), mostly to the soluble form at $25^{\circ}C$. The proteins were easily purified to homogeneity by column chromatography using Ni-NTA resin, and were found to be active. The galactosyltransferases exhibited pH optimum at 6.5-7.0, and had an essential requirement for the $Mn^{+2}$ ions for its action. The $Mg^{+2}$ and $Ca{+2}$ ions showed about half of the galactosyltransferase activities with the $Mn^{+2}$ ion. In the presence of the $Fe^{+2}$ ion, partial activation was observed with the $\beta$-1,4-galactosyltransferase from N. meningitidis(64% of the enzyme activity with the $Mn^{+2}$$Ni^{+2}$, $Zn^{+2}$, and $Cu^{+2}$ ions could not activate the $\beta$-1,4-galactosyltransferase activity. The inhibited enzyme activity with the $Ni^{+2}$ ion was partially recovered with the $Mn^{+2}$$Fe^{+2}$, $Zn^{+2}$, and $Cu^{+2}$ ions, the $Mn^{+2}$$\beta$-1,4-galactosyltransferase activity was 1.5-fold stimulated with the non-ionic detergent Triton X-100 (0.1-5%).

• Journal of Animal Science and Technology
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• v.62 no.5
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• pp.648-658
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• 2020

The objective of the study was to evaluate the effects of Lactobacillus, cellulase, and molasses on chemical composition, fermentation qualities, and microorganism count of sugarcane bagasse silage after 30-days fermentation. The treatments were arranged according to a factorial arrangement (2 × 2 × 2) + 1, in a complete randomized design. The first factor consisted of two levels of Lactobacillus casei TH14 (TH14, 0 and 0.05 g/kg fresh matter; the second factor consisted of two levels of cellulase enzyme (C, 0 and 10⁴ U/kg fresh matter); and the third factor consisted of two levels of molasses (M, 0 and 5 g/ 100 mL distilled water). A treatment (+1) referred to the use of rice straw without any treatments. The result showed that dry matter increased by 4% and neutral detergent fiber decreased by 2% of sugarcane bagasse when ensiled as a combination of additives as compared to untreated sugarcane bagasse. The pH and ammonia nitrogen were significantly dropped to 3.5 and 2.3 g/kg dry matter. Furthermore, lactic acid was increased by 64% when compared to untreated sugarcane bagasse, respectively. Lactic acid bacteria count was increased by 28% as compared to untreated sugarcane bagasse. Based on this experiment, fermenting with L. casei TH14, cellulase, and molasses in combination resulted in the promotion of the best qualities of sugarcane bagasse silage.

• Korean Journal of Microbiology
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• v.46 no.1
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• pp.68-72
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• 2010

The ${\beta}$-1,4-glucanase gene of Bacillus licheniformis B1 was expressed in Esherichia coli BL21, and a protein with a mass of 50 kDa that was soluble was overproduced. A protein with a mass of 37 kDa was secreted from B. licheniformis. It seems that the ${\beta}$-1,4-glucanase produced in E. coli contained the leader peptide and unprocessed carboxy-terminal region, but its processing occurred in the carboxyterminal in Bacillus. The optimal temperature of ${\beta}$-1,4-glucanase was $40^{\circ}C$. The enzyme still had 76% maximal activity at $60^{\circ}C$. The optimal pH of the enzyme was 7. The enzyme retained considerable activities over the weak-acidic, neutral, and weak-basic pH range. Acidic fungal cellulases are used in food, detergent, pulp, paper, textile industries. However, studies about neutral and alkaline cellulase are not enough. The cellulase developed in this study may be useful for industrial applications in the fields of biofuel development.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.8
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• pp.1022-1027
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• 2010

Effects of increasing dietary rice straw on chewing activity, ruminal fermentation, and fibrolytic enzyme activity in growing goats were investigated in a $4{\times}4$ Latin Square experiment. The goats were offered four diets with an increasing proportion of rice straw (i.e. 0.05, 0.10, 0.15 and 0.20, respectively, on dry matter basis). Increasing level of rice straw increased ($P_{linear\;effect}$ <0.05) the time spent on eating, ruminating, and chewing. The ruminal pH and acetate: propionate ratio were increased ($P_{linear\;effect}$ <0.05), while the $NH_3$-N concentration was decreased ($P_{linear\;effect}$ <0.01). Increasing level of rice straw in the diet increased ($P_{linear\;effect}{\leq}0.01$) molar proportion of acetate and isovalerate, and decreased ($P_{linear\;effect}$ <0.01) molar proportion of propionate. The CMCase, xylanase and cellobiase activities in the rumen were decreased ($P_{linear\;effect}$ <0.05) with increasing level of dietary rice straw, whereas the avicelase activity was increased ($P_{linear\;effect}$ <0.01). In summary, increased level of rice straw elevated the dietary neutral detergent fibre (NDF) content in the diet and had a great impact on chewing activity and ruminal fermentation.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.2
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• pp.240-247
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• 2012

Dietary supplementation with conventional linted cottonseed hulls (LCSH) is a common practice in livestock production all over the world. However, supplementation with mechanically delinted cottonseed hulls (DCSH) and cottonseed linter residue (CLR) is uncommon. Cottonseed by-products, including LCSH, DCSH and CLR, were assessed by chemical analysis, an in situ nylon bag technique, an in vitro cumulative gas production technique and in vitro enzyme procedure. The crude protein (CP) content of CLR (302 g/kg dry matter (DM)) was approximately 3 times that of LCSH and 5 times that of DCSH. The crude fat content was approximately 3 times higher in CLR (269 g/kg DM) than in LCSH and 4 times higher than in DCSH. Neutral detergent fibre (311 g/kg DM) and acid detergent fibre (243 g/kg DM) contents of CLR were less than half those of DCSH or LCSH. Metabolisable energy, estimated by in vitro gas production and chemical analyses, ranked as follows: CLR (12.69 kJ/kg DM)>LCSH (7.32 kJ/kg DM)>DCSH (5.82 kJ/kg DM). The in situ degradation trial showed that the highest values of effective degradability of DM and CP were obtained for CLR (p<0.05). The in vitro disappearance of ruminal DM ranked as follows: CLR>LCSH>DCSH (p<0.05). The lowest digestibility was observed for DCSH with a two-step in vitro digestion procedure (p<0.05). The potential gas production in the batch cultures did not differ for any of the three cottonseed by-product feeds. The highest concentration of total volatile fatty acids was observed in CLR after a 72 h incubation (p<0.05). The molar portions of methane were similar between all three treatments, with an average gas production of 22% (molar). The CLR contained a higher level of CP than did LCSH and DCSH, and CLR fermentation produced more propionate. The DCSH and LCSH had more NDF and ADF, which fermented into greater amounts of acetate.

• Applied Biological Chemistry
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• v.38 no.4
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• pp.320-324
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• 1995

Solubilization of microsomal ${\Delta}^{5}-3{\beta}$-hydroxy steroid acyl transfearse(${\Delta}^{5}-3{\beta}$-OH-SAT) of rat brain and its reconstitution into liposomes were investigated. Among the detergents utilized for the solubilization, deoxycholic acid was superior to Tween 80 or Triton X-100 for the reconstituted activity of ${\Delta}^{5}-3{\beta}$-OH-SAT. The enzyme activity was shown to be affected by the nature of phospholipids used for the preparation of the liposome. Phosphatidylcholines from egg yolk and soybean showed the highest activity of ${\Delta}^{5}-3{\beta}$-OH-SAT and phosphatidylethanolamine came next. However phosphatidylserine and phosphatidic acid showed a lower activity than those obtained before the reconstitution. This study suggests that the presence of quaternary ammonium salt or amine group in the phospholipids stimulates the activity of ${\Delta}^{5}-3{\beta}$-OH-SAT. However the presence of a carboxylic group or the absence of the amine group may have an inhibitory effect on the ${\Delta}^{5}-3{\beta}$-OH SAT.