• Animal Bioscience
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• v.34 no.3_spc
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• pp.371-384
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• 2021

Objective: Wheat bran (WB) was co-fermented with Aspergillus oryzae and phytase (Phy) to determine whether co-fermentation improve WB phosphorus and fiber utilization in Isa-brown layers. Methods: A total of 112 Isa brown layer were randomly divided into 7 treatments with 8 replicates per a treatment and 2 hens per a replicate. The treatments included basal diet (control), basal diet supplemented with 250 unit/kg Phy (control+Phy), diet with 10% WB (10% WB), diet with 5% WB and 250 unit/kg Phy (5% WB+Phy) diet with 10% WB and 250 unit/kg Phy (10% WB+Phy), diet with 5% fermented WB supplemented with molasses and phy (PCFWH) and 125 unit/kg Phy (5% PCFWH), and diet with 10% PCFWH (10% PCFWH). The intestinal microbial population, intestinal morphology, serum antioxidant enzyme activities, and excreta phosphorus content were assessed. Results: In PCFWH, spore counts, protease activity, xylanase activity, and ferulic acid were 8.50 log/g dry matter (DM), 190 unit/g DM, 120 unit/g DM, and 127 ㎍/g, respectively. Xylobiose and xylotriose were released in PCFWH, while they were not detectable in WB. Antioxidant capacity was also enhanced in PCFWH compared to WB. The 10% WB+Phy and 10% PCFWH groups produced higher egg mass, but hens fed 5% WB+Phy had the lowest amount of feed intake. Eggs from 10% PCFWH had better eggshell weight, eggshell strength, and eggshell thickness. Birds fed with 10% PCFWH also had higher serum superoxide dismutase and catalase activities. Compare to control, 10% PCFWH significantly reduced excreta phosphorus content. Conclusion: Diet inclusion of 10% PCFWH improved egg quality, antioxidant status, and excreta phosphorus content of laying hens.

• Microbiology and Biotechnology Letters
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• v.18 no.6
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• pp.633-639
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• 1990

The genes for cellulases of Pseudomonas sp. LBC505 and CYC10, potent cellulase complex-producing strains, were cloned in Escherichia coli with pUC19. Recombinant plasmids pLCl and pLC2 were isolated from transformants producing cellulase by Congo red staining, and their genes cloned were 0.7 kb and 4.6 kb HindIII fragments, respectively. The inserts of pLCl and pLC2 were hybridized to chromosomal DNAs digested with HindIII from Pseudomona~ sp. LBC505 and CYC10, respectively. Immunodiffusion assays revealed that pLC1-and pLC2-encoded cellulase showed similarity with that of host strains. About 24% of cellulase activity was observed in the extracellular fraction of E. coli carrying pLC1, and its activity was higher about 1.4 times than that of LBC505. The enzymatic properties of pLC1 and pLC2 encoded cellulase were the same as those of cellulase from host strains. HPLC analysis and substrate specificity showed that cellulases were the same as those of cellulase from host strains. HPLC analysis and substrate specificity showed that cellulases cloned were endocellulase.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.9
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• pp.1354-1360
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• 2006

Maize-soybean meal diets with 0, 100, 200 and 300 g/kg double zero rapeseed meal ('00' RSM) with and without an enzyme mixture (xylanase, pectinase, cellulase) at a level of 1.6 g/kg were evaluated with 624 day-old broiler chicks for 5 weeks. The birds were randomly allocated to eight dietary treatments with three replicates of 26 birds each. Average daily gain (ADG) and feed intake (FI) were recorded weekly and ileal viscosity, organ weights, serum enzyme activity, hormonal profile and hematological parameters were measured at the end of week 5. Average daily gain during the weekly periods was significantly influenced by the dietary level of '00'RSM (p<0.01). Inclusion of '00' RSM improved the ADG up to day 28 with the increased level; beyond that time no improvement was recorded when compared to control groups. However, ADG from 1-35 days was significantly different between 300 g/kg inclusion level of '00' RSM and the control diet. Inconsistent decline in feed intake and feed conversion ratio was observed up to day 21 and the trend was reversed thereafter. The proportion of '00' RSM in the diet had a significant ($p{\leq}0.05$) influence on thyroid weight but had no effect on the relative weights of liver and heart, serum enzyme activities (${\gamma}$-glutamyl transferase, alanine amino transferase and aspartate amino transferase), thyroid hormones ($T_3$ and $T_4$), hemoglobin level and hematocrit. Significant improvement in ADG was recorded during the 2nd week of age with the addition of enzyme, whereas for all other periods, including the whole period of the trial, higher but non-significant ADG was observed. FI and FCR were not affected by the addition of enzyme but there was a numerical reduction in FCR during the whole period. The addition of enzyme reduced the ileal viscosity at all levels of '00' RSM inclusion. The results suggest that '00' RSM can be included up to 300 g/kg in broiler diets without any adverse effects on health and performance. The addition of commercial enzyme mixture containing xylanase, pectinase, cellulase to broiler diets containing '00'RSM has some effect on growth rate and feed conversion efficiency.

• Applied Biological Chemistry
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• v.19 no.3
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• pp.139-144
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• 1976

In order to investigate the properties of enzymes from two strains of mold, reported in the previous paper, (1) studies have been made concerning the characteristics of cellulase of Aspergillus niger-SM6 and Trichoderma viride-SM10, and summarized as follows. 1. In the semi-purification the recovery of ${\beta}-glucosidase$ was the highest when 80-90% ethanol was used and 0.8 saturation of $(NH_4)_2SO_4$. 2. The characteristics of the semi-purified enzyme were as follows. Aspergillus niger-SM6 Trichoderma viride-SM10 Optimum pH 3.5 4.0 pH stability 3.0-6.0 3.0-6.0 Optimum temperature $60^{\circ}C$ $60^{\circ}C$ Heat stability below $60^{\circ}C$ below $50^{\circ}C$ Optimum reaction time 30 min. 60 min. Optimum CMC concentration 3% 3% 3. The Km values of CMCase were 0.8% and 1.01 for Aspergillus niger-SM6 and Trichoderma viride-SM10, respectively. 4. In the strain of Aspergillus niger-SM6, there were high activity of xylanase and pectinase.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.6
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• pp.482-493
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• 2005

Biomass is originally photosynthesized from inorgainic compounds such as $CO_2$, minerals, water and solar energy. Recent studies have shown that anaerobic bacteria have the ability to convert recalcitrant biomass such as cellullosic or chitinoic materials to useful compounds. The biomass containing agricultural waste, unutilized wood and other garbage is expected to utilize as feed, food and fuel by microbial degradation and other metabolic functions. In this study we isolated several anaerobic, cellulolytic and chitinolytic bacteria from rumen fluid, compost and soil to study their related enzymes and genes. The anaerobic and cellulolytic bacteria, Clostridium thermocellum, Clostridium stercorarium, and Clostridium josui, were isolated from compost and the chitinolytic Clostridium paraputrificum from beach soil and Ruminococcus albus was isolated from cow rumen. After isolation, novel cellulase and xylanase genes from these anaerobes were cloned and expressed in Escherichia coli. The properties of the cloned enzymes showed that some of them were the components of the enzyme (cellulase) complex, i.e., cellulosome, which is known to form complexes by binding cohesin domains on the cellulase integrating protein (Cip: or core protein) and dockerin domains on the enzymes. Several dockerin and cohesin polypeptides were independently produced by E. coli and their binding properties were specified with BIAcore by measuring surface plasmon resonance. Three pairs of cohesin-dockerin with differing binding specificities were selected. Two of their genes encoding their respective cohesin polypeptides were combined to one gene and expressed in E. coli as a chimeric core protein, on which two dockerin-dehydrogenase chimeras, the dockerin-formaldehyde dehydrogenase and the dockerin-NADH dehydrogenase are planning to bind for catalyzing $CO_2$ reduction to formic acid by feeding NADH. This reaction may represent a novel strategy for the reduction of the green house gases. Enzymes from the anaerobes were also expressed in tobacco and rice plants. The activity of a xylanase from C. stercorarium was detected in leaves, stems, and rice grain under the control of CaMV35S promoter. The digestibility of transgenic rice leaves in goat rumen was slightly accelerated. C. paraputrificum was found to solubilize shrimp shells and chitin to generate hydrogen gas. Hydrogen productivity (1.7 mol $H_2/mol$ glucos) of the organism was improved up to 1.8 times by additional expression of the own hydrogenase gene in C. paraputrficum using a modified vector of Clostridiu, perfringens. The hydrygen producing microflora from soil, garbage and dried pelletted garbage, known as refuse derived fuel(RDF), were also found to be effective in converting biomass waste to hydrogen gas.

• Journal of Animal Science and Technology
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• v.48 no.2
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• pp.307-314
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• 2006

To search direct fed microbials, we isolated a Candida sp. from kefir grain. The isolated Candida sp. strain showed 99.8% of identity to the species of Candida kefyr by API 20C kit. Enzyme activity of Candida kefyr was higher in amylase (0.33±1.12μmol/min/mg) than that in phytase (0.052±0.98μmol/ min/mg) cellulase(0.051±μmol/min/mg) and xylanase (0.011±0.98mol/min/mg). The maximum numbers of Candida kefyr in growth curve were reached at 30 h fermentation. Candida kefyr showed high resistances to acidic environment, which was not perfectly extincted even at pH 2.0. And it showed high tolerance to bile salt which had almost 97.2% of survival in the presence of 1.0% bile salt.Especially, Candida kefyr showed high heat stability which remained 10% of initial microorganisms at 60℃. Candida kefyr was not generally inhibited by most of 11 antibiotic agent which contained tetracycline groups. These results suggest that the isolated Candida kefyr has a useful properties as probiotics.

• Food Science of Animal Resources
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• v.25 no.3
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• pp.357-364
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• 2005

To search probiotic microorganisms, we isolated Lactobacillus sp. from kefir, The Lactobacillus sp. strain showed $99.5\%$ of identity to species Lactobacillus rhamnosus by API kit. Lactobacillus rhamnosus showed high resistances to acidic environment, which grew well even at pH 2.0 and $1.0\%$ bile salt Enzyme activity of Lactobacillus rhamnosus was higher in amylase ($0.673\;{\mu}mol/min/mg$) than that in xylanase ($0.288\;{\mu}mol/min/mg$), cellulase($0.117\;{\mu}mol/min/mg$) and phytase($0.269\;{\mu}mol/min/mg$). Especially, the Lactobacillus rhamnosus showed high heat stability which remained $1{\times}10^6\;CFU/ml$ at $60^{\circ}C$. The maximum numbers of Lactobacillus rhamnosus on growth owe was reached at 24 h fermentation and pH was decreased to 4.6. The resistances of Lactobacillus rhamnosus to acidic pH and bile salt were better than that of Lactobacillus acidophilus used as control. When Lactobacillus rhamnosus was cultured with E. coli in MRS broth, E. coli was disappeared after 18 h. These result suggest that the isolated Lactobacillus rhamnosus has a useful probiotics properties.

• The Korean Journal of Mycology
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• v.40 no.3
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• pp.152-158
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• 2012

The lignocellulytic enzymes including a-amylase (EC 3.2.1.1), lignin peroxidase (EC 1.11.1.14), laccase (EC 1.10.3.2), xylanase (EC 3.2.1.8), ${\beta}$-xylosidase (EC 3.2.1.37), ${\beta}$-glucosidase (EC 3.2.1.21) and cellulase (EC 3.2.1.4) were extracted from spent mushroom compost (SMC) of Pleurotus eryngii. Different extraction buffers and conditions were tested for optimal recovery of the enzymes. The optimum extraction was shaking incubation (200 rpm) for 2 h at $4^{\circ}C$. ${\alpha}$-Amylase was extracted with the productivity range from 1.20 to 1.6 Unit/SMC g. Cellulase was recovered with the productivity range from 2.10 to 2.80 U/gf. ${\beta}$-glucosidase and ${\beta}$-xylosidase productivities showed lowest recovery producing 0.1 U/g and 0.02 U/g, respectively. The P. eryngii SMCs collected from three different mushroom farms showed different recovery on laccase and xylanse, cellulase. Furthermore, the water extracted SMC was compared to commercial enzymes for its industrial application in decolorization and cellulase activity.

• Journal of Life Science
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• v.17 no.12
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• pp.1660-1668
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• 2007

The non-ionic surfactant (NIS) Tween 80 was evaluated for its ability to influence invitro cumulative gas production, dry matter digestibility, cellulolytic enzyme activities, anaerobic microbial growth rates, and adhesion to substrates by mixed rumen microorganisms on rice straw, alfalfa hay, cellulose filter paper and tall fescue hay. The addition of NIS Tween 80 at a level of 0.05% increased significantly (P<0.05) in vitro DM digestibility, cumulative gas production, microbial growth rate and cellulolytic enzyme activity from all of substrates used in this study. In vitro cumulative gas production from the NIS-treated substrates; rice straw, alfalfa hay, filter paper and tall fescue hay was significantly (P<0.05) improved by 274.8, 235.2, 231.1 and 719.5% compared with the control, when substrates were incubated for 48 hr in vitro. The addition of 0.05% NIS Tween 80 to cultures growing on alfalfa hay resulted in a significant increase in CMCase (38.1%), xylanase (121.4%), Avicelase (not changed) and amylase (38.2%) activities after 36 h incubation. These results indicated that the addition of 0.05% Tween 80 could greatly stimulate the release of some kinds of cellulolytic enzymes without decreasing cell growth rate in contrast to trends reported with aerobic microorganism. Our SEM observation showed that NIS Tween. 80 did not influence the microbial adhesion to substrates used in the study. Present data clearly show that improved gas production, DM digestibility and cellulolytic enzyme activity by Tween 80 is not due to increased bacterial adhesion on the substrates.

• Applied Biological Chemistry
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• v.47 no.1
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• pp.78-84
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• 2004

In order Nuruk to improve the quality of millet wine, a traditional Jeju cereal wine, yeasts and molds were isolated from 35 kinds of Nuruk collected nationwide. Isolated strains were screened for saccharification of starch and brewing of millet wine. Fermentation characteristics of millet wine with different types of Nuruk were also investigated. The average number of microbial populations in the Nuruk were $6.4{\times}10^5{\sim}4.5{\times}10^7\;cfu/g$ for molds and $1.4{\times}10^4{\sim}7.7{\times}10^7\;cfu/g$ for yeasts. Among the 169 strains of molds and 103 strains of yeasts, 16 strains were screened for saccharifying activity on starch as a substrate, and one yeast strain was screened for the brewing of millet wine. A8-3, supposed as Aspergillus sp., showed the highest enzyme activities of glucdamylase, ${\alpha}-amylase$ and xylanase while B23-3 strain, supposed as Rhizopus sp., showed the highest saccharifying activity. A10-4, supposed as Saccharomyces sp., showed the highest level of weight loss from $CO_2$ evolution, sugar and alcohol tolerance during fermentation. When the Nuruk was made after inoculation with the selected strains, saccharifying activity was higher for the co-cultivation of A8-3 and B23-3 than individual cultivation of each strain. Similar saccharifying activities were shown in both disc-type and pellet-type Nuruk. It was suggested that pellet-type Nuruk could improve fermentation yield. The collected Nuruk consisted of $10{\sim}13%$ moisture, $55{\sim}70%$ total sugar, $10{\sim}18%$ crude protein, $0.2{\sim}1.0%$ crude fat and $1.8{\sim}2.1%$ ash. The Nuruk made in this study was composed of $12{\sim}15%$ moisture, $61{\sim}71%$ total sugar, $15{\sim}20%$ crude protein, $0.4{\sim}1.5%$ crude fat and $1.1{\sim}1.5%$ ash.