• Journal of Microbiology and Biotechnology
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• v.31 no.8
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• pp.1123-1133
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• 2021

Biodegradation is the key process involved in natural lignocellulose biotransformation and utilization. Microbial consortia represent promising candidates for applications in lignocellulose conversion strategies for biofuel production; however, cooperation among the enzymes and the labor division of microbes in the microbial consortia remains unclear. In this study, metagenomic analysis was performed to reveal the community structure and extremozyme systems of a lignocellulolytic microbial consortium, TMC7. The taxonomic affiliation of TMC7 metagenome included members of the genera Ruminiclostridium (42.85%), Thermoanaerobacterium (18.41%), Geobacillus (10.44%), unclassified_f__Bacillaceae (7.48%), Aeribacillus (2.65%), Symbiobacterium (2.47%), Desulfotomaculum (2.33%), Caldibacillus (1.56%), Clostridium (1.26%), and others (10.55%). The carbohydrate-active enzyme annotation revealed that TMC7 encoded a broad array of enzymes responsible for cellulose and hemicellulose degradation. Ten glycoside hydrolases (GHs) endoglucanase, 4 GHs exoglucanase, and 6 GHs β-glucosidase were identified for cellulose degradation; 6 GHs endo-β-1,4-xylanase, 9 GHs β-xylosidase, and 3 GHs β-mannanase were identified for degradation of the hemicellulose main chain; 6 GHs arabinofuranosidase, 2 GHs α-mannosidase, 11 GHs galactosidase, 3 GHs α-rhamnosidase, and 4 GHs α-fucosidase were identified as xylan debranching enzymes. Furthermore, by introducing a factor named as the contribution coefficient, we found that Ruminiclostridium and Thermoanaerobacterium may be the dominant contributors, whereas Symbiobacterium and Desulfotomaculum may serve as "sugar cheaters" in lignocellulose degradation by TMC7. Our findings provide mechanistic profiles of an array of enzymes that degrade complex lignocellulosic biomass in the microbial consortium TMC7 and provide a promising approach for studying the potential contribution of microbes in microbial consortia.

• Nutrition Research and Practice
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• v.13 no.4
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• pp.286-294
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• 2019

BACKGROUND/OBJECTIVES: Docosahexaenoic acid (DHA), an n-3 long chain polyunsaturated fatty acid (LCPUFA), is acquired by dietary intake or the in vivo conversion of ${\alpha}$-linolenic acid. Many enzymes participating in LCPUFA synthesis are regulated by peroxisome proliferator-activated receptor alpha ($PPAR{\alpha}$). Therefore, it was hypothesized that the tissue accretion of endogenously synthesized DHA could be modified by $PPAR{\alpha}$. MATERIALS/METHODS: The tissue DHA concentrations and mRNA levels of genes participating in DHA biosynthesis were compared among $PPAR{\alpha}$ homozygous (KO), heterozygous (HZ), and wild type (WT) mice (Exp I), and between WT mice treated with clofibrate ($PPAR{\alpha}$ agonist) or those not treated (Exp II). In ExpII, the expression levels of the proteins associated with DHA function in the brain cortex and retina were also measured. An n3-PUFA depleted/replenished regimen was applied to mitigate the confounding effects of maternal DHA. RESULTS: $PPAR{\alpha}$ ablation reduced the hepatic Acox, Fads1, and Fads2 mRNA levels, as well as the DHA concentration in the liver, but not in the brain cortex. In contrast, $PPAR{\alpha}$ activation increased hepatic Acox, Fads1, Fads2, and Elovl5 mRNA levels, but reduced the DHA concentrations in the liver, retina, and phospholipid of brain cortex, and decreased mRNA and protein levels of the brain-derived neurotrophic factor in brain cortex. CONCLUSIONS: LCPUFA enzyme expression was altered by $PPAR{\alpha}$. Either $PPAR{\alpha}$ deficiency or activation-decreased tissue DHA concentration is a stimulus for further studies to determine the functional significance.

• Journal of Animal Science and Technology
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• v.61 no.2
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• pp.87-97
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• 2019

Phytate induced excessive mineral excretion through poultry litter leads to poor performance and environmental pollution. Exogenous microbial phytase supplementation to poultry diets reduce the environmental excretion of nutrient and improve bird's performance. However, excessive dietary sodium (Na) level may hinder the phytase-mediated phytate hydrolysis and negate the beneficial effects of phytase. Therefore, this experiment was conducted to investigate the effects of different concentration dietary Na on phytase activity and subsequent impact on broiler performance, bone mineralisation and nutrient utilisation. In this study, six experimental diets, consisting of three different levels of Na (1.5, 2.5, or 3.5 g/kg) and two levels of microbial phytase (0 or 500 U/kg) were formulated by using $3{\times}2$ factorial design. The six experimental diets were offered to 360 day-old Ross 306 male chicks for 35 days, where, each experimental diet consisted of 6 replicates groups with 10 birds. Along with growth performance, nutrient utilization, intestinal enzyme activity, dry matter (DM) content of litter and mineral status in bone were analysed. Dietary Na and phytase had no effect on bode weight gain and feed intake. Birds on the low Na diet showed higher (p < 0.05) feed conversion ratio (FCR) than the mid-Na diets. High dietary Na adversely affected (p < 0.001) excreta DM content. Phytase supplementation to the high-Na diet increased (p < 0.01) the litter ammonia content. High dietary Na with phytase supplementation improved ($Na{\times}phytase$, p < 0.05) the AME value and ileal digestibility of Ca and Mg. The total tract retention of Ca, P, and Mg was reduced with high Na diet, which was counteracted by phytase supplementation ($Na{\times}phytase$, p < 0.001). The diets containing mid-level of Na improved (p < 0.001) the function of Na-K-ATPase and Mg-ATPase in the jejunum. The overall results indicate that high dietary Na did not affect phytase activity but influenced the nutrient utilization of birds, which was not reflected in bird overall performance.

• Applied Chemistry for Engineering
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• v.22 no.3
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• pp.336-339
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• 2011

Algae is an abundant and potential fermentation substrate. The enzymatic hydrolysis of algae was investigated by pre-treating an alkaline hydrogen peroxide with commercial cellulase and viscozyme. Algae used in this study was the Ulva pertusa. The evaluated response was the yield of released glucose after the enzymatic hydrolysis. Alkaline hydrogen peroxide containing mixtures of 1 wt% hydrogen peroxide and 1~1.75 wt% sodium hydroxide was also used. The results show that the highest glucose conversion was obtained for Ulva pertusa using 5 wt% hydrogen peroxide at $60^{\circ}C$ for 3 h. The required amount of enzymes after the pre-treatment with alkaline hydrogen peroxide were reduced by far compared to that of untreated Ulva pertusa. Also, the amount of glucose that is released during the enzymatic hydrolysis was increased.

• Journal of Animal Science and Technology
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• v.64 no.5
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• pp.854-862
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• 2022

The purpose of this study is to determine the impact of different levels of crude protein (CP) diets supplemented with dietary protease on the growth performance and nutrient digestibility of the weanling pigs. In a 5-week study, 100 crossbred ([Landrace × Yorkshire] × Duroc) of weaner pigs that have an average initial body weight (BW) of 7.17±1.06 kg were assigned to one of four dietary treatments with 5 replications and 5 pigs (3 gilts and 2 castrated male pigs) per pen in a randomized complete block design. The dietary treatments were as follows: Phase 1: CON: basal diets (20.60% CP); low protein (LP): CON - 0.30% CP; PLP1: (CON - 0.30% CP) + 0.05% protease; PLP2: (CON - 0.50% CP) + 0.05% protease. Phase 2: CON: basal diets (18.88% CP); LP: CON - 0.30% CP; PLP1: (CON - 0.30% CP) + 0.05% protease; PLP2: (CON - 0.50% CP) + 0.05% protease. The addition of protease to low CP diets significantly increased the feed conversion ratio (FCR) (p = 0.039), BW (p = 0.046), average daily gain (ADG) (p = 0.049), and average daily feed intake (ADFI) tended to increase (p = 0.053) in the young pigs during phase 1. However, FCR tended to increase throughout the experiment but did not change during phase 2, whereas BW, ADG, and ADFI stayed unchanged throughout phase 2 and overall. There was no significant difference in dry matter, nitrogen (N), and gross energy of nutrient digestibility in all phases and overall in weaned pigs with low CP when protease was fed. In contrast, adding protease to the low CP diets increased the tendency of N digestibility (p = 0.059) during phase 1. It is concluded that dietary protease supplementation tended to increase N retention during the first phase of the weaning period, hence increasing piglet performance.

• Animal Bioscience
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• v.35 no.9
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• pp.1390-1399
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• 2022

Objective: The objective of this study was to evaluate the effects of dietary supplementation of Schizosaccharomyces pombe (S. pombe) -expressed phytase on growth performance, apparent ileal digestibility, organ indexes, meat quality, toe ash, and footpad lesions score in broiler chicks. Methods: A total of 390 one-day-old broiler chicks were randomly assigned to 5 groups based on the initial body weight (42.15±0.17 g), there were 6 replicate cages per treatment and 13 birds (mixed sex) per cage. The experimental period was 45 days, including 4 periods (starter, days 1 to 10; grower, days 11 to 24; finisher 1, days 25 to 38; finisher 2, days 39 to 45). Dietary treatments were based on a corn-soybean meal-basal diet and supplemented with 500, 750, 1,000, and 1,500 FTU/kg S. pombe-expressed phytase. One phytase unit (FTU) was defined as the amount of enzyme that catalyzes the release of one micromole phosphate from phytate per minute at 37℃ and pH 5.5. Results: The inclusion of increasing levels of phytase in the diet linearly increased the body weight gain during days 1 to 10 (p = 0.001), 25 to 38 (p = 0.016), 39 to 45 (p = 0.018), and 1 to 45 (p = 0.004), feed intake during days 25 to 38 (p = 0.032), feed conversion ratio during days 1 to 10 (p = 0.001), 39 to 45 (p = 0.038), and 1 to 45 (p = 0.012), carcass weight (p = 0.035), toe ash (p<0.001), and apparent ileal phosphorus digestibility (p = 0.049). However, the footpad lesions score (p = 0.040) decreased linearly with the increase in phytase levels in the diet. Conclusion: Dietary supplementation of S. pombe-expressed phytase was beneficial to the growth performance, toe ash, apparent ileal phosphorus digestibility, and footpad lesions of broiler chicks in a dose-dependent manner.

• Journal of Microbiology and Biotechnology
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• v.33 no.11
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• pp.1521-1530
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• 2023

An α-L-rhamnosidase gene from Thermoclostridium. stercorarium subsp. thermolacticum DSM 2910 (TstRhaA) was cloned and expressed. The maximum TstRhaA activity of the protein reached 25.2 U/ml, and the molecular mass was approximately 106.6 kDa. The protein was purified 8.0-fold by Ni-TED affinity with an overall recovery of 16.6% and a specific activity of 187.9 U/mg. TstRhaA activity was the highest at 65℃ and pH 6.5. In addition, it exhibited excellent thermal stability, better pH stability, good tolerance to low concentrations of organic reagents, and high catalytic activity for p-nitrophenyl-α-L-rhamnopyranoside (pNPR). Substrate specificity studies showed that TstRhaA exhibited a high specific activity for rutin. At 60℃, pH 6.5, and 0.3 U/ml enzyme dosage, 60 g/l rutin was converted to 45.55 g/l isoquercitrin within 150 min. The molar conversion rate of rutin and the yield of isoquercitrin were 99.8% and 12.22 g/l/h, respectively. The results suggested that TstRhaA could be used for mass production of isoquercitrin.

• Journal of Plant Biotechnology
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• v.35 no.3
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• pp.169-176
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• 2008

Salicylic acid(SA) is a phytohormone that is related to plant defense mechanism. The SA accumulation is triggered by abiotic and biotic stresses. SA acts as a signal molecular compound mediating systemic acquired resistance and hypersensitive response in plant. Although the role of SA has been studied extensively, an understanding of the SA regulatory mechanism is still lacking in plants. In order to comprehend SA regulatory mechanism, we have been transformed with a SID2 promoter:GUS::LUC fusion construct into siz1-2 mutant and wild plant(Col-0). SIZ1 encodes SUMO E3 ligase and negatively regulates SA accumulation in plants. SID2(SALICYLIC ACID INDUCTION DEFICIENT2) is a crucial enzyme of SA biosynthesis. The Arabidopsis SID2 gene encodes isochorismate synthase(ICS) that controls SA level by conversion of chorismate to isochorismate. We compared the regulation of SID2 in wild-type and siz1-2 transgenic plants that express SID2 promoter:GUS::LUC constructs respectively. The expressions of $\beta$-GLUCURONIDASE and LUCIFERASE were higher in siz 1-2 transgenic plant without any stress treatment. SID2 promoter:GUS::LUC/siz1-2 transgenic plant will be used as a starting material for isolation of siz1-2 suppressor mutants and genes involved in SA-mediated stress signaling pathway.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.3
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• pp.335-343
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• 2006

This study was undertaken to investigate the effects of meal pattern on lipogenesis and activities of lipogenic enzyme in rats epididymal and mesenteric adipose tissues. A high fat diet was fed either ad libitum or in 1 meal during the last 3 h of the dark cycle for 4 weeks. Lipogenesis was measured as glucose conversion to total lipid and activities of glucose-6-phosphate dehydrogenase (G6PDH), 6-phophogluconate dehydrogenase (6PGDH) and NADP-malate dehydrogenase (ME) were determined by measuring NADPH production. Lipoprotein lipase (LPL) activity and serum lipoprotein concentrations were also measured. Meal-fed (3 h) rats had a decreased food intake, body weight and carcass fat compared with rats fed ad libitum. The serum triglyceride concentration of meal-fed rats tended to be higher than that of the ad libitum rats. However, there were no differences between meal-fed group and ad libitum group in serum concentrations of HDL-cholesterol, LDL-cholesterol and total-cholesterol. Rates of lipogenesis in both epididymal and mesenteric adipose tissues were significantly higher in the meal-fed group than that in the ad libitum group. In addition, meal-fed group showed higher G6PDH, 6PGDH and LPL activities in both epididymal and mesenteric adipose tissues, but exerted no significant effect on ME activity. These results suggest that meal-fed rats compared with ad libitum rats have marked lipogenic capacity, although such elevation probably does not result in increase in carcass fat concentration. Thus, meal-fed diet can be an important determinant of the alterations in adipose lipid metabolism.

• ANNALS OF ANIMAL RESOURCE SCIENCES
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• v.28 no.3
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• pp.97-107
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• 2017

The present study was conducted to investigate the effects on growth performance, nutrient digestibility, and gut health of broiler chickens when a dietary supplementation of multienzymes was added to diets, containing different energy levels. A total of 480 broiler chickens of similar body weight (Ross 308, 1-day-old) were randomly subjected to four treatments. The dietary treatments included a corn-soybean meal-based diet supplemented with: multienzyme (amylase+protease+ mannanase+xylanase+phytase), 0.05% enzyme, and different energy levels (3010 and 3060 kcal/kg). The experimental diets were fed to the chicks in a mash form for 35 days in two phases (1-21 d, phase I; and 22-35 d, phase II). During the overall period, chicks fed with diets supplemented with multienzymes had a better weight gain (p<0.05) and feed conversion ratio (FCR) than those fed with diets without enzymes. There was no difference in the growth rate and FCR among the chicks fed with diets supplemented with enzymes, even though the dietary energy levels were different. The apparent fecal and ileal digestibility of dry matter, gross, crude protein, calcium, and phosphorus were significantly enhanced (p<0.05). The population of cecal and ileal Lactobacillus spp. was significantly increased (p<0.05), and Clostridium spp. and coliforms were significantly decreased (p<0.05) in diets supplemented with enzymes. Villus height and villus height to crypt depth ratio in the small intestine was also significantly enhanced (p<0.05) in diets supplemented with enzymes. In conclusion, multienzyme supplementation had positive effects on the weight gain of broilers, FCR, digestibility of nutrients, and on the growth of intestinal microbiota.