• Asian-Australasian Journal of Animal Sciences
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• v.17 no.4
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• pp.523-532
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• 2004

An experiment was conducted to investigate the effect of microbial phytase ($Natuphos^{R}$) supplementation in combination with enzyme complex (composed of enzymes targeted to SBM dietary components such as $\alpha$-galactosides and galactomannans; $Endo-Power^{R}$) to diet with low nutrient levels on growth performance and ileal nutrient digestibility of weaned pigs. A total of 210 crossbred weaned pigs (Landrace$\times$Yorkshire$\times$Duroc), 6.68$\pm$0.98 kg of initial body weight, were randomly allotted to five dietary treatments, based on weight and age, according to a randomized complete block design. There were three pens per treatment and 14 pigs per pen. The dietary treatments were 1) CON (Control diet with no phytase and enzyme complex (EC)), 2) LP+EC 100 (Control diet with 0.15% unit lower available phosphorus (aP) level+0.1% phytase (500 FTU/kg diet) and 0.1% enzyme complex), 3) LP+EC 80 (Control diet with 0.15% unit lower aP level+0.08% phytase (400 FTU/kg diet) and 0.08% enzyme complex, 4) LPEA+EC 100 (Control diet with 0.15% unit lower aP and 3% lower ME and amino acid levels (lysine, methionine, threonine and typtophan)+0.1% phytase (500 FTU/kg diet) and 0.1% enzyme complex), 5) LPEA+EC 80 (Control diet with 0.15% unit lower aP and 3% lower ME and amino acid levels+0.08% phytase (400 FTU/ kg diet) and 0.08% enzyme complex). For the determination of ileal nutrients digestibility, a total of 15 T-cannulated pigs (initial body weight; 7.52$\pm$1.24 kg; 3 replicates per treatment) were used in the present study. Piglets were weighted and allotted into same dietary treatments as one in growth trial and phase I experimental diets were provided for ileal digestibility study. There was no significant difference (p>0.05) in average daily gain (ADG) and average daily feed intake (ADFI) among dietary treatments during the whole experimental period (0 to 5 weeks). However, piglets in LP+EC 100 group had a significantly higher gain/feed ratio (G:F) than piglets had in control (p<0.05). Crude protein, energy and phosphorus digestibilities were significantly improved when both of phytase and enzyme complex were supplemented at the revel of 0.1%, respectively to diets with low nutrient level (aP or (and) ME and amino acids) (p<0.05). Piglets in LP+EC 100 and LPEA+EC 100 groups showed significantly higher phosphorus content (%) in bone than that of piglets in control group (p<0.05). Supplementation of both of phytase and enzyme complex at 0.1%, respectively, to diet with low nutrient levels (aP or (and) ME and amino acids) significantly improved total ileal essential amino acid and nonessential amino acid digestibilities compared to control group (p<0.05). In conclusion, the results from the present study suggest that the simultaneous inclusion of phytase and enzyme complex to diets at recommended level is advantageous with respect to improving growth performance and nutrient digestibility of weaned pigs and may contribute to increased economic return when added to corn-soy based weaned pig diets.

• Journal of Microbiology and Biotechnology
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• v.28 no.7
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• pp.1122-1132
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• 2018

In this study, we attempted to find new and efficient microbial enzymes for producing rare sugars. A ribose-5-phosphate isomerase B (OsRpiB) was cloned, overexpressed, and preliminarily purified successfully from a newly screened Ochrobactrum sp. CSL1, which could catalyze the isomerization reaction of rare sugars. A study of its substrate specificity showed that the cloned isomerase (OsRpiB) could effectively catalyze the conversion of $\text\tiny{L}$-rhamnose to $\text\tiny{L}$-rhamnulose, which was unconventional for RpiB. The optimal reaction conditions ($50^{\circ}C$, pH 8.0, and 1 mM $Ca^{2+}$) were obtained to maximize the potential of OsRpiB in preparing $\text\tiny{L}$-rhamnulose. The catalytic properties of OsRpiB, including $K_m$, $k_{cat}$, and catalytic efficiency ($k_{cat}/K_m$), were determined as 43.47 mM, $129.4sec^{-1}$, and 2.98 mM/sec. The highest conversion rate of $\text\tiny{L}$-rhamnose under the optimized conditions by OsRpiB could reach 26% after 4.5 h. To the best of our knowledge, this is the first successful attempt of the novel biotransformation of $\text\tiny{L}$-rhamnose to $\text\tiny{L}$-rhamnulose by OsRpiB biocatalysis.

• Journal of Life Science
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• v.7 no.1
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• pp.30-38
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• 1997

For screening thermostable $\alpha$-amylase from thermophiles, various samples from extreme environments such as hot spring and sewage near them, and compoat, wereexamined microbial growth in enrichment culture medium at 55$\circ$C on the assumption that enzymes from thermophiles are inevitable thermostable. One strain showing higher $\alpha$-amylase activity was pure cultured and designated as Bacillus sp. TR-25 from the results of morphological, cultural and physiological characteristics. The most important carbon sourses for the enzyme production were soluble starch, dextrin, potato starch and corn starch. Glucose and fructose had a catabolite repression on the enzyme production. The good nitrogen sources for the enzyme production were yeat extract, nutrient broth, tryptone, corn steep liquor and ammonium sulfate. The enzyme production was accelerated by addition of CaCl$_{2}$. $\cdot $ H$_{2}$O. The optimal medium composition for the enzyme production was soluble starch 2.0%, yeast extract 0.55, CaCl$_{2}$ $\cdot $ 2H$_{2}$O 0.015, Tween 80 0.001%, pH8.0, respectively. In jar fermenter culture, this strain shows a rapid growth and required cheaper carbon and nitrogen source. These properties are very useful to fermentation industry. The $\alpha$-amylase of this strain demonstrated a maximum activity at 80$\circ$C, pH 5.0, respectively. And calcium ion did not improve thermostability of the enzyme. At 10$0^{\circ}C$, this enzyme has 235 of relative activity. Transformation was carried out by thermophilic Bacillus sp. TR-25 genomic DNA. As a result, the transformant has increased thermostable $\alpha$-amylase activity.

• 한국균학회소식:학술대회논문집
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• 2015.11a
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• pp.11-11
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• 2015

A total of sixty-six samples of Nuruk, a fermention starter used to make the Korean traditional rice wine, Makgeolli, were collected from central and southern regions of Korea in 2013 and 2014. We classified two groups of the Nuruk samples, "commercial" and "home-made", according to the manufacturing procedure and purpose of use. Commercial Nuruks were made in a controlled environment where the temperature and humidity are fixed and the final product is supplied to Makgeolli manufacturers. Home-made Nuruks were made under uncontrolled conditions in the naturally opened environment and were intended for use in the production of small amounts of home-brewed Makgeolli. We obtained more than five hundred isolates including filamentous fungi and yeasts from the Nuruk samples followed by identification of fungal species. Also we stored glycerol stocks of each single isolate at $-70^{\circ}C$. We identified the species of each isolate based on the sequences of ITS regions amplified with two different universal primer pairs. We also performed morphological characterization of the filamentous fungi and yeast species through observations under the microscope. We investigated the major fungal species of commercial and home-made Nuruks by counting the colony forming units (CFU) and analyzing the occurrence tendency of fungal species. While commercial Nuruks contained mostly high CFU of yeasts, home-made Nuruks showed relatively high occurrence of filamentous fungi. One of the representative Nuruk manufacturers used both domestic wheat bran and imported ones, mainly from US, as raw material. Depending on the source of ingredient, the fungal diversity was somewhat different. Another commercial Nuruk sample was collected twice, once in 2013 and again in 2014, and showed different diversity of fungal species in each year. Nuruks obtained from the southern regions of Korea and Jeju island showed high frequency of yeast such as Saccharomycopsis fibuligera and Pichia species as well as unique filamentous fungus, Monascus species. S. fibuligera was easily found in many Nuruk samples with high CFU. The major filamentous fungi were Aspergillus, Lichtheimia, Mucor and Penicillium species. In order to further our understanding of the isolates and their potential industrial applications, we assayed three enzymes, alpha amylase, glucoamylase and acid protease from 140 isolates out of about five hundred isolates and selected about 10 excellent strains with high enzyme activities. With these fungal isolates, we will perform omics analyses including genomics, transcriptomics, metabolic pathway analyses, and metabolomics followed by whole genome sequencing of unique isolates associated with the basic research of Nuruk and that also has applications in the Makgeolli making process.

• Korean Journal of Food Science and Technology
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• v.27 no.4
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• pp.598-604
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• 1995

The contents in large intestine of Sprague Dawley rats fed polygalae radix(Polygala tennuifolia), onion(Allium cepa) and mugwort(Artemisia asiatica)-supplemented diets for 14 days were analysed for changes of major intestinal microflora, activities of ${\beta}-glucosidase\;and\;{\beta}-glucuronidase$ and amounts of putrefactive products such as indole and volatile basic nitrogen. The rats having ingested $5%{\sim}10%$ mugwort water or ethanol extract-supplemented diets showed a significant increase in intestinal bifido-bacteria and a decrease in clostridia and E. coli (p<0.05). And 10% onion juice group also showed a similar beneficial microflora change. In 5% mugwort powder-supplemented group, ${\beta}-glucosidase\;and\;{\beta}-glucuronidase$ activities in the intestinal contents were lowered, but the changes were not significant. Indole contents and pH in this group were significantly low compared with that of control (p<0.05). However, the activities of ${\beta}-glucosidase$ in 5% polygalae radix water extract and 10% onion juice-supplemented group and ${\beta}-glucuronidase\;in\;5%{\sim}10%$ mugwort water and ethanol extract-supplemented group were significantly higher than those of control (p<0.05). The intestinal indole contents of rats were significantly increased by feeding diet with water extract of polygalae radix and ethanol extract of mugwort which had brought comparatively large amount of protein in intestine (p<0.05). However, polygalae radix, onion, and mugwort-supplemented group had no effect on volatile basic nitrogen.

• Journal of Dairy Science and Biotechnology
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• v.21 no.2
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• pp.97-104
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• 2003

It is extremely important to destroy the antigenicity of milk proteins for dietetic treatment of infants with milk allergy. Enzymatic digestion of milk protein is not only effective for destroying antigenicity, but it also is less liable to alter the nutritive value. Bovine casein was hydrolyzed with eight different commercial proteases derived from bacterias or fungi, either individually or in combination to eliminate protein allergenicity. The average molecular weight of casein hyrdolysates determined by size exclusion chromatography is about 550${\sim}$2,300 dalton range. Antigenicity of the casein hyrdolysates was not detected by heterologous passive cutaneous anaphylaxis in guinea pig-rabbit antiserum system. The inhibition test on the enzyme-linked immunosorbent assay(ELISA) showed that the antigenicity of casein hydrolysates is lowed up to 1/8,000 than that of intact bovine casein. As the enzyme reaction was carried out by the combination of bacterial and fungal protease, casein hydrolysates showed much lower bitterness and antigenicity. It suggests that these hydrolysates will be applied to many kinds of foods including the development of hypo-allergenic infant formula.

• Journal of Soil and Groundwater Environment
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• v.19 no.4
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• pp.62-69
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• 2014

In the results of monitoring nitrate concentration in more than 8,000 groundwater wells around agro-livestock, the average and maximum nitrate concentration was 9.4 mg/L and 101.2 mg/L, respectively. Since about 31% of the monitoring wells was exceed the quality standard for drinking water, nitrate control such as remediation or source regulation is required to conserve safe-groundwater in South Korea. Typical nitrate-treatment technologies include ion exchange, reverse osmosis, and biological denitrification. Among the treatment methods, biological denitrification by indigenous microorganism has environmental and economic advantages for the complete elimination of nitrate because of lower operating costs compared to other methods. Major mechanism of the process is microbial reduction of nitrate to nitrite and nitrogen gas. Three functional genes (nosZ, nirK, nirS) that encode for the enzyme involved in the pathway. In this work, we tried to develop simple process to determine possibility of natural denitrification reaction by monitoring the functional gene. For the work, the functional genes in nitrate-contaminated groundwater were monitored by using PCR with specific target primers. In the result, functional genes (nosZ and nirK) encoding denitrification enzymes were detected in the groundwater samples. This method can help to determine the possibility of natural-nitrate degradation in target groundwater wells without multiplex experimental process. In addition, for field-remediation application we selected nitrate-contaminated site where 200~600 mg/L of nitrate is continuously detected. To determine the possibility of nitrate-degradation by stimulated-natural attenuation, groundwater was sampled in two different wells of the site and nitrate concentration of the samples was 300 mg/L and 616 mg/L, respectively. Fumarate for different C/N ratio was added into microcosm bottles containing the groundwater to examine denitrification rate depending on carbon concentration. In the result, once 1.5 times more than amount of fumarate stoichiometry required was added, the 616 mg/L of nitrate and 300 mg/L of nitrate were completely degraded in 8 days and 30 days. The nitrite, byproduct of denitrification process, was also completely degraded during the experimental period.

• Animal Bioscience
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• v.35 no.10
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• pp.1461-1478
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• 2022

The maintenance of poultry gut health is complex depending on the intricate balance among diet, the commensal microbiota, and the mucosa, including the gut epithelium and the superimposing mucus layer. Changes in microflora composition and abundance can confer beneficial or detrimental effects on fowl. Antibiotics have devastating impacts on altering the landscape of gut microbiota, which further leads to antibiotic resistance or spread the pathogenic populations. By eliciting the landscape of gut microbiota, strategies should be made to break down the regulatory signals of pathogenic bacteria. The optional strategy of conferring dietary fibers (DFs) can be used to counterbalance the gut microbiota. DFs are the non-starch carbohydrates indigestible by host endogenous enzymes but can be fermented by symbiotic microbiota to produce short-chain fatty acids (SCFAs). This is one of the primary modes through which the gut microbiota interacts and communicate with the host. The majority of SCFAs are produced in the large intestine (particularly in the caecum), where they are taken up by the enterocytes or transported through portal vein circulation into the bloodstream. Recent shreds of evidence have elucidated that SCFAs affect the gut and modulate the tissues and organs either by activating G-protein-coupled receptors or affecting epigenetic modifications in the genome through inducing histone acetylase activities and inhibiting histone deacetylases. Thus, in this way, SCFAs vastly influence poultry health by promoting energy regulation, mucosal integrity, immune homeostasis, and immune maturation. In this review article, we will focus on DFs, which directly interact with gut microbes and lead to the production of SCFAs. Further, we will discuss the current molecular mechanisms of how SCFAs are generated, transported, and modulated the pro-and anti-inflammatory immune responses against pathogens and host physiology and gut health.

• Journal of Microbiology and Biotechnology
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• v.34 no.2
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• pp.457-466
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• 2024

Cellobiose dehydrogenases (CDHs) are a group of enzymes belonging to the hemoflavoenzyme group, which are mostly found in fungi. They play an important role in the production of acid sugar. In this research, CDH annotated from the actinobacterium Cellulomonas palmilytica EW123 (CpCDH) was cloned and characterized. The CpCDH exhibited a domain architecture resembling class-I CDH found in Basidiomycota. The cytochrome c and flavin-containing dehydrogenase domains in CpCDH showed an extra-long evolutionary distance compared to fungal CDH. The amino acid sequence of CpCDH revealed conservative catalytic amino acids and a distinct flavin adenine dinucleotide region specific to CDH, setting it apart from closely related sequences. The physicochemical properties of CpCDH displayed optimal pH conditions similar to those of CDHs but differed in terms of optimal temperature. The CpCDH displayed excellent enzymatic activity at low temperatures (below 30℃), unlike other CDHs. Moreover, CpCDH showed the highest substrate specificity for disaccharides such as cellobiose and lactose, which contain a glucose molecule at the non-reducing end. The catalytic efficiency of CpCDH for cellobiose and lactose were 2.05 × 10⁵ and 9.06 × 10⁴ (M^-1 s^-1), respectively. The result from the Fourier-transform infrared spectroscopy (FT-IR) spectra confirmed the presence of cellobionic and lactobionic acids as the oxidative products of CpCDH. This study establishes CpCDH as a novel and attractive bacterial CDH, representing the first report of its kind in the Cellulomonas genus.

• Journal of Korean Society of Environmental Engineers
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• v.29 no.8
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• pp.895-903
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• 2007

As for the increasing demanding on the development of direct-ecological landfill monitoring methods, it is needed for critically defining the condition of landfills and their influence on the environment, quantifying the amount of enzymes and bacteria mainly concerned with biochemical reaction in the landfills. This study was thus conducted to understand the fates of contaminants in association with groundwater quality parameters. For the study, groundwater was seasonally sampled from four closed unsanitary landfills(i.e. Cheonan(C), Wonju(W), Nonsan(N), Pyeongtaek(P) sites) in which microbial diversity was simultaneously obtained by 16S rDNA methods. Subsequently, a number of primer sets were prepared for quantifying the specific gene of representative bacteria and the gene of encoding enzymes dominantly found in the landfills. The relationship between water quality parameters and gene quantification were compared based on correlation factors. Correlation between DSR(Sulfate reduction bacteria) gene and BOD(Biochemical Oxygen Demand) was greater than 0.8 while NSR(Nitrification bacteria-Nitrospira sp.) gene and nitrate were related more than 0.9. A stabilization indicator(BOD/COD) and MTOT(Methane Oxidation bacteria), MCR(Methyl coenzyme M reductase), Dde(Dechloromonas denitrificans) genes were correlated over 0.8, but ferric iron and Fli(Ferribacterium limineticm) gene were at the lowest of 0.7. For MTOT, it was at the highest related at 100% over BOD/COD. In addition, anaerobic genes(i.e., nirS-Nitrite reductase, MCR. Dde, DSR) and DO were also related more than 0.8, which showing anaerobic reactions generally dependant upon DO. As demonstrated in the study, molecular biological investigation and water quality parameters are highly co-linked, so that quantitative real-time PCR could be cooperatively used for assessing landfill stabilization in association with the conventional monitoring parameters.