• 한국초지조사료학회지
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• 제43권3호
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• pp.148-155
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• 2023

This study was aimed to isolate bacterial inoculants producing chitinase and evaluate their application effects on corn silage. Four corn silages were collected from four beef cattle farms to serve as the sources of bacterial inoculants. All isolates were tested against Fusarium graminearum head blight fungus MHGNU F132 to confirm their antifungal effects. The enzyme activities (carboxylesterase and chitinase) were also measured to isolate the bacterial inoculant. Based on the activities of anti-head blight fungus, carboxylesterase, and chitinase, L. buchneri L11-1 and L. paracasei L9-3 were subjected to silage production. Corn forage (cv. Gwangpyeongok) was ensiled into a 10 L mini silo (5 kg) in quadruplication for 90 days. A 2 × 2 factorial design consists of F. graminearum contamination at 1.010⁴ cfu/g (UCT (no contamination) vs. CT (contamination)) and inoculant application at 2.1 × 10⁵ cfu/g (CON (no inoculant) vs. INO (inoculant)) used in this study. After 90 days of ensiling, the contents of CP, NDF, and ADF increased (p<0.05) by F. graminearum contamination, while IVDMD, acetate, and aerobic stability decreased (p<0.05). Meanwhile, aerobic stability decreased (p<0.05) by inoculant application. There were interaction effects (p<0.05) on IVNDFD, NH3-N, LAB, and yeast, which were highest in UCT-INO, UCT-CON, CT-INO, and CT-CON & INO, respectively. In conclusion, this study found that mold contamination could negatively impact silage quality, but isolated inoculants had limited effects on IVNDFD and yeast.

• Journal of Microbiology and Biotechnology
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• 제28권9호
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• pp.1502-1510
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• 2018

Organic solvent-tolerant (OST) enzymes are widely applied in various industries for their activity and stability in organic solvents, for their higher substrate solubility, and for their greater stero-selectivity. However, the criteria for identifying OST enzymes largely remain undefined. In this study, we compared the amino acid composition of 19 OST esterases with that of 19 non OST esterases. OST esterases have increased the ratio of Ala and Arg residues and decreased the ratio of Asn, Ile, Tyr, Lys, and Phe residues. Based on our amino acid composition analysis, we cloned a carboxylesterase (EstSP2) from a psychrophilic bacterium, Sphingomonas glacialis PAMC 26605, and characterized its recombinant protein. EstSP2 is a substrate specific to p-nitrophenyl acetate and hydrolyzed aspirin, with optimal activity at $40^{\circ}C$; at $4^{\circ}C$, the activity is approximately 50% of its maximum. As expected, EstSP2 showed tolerance in up to 40% concentration of polar organic solvents, including dimethyl sulfoxide, methanol, and ethanol. The results of this study suggest that selecting OST esterases based on their amino acid composition could be a novel approach to identifying OST esterases produced from bacterial genomes.

• Environmental Analysis Health and Toxicology
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• 제7권1_2호
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• pp.17-34
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• 1992

The object of this study is to investigate the toxicity of S-biol (d-allethron-d-transchrysanthemate) and the mode of action between other synthetic pyrethroid insecticides and S-biol in rats. Rats were treated daily with S-biol (25 mg/kg, 50 mg/kg, 100mg/kg) by oral administration for 5 weeks. The experimental results were summerized as follows: Biochemical parameters such as LDH and Glucose in serum were much more increased in control groups. No significant hematological change excepts for MCHC in rats treated with S-biol 100 mg/kg were observed in all groups compared to control groups. In animals treated with S-biol for 4∼5 weeks, the levels of cytochrome P-450 in the liver were significantly increased. In renal microsomal fractions, however, no significant changes of cytochrome P-450 contents were observed. The activitis of ATPase in groups treated with S-biol (50 mg/kg, 100 mg/kg) were decreased compared to those in other groups. TBA values and activities of glucose-6-phosphatase in the liver were increased a little in the groups treated with S-biol (100 mg/kg) for 5 weeks. The activities of cholinesterase in hepatic and serum were not significantly changed in all groups but slightly decreased in animals treated with high dose of S-biol (100 mg/kg). The activities of carboxylesterase in serum and in the liver were slightly increased but not significantly changed.

• Parasites, Hosts and Diseases
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• 제56권6호
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• pp.553-558
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• 2018

Pathogenic Acanthamoeba spp. cause granulomatous amoebic encephalitis and keratitis. Acanthamoeba keratitis (AK) is a rare but serious ocular infection that can result in permanent visual impairment or blindness. However, pathogenic factors of AK remain unclear and treatment for AK is arduous. Expression levels of proteins secreted into extracellular space were compared between A. castellanii pathogenic (ACP) and non-pathogenic strains. Two-dimensional polyacrylamide gel electrophoresis revealed 123 differentially expressed proteins, including 34 increased proteins, 7 qualitative increased proteins, 65 decreased proteins, and 17 qualitative decreased proteins in ACP strain. Twenty protein spots with greater than 5-fold increase in ACP strain were analyzed by liquid chromatography triple quadrupole mass spectrometry. These proteins showed similarity each to inosine-uridine preferring nucleoside hydrolase, carboxylesterase, oxygen-dependent choline dehydrogenase, periplasmic-binding protein proteinases and hypothetical proteins. These proteins expressed higher in ACP may provide some information to understand pathogenicity of Acanthamoeba.

• 한국응용곤충학회지
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• 제32권3호
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• pp.348-353
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• 1993

포장의 약제저항성 변동요인을 조사하기 위하여 복수아혹진딧물의 Carboxyl esterase(CE) 활서측정에 의한 저항성도의 변동상황을 조사하였다. 비닐하우스에서 고추육묘중 발생한 복수아혹진딧물의 CE활성 측정 결과 약제저항성도는 무처리구가 40이였으나 아세페이트를 처리한구는 78로 약제처리로 인한 저항성도가 38이 증가하였다. 또한 노천망실에서 약제처리를 하지 않고 재배한 케일에 발생한 복숭아혹진딧물의 CE활성에 의한 저항성도는 7월이 24이었으나, 8, 9월의 활성이 계속 증가하여 10월이 83으로서 최고에 달하였다가 11월에는 다시 81에서 79로 약천 떨어지고 있어 약제이외에 저항성도의 자연변동요인이 있음을 확인하였다. 전국적으로 18개장소에서 채집한 진딧물의 CE활성을 측정한 결과 저항성도의 평균이 여름은 50$\pm$14였으며 늦가을인 11월은 82$\pm$10으로서 여름보다 늦가을인 11월이 32가 높았으며 그중에도 약제를 살포사지 않은 곳 보다 약제살포 회수가 많은 곳일수록 높았다.

• 한국동물학회지
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• 제33권3호
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• pp.337-345
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• 1990

배추흰나비로부터 정제된 3개의 esterase의 특성을 조사하였으며 면역학적 연구를 통해 E6의 생리적 역할을 추정하였다. 억제제의 연구결과, E2, E6, E11 모두 carboxylesterase임이 밝혀졌다. Km 값은 E2, E6 및 E11이 각각 6.89 X 10-$^4$M. 3.19 $\times$ l0-$^4$M, 3.69 X 10-$^4$M이었다. 분자량은 E2, E6 및 E11이 각각 42 KD, 81 KD, 174 KD이었고 등전점은 각각 pH 5.54, pH 5.89, pH 6.50이었다. 변태에 따른 E6의 농도는 5령 말기에서 가장 높았으며 특히 중장조직에서 높게 나타났다. 따라서 E6은 소화효소로 사료된다.

• Journal of Microbiology and Biotechnology
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• 제24권11호
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• pp.1484-1489
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• 2014

A novel esterase gene, est01, was successfully unearthed from a biogas digester microbiota metagenomic library. The 1,194 bp est01 gene encodes a protein of 44,804 Da (designated Est01). The amino acid sequence of Est01 shows only moderate (33%) identity to a lipase/esterase. Phylogenetic analysis and biochemical characterization confirmed that Est01 is a new member of family VIII esterases. The purified Est01 from recombinant Escherichia coli BL21 (DE3) showed high hydrolytic activity against short-chain fatty acid esters, suggesting that it is a typical carboxylesterase rather than a lipase. Furthermore, the Est01 was even active at $10^{\circ}C$ (43% activity remained), with the optimal temperature at $20^{\circ}C$, and had a broad pH range from 5.0 to 10.0, with the optimal pH of 8.0. These properties suggest that Est01 is a cold-adaptive esterase and could have good potential for low-temperature hydrolysis application.

• 농업생명과학연구
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• 제46권3호
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• pp.109-118
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• 2012

The gene encoding an esterase enzyme was cloned from a metagenomic library of cow rumen bacteria. The esterase gene (est2R) was 2,120 bp in length, encoding a protein of 516 amino acid residues with a calculated molecular weight of 57,286 Da. The molecular weight of the enzyme was estimated to be 57,000 Da by SDS-PAGE. Est2R shared 35.6% amino acid identity with esterase (CAH19079) of uncultured prokaryote. The Est2R was most active at $20-40^{\circ}C$, and showed optimum at $30^{\circ}C$ and pH 8.0. The most activity of Est2R for the different chain length of p-nitrophenyl ester group as substrate was p-nitrophenyl acetate. Moreover, the enzyme was found to be most active without organic solvent, followed by 98% active with ethanol, and the enzyme activity was highly affected by the acetonitrile. The enzyme was significantly inhibited by $Zn^{2+}$ but stimulated by $Ca^{2+}$. So, novel esterase gene est2R is likely to obtain from cow rumen metagenome and supposed to use for industrial purpose.

• Journal of Applied Biological Chemistry
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• 제62권1호
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• pp.73-79
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• 2019

A novel esterase gene (est7S) was cloned from an enriched metagenomic library derived from an oil-spill area. The gene encoded a protein of 505 amino acids, and the molecular mass of the Est7S was estimated to be 54,512 Da with no signal peptide. Est7S showed the highest identity of 40% to an esterase from a sludge metagenome compared to the characterized enzymes with their properties, although it showed 99% identity to a carboxylesterase in the genome sequence of Alcanivorax borkumensis SK2. Est7S had catalytic triad residues, Ser183, Glu312, and His420, and the GESAG motif in most family VII lipolytic enzymes. Est7S was purified from the crude extract of clone SM7 using Sephacryl S-200 HR and HiTrap Q column chromatographies. The purified Est7S was optimally active at $50^{\circ}C$ and pH 10.0. Est7S showed a high specific activity of 366.7 U/mg protein. It preferred short length esters, particularly p-nitrophenyl acetate, efficiently hydrolyzed R- and S-enantiomers of methyl-3-hydroxy-2-methylpropionate, and glyceryl tributyrate. These properties of Est7S may provide potential merits in biotechnological applications such as detergent and paper processing under alkaline conditions.

• Asian-Australasian Journal of Animal Sciences
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• 제31권1호
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• pp.138-148
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• 2018

Objective: Fusarium mycotoxins deoxynivalenol (DON) and zearalenone (ZEN), common contaminants in the feed of farm animals, cause immune function impairment and organ inflammation. Consequently, the main objective of this study was to elucidate DON and ZEN effects on the mRNA expression of pro-inflammatory cytokines and other immune related genes in the kidneys of piglets. Methods: Fifteen 6-week-old piglets were randomly assigned to three dietary treatments for 4 weeks: control diet, and diets contaminated with either 8 mg DON/kg feed or 0.8 mg ZEN/kg feed. Kidney samples were collected after treatment, and RNA-seq was used to investigate the effects on immune-related genes and gene networks. Results: A total of 186 differentially expressed genes (DEGs) were screened (120 upregulated and 66 downregulated). Gene ontology analysis revealed that the immune response, and cellular and metabolic processes were significantly controlled by these DEGs. The inflammatory stimulation might be an effect of the following enriched Kyoto encyclopedia of genes and genomes pathway analysis found related to immune and disease responses: cytokine-cytokine receptor interaction, chemokine signaling pathway, toll-like receptor signaling pathway, systemic lupus erythematosus (SLE), tuberculosis, Epstein-Barr virus infection, and chemical carcinogenesis. The effects of DON and ZEN on genome-wide expression were assessed, and it was found that the DEGs associated with inflammatory cytokines (interleukin 10 receptor, beta, chemokine [C-X-C motif] ligand 9, CXCL10, chemokine [C-C motif] ligand 4), proliferation (insulin like growth factor binding protein 4, IgG heavy chain, receptor-type tyrosine-protein phosphatase C, cytochrome P450 1A1, ATP-binding cassette sub-family 8), and other immune response networks (lysozyme, complement component 4 binding protein alpha, oligoadenylate synthetase 2, signaling lymphocytic activation molecule-9, ${\alpha}$-aminoadipic semialdehyde dehydrogenase, Ig lambda chain c region, pyruvate dehydrogenase kinase, isozyme 4, carboxylesterase 1), were suppressed by DON and ZEN. Conclusion: In summary, our results indicate that high concentrations of DON and ZEN suppress the inflammatory response in kidneys, leading to potential effects on immune homeostasis.