• Title/Summary/Keyword: gene conversion

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Isolation and Characterization of Xylanase from a Novel Strain, Penicillium menonorum SP10

  • Thi Thu Huong Luong;Supattra Poeaim ;Narumon Tangthirasunun
    • Mycobiology
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    • v.51 no.4
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    • pp.239-245
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    • 2023
  • Xylanase has been applied in various sectors, such as biomass conversion, paper, pulp, textiles, and pharmaceutical industries. This study aimed to isolate and screen potential xylanase-producing fungi from the soil of Suphan Buri Province, Thailand. Fifteen fungi were isolated, and their xylanase activities were tested by the qualitative method. The result showed that isolate SP3, SP10 and SP15 gave high xylanase activity with potency index (PI) of 2.32, 2.01 and 1.82, respectively. These fungi were selected for the xylanase quantitative test, isolate SP10 performed the highest xylanase activity with 0.535 U/mL. Through molecular methods using the 𝛽-tubulin gene, isolate SP10 was identified as Penicillium menonorum. The xylanase characteristics from P. menonorum SP10 were determined, including the xylanase isoforms and the optimum pH and temperature. The xylanase isoforms on SDS-PAGE indicated that P. menonorum SP10 produced two xylanases (45 and 54 kDa). Moreover, its xylanase worked optimally at pH 6 and 55 ℃ while reaching 61% activity at 65 ℃. These results proposed P. menonorum SP10 as a good candidate for industrial uses, especially in poultry feed and pulp industries, to improve yield and economic efficiency under slightly acidic and high-temperature conditions.

Cloning and Characterization of Ginsenoside-Hydrolyzing β-Glucosidase from Lactobacillus brevis That Transforms Ginsenosides Rb1 and F2 into Ginsenoside Rd and Compound K

  • Zhong, Fei-Liang;Ma, Rui;Jiang, Mingliang;Dong, Wei-Wei;Jiang, Jun;Wu, Songquan;Li, Donghao;Quan, Lin-Hu
    • Journal of Microbiology and Biotechnology
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    • v.26 no.10
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    • pp.1661-1667
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    • 2016
  • The ginsenoside-hydrolyzing β-glucosidase gene (bgy2) was cloned from Lactobacillus brevis. We expressed this gene in Escherichia coli BL21(DE3), isolated the resulting protein, and then utilized the enzyme for the biotransformation of ginsenosides. The bgy2 gene contains 2,223 bp, and encodes a protein of 741 amino acids that is a member of glycosyl hydrolase family 3. β-Glucosidase (Bgy2) cleaved the outer glucose moieties of ginsenosides at the C-20 position, and the inner glucose at the C-3 position. Under optimal conditions (pH 7.0, 30℃), we used 0.1 mg/ml Bgy2 in 20 mM sodium phosphate buffer (PBS) for enzymatic studies. In these conditions, 1.0 mg/ml ginsenoside Rb1 and ginsenoside F2 were converted into 0.59 mg/ml ginsenoside Rd and 0.72mg/ml compound K, with molar conversion productivities of 69% and 91%, respectively. In pharmaceutical and commercial industries, this recombinant Bgy2 would be suitable for producting ginsenoside Rd and compound K.

Tamoxifen Resistance and CYP2D6 Copy Numbers in Breast Cancer Patients

  • Motamedi, Sahar;Majidzadeh, Keivan;Mazaheri, Mahta;Anbiaie, Robab;Mortazavizadeh, Seyed Mohammad Reza;Esmaeili, Rezvan
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.12
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    • pp.6101-6104
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    • 2012
  • Background: Breast cancer accounts about one million from total annual ten million new diagnosed cases of neoplasia worldwide and is the main cause of death due to cancer in women. Tamoxifen is the most popular selective estrogen receptor modulator used in anti estrogen treatments. Tamoxifen must be converted into its metabolite endoxifen for biologic effects; this conversion process is catalysed by highly polymorphic cytochrome P450 2D6 (CYP2D6). This study surveyed copy number variation of the CYP2D6 gene and its possible correlation with Tamoxifen resistance in breast cancer patients. Methods: This case control study was performed on samples taken from 79 patients with breast cancer who used tamoxifen in Yazd and Tehran Cities, Iran. Real time reactions were conducted for 10 healthy samples using the comparative $C_t$ (Cycles threshold) method, each pair of genes being compared and samples with ratios around 1 were taken as control samples. Proliferation reactions were done by Real-Time PCR ABI Prism 7500. All registered data were transformed into SPSS 15 program and analyzed. Results: Efficiency of PCR for both CYP2D6 and ALB genes was 100%. From all 23 drug resistant patients 21.7% had one copy, 47.8% two copies and 30.4% had three copies. Also from all 56 drug sensitive patients, 26.8% had one copy, 51.8% two copies and 21.4% had three copies. The percentage of patients with one and two copies was similar between two groups but patients with three copies were more likely to belong to the drug resistant group more. Odd ratios for one and two copies were 0.759 and 0.853 respectively, indicating possible protective effects while that for three copies was 1.604. Conclusions: Based on our study there is no significant link between CYP2D6 gene copy numbers and tamoxifen resistance in women with breast cancer. But more studies considering other influencing factors appear warranted.

Effect of the Transformed Lactobacillus with Phytase Gene on Pig Production Performance, Nutrient Digestibility, Gut Microbes and Serum Biochemical Indexes

  • Yin, Q.Q.;Chang, J.;Zuo, R.Y.;Chen, L.Y.;Chen, Q.X.;Wei, X.Y.;Guan, Q.F.;Sun, J.W.;Zheng, Q.H.;Yang, X.;Ren, G.Z.
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.2
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    • pp.246-252
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    • 2010
  • In order to improve the availability of phytase and probiotics together, a phytase gene from Aspergillus ficuum has been expressed in Lactobacillus. In this study, the transformed Lactobacillus with phytase gene was fed to pigs to determine its effect on pig production, feed conversion and gut microbes. Forty eight, 60-day-old, castrated pigs (Duroc${\times}$Landrace${\times}$Pietrain) were assigned to 6 groups, 8 pigs for each group. Group 1 was the control, group 2 was added with chlortetracycline (500 mg/kg), group 3 was added with the transformed Lactobacillus (500 mg/kg) with 20% (w/w) of calcium monohydrogen phosphate (CMP, $CaHPO_{4}$) removed, group 4 was added with the natural Lactobacillus (500 mg/kg) with 20% (w/w) of CMP removed, group 5 was added with the transformed Lactobacillus (500 mg/kg) with 40% (w/w) of CMP removed, group 6 was added with phytase (500 mg/kg) with 40% (w/w) of CMP removed. The results showed: i) the average daily gain (ADG) was improved in groups 2, 3 and 4 (p<0.05); ii) the diarrhea rates in the groups added with Lactobacillus were lower than in the other groups (p<0.05), in which the transformed Lactobacillus had more effect on reducing digestive disease; iii) the transformed Lactobacillus was most effective in improving the digestibilities of crude protein (CP), calcium (Ca), phosphorus (P), compared with the other groups (p<0.05); iv) Lactobacillus could increase lactic acid bacterium number and ammonia concentrations, and decrease pH values and E. coli number in pig feces (p<0.05); v) the phytase activity in the feces of pigs fed with the transformed Lactobacillus was 133.32 U/g, which was higher than in group 4 (9.58 U/g, p<0.05), and was almost the same as group 6 (135.94 U/g); vi) the transformed Lactobacillus could increase serum concentrations of IgA, triglyceride, and glutamic oxaloacetic transaminase activity (p<0.05), and had no significant effect on other serum indexes (p>0.05).

Polymorphisms in the uncoupling protein 3 gene and their associations with feed efficiency in chickens

  • Jin, Sihua;Yang, Lei;He, Tingting;Fan, Xinfeng;Wang, Yiqiu;Ge, Kai;Geng, Zhaoyu
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.9
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    • pp.1401-1406
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    • 2018
  • Objective: The uncoupling protein 3 (UCP3) is a member of the mitochondrial anion carrier superfamily and has crucial effects on growth and feed efficiency in many species. Therefore, the objective of the present study was to examine the association of polymorphisms in the UCP3 gene with feed efficiency in meat-type chickens. Methods: Six single nucleotide polymorphisms (SNPs) of the UCP3 gene were chosen to be genotyped using matrix-assisted laser desorption-ionization time-of-flight mass spectrometry in meat-type chicken populations with 724 birds in total. Body weight at 49 (BW49) and 70 days of age (BW70) and feed intake (FI) in the interval were collected, then body weight gain (BWG) and feed conversion ratio (FCR) were calculated individually. Results: One SNP with a low minor allele frequency (<1%) was removed by quality control and data filtering. The results showed that rs13997809 of UCP3 was significantly associated with BWG and FCR (p<0.05), and that rs13997811 had significant effects on BW70 and BWG (p<0.05). Rs13997812 of UCP3 was strongly associated with BW70, FI, and FCR (p<0.05). Furthermore, individuals with AA genotype of rs13997809 had significantly higher BWG and lower FCR (p<0.05) than those with AT genotype. The GG individuals showed strongly higher BW70 and BWG than AA birds in rs13997811 (p<0.05). Birds with the TT genotype of rs13997812 had significantly greater BW70 and lower FCR compared with the CT birds (p<0.05). In addition, the TAC haplotype based on rs13997809, rs13997811, and rs13997812 showed significant effects on BW70, FI, and FCR (p<0.05). Conclusion: Our results therefore demonstrate important roles for UCP3 polymorphisms in growth and feed efficiency that might be used in meat-type chicken breeding programs.

Effects of feeding corn naturally contaminated with aflatoxin on growth performance, apparent ileal digestibility, serum hormones levels and gene expression of Na+, K+-ATPase in ducklings

  • Abbasi, Farzana;Liu, Jingbo;Zhang, Hongfu;Shen, Xiaoyun;Luo, Xuegang
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.1
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    • pp.91-97
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    • 2018
  • Objective: A 14-d trial was conducted to determine the effects of feeding corn naturally contaminated with aflatoxin $B_1$ ($AFB_1$) on growth performance, apparent ileal digestibility, serum hormones levels and gene expression of $Na^+$, $K^+-ATPase$ in ducklings. Methods: A total of 704 ducklings were blocked on the basis of sex and body weight (BW), and then allocated randomly to one of the following two treatments: i) CON, basal diet and ii) $AFB_1$, diets with 100% of normal corn replaced with $AFB_1$ contaminated corn. There were 22 pens per treatment and 16 birds per pen. The concentration of $AFB_1$ was 195.4 and $124.35{\mu}g/kg$ in the contaminated corn and $AFB_1$ diet, respectively. Results: The $AFB_1$ decreased average daily gain, average daily feed intake, d 7 BW, final BW in the whole trial, and feed conversion ratio (FCR) during d 8 to 14 and d 1 to 14 by 10% to 47% (p<0.05), while FCR during d 1 to 7 was increased (p<0.05). $AFB_1$ did not affect mortality to 7 d of age, and then increased to 5.8% from 8 to 14 d of age (p<0.01). Apparent ileal gross energy digestibility was reduced by $AFB_1$, whereas apparent ileal digestibility of dry matter, nitrogen, and amino acid was improved (p<0.01). Feeding $AFB_1$ diets increased serum concentration of leptin and insulin-like growth factors-1 (IGF-1) (p<0.05), but had no effect on neuropeptide Y, ghrelin, cholecystokinin-8 or insulin (p>0.05). Dietary treatments did not influence relative expression of jejunal $Na^+$, $K^+-ATPase$ gene (p>0.05). Conclusion: Taken together, feeding corn naturally contaminated with $AFB_1$ reduced growth performance, improved apparent ileal digestibility, and affected serum leptin and IGF-1 in ducklings from d 1 to 14.

Investigation of Porcine Melanocortin-4 Receptor (MC4R) Polymorphism on Economic Traits (돼지 melanocortin-4 receptor (MC4R) 유전자의 경제형질과의 연관성에 관한 연구)

  • Kim Kwan-Suk;Shin Hee Young;Lee Joong-Jae;Hong Sung-Kwang;Choi Bong-Hwan;Kim Tae-Hun;Lee Hak-Kyo;Cho Byung-Wook
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.968-971
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    • 2005
  • The Melanocortin-4 receptor gene (MC4R) regulates the energy balance and the genetic basis of obesity. A polymorphism in the porcine melanocortin-4 receptor has previously shown to be associated with growth, fat deposition and feed intake. In this study, the polymorphism of the gene was studied in several pig breeds of Duroc, Landrace, Berkshire, and Yorkshire. The results showed that the frequencies of MC4R genotype varied among those breeds. Association analyses were also performed between the MC4R polymorphism and average daily gain, feed conversion ratio, backfat thickness and lean percentage phenotypes. The results strongly support that the MC4R polymorphism can be used DNA marker selection indicator for economically important traits for pig breeding program in Korea.

Effects of Corn Silage on Growth Performance, Carcass Characteristics, and Gene Expression of Longissimus dorsi Muscle Biopsy of Hanwoo Steers (옥수수 사일리지가 거세한우의 성장, 도체특성 및 등심 유전자 발현에 미치는 영향)

  • Kim, U.H.;Kang, D.H.;Park, B.H.;Jang, S.S.;Chung, K.Y.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.24 no.1
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    • pp.41-48
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    • 2022
  • This study was conducted to investigate effects of corn silage on final fattening performance, carcass characteristics, and gene expression of Hanwoo Longissimus dorsi muscle biopsy. Twenty one steers with initial body weight of control 291.5±6.5kg, corn silage 291.7.0±17.3kg were used for 18 months of fattening period. Average daily gain of corn silage tended to increase compared to control in early fattening period(p=0.092). Feed conversion ratio of corn silage was higher than control in early fattening(p=0.005). The animals in corn silage increased A grade 23% in meat quantity than the control. Myogenic gene expression on the Longissimus dorsi biopsy were compared between corn silage and control. The level of myosin heavy chain(MHC) I, IIX mRNA were greater than the control in the whole period(p<0.05). The level of Peroxisome proliferator-activated receptor γ (PPAR γ) mRNA was greater than the corn silage(p<0.05). Inconclusion, corn silage will be possible to use as an alternative concentrate feeding system.

Estimation of Anti-proliferative Activity of Saccharin against Various Cancer Cell Lines and MSCs (다양한 암세포 주와 MSCs에 대한 Saccharin의 항증식성 평가)

  • Choi, Jeong Su;Park, Sang Yong;Yang, Man Gil;Lee, Dong Beom;Lee, Tae Bok;Heo, Ji Hye;Lee, Min Woo;Kim, Suhng Wook
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.3
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    • pp.169-175
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    • 2016
  • Saccharin (o-benzoic sulfimide) is the first artificial and non-caloric sweetener that was first synthesized in 1879. In this study, we examined the biological activity of saccharin against various human cancer cell lines and human bone marrow-derived mesenchymal stem cells. A viability assay based on the conversion of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was performed to test for the cytotoxicity of saccharin about the four human cancer cell lines (H460, H157, A549 and SKOV3), one murine cancer cellline (Raw264.7), and MSCs. In order to find the differentially expressed gene in saccharin-treated MSCs against untreated MSCs, we performed annealing control primer (ACP)-based differential display reverse transcriptionp-olymerase chain reaction (DDRT-PCR). All tested cells were treated with saccharin at various concentrations (0.0, 4.8, 7.2, 9.6, 12.0, 14.4 mg/mL) for 48 hr. The number of metabolically active cancer cells decreased when treated with the saccharin at various concentrations for 48 hr as compared with the untreated cells. The decrease in cell survival was more evident with increasing concentrations of saccharin. Moreover, novel candidate genes, which were differentially expressed in MSCs in response to saccharin, were identified in 16 bands on 2% agarose gel. This revealed 16-7 up-regulated and 9 down-regulated-differentially expressed genes indicated by arrows. One of these candidate genes was a FK506-binding protein gene. The functional roles of FK506 binding proteins, with respect to the activities of stem cell proliferation, were not characterized. Further studies are required to get a better understanding of FK506-binding proteins in its roles in increasing stem cell proliferative activities from using saccharin.

Conversion of Ginsenoside $Rb_1$ by Ginseng Soil Bacterium Cellulosimicrobium sp. Gsoil 235 According to Various Culture Broths (인삼 토양 미생물 Cellulosimicrobium sp. Gsoil 235의 배지조성에 따른 Ginsenoside $Rb_1$ 전환)

  • Na, Ju-Ryun;Kim, Yu-Jin;Kim, Se-Hwa;Kim, Ho-Bin;Shim, Ju-Sun;Kim, Se-Young;Yang, Deok-Chun
    • Microbiology and Biotechnology Letters
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    • v.37 no.1
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    • pp.55-61
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    • 2009
  • Ginseng saponins (a secondary metabolite, termed ginsenosides) are the principal bioactive ingredients of ginseng, and modification of the sugar chains may markedly change the its biological activity. One of soil bacteria having $\beta$-glucosidase (to transform ginsenoside $Rb_1$) activity was isolated from soil of a ginseng field in Daejeon. 16S rRNA gene sequence analysis revealed that the isolate belonged to the genus Cellulosimicrobium, with highest sequence similarity (99.7%) to Cellulosimicrobium funkei ATCC BAA-$886^T$. The strain, Gsoil 235, could transform ginsenoside $Rb_1$ into Rd, $Rg_3$ and 3 of un-known ginsenosides by the analyses of TLC, HPLC. By investigating its deglycosylation progress, the optimal broth for, $\beta$-glucosidase was nutrient broth (In 48 hours, almost ginsenoside $Rb_1$ could be transformed into minor ginsenosides). On the contrary, the optimal broth for growth was determined as trypic soy broth (TSB).