• Journal of Radiation Protection and Research
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• v.31 no.2
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• pp.65-68
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• 2006

In this study, the extent of plasmid DNA damage was observed according to concentration of BSH(Boron Sulfhydryl Hydride) and irradiation doses of neutron and ${\gamma}$-ray. The plasmid used was both pBR 322 (2870 bp) and ${\Phi}X174$ RF(5386 bp) DNA. Plasmid DNA damage by irradiation in the presence of BSH was analyzed by agarose gel electrophoresis. In the neutron experiment, DNA damage of both plasmid DNAs was increased according to increasing the concentration of BSH and neutron doses. But in the ${\gamma}$-ray experiment, there appeared no dose dependency as compared to the neutron experiment. The extent of the plasmid DNA damage in the presence of BSH was somewhat different according to irradiation by neutron or ${\gamma}$-ray.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.10
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• pp.1505-1512
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• 2005

Bile salt deconjugation is the most biologically significant reaction among the bacterial alterations of bile acids in the gastrointestinal tract of human and animal. The responsible enzyme, bile salt hydrolase (BSH), catalyzes the hydrolysis of glycineand/or taurine-conjugated bile salts into amino acid residues and deconjugated bile acids. Herein we review current knowledge on the distribution of BSH activity among various microorganisms with respect to their biochemical and molecular characteristics. The proposed physiological impact of BSH activity on the host animal as well as on the BSH-producing bacterial cells is discussed. BSH activity of the probiotic strains is examined on the basis of BSH hypothesis, which was proposed to explain cholesterol-lowering effects of probiotics. Finally, the potential applications of BSH research are briefly discussed.

• Journal of Dairy Science and Biotechnology
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• v.29 no.1
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• pp.49-54
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• 2011

Bile salt hydrolase (BSH, EC 3.5.1.24) activity, which cleaves amide bond between carboxyl group (bile acid) and amino group (glycine or taurine), is commonly detected in gut-associated species of human and animal. During the screening of BSH active strains from the fecal samples of elderly human volunteers, strain CU30-2 was isolated on the basis of the highly active BSH producing activity. A bsh gene of the isolate was cloned into the pET22b expression vector and overexpressed in Escherichia coli BL21 (DE3) Gold by induction with 1mM IPTG. The overexpressed BSH enzyme with 6x His-tag was purified with apparent homogeneity using a $Ni^+$-NTA agarose column and characterized. The BSH enzyme of E. faecalis CU30-2 exhibited approximately 50 times higher activity against glycol-conjugated bile salts than tauro-conjugated bile salts having the highest activity against glycocholic acid. Considering the prevalence of E. faecalis strains in the human GI tract and glycol-conjugates dominated bile acid composition of human bile, further study is needed to investigate the impact of the BSH activity exerted by E. faecalis strains to the host as well as to the BSH producing strains.

• Journal of Microbiology and Biotechnology
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• v.18 no.3
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• pp.449-456
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• 2008

Phenotypic screening for bile salt hydrolase (BSH) activity was performed on Lactobacillus acidophilus PF01 isolated from piglet feces. A gene encoding BSH was identified and cloned from the genomic library of L. acidophilus PF01. The bsh gene and surrounding regions were characterized by nucleotide sequence analysis and were found to contain a single open reading frame (ORF) of 951 nucleotides encoding a 316 amino acid protein. The potential bsh promoter region was located upstream of the start codon. The protein deduced from the complete ORF had high similarity with other BSHs, and four amino acid motifs located around the active site, FGRNXD, AGLNF, VLTNXP, and GXGXGXXGXPGD, were highly conserved. The bsh gene was cloned into the pET21b expression vector and expressed in Escherichia coli BLR(DE3) by induction with 0.1mM of isopropylthiogalactopyranoside. The BSH enzyme was purified with apparent homogeneity using a $Ni^{2+}$-NTA agarose column and characterized. The overexpressed recombinant BSH enzyme of L. acidophilus PF01 exhibited hydrolase activity against tauroconjugated bile salts, but not glycoconjugated bile salts. It showed the highest activity against taurocholic acid. The maximum BSH activity occurred at approximately $40^{\circ}C$. The enzyme maintained approximately 70% of its maximum activity even at $60^{\circ}C$, whereas its activity rapidly decreased at below $37^{\circ}C$. The optimum pH was 6, and BSH activity was rapidly inactivated below pH 5 and above pH 7.

• Proceedings of the Korean Nuclear Society Conference
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• 2004.10a
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• pp.1212-1213
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• 2004

The plasmid was used pBR 322 and ${\varphi}X174$ RF DNA. In neutron experiment, damage of pBR 322 and ${\varphi}X174$ RF DNA were observed according to increasing concentration of BSH and neutron dose. Damage of plasmid DNA appeared obvious by increasing of BSH and neutron irradiation. In ${\gamma}-$ radiation experiment, it was carried out like above neutron experiment but damages of two plasmid appeared no differences from the control unlike neutron result.

• Journal of Microbiology and Biotechnology
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• v.21 no.8
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• pp.838-845
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• 2011

The present work describes the identification, purification, and characterization of bile salt hydrolase (BSH) from Bifidobacterium animalis subsp. lactis. The enzyme was purified to electrophoretic homogeneity by hydrophobic chromatography, ion-exchange chromatography and ultrafiltration. SDS-PAGE analysis of putative BSH and gel filtration revealed that the analyzed protein is presumably a tetramer composed of four monomers each of about 35 kDa. The purified enzyme was analyzed by liquid chromatography coupled to LTQ FT ICR mass spectrometry and unambiguously identified as a bile salt hydrolase from B. animalis. The isoelectric point of the studied protein was estimated to be around pH 4.9. The pH optimum of the purified BSH is between 4.7 to 6.5, and the temperature optimum is around 50oC. The BSH of B. animalis could deconjugate all tested bile salts, with clear preference for glycine-conjugated bile salts over taurine-conjugated forms. Genetic analysis of the bsh showed high similarity to the previously sequenced bsh gene from B. animalis and confirmed the usefulness of bile salt hydrolase as a genetic marker for B. animalis identification.

• Journal of Microbiology and Biotechnology
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• v.25 no.12
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• pp.2100-2105
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• 2015

Probiotic bacteria must have not only tolerance against bile salt but also no genes for antibiotic resistance. Leuconostoc citreum is a dominant lactic acid bacterium in various fermented foods, but it is not regarded as a probiotic because it lacks bile salt resistance. Therefore, we aimed to construct a bile salt-resistant L. citreum strain by transforming it with a bile salt hydrolase gene (bsh). We obtained the 1,001 bp bsh gene from the chromosomal DNA of Lactobacillus plantarum and subcloned it into the pCB4170 vector under a constitutive P710 promoter. The resulting vector, pCB4170BSH was transformed into L. citreum CB2567 by electroporation, and bile salt-resistant transformants were selected. Upon incubation with glycodeoxycholic acid sodium salt (GDCA), the L. citreum transformants grew and formed colonies, successfully transcribed the bsh gene, and expressed the BSH enzyme. The recombinant strain grew in up to 0.3% (w/v) GDCA, conditions unsuitable for the host strain. In in vitro digestion conditions of 10 mM bile salt, the transformant was over 67.6% viable, whereas only 0.8% of the host strain survived.

• Journal of Plant Biology
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• v.38 no.4
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• pp.365-371
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• 1995

Binary vectors, pBI-ActR1, pBI-ActF1 and pBSH-ActR1, were constructed using pGA642, the replication origin of pTi12 and the rice actin promoter. The sizes of pBI-ActR1, pBI-ActF1 and pBSH-ActR1 were 12.9 kb, 13.2 kb and 11.95 kb, respectively. These vectors containing a rice actin promoter followed by a GUS structural gene could induce stronly the expression of GUS gene in transformed rice cells. Rice explants from 3-4 day old seedlings after germinatin were cocultured with A. tumefaceins harboring pBI-ActR1, pBI-ActF1 or pBSH-ActR1, and then GUS expression in the explants was assayed. Transformation of rice explants by these binary vectors was tissue-specific, such that the meristematic regions of shoot apex, root and hypocotyl were transformed by these binary vectors.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.52 no.4
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• pp.469-473
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• 2007

With the aim to analyze stability performance of six promising barley genotypes, eleven yield related characters were evaluated employing varied irrigation treatments under the tropical climate of Northern part in Bangladesh. Analysis of variance(ANOVA), phenotypic index, regression co-efficient(bi) and deviation from regression($s^2_d$) of the individual genotypes were estimated to evaluate the stable performance of the genotypes. A significant interaction was observed between the genotypes and irrigation period($G{\times}T$). Among all the genotypes, BSH-2 showed stable performance for plant height under different irrigation period, where $P>\bar{X},\;bi{\sim}1\;and\;s^2_d{\sim}0$. High phenotypic index, lower bi value and low deviations from regression were observed in case of spikelet number per spike and grain number per spike for genotype BSH-2 and plant height, spike length and harvest index per plant for BB-2 which suggest that those parameters were not usually affected by irrigation. On the other hand the genotype BSH-2 for tiller number and BB-1 for the fertile tiller number were not suitable for favorable moisture content, where $P<\bar{X},\;bi>1.0\;and\;low\;s^2_d$. Thus we suggest that genotype BSH-2 might have transmit high mean and increased phenotypic stability to the next progenies, which may consider as an ideal genotype for developing improved barely cultivars.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.3
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• pp.309-318
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• 2009

This study was conducted to investigate the cholesterol lowering and anti-obesity effects of an ethanol extract of broccoli sprouts (BS) in rats fed high fat diet. Male Sprague-Dawley rats weighing $150{\sim}155g$, were divided into 6 groups; a normal diet group (ND), a high fat diet group (HFD), a normal diet and BS with 200 mg/kg treated group (ND-BSL), a normal diet and BS with 400 mg/kg treated group (ND-BSH), a high fat diet and BS with 200 mg/kg treated group (HFD-BSL), and a high fat diet and BS with 400 mg/kg treated group (HFD-BSH). The body weight gain and mesenteric adipose tissue weight were increased by high fat diet, but gradually decreased to the corresponding level of ND group after administration of BS extract. The liver and epididymal adipose tissue weights of HFD group were the highest among the six groups, although the difference was not significant. Food intake was lower in high fat diet groups compared with normal diet groups. The serum ALT and AST activities that were elevated by the high fat diet were significantly decreased after BS administration. Levels of serum total cholesterol, LDL-cholesterol, triglyceride and the atherogenic index tended to be decrease in the BS administered groups compared with HFD group. However, HDL-cholesterol level in serum decreased in HFD group and markedly increased in BS administered groups. There were no differences in the contents of serum triglyceride, LDL-cholesterol and HDL-cholesterol between normal diet groups. Levels of total cholesterol and triglyceride in liver and adipose tissues were also lower in BS administrated groups than in HFD group. The activities of heparin-releasable lipoprotein lipase (HR-LPL) and total-extractable LPL (TE-LPL) in adipose tissue were increased in HFD group compared with the BS administered groups, but those of the ND-BSL group and ND-BSH group were similar to ND group. These results suggest that BS ethanol extract may exert cholesterol-lowering effect and potentially reduce lipid storage.