Park, Sung-Yong;Bang, Mi-Ae;Oh, Boung-Jun;Park, Jeong-Hoon;Song, Won-Seob;Choi, Kyung-Min;Choung, Eui-Su;Boo, Hee-Ock;Cho, Seung-Sik
Korean Journal of Microbiology
/
v.49
no.3
/
pp.262-269
/
2013
The object of this study was to improve the quality of Cheonggukjang with new starter, Bacillus subtilis BC-P1. Twenty strains were isolated from the commercial cheonggukjang and 1 Bacillus strain (BC-P1) with protease activity was selected. The 16S rRNA gene sequence revealed that the BC-P1 was closely related to B. subtilis with 99% homology. The quality characteristics of chunggukjang fermented with B. subtilis BC-P1, Bacillus nato (PC) and commercial chunggukjang (NC) were investigated. The characteristics of fermentation were determined by protease, lipase, xylanase, chitinase, and fibrinolytic activities, reducing sugar, nutrient composition and amino acid contents of cheonggukjang sample. Cheonggukjang fermented with B. subtilis BC-P1 showed the strongest fibrinolytic, xylanase, and chitinase activities. Reducung sugar contents of Cheonggukjang samples were $30.16{\pm}2.11$ mg/g (NC), $28.56{\pm}1.52$ mg/g (PC), $32.39{\pm}1.87$ mg/g (BC-P1). And their total amino acid contents were 338.99 mg% (NC), 445.19 mg% (PC), 741.35 mg% (BC-P1). These results suggested that B. subtilis BC-P1 was suitable to be used as a starter to enhance the quality and effects of cheonggukjang.
The evolution dynamics of nanoscale Ge islands on both Si (001) and (113) surfaces is explored using ultraviolet photoelectron emission microscopy (UV-PEEM). Real-time monitoring of the in-situ growth of the Ge island structures can allow us to study the variation of the size, the shape and the density of the nanostructures. For Ge depositions greater than ${\sim}4$ monolayer (ML) with a growth rate of ${\sim}0.4\;ML/min$ at temperatures of $450-550^{\circ}C$, we observed island nucleation on both surfaces indicating the transition from strained layer to island structure. During continuous deposition the circular islands grew larger via ripening processes. AFM measurements showed that the islands grown on Si (001) were dome-shaped while the islands on Si (113) were multiple-side faceted with flat tops of (113)-orientation. In contrast, for Ge deposition with a lower growth rate of ${\sim}0.15\;ML/min$ on Si(113), we observed the shape transition from circular into elongated island structures. The elongated islands grew longer along the [$33\bar{2}$] during continuous Ge deposition. The shape evolution of the islands is discussed in terms of strain relaxation and kinetic effects.
The Photosynthetic bacterium, Rhodopseudomonas sphaeroides strain B6 was irnmobilized on agar gel. The optimum concentration of agar for hydrogen production was 2% (w/v). Maximum rates of hydrogen production by immobilized (300 ml of gel; 2.85 rng dry cells/ml) and free cells (1l culture; 0.87 mg dry cells/ml) were 47.5 and 48.0 ml/hr/culture, respectively. However, when both cultures were fed by 10 mmoles of lactate as limited electron donor at the later period of incubation, the activity of hydrogen production by free cells was significantly decreased but, immobilized cells continued hydrogen production with almost the same initial rate. Wc examined hydrogen production by immobilized cells of strain B6 under periodic illumination for 12 hr-intervals. When the culture was periodically fed by basal medium containing 9.3 rnmoles of DL-lactate and 1.86 mmoles of L-glutamate as consumed electron donor and nitrogen source, respectively, for every one liter of hydrogen produced, hydrogen was evolved continuously with the average rate of 510 ml/day/300 ml gel (2.9 rng dry cellslml) during the incubation time for 228 hr.
Heme oxygenase-1 (HO-1) is the inducible from of the rate-limiting enzyme of heme degradation; it regulates the cellular contents of heme. HO-1 is up-regulated by various stimuli including oxidative stress so that it is thought to participate in general cellular defense mechanisms against oxidative stress in mammalian cells. To investigate the role of the cAMP-dependent protein kinase A (PKA) signaling pathway on nitrogen oxidative stress-induced HO-1 gene expression, RAW 264.7 cell cultures were treated with sodium nitroprusside (SNP). SNP increased the expression of HO-1 mRNA and protein, time- and concentration-dependently. Treatment with H89, PKA inhibitor, but not LY83583, guanylate cyclase inhibitor, significantly diminished the HO-1 expression by SNP, indicating that cAMP plays a crucial role in the induction of HO-1. Incubation with cAMP-elevating agents, such as forskolin or isoproterenol resulted in up-regulation of the expression of HO-1. Forskolin-induced expression of HO-1 was inhibited by H89. Furthermore, propranolol, $\beta$-adrenoceptor blocker, inhibited the isoproterenol-induced HO-1 expression, supporting the importance of cAMP in the induction of HO-1 expression. Higenamine-S, but not higenamineR, enhanced the HO-1 expression induced by SNP. Furthermore, cellular toxicity induced by hydrogen peroxide was attenuated by the presence of SNP, which was further increased by the presence of ZnPPIX, HO-1 inhibitor. Collectively, these results strongly suggest that up-regulation of HO-1 expression in RAW 264.7 cells involves PKA signal pathway.
Expression of f3 ~agarase Gene and Catabolite Repression in Escherichia coli by the Promoter of Alginate Lyase Gene Isolated from Marine Pseudomonas sp. Jin, Cheal~Ho, J~Hyeon Park, Jeong-Hyun Han, YoonM Hyeok Chae, Jong~Hee Lee, Jung-Kee Lee!, and In-800 Kong*. Faculty of Food Science and Biotechnology, Pukyong National UniversitYt Pusan 608-737, Korea, llnBioNet Co. 1690-3 Taejon 306-230, Korea - Promoter is a key factor for expression of the recombinant protein. There are many promoters for overexpression of protein in various organisms. The aly promoter of Pseudomonas sp. W7 isolated from marine environment was known to be a constitutive expression promoter of the alginate lyase gene, and it's promoter activity is repressed by glucose in Escherichia coli. To investigate the catabolite repression of the aly promoter ~md association between the promoter mutants, f3 agarase gene, which was also cloned from Pseudomonas sp. W7 was connected to the aly promoter with the sequence the coding 46 N-terminal amino acids ofthe alginate lyase gene. The constructed plasmid was introduced into E. coli and the agarase activity was measured. Fourty six amino acids of the alginate lyase gene was serially deleted using peR to the direction of 5' upstream region and subcloned. The agarase was overexpressed by the aly promoter and the production of agarase was repressed by the addition of glucose into culture media. Fourty six amino acids of alginate lyase did not affect the production of agarase at all. The deletion of a putative stem-loop structure in the aly promoter induced the decrease of f3 -agarase productivity.
The aim of this modelling study was to investigate the effect of large herd size (and land areas) on walking distances and milking interval (MI), and their impact on milk yield and economic penalties when 50% of the total diets were provided from home grown feed either as pasture or grazeable complementary forage rotation (CFR) in an automatic milking system (AMS). Twelve scenarios consisting of 3 AMS herds (400, 600, 800 cows), 2 levels of pasture utilisation (current AMS utilisation of 15.0 t dry matter [DM]/ha, termed as 'moderate'; optimum pasture utilisation of 19.7 t DM/ha, termed as 'high') and 2 rates of incorporation of grazeable complementary forage system (CFS: 0, 30%; CFS = 65% farm is CFR and 35% of farm is pasture) were investigated. Walking distances, energy loss due to walking, MI, reduction in milk yield and income loss were calculated for each treatment based on information available in the literature. With moderate pasture utilisation and 0% CFR, increasing the herd size from 400 to 800 cows resulted in an increase in total walking distances between the parlour and the paddock from 3.5 to 6.3 km. Consequently, MI increased from 15.2 to 16.4 h with increased herd size from 400 to 800 cows. High pasture utilisation (allowing for an increased stocking density) reduced the total walking distances up to 1 km, thus reduced the MI by up to 0.5 h compared to the moderate pasture, 800 cow herd combination. The high pasture utilisation combined with 30% of the farm in CFR in the farm reduced the total walking distances by up to 1.7 km and MI by up to 0.8 h compared to the moderate pasture and 800 cow herd combination. For moderate pasture utilisation, increasing the herd size from 400 to 800 cows resulted in more dramatic milk yield penalty as yield increasing from c.f. 2.6 and 5.1 kg/cow/d respectively, which incurred a loss of up to $AU 1.9/cow/d. Milk yield losses of 0.61 kg and 0.25 kg for every km increase in total walking distance (voluntary return trip from parlour to paddock) and every one hour increase in MI, respectively. The high pasture utilisation combined with 30% of the farm in CFR in the farm increased milk yield by up to 1.5 kg/cow/d, thereby reducing loss by up to $0.5/cow/d (c.f. the moderate pasture and 800 cow herd scenario). Thus, it was concluded that the successful integration of grazeable CFS with pasture has the potential to improve financial performance compared to the pasture only, large herd, AMS.
The aim of this study was to screen lactic acid bacteria for the fermentation of garlic and to assess the increase in inhibitory activity of garlic fermented against antibiotic-resistant pathogens for use as an animal feed supplement. We screened 45 strains of lactobacillus for the fermentation of garlic. Of these strains, 23 showed similar growth rates with or without allicin. Cultures of the 23 strains were mixed with an equivalent amount of garlic juice and incubated overnight at $37^{\circ}C$. The three strains with the lowest pH values were Lactobacillus paracasei KCTC 3169, L5 strain, and L. reuteri SW. Garlic juice fermented by the L5 strain more strongly inhibited antibiotic-resistant pathogenic bacteria than L. paracasei KCTC 3169, L. reuteri SW, or garlic juice itself. By examining carbohydrate utilization, morphologic properties and 16S rRNA gene sequences, we identified the L5 strain as Pediococcus pentosaceus and deposited it in the name of P. pentosaceus KACC 91419 into the Korea Agricultural Culture Collection. To identify the antimicrobial compound from the garlic filtrate fermented by P. pentosaceus KACC 91419, we fractionated P. pentosaceus KACC 91419 culture on a C18 column and checked the antimicrobial activity of fractions A6 to A10. Only fraction A9 showed inhibitory activity on Staphylococcus aureus. Comparing the mass spectra of the fractions with and without antimicrobial activity, we observed a single dominant product ion (m/z 157.99) from the fraction showing antimicrobial activity. Its molecular mass (157.99) was 2 atomic mass units less than that of allicin (162.02). This suggests that allicin might be converted to its derivative, which has antimicrobial activity, during fermentation by P. pentosaceus KACC 91419.
For screening of useful enzymes producing microorganisms from Meju, we isolated high lipase producing strains and their lipolytic enzyme activities were then tested. The lipolytic enzyme activities of isolated microorganisms were therefore tested on the Y124 strain. The gene sequence analysis of ITS from Y124 strain revealed Yarrowia lipolytica. Lipase production by the Y124 strain was studied in media containing various carbon sources. The Y124 strain drastically increased lipolytic enzyme activity in YPO media containing olive oil, as well as in YPDO media containing both olive oil and glucose. Maximal lipase production was achieved in YPD (yeast extract-peptone-D-glucose) media containing 0.7% olive oil when cultured at $30^{\circ}C$ for 8 hrs. The lipase produced from the Y124 strain showed the highest activity in p-NPO (p-nitrophenyl octanoate ($C_8$)), amongst the various p-nitrophenyl esters.
The purpose of this study was to analyze the asymmetry of the reaction forces on lower limbs between dominant and nondominant sides during sit-to-stand movement in normal subjects. Fourteen normal subjects ($22.6{\pm}2.3yrs$, all men) participated in this study. To measure the reaction forces during sit-to-stand movement, two force plates were mounted on the ground and one dual top force plate was mounted on a chair. Five events(movement onset, max thigh reaction force, transition, max hip angle, seat off) were determined from the reaction force and joint angle trajectories. For each of thigh, foot, and total reaction forces, two-way ANOVA was performed with the events and sides as factors. Also investigated was the leg asymmetry expressed as the ratio of the reaction forces of dominant and nondominant sides. The significance of asymmetry was investigated and two-way ANOVA was performed with the events and body parts(foot, thigh and total) as factors. Thigh reaction force and total reaction force showed interaction of events and leg sides(p < 0.01). Post-hoc comparisons showed they were different between sides at the latter stage(transition, max hip angle, and seat off events) (p < 0.01). Asymmetry was also significant at the latter stage(transition, max hip angle, and seat off events) (p < 0.01). Interaction of events and body parts on asymmetry was significant(p < 0.01) and asymmetry was greater in thigh reaction forces than total and foot reaction forces at the events of the latter stage(p < 0.01). The results suggest that asymmetry exist in normal subject and should be fully considered when investigating sit-to-stand strategy of patients.
The present study was undertaken to study the effect of thermal stress on sexual behaviour of superovulated ewes. Fourteen adult Bharat Merino ewes with an average body weight of 29.4${\pm}$2.34 kg were randomly allocated into two groups of 7 each. All the animals were grazed on natural pasture in the morning and evening hours and housed in shed during night. Animals of Group-1 were housed in shed from 10:00 to 16:00 h while the animals of Group-2 were exposed to thermal stress in a hot chamber ($40^{\circ}C$ /6 h/day). All the animals were offered drinking water once a day at 16:30 h. Meteorological observations i.e. dry bulb, wet bulb, minimum and maximum temperature were recorded daily inside the shed as well as in hot chamber throughout the experimental period. For superovulation of animals, standard protocol developed at the Institute, using FSH (Ovagen 5.4 mg in eight injections) and PMSG (200 IU) was followed. Various sexual behaviour parameters (circling, tail fanning, head turning, standing and approaching to ram) and estrus incidence (onset of estrus and estrus duration) were observed in both the groups. The different estrus symptoms were graded subjectively on arbitrary scale of 0-5 where 0 representing no sexual behaviour (0%) and 5 representing maximum intensity in sexual behaviour (100%). Estrus was detected with the help of a marked aproned ram of proven vigor at six hourly intervals. The average percent values for sexual behaviour parameters recorded in Group-1 and Group-2 animals were 53.7${\pm}$3.76 vs. 41.1${\pm}$2.18 for circling, 71.8${\pm}$5.42 vs. 49.0${\pm}$4.41 for tail fanning, 64.7${\pm}$3.30 vs. 44.5${\pm}$4.34 for head turning, 90.1${\pm}$3.16 vs. 75.8${\pm}$4.02 for standing and 63.8${\pm}$4.8 vs. 41.9${\pm}$4.58 for approaching to ram. Animals exposed to thermal stress had significantly lower values of these sexual behaviour parameters. The animals kept in shed exhibited estrus earlier (25.4${\pm}$1.12 h) and duration was higher (37.7${\pm}$1.59 h) as compared to animals exposed to thermal stress i.e. 30.6${\pm}$1.16 h and 31.7${\pm}$3.57 h, respectively. The findings of the study indicate that thermal stress reduces the intensity of sexual behaviour in ewes and may result in failure of the animal to mate and conceive.
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