• 한국미생물·생명공학회지
• /
• 제46권1호
• /
• pp.9-17
• /
• 2018

The aims of this work were to characterize and determine bioactivities of crude exopolysaccharide (EPS) extract from Bacillus tequilensis PS21 isolated from milk kefir from Kampaeng Petch, Thailand. B. tequilensis PS21 produced 112.1 mg dried EPS/l from initial 80 g/l lactose in modified TSB media at 52 h, with EPS product yield of 8.9 mg EPS/g lactose and specific product yield of 0.3 mg EPS/mg biomass. The FTIR result confirmed EPS to be a protein-bound polysaccharide and SEM analysis showed the morphology to be a grainy appearance with an uneven surface, covered with pores. HPLC analysis determined EPS as a heteropolysaccharide consisting of five sugar units with the following molar ratios; xylose (17.65), glucose (2.54), ribose (1.83), rhamnose (1.23), and galactose (1). Chemical components of this EPS were predominantly carbohydrate at 697.8 mg/g EPS (65%), protein 361.4 mg/g EPS (34%), and nucleic acid 12.5 mg/g EPS (1%). The EPS demonstrated antioxidant activities at 57.5% DPPH scavenging activity, $37.2{\mu}M\;Fe(II)/mg$ EPS and $34.9{\mu}M\;TEAC\;{\mu}M/mg$ EPS using DPPH, FRAP and ABTS assays, respectively. EPS also exhibited anti-tyrosinase activity at 34.9% inhibition. This work represents the first finding of EPS produced by Bacillus sp. from Thai milk kefir which shows potential applications in the production of antioxidant functional foods and whitening cosmetics. However, optimization of EPS production for industrial exploitation requires further study to ascertain the economic potential.

• 한국환경독성학회:학술대회논문집
• /
• 한국환경독성학회 2003년도 춘계학술대회
• /
• pp.195-195
• /
• 2003

It is well known that diesel exhaust particulate matter contains mutagenic PAHs, such as benzo[${\alpha}$]pyrene, benz[${\alpha}$]anthracene, chrysene, etc. Therefore it is suspected that these chemicals act on estrogen receptor and reveal endocrine-disrupting effects. Recent attention has focused on causative chemicals of endocrine-disrupting effects. We examined the estrogenic activity of respirable diesel exhaust particulate matter derived from diesel powered vehicle. PM2.5 diesel exhaust of vehicle was collected using a high volume sampler equipped with a cascade impactor. Diesel exhaust samples were fractionated according to EPA methods. The presence of estrogenic and antiestrogenic chemicals in PM 2.5 diesel exhaust was determined using E-screen assay. To quantitatively assess the estrogenic and antiestrogenic activities in diesel exhaust particulate matter, estradiol equivalent concentration (bio-EEQ) was calculated by comparing the concentration response curve of the sample with those of the estrogen calibration curve. Weak estrogenic activities and strong antiestrogenic activities were detected in the crude extract and moderately polar fractions. Higher antiestrogenic potency was observed with higher EROD activities in aliphatic and aromatic compounds fraction. In conclusion, estrogenic/antiestrogenic-like activities were present in diesel exhaust particulate matter. However, the health consequences of this observation was unknown, the presence of these activities may contribute to and exacerbate adverse health effect evoked by diesel exhaust particulate matter.

• 한국환경독성학회:학술대회논문집
• /
• 한국환경독성학회 2003년도 춘계학술대회
• /
• pp.194-194
• /
• 2003

The International Agency for Research on Cancer (IARC, 1989) has classified whole diesel exhaust as probably carcinogenic to humans. Diesel exhaust particulate matter (DPM) adsorbs different chemical substances including PAHs and nitroarenes. DPM is emphasized because it is a major component of diesel exhaust, it is suspected of contributing to a health hazard. Diesel exhaust is a complex mixture of carbon particles and associated organics and inorganics, and it is not known what fraction or combination of fractions cause the health effects [cancer effects, noncancer effects (respiratory tract irritation/inflammation and changes in lung function)] that have been observed with exposure to diesel exhaust. In order to identify which chemical classes are responsible for the majority of the observed biological activities, we performed a particular biological/chemical analysis. Respirable particulate matter (PM2.5: <2.5mm) was collected from diesel engine exhaust using a high-volume sampler equipped with a cascade impactor. Particulate oganic matter was extracted by the dichloromethane/sonication method and the crude extract was fractionated according to EPA recommended procedure into seven fractions by acid-base partitioning and silica gel column chromatography. We examined genotoxic potentials of diesel exhaust particulate matter using novel genotoxicity tests, which are rapid, simple and sensitive methods for assessing DNA-damage at the DNA and chromosomal level (comet assay, in vitro MN test and Ames test). Higher genotoxic potency was observed in non polar fractions and several PAHs were detected by GC-MS, such as 1,2,5,6 dibenzanthracene, chrysene, 1,2-benzanthracene, phenanthrene and fluoranthene.

• Applied Biological Chemistry
• /
• 제36권2호
• /
• pp.121-126
• /
• 1993

참깨박에 함유되어 있는 불용성 형태의 단백질을 가용성 형태의 단백질로 용출시키기 위하여 Bacillus sp. CW-1121이 생성하는 효소를 작용시켰다. 이때 참깨박의 단백질 용출을 위한 최적 pH는 7.5였으며, 최적 온도는 $40^{\circ}C$였고 최적 조건하에서 2시간의 소화로 약 60%의 용해도를 보였다. 단백질의 용해도는 참깨 단백질이 pH4.5에서 가장 낮아 등전점을 보였고, pH3.0과 pH6.0 이상에서는 크게 증가하였다. 효소처리 참깨박 단백질을 전기영동한 결과 수용성 단백질은 4개, 염용성 단백질은 2개의 밴드가 관찰되었다. 분리 단백의 아미노산 조성은 수용성 단백질의 경우 serine이 17.24 mg/g, 염용성 단백질은 glutamic acid가 10.77 mg/g, 유리아미노산은 glutamic acid가 6.55 mg/g으로 가장 많이 함유되어 있었으며 특히 필수아미노산의 조성도 상당히 좋은 편이었다.

• Applied Biological Chemistry
• /
• 제45권4호
• /
• pp.190-194
• /
• 2002

Bacillus subtilis의 glutamyl-tRNA synthetase(GluRS)는 대장균에서 발현될 때 숙주세포의 $tRNA_1^{Gln}$에 glutamate를 잘못 아실화하여 독성을 나타내는 것으로 추정되고 있다. 이러한 B. subtilis GluRS를 대장균에서 과발현 시키기 위하여 B. subtilis 168 균주의 chromosomal DNA에서 GluRS의 유전자(gltX)를 PCR을 이용하여 증폭하고 T7 promoter에 의해 발현이 조절되는 pET11a expression vector에 클로닝하였다. 이 재조합된 pEBER plasmid DNA로 T7 RNA polymerase를 갖는 대장균 NovaBlue(DE3)에 형질전환하였다. 형질전환된 대장균에 IPTG를 처리하여 과량 생성된 GluRS 단백질은 ammonium sulfate 분별침전 후 EPLC를 이용한 Source Q column anion exchange chromatography, Superdex 200 column gel filtration, Mono Q column anion exchange chromatography로 정제하였다. 정제된 B. subtilis의 GluRS 분자량은 약 55 kDa이었으며 효소의 활성도는 조효소액에 비해 18배로 증가하였다.

• Asian-Australasian Journal of Animal Sciences
• /
• 제13권10호
• /
• pp.1422-1428
• /
• 2000

This investigation was conducted to evaluate the shrimp flesh (SF) and shrimp by-product (SB) of the most abundant shrimp species (Metapenaeus affinis, Penaeus semisulcatus and Penaeus monodon) caught in Central Vietnam, with the emphasis on yield, gross and amino acid (AA) composition and effect of heat treatment. The results showed that the mean edible SF and SB (head and shells with tail) yields of the three shrimp species averaged 56.7 and 43.3%, respectively, of the total wet body weight, with the M. affinis generating the highest by-product yield (45.7%) and P. semisulcatus (40.6%) the lowest. Significant differences in dry matter (DM), crude protein (CP) and ash content were found between SF and SB. The DM content of SF (21.5%) was lower than of SB (24.9%) and the ash content (on a DM basis) of the SB in all shrimp species was more than three times that of the SF (p<0.05), whereas the CP content was almost twice as high in the SF as compared with the SB (p<0.05). The SB of the three species contained (on a DM basis) between 44.0 and 49.8% CP (p<0.05) and between 13.5 and 18.1% chitin (p<0.05). The Ca content of SB differed also between species (p<0.05). On average, the sum of AA in SB corresponded to 89.3% of the CP and essential AA accounted for about 50% of the total AA. The most abundant AA were arginine, aspartic and glutamic acids, which accounted for 33% of the total AA. Minor, but significant differences in some AA concentrations of SB between species were observed (p<0.05). With the exception of the DM and ether extract content, all other chemical constituents of entire shrimp, SF and SB were not significantly affected by heat treatment (p>0.05).

• Asian-Australasian Journal of Animal Sciences
• /
• 제16권5호
• /
• pp.665-671
• /
• 2003

A factorial experiment was conducted to assess nutrient utilization by growing lambs maintained on three levels each of digestible energy (high: HE, medium: ME, low: LE) and protein (high: HP, medium: MP, low: LP) in nine combinations (HEHP, HEMP, HELP, MEHP, MEMP, MELP, LEHP, LEMP, LELP). The experiment was conducted during the hot season in a semiarid location. Daily dry matter intake (DMI) was similar in all the groups in terms of unit body weight or metabolic body size. Digestibility of DM and nitrogen free extract increased (p<0.01) from low to medium and high energy regimen while the CF digestibility followed a reverse trend. The digestibility of crude protein (CP) decreased from high to medium and low protein regimens while it was similar in terms of energy variation. Nitrogen intake was higher in high followed by medium and low protein regime while fecal and urinary nitrogen loss were similar in all the treatment groups. Lambs in all the three levels of protein were in positive N balance and percent N retention was higher (p<0.01) in high followed by medium and low protein levels whereas it was similar in terms of energy variation. Initial body weight was similar in all the groups while final weight, total gain in the experiment and average daily gain (ADG) were higher in high than medium and low energy regimens. It is concluded that crossbred lambs required 75.1 g DM, 9.6 g CP, 6.3 g DCP and 711 KJ DE/kg W $^{0.75}$or 11.0 g CP/MJ DE or 7.2 g DCP/MJ DE for 93 g average daily gain in a hot semiarid environment.

• Asian-Australasian Journal of Animal Sciences
• /
• 제30권1호
• /
• pp.51-56
• /
• 2017

Objective: The objective of this study was to evaluate the intake and nutrient digestibility, nitrogen balance and ruminal ammonia nitrogen in lambs of diets containing different levels of residual frying oil. Methods: Levels of 0, 20, 40, 60, and 80 g/kg dry matter (DM) base of residual frying oil in the diets of lambs were evaluated. Five castrated lambs with initial body weights of $36.8{\pm}3.3kg$, distributed in a Latin square ($5{\times}5$) design, were used. Results: There was a decreasing linear effect on the intake of DM, organic matter (OM), crude protein (CP), neutral detergent fiber (NDF), total carbohydrates (TCH), and nonfibrous carbohydrates (NFC). There was an increased linear intake of ether extract (EE). The apparent digestibility of DM, OM, CP, NDF, TCH, and NFC, as well as urine nitrogen excretion, nitrogen balance and ruminal parameters, were not influenced by different levels of residual frying oil in the diet. EE digestibility presented a crescent linear effect. Conclusion: It can be concluded that the addition of residual frying oil to the diets of sheep can affect nutrient intake without affecting the digestibility of most nutrients (with the exception of EE), nitrogen balance and ruminal ammonia nitrogen concentration.

• Journal of Microbiology and Biotechnology
• /
• 제13권1호
• /
• pp.104-109
• /
• 2003

Enhancement of the production of benzoylformate reductase (BFR) was attempted under oxidative stress established by natural electron carriers. -lipoic acid (LA), flavin adenine dinucleotide (FAD), and ubiquinone (UQ) did not inhibit growth of E. faecalis when their concentrations were as high as $10{\mu}M$, while $H_2O_2$ and methyl viologen ($MV^2+$) inhibited the bacterial growth. BFR activity in the bacterial extract had increased rapidly after 1 h of cultivation after the addition of $4{\mu}M$ of natural electron carriers, and the activity was maintained during further cultivation. BFR activity of the cells treated with the natural electron carriers was $40\%$ higher than that of the control. In the presence of $4{\mu}M\;H_2O_2\;and\;MV^2+$, BFR activity increased, reaching the highest activity at about 5 h cultivation, and then decreased with further cultivation. It seems that natural electron carriers not only stimulate the induction of BFR, but also stabilize the enzyme. BFR was hardly affected by LA, FAD, and UQ, while $H_2O_2\;and\;MV^2+$ inactivated the crude enzyme. The decrease of BFR activity in the presence of $H_2O_2\;and\;MV^2+$ might be ascribed to inactivation of the enzyme by the oxidants.

• Applied Biological Chemistry
• /
• 제36권3호
• /
• pp.172-177
• /
• 1993

참깨박에 함유되어 있는 불용성 형태의 단백질을 Bacillus sp. CW-1121이 생성하는 효소를 이용하여 가용성 형태의 단백질로 용출시키기 위하여 효소를 작용시켰다. 효소 처리 참깨박 단백질의 기포 형성력은 염용성 단백질이 수용성 단백질에 비해 현저히 낮았다. 기포 안정성은 염용성 단백질이 10분 이내 크게 감소된 반면 수용성 단백질의 기포 안정성은 30분 까지 완만하게 감소하였다. 유화력은 각 단백질의 등전점 부근에서 가장 낮았으며, 염용성 단백질이 수용성 단백질에 비해 낮았다 유화 안정성은 유화력과 비슷한 크기로 나타나 $80^{\circ}C$에서의 30분 가열에서도 이들 단백질이 열에 안정함을 보였다. 유지 및 수분 흡착력은 염용성 단백질이 수용성 단백질 보다 높은 값을 나타내었다.