• 한국미생물·생명공학회지
• /
• 제34권1호
• /
• pp.40-46
• /
• 2006

단위 동물의 사료 내 phytate 인의 효율적 이용으로 배출되는 인의 양을 줄여 환경오염을 감소시키기 위해 phytate의 분해활성이 뛰어난 phytase 효소 분비 미생물을 탐색하기 위하며 우분으로부터 phytate 분해활성이 뛰어난 phytase를 생산하는 균주를 분리하였다. 분리균주를 동정한 결과, API 50 CHB test에 의해 B. circulans로 분류되었고, 16S rRNA sequencing 결과, B. subtilis로 동정되어 Bacillus sp. CF 5-26으로 명명하였다. 효소생산을 위한 최적배지 조성은 10% rice bran extract, 0.1%, whey protein powder, $0.01%\;CaCl_{2},\;0.01%\;KH_{2}PO_4$ 이었다. Bacillus sp. CF 5-26이 생산하는 phytase는 ethanol 침전, Sephadex G-100, CM Sepharose CL-6B, Sephacryl S-100-HR column chromatography를 통하여 정제도 20.3배, 수율 5.6% 정제되었고, SDS-PAGE에서 분자량 66 kDa의 단일 band를 확인하였다. 정제된 phytase는 pH 5.0, 7.0, 11.0에서 안정하였으며, $100^{\circ}C$에서 1시간 처리하였을 때 50%의 잔존활성을 보였다. 기질 특이성은 inositol polyphosphate인 sodium phytate 분해활성이 뛰어났으며, tripolyphosphate와 pyrophosphate에도 약간의 활성을 보였다. Bacillus sp. CF 5-26이 생산하는 phytase의 sodium phytate에 대한 Km은 0.64 mM 이었고, Vmax는 $4.41{\mu}mol/min$ 이었다.

• Journal of Nutrition and Health
• /
• 제30권4호
• /
• pp.406-414
• /
• 1997

It has been known that dietary phytate decreases the absorption of body zinc pool which is composed of the dietary and endogenous zinc in the body. The purpose of this study was to examine the effect of phytate on the absorption of total bodyzinc in Zn-depleted rats. Rats were Zn-depleted with either low(0.8%) or high(1.6%) Ca diet containing sodium phytate for 4 weeks. After zinc depletion, rats were assigned into phytate or non-phytate dietary groups within each low-or high-Ca dietary group. ant feces were collected for 2 weeks of the initial collection and 1 week after dietary crossover, during which the phytate and the non-phytate diet was switched over within the same Ca group. The content of Zn and Ca measured by atomic absorption spectrophotometer and phytate content was analyzed. food intake was higher in the high Ca group than in the low Ca group(p <0.0001), and was also higher in the non-phytate group than in the phytate group(p <0.0001). Food intake and phytate level affected body weight gain in rats(p <0.0001). Zinc excretion in the total feces was higher in the phytate group than in the non-phytate group at both low and high Ca level(p <0.0001), except during the crossover collection period in high Ca group. Calcium, however, didn't show any synergistic effect on phytate effect(p <0.05). This study showed that phytate decreased the absorption of total body zinc at both low and high Ca levels in Zn-depleted rats. A large portion of total body zinc originated from the endogenous zinc pool in these rats. The results of the present study showed the same effect of phytate on the endogenous zinc in Zn-depleted rats as in a previous study, confirming that phytate adversely affects zinc bioavailability, especially under marginal and poor zinc nutrition.

• Journal of Nutrition and Health
• /
• 제30권4호
• /
• pp.394-405
• /
• 1997

Endogenous zinc is important for maintaining zinc homeostasis because the size of endogenous zinc pool is almost 3-4 times bigger than that of dietary zinc. The purpose of this study was to examine the phytate effect on the reabsorption of endogenous zinc and the additional Ca effect on the phytate effect. Rats were fed a casein-based diet with added sodium phytate containing either high(1.6%) or low(0.8%) Ca concentrations for 4 weeks to reduce the body zinc pool. After the depletion period, $^{65}$ Zn was given by intraperitoneal injection to label the endogenous zinc pool. Rats were then assigned into phytate or non-phytate group within the same Ca group. feces were collected for 2 weeks of the initial collection period and 1 week after dietary crossover. The ratios of excreted fecal $^{65}$ Zn radioactivity of phytate group non-phytate group were determined as a measure of the phytate effect on the endogenous zinc. Mean fecal $^{65}$ Zn radioactivity was higher in the phytate group than in the non-phytate group during the entire 3 weeks of the collection period in the low Ca group, and during the initial collection period in the high Ca group(p <0.0001). This study showed an adverse phytate effect on endogenous zinc at both high and low dietary Ca levels. Elevated dietary Ca levels showed a synergistic effect on the phytate effect on endogenous zinc(p <0.05). These results imply greater phytate effect on zinc homeostasis rather than on zinc bioavailability through complexing with the endogenous zinc which is larger portion than the dietary zinc on zinc homeostasis.

• 한국식품과학회지
• /
• 제35권3호
• /
• pp.510-518
• /
• 2003

자외선에 노출된 피부에서 생성된 활성산소종에 의한 산화적 스트레스는 세포 구성 성분들을 손상시키고 궁극적으로는 피부 광노화 및 발암을 야기시킬 수 있다. 이 연구에서는 대두(Glycine max MERRILL)의 성분인 genistein(4',5,7-trihydroxyisoflavone)을 포함하는 몇 가지 천연 황산화제들에 대하여 로즈벵갈로 증감된 사람 적혈구의 광용혈에 대한 보호효과를 조사하였다. Genistein$(10-100\;{\mu}M)$은 농도-의존적으로 광용혈을 억제하였다. 그리고 지질 과산화 연쇄반응의 차단제인 ${\alpha}$-tocopherol보다도 더 효과적이었다. 그러나, Genitein의 배당체인 genistin, 소용성 항산화제인 L-ascorbate, 철-킬레이트제인 sodium phytate는 광용혈에 대하여 보호 효과를 나타내지 않았다. 오히려 보다 높은 농도$(500\;{\mu}M)$에서는 L-ascorbate와 sodium phytate 모두 광용혈을 촉진시켰다. 반면, $^1O_2$ 소광제로 알려진, ${\alpha}$-carotene 3,3'-diol은 현저한 보호 효과를 보여주었다. 이 결과는 로즈벵갈로 증감된 광용혈에 있어서 $^1O_2$이 중요함을 가리킨다. Luminol-의존성 화학발광법을 이용하여, $Fe^{3+}-EDTA/H_2O_2$ 계에서 생성된 ROS에 대한 몇가지 천연 항산화제들의 ROS 소거활성을 측정하였다. 활성 산소 소거활성$(OSC_{50})$의 크기는 sodium phytate > L-ascorbate>${\alpha}$-tocopherol > genistein > genistin순이었다. Gnistein, genistin, ${\alpha}$-tocopherol, L-ascorbate 및 sodium phytate의 활성산소 소거활성$(OSC_{50})$은 각각 41.0, 109.0, 9.0, 5.2, 및 $0.56{\mu}M$이었다. Free radical 소거활성$(FSC_{50})$의 크기는 L-ascorbate > ${\alpha}$-tocopherol > genistein > genistin순으로 나타났다. 이상의 결과들은 genistein이 $^1O_2$ 혹은 다른 ROS를 소거함으로써 그리고 ROS에 대항하여 세포막을 보호함으로써 생체계, 특히 태양 자외선에 노출된 피부에서 항산화제로서 작용할 수 있음을 가리킨다.

• Journal of Microbiology and Biotechnology
• /
• 제15권4호
• /
• pp.745-748
• /
• 2005

A phytase from Aeromonas sp. LIK 1-5 was partially purified by ammonium sulfate precipitation and DEAE-Sephacel column chromatography. Its molecular weight was 44 kDa according to SDS-PAGE gel. Enzyme activity was optimal at pH 7 and at $50^{\circ}C$. The purified enzyme was strongly inhibited by 2 mM EDTA, $Zn^{2+},\;Co^{2+},\;or\;Mn^{2+}$, and activated by 2 mM $Ca^{2+}$. The K_m value for sodium phytate was 0.23 mM, and the enzyme was resistant to trypsin. The N-terminal amino acid sequence of the phytase was similar to that of other known alkaline phytases. The phytase was specific for ATP and sodium phytate, which is different from other known alkaline phytases. Based on the substrate specificity, the phytase may therefore be a novel alkaline phytase.

• 한국미생물·생명공학회지
• /
• 제29권2호
• /
• pp.78-83
• /
• 2001

본 연구는 phytic acid를 myo-inositol과 무기태인으로 분해시키는 효소인 phytase 생산균주의 분리 및 효소생산의 배지최적하에 관한 것이다. 1차로 calcium phytate를 기질로 함유한 phytase screening 배지를 이용하여 phytase 생산을 나타내는 균주 35가지를 이용하여 분리한 후, sodium phytate를 기질로 하여 재현성 있는 phytase 활성을 나타내는 균주 12가지를 선발하였다. 12개의 균주 중 BHI broth에서 배양한 조효소액의 phytase 활성이 가장 우수한 것으로 나타난 YH100을 선발하여 주사현미경 관찰, 16S rRNA sequence 분석, GC conten(mol%) 조성, 지방산 분석, API 20E kit를 이용한 test 결과 Enterobacter cloacae로 동정되어, 이 균주를 Enterobacter cloacae YH100이라 명명하였다. Enterobacter cloacae YH100에 의한 phytase 생산을 위한 최적 배지 조성을 파악한 결과 glucose 2.0%(w/v), peptone 1.0%(w/v), beef extract 1.0%(w/v), KCI 0.1%(w/v), sodium phytate 0.1%(w/v)로 나타났다.

• Applied Biological Chemistry
• /
• 제39권1호
• /
• pp.32-38
• /
• 1996

대두에 함유된 영양저해인자의 하나인 phytic acid를 감소시키기 위해 일정시간 침지시킨 대두를 microwave로 가열처리하여 phytic acid 및 phosphorus의 함량 변화를 관찰하였다. 단원콩, 만리콩 및 Amsoy의 phytic acid 함량은 각각 탈지대두 1 g중에 19.19 mg, 18.38 mg 및 16.73 mg 함유되어 있었다. 가열시간이 길어짐에 따라 콩에 함유된 phytic acid 및 phytate phosphorus 함량은 감소하는 경향을 보여서, 5분 가열시에 단원콩, 만리콩 및 Amsoy의 phytic acid 감소율은 각각 26.29%, 24.85%, 23.72%이었다. 단원콩을 12에서 30분간 autoclaving 처리하였을때 phytic acid 및 phytate phosphorus의 감소율은 18.62%와 16.95%로 microwave로 5분 처리하였을 때가 훨씬 효과적이었다. 대두의 수분함량과 microwave 가열효과와의 관계는 수분함량이 낮을수록 phytic acid의 감소 효과가 컸다. 몇가지 염용액에 12시간 침지시킨후 4분간 가열처리시 phytic acid의 감소율은 2.5% NaCl에서 24.52%, 2% $NaHCO_3$에서 26.62%, 혼합 염용액(M.S.S.)에서 28.47%로 증류수의 21.53%보다 약간 높게 나타났으며, phytate phosphorus 감소율 또한 위의 세가지 염용액에서 26.17%, 24.64% 및 26.60%로서 증류수의 19.73%보다 약간 더 효과가 있는 것으로 나타났다.

• Asian-Australasian Journal of Animal Sciences
• /
• 제25권10호
• /
• pp.1466-1472
• /
• 2012

A bacterial isolate derived from soil samples near a cattle farm was found to display extracellular phytase activity. Based on 16S rRNA sequence analysis, the strain was named Bacillus sp. T4. The optimum temperature for the phytase activity toward magnesium phytate (Mg-$InsP_6$) was $40^{\circ}C$ without 5 mM $Ca^{2+}$ and $50^{\circ}C$ with 5 mM $Ca^{2+}$. T4 phytase had a characteristic bi-hump two pH optima of 6.0 to 6.5 and 7.4 for Mg-$InsP_6$. The enzyme showed higher specificity for Mg-$InsP_6$ than sodium phytate (Na-$InsP_6$). Its activity was fairly inhibited by EDTA, $Cu^{2+}$, $Mn^{2+}$, $Co^{2+}$, $Ba^{2+}$ and $Zn^{2+}$. T4 phytase may have great potential for use as an eco-friendly feed additive to enhance the nutritive quality of phytate and reduce phosphorus pollution.

• 대한수의학회지
• /
• 제47권4호
• /
• pp.449-456
• /
• 2007

The approval of use of certain food-grade phosphates as food additives in a wide variety of meat products greatly stimulated research on the applications of phosphates in foods. Although phosphates have never been classified as antimicrobial agents, a number of investigators have reported that phosphates have antimicrobial activities. Phytic acid is a natural plant inositol hexaphosphate constituting 1-5% of most cereals, nuts, legumes, oil seeds, pollen, and spores. In this study, we investigated antibacterial activities of sodium phytate (SPT), sodium pyrophosphate (SPP), sodium tripolyphosphate (STPP) on Salmonella typhimurium in tryptic soy broth and in row meat media including chicken, pork and beef. SPY, SPP and STPP at the concentrations of 0.5 and 1% dose-dependently inhibited the growth of S. typhimurium in tryptic soy broth at various pHs. The antibacterial activities of SPT and STPP were the stronger than that of SPP. In chicken, pork, and beef, SPT, SPP and STPP at the concentrations of 0.1, 0.5 and 1.0% significantly inhibited the bacterial growth in a dose-dependant manner (p < 0.05). The antibacterial activities of SPT, SPP, and STPP were more effective in chicken than beef. SPT and STPP at the concentration of 1% reduced the bacterial count by about 2 log units. The addition of SPT, SPP and STPP at the concentration of 0.5% in meats increased the meat pHs by 0.28-0.48 units in chicken, pork, and beef. These results suggest that SPT and STPP were equally effective for the inhibition of bacterial growth both in TSB and meat media and that SPT can be used as an animal food additive for increasing shelf-life and functions of meats.

• 공업화학
• /
• 제29권6호
• /
• pp.740-745
• /
• 2018

본 연구에서는 비정질 칼슘 포스페이트(ACP) 나노 입자의 합성과 특성 분석을 진행하였다. 염화칼슘(calcium chloride ($CaCl_2$))과 아데노신 인산나트륨(disodium adenosine triphosphate ($Na_2ATP$)) 그리고 피트산 나트륨(sodium phytate) 첨가제를 열수 반응을 통해 상대적으로 단분산된 100 nm 크기 이하의 ACP 나노 입자를 성공적으로 합성하였고 나노 입자의 화학적 조성과 구조를 재료 분석을 통해 확인하였다. 피트산 나트륨 첨가제의 사용을 통해 얻은 ACP 나노 입자는 비정질성을 유지하고 결정성 하이드록시아파타이트(HAP)로의 전환을 방지하는 안정성이 향상되었음을 발견하였다. 본 연구를 통해 발견된 향상된 안정성을 가지는 ACP 나노 입자는 재생 의학 분야에서의 생체 적합 물질로의 응용에 중요한 잠재적 용도가 있을 것이라 사료된다.