This study was to investigate whether the bioactivity of L. plantarum L67 can be enhanced by treatment with Zanthoxylum piperitum DC (ZPDC) glycoprotein or compounds of similar molecular weights (24 kDa). When L. plantarum L67 was treated with ZPDC glycoprotein of various concentrations, the intensities of a few SDS-PAGE bands were slightly changed. The amount of a 23-kDa protein was increased upon treatment with increasing concentrations of ZPDC glycoprotein. The activities of antioxidant enzymes (SOD, GPx, and CAT), growth rate, and β-galactosidase activity were more increased at treatment with ZPDC glycoprotein than at treatment of L. plantarum L67 only, whereas the ATPase activity was decreased. Interestingly, it did not affect scavenging activity for the DPPH. The founding result suggest that, one of beneficial strategies for increase the bioactivity of L. plantarum L67 might be to take ZPDC.

Obesity is the most common nutritional disorder in the developed world and has become a global epidemic in recent years. In this study, Zanthoxylum planispinum extracts (ZPE) were evaluated on the effect on inhibition of pancreatic lipase and lipid metabolism by oral treatment for 2 months in high-fed diet obesity-induced Balb/c mice. The ZPE showed pancreatic lipase inhibitory activity with IC50 of 0.3 ㎍/㎖. No significant difference in feed intake was observed among the groups. The high-fat diet-treated Z. planispinum extracts groups (HFD+ZPE, 100 ㎎/㎏) significantly decreased body weight compared to the high-fat diet vehicle groups (HFD, p<0.05). The high-fat diet-treated XenicalⓇ groups (HFD+Xenical, n=10, 30 ㎎/㎏) also showed a significant reduction of body weight compared to HFD (p<0.05). Biochemical parameters (triglyceride, total cholesterol, and high-density lipoprotein cholesterol) in HFD plus ZPE diet groups were significantly lower than those of the HFD groups (p<0.05). These results indicated that ZPE more effectively suppressed the effects of HFD on body fat gain with the inhibitory effect on pancreatic lipase.

Electroantennogram techniques (EAGs) were employed to record olfactory responses in the antennae of the adult female stable fly, Stomoxys calcitrans (Diptera: Muscidae), to Zanthoxylum piperitum pericarp steam distillate (ZP-SD), Z. armatum seed oil (ZA-SO) and their 29 volatile constituents alone as well as binary mixture of the ZP-SD and ZA-SO with 1-octen-3-ol. The electrophysiological responses of the test materials were compared with those of DEET and 1-octen-3-ol. At concentration of 10-1 (v/v) in mineral oil, ZP-SD, ZA-SO, and all volatiles elicited EAG responses in the fly antennae except for DEET, which is extremely low volatile. ZP-SD, ZA-SO, and some of the test volatiles elicited EAG responses rather lower than 1-octen-3-ol, which showed vapor phase repellency and toxicity to the fly in our previous behavioral research. This suggests that the stable fly possesses olfactory receptor neurons responding to given repellent compounds per se. In binary mixture with 1-octen-3-ol at concentration of 10-1, ZP-SD and ZA-SO elicited 16±0.55 and 18±0.63 mV while ZP-SD, ZA-SO, and 1-octen-3-ol alone elicited responses of 14±0.45, 15.8±0.37 and 16.2±0.58 mV, respectively. Among the volatile compounds, terpinen- 4-ol, β-myrcene, α-phellanderene, citronellal, and limonene oxide elicited 80 – 96% relative EAG amplitude compared to 1-octen-3-ol as a reference (100%). Based on structure-activity relationships, constituents having aldehyde moiety elicited bigger EAG responses than those of alcohol or ether one. The perception in the fly antenna to plant volatile compounds exhibited complicated patterns of olfactory responses. Current and future directions of this study on sensory processing mechanisms underlying repellent behaviors will be discussed.

The vapor phase repellency and toxicity of Zanthoxylum piperitum pericarp steam distillate (ZP-SD), Z. armatum seed oil (ZA-SO), and their 29 constituents to the adult female stable fly, Stomoxys calcitrans (Diptera: Muscidae), were examined using filter paper fumigation bioassay. Results were compared with those of the currently used repellent: DEET. Both of ZP-SD and ZA-SO exhibited vapor phase repellency and toxicity to female flies at 5 to 40 mg/filter paper (0.23 to 1.82 mg/cm3 air) during a 120-min exposure, whereas DEET exhibited neither repellency nor toxicity to the stable fly. At 5 mg/filter paper, vapor phase of ZP-SD and ZA-SO repelled 50 to 67% and 61 to 51% flies, respectively, to control area during 30 to 120 min. At 40 mg/filter paper, vapor phase of ZP-SD and ZA-SO caused 100% and 81% mortality, respectively, after 120 min of exposure. Among the tested volatile constituents, cuminaldehyde was the strongest repellent and toxic compound to the fly and was four times more toxic than ZP-SD and ZA-SO. Cuminaldehyde vapor phase repelled 62% flies to control area after 30 min at 2.5 mg/filter paper and caused 100% mortality after 120 min at 10 mg/filter paper. Based on the structure-activity relationships, the toxicity and repellency of constituents having aldehyde moiety were better than those of alcohol or ether one. Overall, ZP-SD, ZA-SO, and their bioactive constituents could be useful as potential vapor repellents to control stable fly population.

Gochujang samples with essential oils were evaluated for sensory including taste and, smell during various storage periods with the aim of developing improved high value-added products. The sensory testing was performed on Gochujang with 0.005% Zanthoxylum piperitum and Chrysanthemum indicum essential oils, extracted through steam distillation. During storage at 4 and 20℃ for 8 weeks, sensory testing was performed at 0, 1, 2, 4 and 8 weeks by trained panelists. Overall quality decreased in Gochujang samples containing Z.piperitum essential oil and increased in samples containing Cindicum essential oil, at 4 and 20℃. Samples stored at 4℃ were quality more then those stored at 20℃. Sensory evaluation was highest for Gochujang with Cindicum essential oil stored at 4℃. It is concluded that functional Gochujang can be developed by adding small quantities of essential oils. Increasing the amount of essential oil added is likely to affect the innate taste and flavor of Gochujang.

The repellent constituents of essential oils from Zanthoxylum armatum (Rutaceae) seed and Zanthoxylum piperitum fruit against female Stomoxys calcitrans were analyzed by gas chromatography and gas chromatography-mass spectrometry. The repellency of two Zanthoxylum essential oils and 11 oil compounds against female S. calcitrans was examined using a skin bioassay. Results were compared with those of the most widely used repellent DEET. Both Z. armatum seed and Z. piperitum fruit oils provided complete and good protection from fly bites for 30 min at 0.4 and 0.2 mg/cm2, respectively. At 0.05 mg/cm2, both oils gave ca 70 and 40% repellency at 30 and 60 min post-treatment, respectively, whereas DEET provided 83 and 73% protection. Of test compounds, cuminaldehyde at 0.05 mg/cm2 gave 92 and 69% repellency at 10 and 30 min post-treatment, respectively, whereas 1,8-cineole, and citronellal provided approx. 76% repellency at 10 min. The Zanthoxylum oils and their constituents, particularly cuminaldehyde, merit further study as potential insect repellents for the protection from humans and domestic animals from biting and nuisance caused by S. calcitrans.

Compositional changes In essential oil of Zanthoxylum piperitum A.P. DC. were investigated under six different storage conditions for 3 months. Essential oil from Zanthoxylum piperitum was collected by steam distillation method and analyzed by gas chromatography-mass selective detector (GC-MSD). Forty-one volatile compounds, consisting of 12 hydrocarbons, 11 alcohols, 8 aldehydes, 3 oxides, 3 esters, 3 ketones and 1 acid were identified from the fresh essential oil of Zanthoxylum piperitum. In essential oils, compositional changes occurred in particularly monoterpene hydrocarbons. Total levels of ketones, esters, oxides and alcohols increased during storage. Moreover, aerobic condition caused decrease in a few constituents duringstorage even at low temperature.

경상북도 상주시 청화산 일대에서 채집한 야생 초피잎을 음지에서 자연 건조한 것과 blanching한 후 건조한 것을 분쇄하여 기계적으로 분석한 결과 잎을 자연 건조한 것에서는 55개의 Peak가 확인되었으며 blanching한 후 건조한 시료에서는 35개의 peak가 확인되었다. 그리고 자연 건조한 시료 중에서 검출된 55 개의 물질들 중에는 hydrocarbon 계통의 물질들이 decane, dodecane, sabinene, myrcene등을 비롯한 23개의 물질이 확인되었고 alcohol 계통의 물질들은 isobutylalcohol, cis-pentenol, 1-ponton-3-ol등 10개의 물질이 확인되었다. 그리고 aldehyde로는 3-methylbutanal, hexanal. 2, 6-dimethyl hept-5-al등 7개 물질이 확인되었으며 ketone 화합물은 3-hydroxy-2-butanone. 2-nonanone, 2-undecanone. 2-tridecanone이 확인되었다. cis-3-hexenyl acetate, linalyl acetate. citronellyl, acetate, neryl acetate 등 6개의 ester류가 확인되었다. 그리고 기타 물질로 3-cyano-2, 5-dimethylpyrazine, dimethyl sulfide, chloroform, 1, 8 cineole, dill ether 등이 검출되었다. 그리고 초피의 생잎을 blanching한 후 건조하여 전처리를 거쳐 휘발성 물질을 확인한 결과 35개의 peak를 확인하였는데 그중 hydrocarbon 계통의 물질들이 20개나 확인되었다. 그 내용으로는 1, 1`-oxybis-ethane. α-pinene, camphene, myrcene, β-phellandrene. β-caryophyllene 등이다. 그리고 alcohol 류로는 L-linalool, (-)-isopulgerol. α-terpineol. citronellol이며 aldehyde류로는 nonanal, citronellal이다. ketone 화합물은 2-undecanone, 2-tridecanone등이며 그리고 ester류로는 citronellyl acetate, cis-3-hexenyl acetate. neryl acetate 등이다. 기타물질로서는 3-cyano-2, 5-dimethylpyrazine. chloroform 등이다. 그리고 α-pinene, myrcene, β-phellandrene. L-linalool, citronellal, citronellyl acetate. β-caryophyllene이 다른 휘발성 물질보다 비교적 많은 양으로 검출되었다.

Zanthoxylum piperitum D.C. (ZP) peels has been used as a natural spice and herb medicine for hypertension reduction, for strokes, and for its anti-bacterial and anti-oxidant activity. However, the anti-inflammatory mechanisms employed by ZP have yet to be completely understood. In this study, we elucidate the anti-inflammatory mechanism of ZP in lipopolysaccharide (LPS)-induced RAW264.7 cells. We evaluated the effects of ZP in LPS-induced levels of inflammatory cytokines, prostaglandin E2 (PGE2), and caspase-1 using ELISA. The expression levels of inflammatoryrelated genes, including cyclooxygenase (COX)-2 and inducible nitric oxide synthase (iNOS), were assayed by Western blot analysis. We elucidated the effect of ZP on nuclear factor (NF)-κB activation by means of a luciferase activity assay. The findings of this study demonstrated that ZP inhibited the production of inflammatory cytokine and PGE2 and inhibited the increased levels of COX-2 and iNOS caused by LPS. Additionally, we showed that the anti-inflammatory effect of ZP arises by suppressing the activation of NF-κB and caspase-1 in LPS- induced RAW264.7 cells. These results provide novel insights into the pharmacological actions of ZP as a potential candidate for development of new drugs to treat inflammatory diseases.

자외선은 피부 노화를 가속화하여 피부 광노화를 유발하고, 일광 화상, 피부암 등을 유발한다. 자외선 차단제를 사용하더라도 일부 자외선에 의하여 피부 손상은 유발될 수 있기 때문에 자외선에 대한 피부 자체의 방어력을 올려주는 것이 필요하다. 최근, 식물에서 자외선 보호 기능을 하는 것으로 알려진 COP1이 사람의 피부 에서도 자외선에 대한 반응들을 조절한다고 새롭게 밝혀졌다. 본 연구에서는, COP1과 그의 결합 단백질 DET1 이 사람 피부의 각질형성세포에서 자외선에 대한 시그날 조절 물질인 c-Jun 단백질 양을 조절하는 것을 확인하 였다. 자외선에 노출 시 COP1과 DET1 발현이 감소하였고, 그 영향으로 c-Jun 단백질이 증가하였다. 반대로 COP1과 DET1을 발현하는 DNA를 transfection 시켜줄 경우 c-Jun 단백질 양이 감소하였다. 피부 각질형성 세포에서 COP1과 DET1의 발현을 조절할 수 있는 물질을 탐색한 결과, 초피나무 열매 추출물이 COP1과 DET1의 발현을 증가시켜 주었다. 초피나무 열매 추출물은 c-Jun 시그날에 의해서도 조절되는 MMP1이 자외 선에 의해 유도되는 것을 억제하였다. 뿐만 아니라, 초피나무 열매 추출물 PPAR-α 활성이 있어 장벽강화를 통한 피부 보호 효과가 있는데, 자외선에 의하여 염증 유발 물질인 IL-6와 IL-8의 발현이 증가하는 것도 억제 하였다. 사람의 팔에 자외선을 쪼여 준 경우에도 초피나무 열매 추출물이 홍반이 생기는 것을 억제하고 홍반에 의한 색소침착도 억제하였다. 종합적으로, 초피나무 열매 추출물은 다양한 메카니즘을 통하여 자외선으로부터 피부를 보호해 줄 것으로 기대된다.

The morphological characteristics and genetic relationships among 32 germplasms of Zanthoxylum schinifolium and Zanthoxylum piperitum collected from two farms in Korea were investigated. The traits with the most variability were seed color, leaf size, and spine size. The intraspecific polymorphism of Z. schinifolium and Z. piperitum was 96.5% and 60.3%, respectively. The genetic diversity and Shannon’s information index values ranged from 0.11 to 0.33 and 0.19 to 0.50, with average values of 0.26 and 0.42, respectively. Two ISSR primers (UBC861 and UBC862) were able to distinguish the different species. The genetic similarity matrix (GSM) revealed variability among the accessions ranging from 0.116 to 0.816. The intraspecific GSM for Z. schinifolium and Z. piperitum was 0.177–0.780 and 0.250–0.816, respectively. The GSM findings indicate that Z. schinifolium and Z. piperitum accessions have high genetic diversity and possess germplasms qualifying as good genetic resources for cross breeding. The clustering analysis separated Z. schinifolium and Z. piperitum into independent groups, and all accessions could be classified into three categories. Z. Schinifolium var. nermis belonged to independent groups. Comparison of the clusters based on morphological analysis with those based on ISSR data resulted in an unclear pattern of division among the accessions. The study findings indicate that Z. schinifolium and Z. piperitum accessions have genetic diversity, and ISSR markers were useful for identifying Z. schinifolium and Z. piperitum.

생강나무 줄기와 초피 과피의 60% ethanol과 열수 추출물의 식중독 균에 대한 항균 활성과 추출물의 열안정성에 대하여 알아보았다. 100 mg/mL의 농도에서 생강나무 줄기의 60% ethanol 추출물은 MRSA에 대하여 31.50 mm, 열수추출물은 S. aureus에 대해 25.5 mm의 생육 저해환을 보이며 가장 높은 항균활성을 보였으며, B. cereus에 대해서는 가장 낮은 항균활성을 보였다. 같은 농도에서 초피 과피의 60% ethanol 추출물의 항균효과는 S. aurues와 B. cereus에 대해 25 mm의 생육 저해환을 나타냈으며, 열수추출물은 S. aurues 에 대해 22 mm의 생육저해환을 나타내 항균 효과가 가장 크게 나타났으며, E. coli 에 대해서는 낮은 항균활성을 나타내었다. 생강나무 줄기와 초피 과피의 60% ethanol, 열수 추출물 모두 농도가 낮아짐에 따라 생육 저해환의 크기가 작아지는 현상을 보였으며, ethanol추출물이 열수 추출물에 비해 다소 높은 항균력을 나타냈다. 또 추출물의 열안정성 실험에서 열처리군과 비열처리군(대조구)을 비교했을 때 항균 효과가 감소하지 않는 것으로 보아 생강나무 줄기와 초피 과피 추출물은 열에 안정하다고 판단된다. 본 연구 결과 생강나무 줄기와 초피 과피 추출물에 미생물 부패 방지에 효과적인 항균 물질이 함유되어 있으며 비교적 열에 안정하므로 천연보존료로서 산업적 응용이 가능할 것이라 사료된다.

A protocol is described for rapid multiplication of Zanthoxylum piperitum DC. (Rutaceae), an important aromatic and medicinal plant, through shoot-tip explant cultures. Murashige and Skoog (MS) medium supplemented with various concentrations of N-6-benzyladenine (BA), N-6-benzylaminopurine (BAP) and thidiazuron (TDZ), in single or in combination with α-naphthaleneacetic acid (NAA), was used to determine the rate of shoot proliferation. N-6-benzyladenine (BA) used at 0.5mg/l, was the most effective in initiating multiple shoot proliferation at the rate of 23 microshoots per shoot-tip explants after 40 days of culture. Shoot multiplication increased 1.2-fold in each successive subculture. Induction of rooting (98%) was achieved by transferring the shoots to the same basal medium containing 2 mg/l indole-3-butyric acid (IBA). Plantlets went through a hardening phase in a controlled growth chamber, prior to in vivo transfer. These results represented that possible application for the mass production of plantlets through in vitro culture system of Zanthoxylum piperitum DC.

초피나무 종자의 전처리가 포장에서의 출아에 미치는 영향을 구명 하고자 노천매장과 NaClO 10% 등 7처리를 두어 생육 특성 을 분석하였다. 출아율은 노천매장 17.3%에 비해 GA3100 ppm과 NaClO 10% 처리 후 노천매장에서 각각 30.3%, 22.7%로 출아율이 향상되었으며 , 파종 후 26∼33일 사이에서 가장 높은 출아를 보였고, 이후 미미한 증가를 보였다. T/R율은 GA3 100 ppm전처리 후 노천매장에서 뿌리의 발달이 가장 양호하였다.

산초나무는 잎에서 총 52개, 열매에서 48개 향기 성분이 동정되었으며, 4개 지역에서 잎의 주요 공통 성분은 (E)-2-hexenal, α-pinene, (Z)-ocimene+limonene, estragole, germacrene-d 이였으며, 열매에서는 estragole이였다. Hexanal, (Z)-3-hexenol, (E)-2-hexenal, n-hexanol 성분은 잎에서만 검출되었고, undecanone 성분은 열매에서만 나타났다. 지역간 큰 차이를 보인 잎의 정유성분은 hexanal, azulene이었고, 열매에서는 (Z)-ocimene+limonene이였다. 초피나무 잎에서 총 30개, 열매에서 27개 향기 성분이 동정되었으며, 잎의 주요 공통 성분은 백양사와 내장사지역의 성분이 α-pinene, β-phellandrene, 1,8-cineole, citronellal이였고, 통도사 지역은 (Z)-3-hexenol, (E)-2-hexenal, α-pinene, myrcene이 주요성분 이였다. 백양사와 내장사에서 채취한 열매의 공통 성분은 myrcene, (Z)-ocimene+limonene, β-phellandrene이였고, 통도사에서는 β-phellandrene, citronellal, geranyl acetate이 주성분이였다.

초임계추출(SFE)를 이용한 초피나무 과피 중 신미성분의 최적추출조건을 조사하였다. 최적 추출 압력은 300kg/cm2 이었으며 온도는 60℃, 추출 부피는 12 ml, flow rate는 3m/min 그리고 modifier로 사용한 methanol의 함량비율은 20%였다. 추출시간은 초반 20분 안에 대부분의 신미성분이 추출되었다. 또한 용매추출법과 비교한 결과 추출효능이 약 40%정도 향상되었고 시간은 20분으로 단축되었으며 유기용매 methanol은 12 ml밖에 소요되지 않을 뿐만 아니라 주요한 추출용매로 무독성 액화CO2를 사용하기에 독성 유기용매의 다량 사용을 피할 수 있는 친환경적이고 경제적인 방법으로 생각되어 진다.

산초에서 시기에 따라 차이가 크게 나타나는 주요성분은 hexanal, (E)-2-hexenal, α-pinene, myrcene, (Z)-ocimene+limonene, (E)-β-ocimene, linalool, citronellal, estragole 이었으며, 특히 estragole은 모든 지역에서 공통적으로 나타났다. 초피에서 시기에 따라 차이를 보이는 성분은 hexanal, (Z)-3-hexenol, n-hexanol, α-pinene, limonene, 1,8-cineole, citronellal, estragole, citronellol 이었으며, 이중 (Z)-3-hexenol α-pinene, limonene, citronellal이 모든 지역에서 공통으로 나타났다. 수목원에서 월별 변화를 보이는 성분을 관찰한 결과. 산초는 α-pinene, myrcene, (Z)-3-hexenyl acetate, α-phellandrene, (Z)-ocimene+limonene, β-phellandrene, linalool, geranyl acetate 이였으며, 초피는 hexanal, (Z)-3-hexenol, (E)-2-hexenal, n-hexanol, α-pinene, (Z)-ocimene, limonene, citronellal, geranyl acetate, β-caryophyllene 이였다. 그러나 지역별 채집한 개체에 나타났던 estragole이 수목원에 식재된 산초나무 잎에서 미량으로만 존재한 이유는 열매를 맺지 못할 정도의 어린나무였기 때문이라고 생각되어 estragole 성분은 열매를 맺는 시기에 증가되는 성분이라 사료되었다.

간편하고 효과적인 정유성분 분석을 위해 SPME fiber를 선별하고 산초와 초피의 개체 정유성분 분석에 적합한 SPME fiber를 찾고자 실험한 결과는 다음과 같다. 단일물질별 흡착율은 PDMS, PDMS/DVB, CAR/PDMS 가 우수했다. 10종의 단일물질 표준품 혼합액 흡착 실험에서 직접주입 했을때와 가장 유사한 결과를 보인 것은 PDMS, PDMS/DVB fiber이였다. 최종 선발된 PDMS와 PDMS/DVB를 이용하여 24종의 단일물질 표준품 흔합액 흡착과 직접주입시를 비교한 결과 이들 간에 차이가 없었다. 또한 GC주입구의 split ratio별 성분의 검출에서도 차이가 없었다. 흡착시간이 길어질수록 분자량이 큰 성분의 흡착율이 높아지는 경향을 보였지만, 직접주입시와 비교해 볼 때 적절한 흡착시간은 30-40분이였다. SDE 추출물을 이용한 산초와 초피나무 개체 정유성분 분석에는 산초와 초피 모두 PDMS/DVB가 흡착시간 30-40분에서 분자량이 큰 sesquiterpene류까지 흡착하는데 유리하였다.

Yields of various solvent extracts for Zanthoxylum schinifolium and Z. piperitum leaves were higher in ethanol extract layer. Ethanol Extract in sancho and chopi leaves was fractionated with different solvents, such as hexane, chloroform, ethyl acetate, butanol and water, yields of hexane fraction was higher. In the solvent extracts using methanol, ethanol, hexane, chloroform, and water, ethanol extract showed the most effective antimicrobial activities. Ethanol extract in sancho and chopi leaves was fractionated, the most antimicrobial activities of sancho leaf were ethyl acetate fraction, and chopi leaf were chloroform fraction.