• Bulletin of the Korean Chemical Society
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• 제22권1호
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• pp.68-76
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• 2001

We have employed chemical modification to identify amino acids essential for the catalytic activity of alliinase (EC 4.4.1.4) from garlic (Allium sativum). Alliinase degrades S-alkyl-L cysteine sulfoxides, causing the characteristic odor of garlic. The activity of alliinase was rapidly and completely inactivated by N-bromosuccinimide(NBS) and slightly decreased by succinic anhydride and N-acetylimidazole. These results indicate that tryptophanyl, lysyl, and tyrosyl residues play an important role in enzyme catalysis. The reaction of alliinase with NBA yielded a characteristic decrease in both the absorbance at 280 nm and the intrinsic fluorescence at 332 nm with increasing reagent concentration of NBS, consistent with the oxidation of tryptophan residues. Kinetic analysis, fluorometric titration of tryptophans and correlation to residual alliinase activity showed that modification of only one residue present on alliinase led to complete inhibition of alliinase activity. To identify this essential tryptophan residue, we employed chemical modification by NBS in the presence and absence of the protecting substrate analogue, S-ethyl-L-cysteine (SEC) and N-terminal sequence analysis of peptide fragment isolated by reverse phase-HPLC. A fragment containing residues 179-188 was isolated. We conclude that Trp182 is essential for alliinase activity.

• 한국식품영양학회지
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• 제12권1호
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• pp.55-62
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• 1999

Changes in the alliinase activity during the aging of pickled garlic samples prepared by the several methods were investigated. The activity of alliinase in raw garlic was 8,37 units/mg protein. The ac-tivity in the garlic pickled with swoy sauce was reduced to 4.57 units/mg with 52% remaining by 1st week of pickling and to 1.05 units/mg protein with 12% remaining by 2nd week of pickling. The ac-tivity of alliinase in the garlic pickled with vinegar was 2.79 units/mg protein with 32% remaining by 1st week of pickling and was 0.26 units/mg protein only with 3% remaining by 2nd week of pickling. The activity of alliinase in the garlic pickled with 10% salt solution was 5.06 units/mg protein with 58% remaining by 1st week of pickling. After one week pickling the juice of pickled garlic was removed. Then garlics were pickled again with vinegar. The allinase acting in was reduced to 0.85 units/mg pro-tein with 10% remaining by 2nd week of pickling. The activity of alliinase in the garlic pickled with vin-egar was 2.79 units/mg protein with 32% remaining by 1st week of pickling. The juice of pickled garlic was removed after one week. Then the galics were again pickled with saysauce. The allinase activity in the garlic the garlic pickled again with soy sauce was reduced to 0.43 units/mg protein with 5% remain-ing by 2 week of pickling.

• 환경위생공학
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• 제22권1호
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• pp.57-66
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• 2007

Changes in the alliinase activity during the hot air drying and vacuum freeze drying of garlic samples treated as the whole, sliced and crushed state were investigated. The specific activity of alliinase in raw garlic was 8.52 units/mg protein. The activity in the whole garlic prepared by the hot air drying for 8 hrs was reduced remarkably to 5.22 units/mg protein with 61% remaining and to 4.25 units/mg protein with 50% remaining for 36 hrs. The activity in the sliced garlic prepared by the hot air drying for 36 hrs was reduced to 3.55 units/mg protein with 42% remaining and the activity in the crushed garlic prepared by the hot air drying for 36 hrs was reduced to 3.12 units/mg protein with 37% remaining. The garlic sample sliced or crushed was higher than the whole state in the efficiency of drying but was lower in the remaining activity of alliinase. The activity in the whole garlic prepared by the vacuum freeze drying for 8 hrs was reduced to 7.21 units/mg protein with 85% remaining and to 5.53 units/mg protein with 65% remaining for 36 hrs. The activity in the sliced garlic prepared by the vacuum freeze drying for 36 hrs was reduced to 4.55 units/mg protein with 53% remaining and the activity in the crushed garlic prepared by the vacuum freeze drying for 36 hrs was reduced to 4.16 units/mg protein with 49% remaining. The remaining activity of alliinase in the garlic samples prepared by the vacuum freeze drying was higher than the remaining activity in the garlic samples prepared by the hot air drying.

• 한국조리학회지
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• 제14권1호
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• pp.144-151
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• 2008

양파를 원형 그대로, 절편으로 하여 그리고 마쇄한 것을 진공 동결 건조 처리하여 각각 건조 과정 중 allicin 및 diallyl disulfide의 함량과 alliinase 활성 변화를 측정하였다. 생양파의 allicin 및 dia-llyl disulfide의 함량은 각각 26.40 ppm, 2.78 ppm이었다. 양파를 원형 그대로, 절편으로 하여 그리고 마쇄한 시료의 allicin 및 diallyl disulfide의 함량은 진공 동결 건조가 진행함에 따라 증가되었으며, 증가 정도는 시료의 형태에 따라 차이를 나타내었다(p<0.05). 생양파의 alliinase 비활성은 7.536 units/mg protein이었으며, 양파를 원형 그대로, 절편으로 하여 그리고 마쇄한 시료의 alliinase 활성은 진공 동결 건조가 진행함에 따라 감소되었다. 양파를 원형 그대로 8시간 동안 진공 동결 건조하였을 때 효소의 비활성은 5.516 units/mg pro-tein로 감소되었으며 73.2%의 잔존율을 나타내었고, 36시간 동안 진공 동결 건조하였을 때 효소의 비활성은 3.304 units/mg protein로 감소되었으며 43.8%의 잔존율을 나타내었다. 양파를 절편으로 하여 36시간 동안 진공 동결 건조하였을 때 효소의 비활성은 2.366 units/mg protein로 감소되었으며 31.4%의 잔존율을 나타내었고, 양파를 마쇄하여 36시간 동안 진공 동결 건조하였을 때 효소의 비활성은 2.232 units/mg protein로 감소되었으며 29.6%의 잔존율을 나타내었다. 양파를 원형 그대로 진공 동결 건조하였을 때 alliinase 활성에 있어서 가장 높은 잔존율을 나타내었다.

• 한국식품조리과학회지
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• 제10권4호
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• pp.376-380
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• 1994

마늘의 alliinase를 ammonium sulfate 분획과 hydroxylapatite chromatography, concanavalin A-Sepharose affinity chromatography에 의해 정제하여, 23% 의 회수율과 7.6배 정제도를 나타내었다(specific activity 116.6 units/mg). SDS-polyacrylamide gel electrophoresis에서 단일 band를 나타내므로 순수한 aliinase 로 추측되며 이 효소의 분자량은 42K 로 추정된다. 기질로서 S-ethyl-L-cysteine sulfoxide를 사용한 이 효소의 $V_max$값은 2.27${\mu}$monl/mg.min이고 $K_m$은 1O mM이다 . 정제효소의 optimum pH는 6.5 phosphate buffer이며, 40$^{\circ}C$에서 최대활성을 나타내었다. Activation energy value($E^*$)는 4.6Kcal/mole로 추정된다.

• Journal of Crop Science and Biotechnology
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• 제10권1호
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• pp.19-26
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• 2007

Glutathione S-transferases(GSTs, EC 2.5.1.18), glyoxalase-I(EC 4.4.1.5) and alliin lyase(alliinase, EC 4.4.1.4) are important enzyme systems in plant bodies. The first two are mainly detoxifying enzymes that utilize glutathione(GSH) in the defense mechanism, and the last one is mainly involved in secondary metabolism and relevant to sulfur compounds derived from GSH. The activities of the three enzymes have been investigated in soluble extracts of vegetable crops, including pumpkin, cabbage, broccoli, radish, carrot, potato, sweet potato, mungbean, and onion. GST activities were detected in all of the vegetables, and the extract of onion bulb exhibited the highest specific activity(648 nmol/min/mgP). The putative GSTs of most of the vegetables were found to be induced by ethanol. The activities of GSTs in onion bulb were found to be markedly inhibited by S-hexyl glutathione and were also inhibited by S-butyl glutathione and S-propyl glutathione. The anti-CmGSTF1 antiserum recognized a thick band for putative onion GST. The estimated glyoxalase-I activity level was also high in onion bulb(4540 nmol/min/mgP), indicating that the thick band detected by Western blot analysis might result from partial recognition of glyoxalase-I by the antiserum. The specific activities for glyoxalase-I were moderate in radish and carrot, and the extracts of other vegetables had rather low levels of activities. The extract of onion also showed the highest specific activity level for alliinase(2069nmol pyruvate/mgP). The extracts of other vegetables also had alliinase activities, although the estimated values were much lower than that of onion.

• Food Science and Biotechnology
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• 제15권6호
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• pp.838-843
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• 2006

The chemical processes involved in the formation of green pigment in crushed garlic cloves were investigated based on the principle of pink pigmentation in macerated onions. Intact greening and non-greening garlic cloves were either left untreated or heated at $90^{\circ}C$ for 3 min to inactivate enzyme activities. First, a colorless ether soluble compound referred to as color developer reacted with glycine (among all free amino acids) in garlic to form a second compound insoluble in ether. The latter compound then reacted with formaldehyde to yield the green colored pigment. Alliinase activity was necessary for the production of color developer and for the development of green pigment. In greening garlic that had been heat treated, green pigmentation did not proceed due to the heat-inactivation of alliinase, but the addition of alliinase solution into the garlic homogenates restored the pigmentation. However, this phenomenon was not observed in non-greening garlic with or without heat treatment. Finally, the mechanism of green pigment formation in crushed garlicis similar to that of pink pigment formation in macerated onions.

• Journal of Microbiology and Biotechnology
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• 제12권2호
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• pp.331-335
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• 2002

Aqueous extracts of garlic (Allium sativum) preparation were prepared after the samples were exposed to various heat treatments. A quantitative assessment of antimicrobial activities was carried out by determining the minimum inhibitory and microbicidal concentrations (MICs and MMCs) of the various extracts against some selected bacteria and fungi. The antimicrobial activity of garlic decreased as the heating temperature increased. This fact implies that alliinase may be the most critical rate-determinant to produce the activity when garlic is heated.

• 한국식품영양과학회지
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• 제41권5호
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• pp.655-660
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• 2012

밭마늘과 논마늘(영천산)의 매운맛 차이를 알아보고자, 마늘의 매운맛 생성에 관여하는 주요 인자들인 alliin과 allcin 그리고 SAC의 함량을 HPLC로 분석하고, 이들 함함성분의 생성에 관여하는 효소인 alliinase와 GTPase(${\gamma}$-glutamyl trans peptidase) 활성 및 pyruvate 생성량과 일반성분을 분석하였다. 수분함량은 논마늘이 밭마늘에 비하여 높았으며, 조회분, 조지방, 조단백, 탄수화물 함량 역시 밭마늘이 논마늘에 비하여 높았다. 마늘의 주요 매운맛 성분인 allicin 함량은 밭마늘이 $2.83{\pm}0.03$ mg/g, 논마늘이 $2.22{\pm}0.02$ mg/g을 나타내어 밭마늘이 논마늘에 비하여 유의적으로 높았다. Allicin의 전구체인 alliin의 함량은 밭마늘이 $8.97{\pm}0.09$ mg/g, 논마늘이 $8.22{\pm}0.08$ mg/g으로 나타났다. SAC 함량은 밭마늘이 $1.74{\pm}0.17$ mg/g으로, 논마늘의 $1.04{\pm}0.14$ mg/g에 비하여 유의적으로 높았다. 한편, 효소 alliinase 활성도는 밭마늘이 $285{\pm}43{\mu}mole/mg$ protein, 논마늘이 $239{\pm}34{\mu}mole/mg$ protein을 나타내어 유의적인 차이는 없었으나, GTPase 활성은 밭마늘이 86.3 ${\mu}mole/mg$ protein으로 논마늘의 78.4 ${\mu}mole/mg$ protein에 비하여 높게 나타내었다. 이상의 결과를 토대로 밭마늘이 매운 맛이 강한 것은 본 연구결과로부터 allicin 함량이 논마늘에 비하여 높은 데 기인된 것으로 파악되었다. 밭마늘은 논마늘에 비하여 alliinase 활성은 유사하였으나 alliin의 함량이 높아 allicin 생성량이 많았으며, SAC가 높고 GTPase의 활성이 높아 alliin의 함량이 높은 것으로 생각되었다. 따라서, 마늘을 기능성 식품소재로의 활용을 극대화하고자 한다면 밭마늘이 논마늘보다 더 적합한 것으로 생각된다.

• 한국약용작물학회:학술대회논문집
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• 한국약용작물학회 2008년도 추계학술발표회
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• pp.291-292
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• 2008