Bacillus cereus 1-1 균주의 5-Aminolevulinic Acid (ALA) 생산

안경준,Ahn, Kyung-Joon 한국미생물학회 2007 미생물학회지 Vol.43 No.4

Bacillus cereus 1-1 균주는 광이 없는 호기적 환경에서 levulinic acid와 같은 저해제 처리 없이도 5-aminolevulinic acid (ALA)를2 mM까지 생산하였다. B. cereus 1-1 균주는 TCY 배지에서 전 배양과 본 배양을 18시간 동안 지속하고, 배지의 pH가 6.8에 도달하는 대수기 후기에 acetic acid를 비롯한 유기산들을 16 mM 첨가하였을 때 많은 ALA를 생산하였으며, 본 배양 시작시 0.3% glucose를 첨가하는 것이 효과적이었다. Acetic acid 대신 glutamic acid를 첨가하였을 때 ALA 생산이 8시간 이상 지속되었고, $40\;{\mu}M$의 gabaculine을 첨가하면 생산이 현저히 저해되는 것으로 보아 B. cereus 1-1 균의 ALA 생산은 C-5 경로에 의함을 알 수 있었다. Bacillus cereus 1-1 strain produced 2 mM of ALA in the aerobic dark condition without any inhibitor like levulinic acid. The optimum culture conditions for the ALA production were that preculture and main culture were continued for 18 hr in TCY medium, and 16 mM of organic acids like acetic acid were added at the late log phase when the pH was 6.8. And the addition of 0.3% glucose was effective at the beginning of the main culture. ALA production was continued for more than 8 hr by the addition of glutamic acid instead of acetic acid, and was inhibited by addition of $40\;{\mu}M$ gabaculine seriously. These results confirmed that B. cereus 1-1 strain produced ALA through C-5 pathway.

Rhodobacter sphaeroides에서 5-aminolevulinic acid 생산에 대한 850 nm 근적외선 발광다이오드 조사 효과

모상준 ( Sangjoon Mo ) 한국응용생명화학회 2021 Journal of Applied Biological Chemistry (J. Appl. Vol.64 No.3

광감각제를 이용한 광역학 치료는 필요한 특정 부위에만 빛을 조사하여 치료 효과를 나타내는 부작용이 적은 방법이다. 5-aminolevulinic acid (ALA)는 다양한 생물체에서 합성되는 대표적 광감제로 암진단과 치료를 포함하는 다양한 분야에서 사용되고 있다. 본 연구에서는 다양한 파장의 LED, 유기산 전구체 및 glucose 농도 변화를 통한 Rhodobacter sphaeroides의 최적 성장 조건과 ALA 생산 조건을 확립하기 위한 실험을 진행하였다. 백열등과 동일한 광도 아래에서 Rhodobacter sphaeroides에 850 nm LED 빛을 조사하면 대조군 대비 균주의 성장과 ALA의 생산 농도를 각각 1.5배 및 1.8배 증가시킬 수 있고, 전구체로 pyruvic acid를 첨가한 경우 850 nm 파장의 LED만 조사한 경우 보다 ALA의 생산 농도를 약 2.8배 증가 시켰으며 동일 배양 조건에 40 mM glucose를 첨가하여 배양한 결과 Rhodobacter sphaeroides의 성장은 850 nm 파장의 LED 조사와 pyruvic acid를 첨가한 것에 비해 약 2.9배, ALA의 생산 농도는 약 3.4배 (20 mM) 증가되었다. 건조체 질량당 ALA의 생산은 20 mM과 40 mM glucose에서 대조군 대비 각각 약 1.4배 높은 결과를 나타냈다. 결론적으로 다양한 파장의 LED 중 850 nm 파장의 LED가 Rhodobacter sphaeroides의 성장률 및 ALA의 생산을 최대로 높였으며, 5 mM pyruvic acid와 40 mM glucose의 농도에서 최적의 Rhodobacter sphaeroides 성장과 ALA 생산을 확인하였다. 5-aminolevulinic acid (ALA) is a representative photosensitizer used in numerous fields including cancer diagnosis and treatment. In this study, experiments were conducted to optimize the growth of Rhodobacter sphaeroides and production of ALA through LED irradiation of various wavelengths, addition of organic acid precursors of ALA, and changes in glucose concentration. After 72 h cultivation, the 850 nm wavelength LED irradiated at the same light intensity as the incandescent lamp increased the growth of R. sphaeroides and the production of ALA about 1.5- and 1.8-fold as compared with the control, respectively (p <0.0001 and p <0.0001). As a result of culturing R. sphaeroides by irradiating an LED with a wavelength of 850 nm after adding organic acid to the final concentration of 5 mM in culture medium, the production of ALA was increased about 2.8- fold in medium supplemented with pyruvic acid compared with the control (p <0.0001). In addition, the growth of the strain and the production of ALA were increased about 2.9- and 3.4-fold in medium supplemented with 40 mM glucose compared to the control which added only 5 mM pyruvic acid, respectively (p <0.0001 and p <0.0001). The yield of ALA per cell dry mass was about 1.4 folds higher than that of the control in 20 and 40 mM glucose, respectively (p <0.001). In conclusion, the growth of R. sphaeroides and production of ALA were increased by 850 nm wavelength LED irradiation. It also optimized the growth of R. sphaeroides and production of ALA through organic acid addition and glucose concentration changes.

5-Aminolevulinic acid의 국소적 투여에 의한 자궁경부암의 광역학치료에 대한 연구

안진철 ( Jin Chul Ahn ),조현철 ( Hyun Chul Cho ),박진완 ( Jin Wan Park ),박충학 ( Choong Hak Park ),윤문희 ( Moon Hee Youn ) 대한산부인과학회 2010 Obstetrics & Gynecology Science Vol.53 No.12

목적: 자궁경부암에서 5-aminolevulinic acid (5-ALA)의 국소적 투여에 의한 광역학치료의 항암치료효과를 알아보고자 하였다. 연구 방법: Nude mouse에 인체의 자궁경부암세포인 hella cell을 이종이식 후에 일정크기의 종양이 형성되면 20% 5-ALA cream을 만들어 종양에 국소도포하고 밀봉하여 3시간마다 532 nm diode laser를 종양에 조사하여 발광되는 정도를 관찰하여 5-ALA의 생체 내 분포와 최대농도로 종양에 분포되는 시간을 알아보았다. 이렇게 알아낸 시간을 치료를 위한 최적의 투여시 간으로 간주하고 같은 방법으로 이번에는 632 nm diode laser를 이용하여 480 J/cm2의 에너지로 광역학치료를 시행하고 30일간 3일마다 그 크기를 측정하여 치료효과를 판별하였다. 이종이식된 25마리의 nude mouse를 대조군, 5-ALA만 투여한 군, laser만 조사한 군, 광역학치료 군으로 나누어 서로 비교분석하여 종양의 치료효과를 알아보았다. 결과: 5-ALA 국소투여 후 9시간 후에 가장 많이 발광되었다. 광역학치료 군에서 40% (4/10)는 종양의 크기가 감소하였지만, 이를 제외한 나머지 실험군에서는 지속적인 종양 크기 증가가 관찰되었다. 결론: 본 연구에서는 5-ALA의 국소투여 후 9시간이 치료에 가장 적절한 시간임을 알 수 있었다. 5-ALA 국소투여에 의한 광역 학치료는 대조군에 비해 유의한 종양크기의 감소가 없었으나, 반응군(4/10)의 경우는 대조군에 비해 유의한 종양의 크기 감소가 관찰되었다(P<0.05). Objective: The purpose of this study was to investigate the treatment effects of a topical application of 5-aminolevulinic acid (5-ALA) for photodynamic therapy (PDT) to treat cervical cancer. Methods: We first investigated the effects of 5-ALA cream according to application time. And to find the effective 5-ALA concentration and the distribution times in vivo, 20% 5-ALA cream was topically applied to the tumor of the nude mouse. We then observed the distribution of 5-ALA via fluorescence measurement with using a 532 nm diode laser. 25 nude mice were divided into Control, ALA, Laser, and PDT group. To evaluate the PDT effect at cancer lesion, we applied 20% 5-ALA cream to the tumor by the same method, and the PDT was done by using a 632 nm diode laser at the time of the peak level of fluorescence. We checked the changes of the volume of cancer for 30 days, and then biopsy was done. Results: The effective post-irradiation time after topical ALA application was 9 hours. In the PDT group, 40% (4/10) of the mice showed decreased tumor size. Conclusion: The maximum PpIX fluorescence at 9 hours after local applicationof 5-ALA cream was checked. And PDT group did not show any statistical difference than control group in the growth of tumor size than control group. However responding cases (4/10) of PDT group showed the meaningful decrease of tumor size than control group (P<0.05).

Rhodopseudomonas sp. KCTC1437에서의 Polyhydroxyalkanoates와 5-Aminolevulinic Acid의 생합성

이영하,기형석,최강국,문명님,양영기 한국미생물학회 2002 미생물학회지 Vol.38 No.2

Rhodopseudomonas sp. KCTC1437균주를 이용하여 Polyhydroxyalkanoate (PHA)와 5-aminolevulinic acid (ALA)를 생산하기 위한 배양조건과 이들 생합성 조건의 상호관련성에 대하여 조사하였다. PHA생합성을 위한 탄소원으로는 acetic acid가 가장 효과적이었으나, succinic acid를 보조 탄소원으로 사용하였을 때의 세포건체량은 2.5g/ι, PHA함량은 건체량의 73%로서 주 탄소원만을 사용할 때에 비하여 크게 증가하였다. 조사된 탄소원으로부터 생합성된 PHA는 모두 polyhydroxybutyrate 단일중합체 이었으나, valeric acid로부터는 3-hydroxybutyrate와 3-hydroxyvalerate로 구성된 공중합체가 생산되었다. ALA의 생합성을 위하여서는 환원제 인 sodium thioglycolate를 첨가하여 혐기적 조건을 만들고, 탄소원인 acetic acid와 propionic acid 이외에 전구물질인 levulinic acid, succinic acid와 glycine을 반복적으로 공급해주었을 때 가장 좋았으며, 약 400 mg/ι의 ALA를 생산할 수 있었다. 그러나 ALA 생합성의 필수물질인 glycine, levulinic acid와 환원제는 세포생장과 PHA의 생합성을 저해하는 것으로 나타났다. 이러한 실험 결과, Rhodopseudomonas sp. KCTC1437균주로부터 PHA와 ALA를 동시에 생산할 수 있으나, 두 가지 유용산물을 효율적으로 생합성하기 위한 각각의 배양조건이 상호 배타적임을 확인하였다. For elucidating the relationship between the biosynthetic pathways for polyhydroxyslkanoates (PHAs) and 5-aminolevulinic acid (ALA), culture conditions for the production of these two biomaterials by Rhodopseudomonas sp. KCTC 1437 were investigated. Of the carbon substrates tested, acetic acid was the best carbon source for cell growth and PHA biosynthesis. When succinic acid was added as a co-substrate into culture medium, cell growth and PHA production were greatly increased up to 2.5 g/ι and 73% of dry cell weight, respectively. The PHA obtained from the carbon substrates tested was homopolyester of 3-hydroxybutyrate, while valeric acid was only effective for the production of copolyester consisting of 3-hydroxybutyrate and 3-hydroxyvalerate. Anaerobic light culture condition was better for PHA production and cell growth than anaerobic dark or aerobic dark culture condition. The organism was capable of synthesizing ALA when glycine and succinic acid were added to the culture medium. ALA was produced to ca.400 mg/ι when levulinic acid, soccinic acid, and glycine were repeatedly added with a reductant (sodim thioglycolate). However, the presence of glycine, levulinic acid and sodium glycolate inhibited the cell growth and the conversion of carbon substrates to PHA. From these results it is apparent that the production yields of PHA and ALA could not be increased simultaneously because the optimal conditions for the production of PHA and ALA are opposed to each other.

Rhodocyclus gelatinosus KUP-74에 의한 δ-Aminolevulinic acid 생합성의 C_5-전구물질의 영향

최경민,임왕진,황세영 한국산업미생물학회 1993 한국미생물·생명공학회지 Vol.21 No.6

광합성 세균인 Rhodocyclus gelatinosus KUP-74에 의한 δ-aminolevulinic acid(ALA)의 생산을 위하여 C_4와 C_5 ALA 생합성 경로가 활성형으로써, C_5 경로가 C_4 경로에 우선하였다. C_5 경로 전구물질들의 세포막 투과 속도와 이들에 의한 균체의 ALA 생산성의 상관관계를 in vitro 및 in vivo계를 이용하여 검토한 결과, 본 균주의 C_5 경로를 이용한 ALA 생합성을 위하여 L-glutamine이 L-glutamate보다 효과적인 전구물질로 판단되었다. Aminolevulinic acid (ALA) was shown to be synthesized via active pathways of either C_4 or C_5 ALA biosynthesis in cells of a photosynthetic bacterium, Rhodocyclus gelatinosus KUP-74, where the C_5 pathway was appeared to be preferentially expressed in the cells. It was strongly suggested that L-glutamine might be utilized more effectively than L-glutamate to synthesize ALA via C_5 pathway in this bacterium from the fact of relationship between the cellular uptake rates of glutamate and its γ-derivatives and corresponded ALA productivities in vitro and in vivo.

병원성세균에 대한 5-Aminolevulinic Acid로 중재된 광역학치료의 항균효과

권필승 대한임상검사과학회 2022 대한임상검사과학회지(KJCLS) Vol.54 No.4

This study evaluates the improved effect of photodynamic therapy (PDT) by subjecting pathogenic bacteria to a combination of 630 nm light-emitting diode (LED) and 5-aminolevulinic acid (ALA). Bacterial suspensions of 1.5×104 cells/mL were diluted and exposed to ALA concentrations of 10, 5, 2.5, 1.25, and 0.625 mg/mL, incubated for 30 minutes, followed by irradiation with 630 nm LED (18 J/cm2). The non-irradiated P. aeruginosa group and the group administered only LED light averaged 415 and 245 colonies, respectively. Conversely, the PDT group showed an average of 109, 225, 297, and 285 colonies at concentrations of 10, 5, 2.5, and 1.25 mg/mL of ALA. Evaluating the effect on E. faecalis revealed an average of 8,750 and 8,000 colonies in the group that did not receive the control photosensitizer and the group exposed to light alone, respectively. However, an average of 0, 2350, 4825, and 7475 colonies at concentrations of 5, 2.5, 1.25, and 0.625 mg/mL ALA were determined for the PDT groups. In conclusion, better inhibitory effects were observed for E. faecalis than for P. aeruginosa. Moreover, our results validate the possibility of improved PDT efficacy using a combination of ALA and 630 nm LED.

RAW 264.7 대식세포에서 lipopolysaccharide 자극에 의한 염증성 및 산화적 스트레스에 미치는 5-aminolevulinic acid phosphate의 영향

지선영(Seon Yeong Ji),김민영(Min Yeong Kim),황보현(Hyun Hwangbo),이혜숙(Hyesook Lee),홍수현(Su Hyun Hong),차희재(Hee-Jae Cha),김희수(Heui-Soo Kim),김석만(Suhkmann Kim),최영현(Yung Hyun Choi) 한국생명과학회 2021 생명과학회지 Vol.31 No.9

5-ALA-p는 천연 아미노산인 5-ALA를 암모니아수로 용출하고 인산과 아세톤을 첨가하여 광역학 요법에 적합한 특성을 갖도록 개발된 물질이다. 그러나 항산화 및 항염증에 대한 잠재적인 기전을 포함한 약리학적 효능은 아직 명확하지 않다. 본 연구에서는 LPS로 자극된 RAW 264.7 세포에서 산화적 및 염증성 반응에 대한 5-ALA-p의 효과를 평가하였다. 본 연구의 결과에 의하면, 5-ALA-p는 LPS에 의한 RAW 264.7 세포의 과도한 식균 활성을 유의하게 억제하였고 산화적 스트레스를 약화시켰다. 5-ALA-p는 또한 LPS에 의해 감소된 미토콘드리아 생물 발생을 개선하였으며, 이는 5-ALA-p가 LPS로 인한 미토콘드리아 손상을 복원시켰음을 시사한다. 아울러 5-ALA-p는 NO와 TNF-α, IL-1β 및 IL-6과 같은 염증성 사이토카인의 생성을 현저히 억제하였으며, 이는 iNOS 및 각 사이토카인의 발현 감소와 연관성이 있었다. 나아가 5-ALA-p는 NF-κB의 핵 전이를 감소시키고 MAPKs의 인산화를 억제하여 5-ALA-p의 항염증 효과가 이들 신호전달 경로의 활성 억제와 매개되었음을 보여주었다. 이러한 결과들은 5-ALA-p가 산화적 및 염증성 스트레스를 줄이는 잠재적인 후보 약물로 적용될 수 있음을 의미한다. 5-Aminolevulinic acid phosphate (5-ALA-p) is a substance obtained by eluting 5-ALA (a natural delta amino acid) with aqueous ammonia, adding phosphoric acid to the eluate, and then adding acetone to confer properties suitable for use in photodynamic therapy applications. However, its pharmacological efficacy, including potential mechanisms of antioxidant and anti-inflammatory reactions, remains unclear. This study aimed to investigate the effects of 5-ALA-p on oxidative and inflammatory stresses in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Our data showed that 5-ALA-p significantly inhibited excessive phagocytic activity via LPS and attenuated oxidative stress in LPS-treated RAW 264.7 cells. Furthermore, 5-ALA-p improved mitochondrial biogenesis reduced by LPS, suggesting that 5-ALA-p restores mitochondrial damage caused by LPS. Additionally, 5-ALA-p significantly suppressed the release of nitric oxide (NO) and pro-inflammatory cytokines, such as tumor necrosis factor α (TNF-α), interleukin (IL)-1β, and IL-6, which are associated with the inhibition of inducible NO synthase and respective cytokine expression. Furthermore, 5-ALA-p reduced the nuclear translocation of nuclear factor-kappa B (NF-κB) and inhibited phosphorylation of mitogen-activated protein kinases (MAPKs), indicating that the anti-inflammatory effect of 5-ALA-p is mediated through the suppression of NF-κB and MAPK signaling pathways. Based on these results, 5-ALA-p may serve as a potential candidate to reduce inflammation and oxidative stress.

5-Aminolevulinic acid improves chicken sperm motility

Taniguchi Shin,Zhu Zhendong,Matsuzaki Mei,Tsudzuki Masaoki,Maeda Teruo 아세아·태평양축산학회 2021 Animal Bioscience Vol.34 No.12

Objective: This study investigated the effects of 5-aminolevulinic acid (5-ALA) on the motility parameters, mitochondrial membrane depolarization, and ATP levels in chicken sperm. Methods: The pooled semen from Barred Plymouth Rock males was used. In the first experiment, the semen was diluted 4-times with phosphate-buffered saline (PBS (-)) containing various concentrations (0, 0.01, 0.05, and 0.1 mM) of 5-ALA, and then the sperm motility parameters after incubation were evaluated by computer-assisted sperm analysis (CASA). In the second experiment, the semen was diluted 4-times with PBS (-) containing 0.05 mM 5-ALA, and then sperm mitochondrial membrane depolarization and ATP levels after 1.5 h of incubation were analyzed with the MitoPT® JC-1 Assay and ATP Assay kits, respectively. In the third experiment, the semen was removed from the seminal plasma and resuspended with the mediums of PBS (-), PBS (-) supplemented with CaCl2 and MgCl2 (PBS (+)) + 5-ALA, PBS (+) + caffeine, and PBS (+) + caffeine + 5-ALA. Then, the sperm motility parameters after incubation were evaluated by CASA. In the last experiment, the semen was treated with the mediums of PBS (-), PBS (-) + 5-ALA, 5.7% glucose, 5.7% glucose + 5-ALA after removing the seminal plasma, and then the sperm motility parameters were evaluated by CASA. Results: The addition of 0.05 mM 5-ALA significantly increased the chicken sperm motility, progressive motility, linearity, average path velocity, curvilinear velocity, straight-line velocity, and the wobble. The sperm mitochondrial membrane depolarization was also increased by the 5-ALA treatment. The 5-ALA treatment decreased the sperm ATP levels. Both the caffeine treatment and glucose treatment decreased the sperm motility during incubation period. Conclusion: 5-ALA might increase sperm mitochondrial membrane depolarization to utilize the ATP for enhancing sperm movement. Objective: This study investigated the effects of 5-aminolevulinic acid (5-ALA) on the motility parameters, mitochondrial membrane depolarization, and ATP levels in chicken sperm.Methods: The pooled semen from Barred Plymouth Rock males was used. In the first experiment, the semen was diluted 4-times with phosphate-buffered saline (PBS (-)) containing various concentrations (0, 0.01, 0.05, and 0.1 mM) of 5-ALA, and then the sperm motility parameters after incubation were evaluated by computer-assisted sperm analysis (CASA). In the second experiment, the semen was diluted 4-times with PBS (-) containing 0.05 mM 5-ALA, and then sperm mitochondrial membrane depolarization and ATP levels after 1.5 h of incubation were analyzed with the MitoPT^® JC-1 Assay and ATP Assay kits, respectively. In the third experiment, the semen was removed from the seminal plasma and resuspended with the mediums of PBS (-), PBS (-) supplemented with CaCl2 and MgCl2 (PBS (+)) + 5-ALA, PBS (+) + caffeine, and PBS (+) + caffeine + 5-ALA. Then, the sperm motility parameters after incubation were evaluated by CASA. In the last experiment, the semen was treated with the mediums of PBS (-), PBS (-) + 5-ALA, 5.7% glucose, 5.7% glucose + 5-ALA after removing the seminal plasma, and then the sperm motility parameters were evaluated by CASA.Results: The addition of 0.05 mM 5-ALA significantly increased the chicken sperm motility, progressive motility, linearity, average path velocity, curvilinear velocity, straight-line velocity, and the wobble. The sperm mitochondrial membrane depolarization was also increased by the 5-ALA treatment. The 5-ALA treatment decreased the sperm ATP levels. Both the caffeine treatment and glucose treatment decreased the sperm motility during incubation period.Conclusion: 5-ALA might increase sperm mitochondrial membrane depolarization to utilize the ATP for enhancing sperm movement.

Methyl 5-aminolaevulinic Acid 국소 광역동 요법을 이용한 광선각화증의 치료

박현선 ( Hyun Sun Park ),진선필 ( Seon Pil Jin ),조광현 ( Kwang Hyun Cho ) 대한피부과학회 2010 大韓皮膚科學會誌 Vol.48 No.10

Background: Photodynamic therapy (PDT) is a treatment modality involving the use of a photosensitizer, oxygen and a light source to induce selective targeted cell death. It is used for various skin conditions, including actinic keratosis (AK). Both 5-aminolevulinic acid and methyl aminolevulinic acid (MAL) are currently available as photosensitizers. Although there are several studies on the treatment of AK using 5-aminolevulinic acid-PDT in Korea, there are few studies on using MAL-PDT for the treatment of AK. Objective: The purpose of this study is to evaluate the efficacy and safety of MAL-PDT for the treatment of AK. Methods: We performed a retrospective study and reviewed 64 AK lesions from 28 patients who were treated by MAL-PDT between January 2008 and April 2010. The data was collected through the medical records, the clinical photographs and the biopsy specimens. Results: The patients were treated with either a single treatment or double treatments 1 week apart. The treatment results were assessed after 12 weeks. Overall, complete remission was achieved in 42/64 lesions (65.6%). Although a single treatment was effective for thin lesions, the complete response rates were significantly lower for the moderately thick and severely thick lesions (100% vs 71.3% vs 22.2%, respectively). Repeated treatment tended to improve the complete response rate of the severely thick lesions. A favorable cosmetic outcome was achieved and only tolerable local side effects were reported after MAL-PDT. The patients were followed up for an average period of 6 months and 4 lesions recurred. Conclusion: MAL-PDT is an effective and well-tolerated treatment option for thin and moderately thick AK. However, further study is required for determining the optimal regimen for thicker lesions and the long-term treatment results of MAL-PDT. (Korean J Dermatol 2010;48(10):837~843)

녹두 잎에서 암 처리에 의한 5-aminolevulinic acid dehydratase 활성의 증가

이성배,오동원,김수현,이인철 대전대학교 기초과학연구소 2013 自然科學 Vol.24 No.-

5-aminolevulinic acid dehydratase(ALAD) is a important enzyme in biosynthetic pathway of tetrapyrrole ring. In this study we examined the changes of the enzyme activity by the growth conditions, especially by the light. Enzyme activity increased gradually according to the leaf development in both group whether grown under light or dark conditions. The ALAD activity, however, always exhibited higher in dark grown etiolated leaves than that of normal leaves grown under light. Enzyme activity in leaves of the seedlings which were transferred from dark conditions to the light was significantly decreased compared with that which were staying under dark conditions. Similar with the above result, ALAD activity was increased in leaves which transferred to the dark conditions from light. These results showed that activity of ALAD was enhanced under the dark conditions. 5-aminolevulinic acid dehydratase(ALAD)는 엽록소나 heme 구조의 tetrapyrrole ring 생합성 과정에서 중요한 효소이다. 본 연구에서는 녹두의 성장 조건, 특히 빛에 의한 효소활성의 변화를 조사해 보았다. ALAD 활성은 잎에 집중적으로 나타나는 조직학적 특성을 보였다. 잎의 효소활성은 개체 발생의 단계에 따라 점차적으로 증가하였다. 그러나 암 상태에서 계속 성장된 황백화된 잎의 효소활성이 주기적인 빛에 조사에 의하여 정상적으로 성장된 잎의 효소 활성에 비하여 높았다. 암 상태에서 빛이 있는 상태로 성장환경이 변화된 잎의 효소 활성은 빛이 조사된 시점부터 현저하게 감소하는 추세를 보였고, 정상상태에서 암 상태로 성장환경이 전환된 잎에서의 효소활성은 서서히 증가하는 양상을 보였다. 이러한 결과는 5-aminolevunic acid dehydratase의 효소활성이 암 상태에서 더 높게 유지됨을 보여 주고 있다.