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Anti-microbial activity of fermented papaya extract against Colletotrichum gloeosporioides (causing anthracnose), Aspergillus flavus (causing aflatoxin), and Lasiodiplodia theobromae (causing black rot)
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Abstracts / Journal of Bioscience and Bioengineering 108 (2009) S21–S28
sequence of fluorescent protein expression DNA and photo-induced activation of protein expression in living cells were evidenced. References 1. Tang, X. J., Swaminathan, J., Gewirtz, A. M., and Dmochowski, I. J.: Regulating gene expression in human leukemia cells using light-activated oligodeoxynucleotides., Nuc. Aci. Res., 1-11 (2007). 2. Mayer, G. and Heckel, A.: Biologically active molecules with a “light switch”., Angew. Chem. Int. Ed., 45, 4900-4921 (2006). 3. Monroe, W. T., Mcquain, M. M., Chang, M. S., Alexander, J. S., and Haselton, F. R.: Targeting expression with light using caged DNA., J. Biol. Chem., 274, 20895-20900 (1999).
doi:10.1016/j.jbiosc.2009.08.105
BP-O8 Anti-microbial activity of fermented papaya extract against Colletotrichum gloeosporioides (causing anthracnose), Aspergillus flavus (causing aflatoxin), and Lasiodiplodia theobromae (causing black rot) Evelyn B. Taboada, and May V. Tampus Department of Chemical Engineering, University of San Carlos, Cebu City, Philippines This work aims to investigate the anti-microbial activity of a fermented papaya extract called Antica® against the disease-causing micro-organisms in agricultural fruits and crops such as Colletotrichum gloeosporioides (causing anthracnose), Aspergillus flavus (producing aflatoxin), and Lasiodiplodia theobromae (causing black rot). This fermented papaya extract is mainly composed of alpha-hydroxy acids (AHAs) as the active ingredients. Such a study is very beneficial in the agricultural sector as this leads to the alternative use of safe, organic, and environmentally-friendly pesticide. Results show that unfiltered Antica® gives a consistently strong inhibition against Colletotrichum gloeosporioides, which causes anthracnose. Its zone of inhibition reaches an average of 24.19 mm during the 5day incubation period and is well-sustained during an extended period of 10 incubation days. This indicates that the unfiltered and undiluted Antica® can inhibit the growth of Colletotrichum gloeosporioides, which causes anthracnose in agricultural fruits and crops. A concentrated sample of Antica® (coded WT002) shows total inhibition against Colletotrichum gloeosporioides as no growth of the microorganism was observed during the extended 10-day incubation period. These results are comparatively superior with bioassays using a known chemical pesticide against anthracnose. On the other hand, unfiltered Antica® exhibited negative inhibition against A. flavus and L. theobromae, with zones of inhibition equal to 6.7 mm during the entire incubation period. doi:10.1016/j.jbiosc.2009.08.106
BP-P1 Production of novel tyrosinase inhibitor from Zygomycota Yukiko Mita, Maki Takano, and Kazuhiro Hoshino University of Toyama, Toyama, Japan
Tyrosinase inhibitor, such as arbutin, cysteine, etc, was used as wellknown components of whitening cosmetics. However, these compounds have serious problems concerning melanin synthesis inhibiting activity and toxicity for skin cells. Therefore, we paid attention to Zygomycota (mainly, Mucor genus) as a new resource for the production of tyrosinase inhibitor. In this research, we attempted to screen for a new tyrosinase inhibitor from this genus. First, the productions of tyrosinase inhibitors were estimated by screening 82 kinds of fungi. When the monophenolase activity and the diphenolase activity for tyrosinase were assayed by using L-tyrosine and L-DOPA as substrates, respectively. As these results, the broth obtained by cultivating Mucor subtilissimus NBRC 6755 for 72 h with glucose as a substrate has higher inhibitor activities. In the monophenolase activity, its inhibition value was markedly increased from 119 s to 294 s. Further, the broth showed an inhibition effect of 21.8% in a diphenolase activity compared with the control. To purify the inhibitor from the broth, the broth was concentrated, extracted by methanol, and fractionated by HPLC, and then the inhibition material was obtained. The material was showed as a mixed-type inhibition for diphenolase activity, and the inhibition constant Ki on DOPA was 0.35 g/L. Moreover, the inhibitor showed an inhibition effect on tyrosinase after heat treatment for 5 min at 100 °C. Next, to estimate the ability of the inhibitor in vivo the inhibition assay for melanin synthesis and the toxicity assay were performed by using mouse B16 melanoma cell. As these results, the melanin synthesis amount decreased up to 37% at the inhibitor concentration of 10 mg/L and further the toxicity was not admitted. doi:10.1016/j.jbiosc.2009.08.487
BP-P2 Two cytotoxic sesquiterpenes from hairy root cultures of Artemisia annua L. induced apoptosis of highly metastatic lung carcinoma cell line 95-D Zhai Dandan,1 and Zhong Jianjiang2 State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China 1 and School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, China 2 Artemisia annua L. is well known as a rich source of many biologically active substances such as sesquiterpenoids, coumarins, and flavonoids (1). Hairy root cultures serve as a good choice for the production of biologically active natural products due to their fast growth rate (2). Recently we isolated a new sesquiterpene, (Z)-7acetoxy-methyl-11-methyl-3-methylene-dodeca-1,6,10-triene (AMDT), along with another known compound drimartol A (DA) from hairy root cultures of A. annua. AMDT and DA were found to have moderate cytotoxic activities against the human cancer cell lines HO8910 (ovary), 95-D (lung), QGY (liver), and HeLa (cervix) cells as assayed by MTT method, and their IC50 values ranged between 17.9 and 73.3 μM. Their induction of apoptosis in highly metastatic lung carcinoma cell line 95-D was investigated in detail. The cell cycle of 95-D cells was arrested in the G1 and G2 phases by AMDT and DA, respectively. The apoptotic rate of 95-D cells was increased with an increase of treatment amount of each compound in a dose-dependent manner. Both AMDT and DA increased the expression of caspase-9 and caspase-3 and lowered the mitochondrial membrane potential. Those findings revealed that AMDT and DA could efficiently induce 95-D cells apoptosis through a mitochondrial-dependent pathway and through the activation of the caspase cascade, and they may be potential chemotherapeutic agents.
Abstracts / Journal of Bioscience and Bioengineering 108 (2009) S21–S28
sequence of fluorescent protein expression DNA and photo-induced activation of protein expression in living cells were evidenced. References 1. Tang, X. J., Swaminathan, J., Gewirtz, A. M., and Dmochowski, I. J.: Regulating gene expression in human leukemia cells using light-activated oligodeoxynucleotides., Nuc. Aci. Res., 1-11 (2007). 2. Mayer, G. and Heckel, A.: Biologically active molecules with a “light switch”., Angew. Chem. Int. Ed., 45, 4900-4921 (2006). 3. Monroe, W. T., Mcquain, M. M., Chang, M. S., Alexander, J. S., and Haselton, F. R.: Targeting expression with light using caged DNA., J. Biol. Chem., 274, 20895-20900 (1999).
doi:10.1016/j.jbiosc.2009.08.105
BP-O8 Anti-microbial activity of fermented papaya extract against Colletotrichum gloeosporioides (causing anthracnose), Aspergillus flavus (causing aflatoxin), and Lasiodiplodia theobromae (causing black rot) Evelyn B. Taboada, and May V. Tampus Department of Chemical Engineering, University of San Carlos, Cebu City, Philippines This work aims to investigate the anti-microbial activity of a fermented papaya extract called Antica® against the disease-causing micro-organisms in agricultural fruits and crops such as Colletotrichum gloeosporioides (causing anthracnose), Aspergillus flavus (producing aflatoxin), and Lasiodiplodia theobromae (causing black rot). This fermented papaya extract is mainly composed of alpha-hydroxy acids (AHAs) as the active ingredients. Such a study is very beneficial in the agricultural sector as this leads to the alternative use of safe, organic, and environmentally-friendly pesticide. Results show that unfiltered Antica® gives a consistently strong inhibition against Colletotrichum gloeosporioides, which causes anthracnose. Its zone of inhibition reaches an average of 24.19 mm during the 5day incubation period and is well-sustained during an extended period of 10 incubation days. This indicates that the unfiltered and undiluted Antica® can inhibit the growth of Colletotrichum gloeosporioides, which causes anthracnose in agricultural fruits and crops. A concentrated sample of Antica® (coded WT002) shows total inhibition against Colletotrichum gloeosporioides as no growth of the microorganism was observed during the extended 10-day incubation period. These results are comparatively superior with bioassays using a known chemical pesticide against anthracnose. On the other hand, unfiltered Antica® exhibited negative inhibition against A. flavus and L. theobromae, with zones of inhibition equal to 6.7 mm during the entire incubation period. doi:10.1016/j.jbiosc.2009.08.106
BP-P1 Production of novel tyrosinase inhibitor from Zygomycota Yukiko Mita, Maki Takano, and Kazuhiro Hoshino University of Toyama, Toyama, Japan
Tyrosinase inhibitor, such as arbutin, cysteine, etc, was used as wellknown components of whitening cosmetics. However, these compounds have serious problems concerning melanin synthesis inhibiting activity and toxicity for skin cells. Therefore, we paid attention to Zygomycota (mainly, Mucor genus) as a new resource for the production of tyrosinase inhibitor. In this research, we attempted to screen for a new tyrosinase inhibitor from this genus. First, the productions of tyrosinase inhibitors were estimated by screening 82 kinds of fungi. When the monophenolase activity and the diphenolase activity for tyrosinase were assayed by using L-tyrosine and L-DOPA as substrates, respectively. As these results, the broth obtained by cultivating Mucor subtilissimus NBRC 6755 for 72 h with glucose as a substrate has higher inhibitor activities. In the monophenolase activity, its inhibition value was markedly increased from 119 s to 294 s. Further, the broth showed an inhibition effect of 21.8% in a diphenolase activity compared with the control. To purify the inhibitor from the broth, the broth was concentrated, extracted by methanol, and fractionated by HPLC, and then the inhibition material was obtained. The material was showed as a mixed-type inhibition for diphenolase activity, and the inhibition constant Ki on DOPA was 0.35 g/L. Moreover, the inhibitor showed an inhibition effect on tyrosinase after heat treatment for 5 min at 100 °C. Next, to estimate the ability of the inhibitor in vivo the inhibition assay for melanin synthesis and the toxicity assay were performed by using mouse B16 melanoma cell. As these results, the melanin synthesis amount decreased up to 37% at the inhibitor concentration of 10 mg/L and further the toxicity was not admitted. doi:10.1016/j.jbiosc.2009.08.487
BP-P2 Two cytotoxic sesquiterpenes from hairy root cultures of Artemisia annua L. induced apoptosis of highly metastatic lung carcinoma cell line 95-D Zhai Dandan,1 and Zhong Jianjiang2 State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China 1 and School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, China 2 Artemisia annua L. is well known as a rich source of many biologically active substances such as sesquiterpenoids, coumarins, and flavonoids (1). Hairy root cultures serve as a good choice for the production of biologically active natural products due to their fast growth rate (2). Recently we isolated a new sesquiterpene, (Z)-7acetoxy-methyl-11-methyl-3-methylene-dodeca-1,6,10-triene (AMDT), along with another known compound drimartol A (DA) from hairy root cultures of A. annua. AMDT and DA were found to have moderate cytotoxic activities against the human cancer cell lines HO8910 (ovary), 95-D (lung), QGY (liver), and HeLa (cervix) cells as assayed by MTT method, and their IC50 values ranged between 17.9 and 73.3 μM. Their induction of apoptosis in highly metastatic lung carcinoma cell line 95-D was investigated in detail. The cell cycle of 95-D cells was arrested in the G1 and G2 phases by AMDT and DA, respectively. The apoptotic rate of 95-D cells was increased with an increase of treatment amount of each compound in a dose-dependent manner. Both AMDT and DA increased the expression of caspase-9 and caspase-3 and lowered the mitochondrial membrane potential. Those findings revealed that AMDT and DA could efficiently induce 95-D cells apoptosis through a mitochondrial-dependent pathway and through the activation of the caspase cascade, and they may be potential chemotherapeutic agents.