• Title/Summary/Keyword: PAL enzyme activity

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Evidence of an Alternative Route of Cellobiase Secretion in the Presence of Brefeldin A in the Filamentous Fungus Termitomyces clypeatus

  • Banik, Samudra Prosad;Pal, Swagata;Chowdhury, Sudeshna;Ghorai, Shakuntala;Khowala, Suman
    • Journal of Microbiology and Biotechnology
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    • v.21 no.4
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    • pp.412-420
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    • 2011
  • Secretion of cellobiase occurred in a brefeldin A (BFA) uninhibited manner in the filamentous fungus Termitomyces clypeatus. Fluorescence confocal microscopy revealed that application of the drug at a concentration of 50 ${\mu}g$/ml caused arrest of Spitzenkorper assembly at the hyphal tip. This resulted in greater than 30% inhibition of total protein secretion in the culture medium. However, the cellobiase titer increased by 17%, and an additional 13% was localized in the vacuolar fraction en route secretion. The secretory vacuoles formed in the presence of the drug were also found to be bigger (68 nm) than those in the control cultures (40 nm). The enzyme secreted in the presence and absence of BFA revealed a single activity band in both cases in native PAGE and had similar molecular masses (approx. 120 kDa) in SDS-PAGE. The BFA enzyme retained 72% of native glycosylation. It also exhibited a higher stability and retained 98% activity at $50^{\circ}C$, 93.3% activity at pH 9, 63.64% activity in the presence of 1M guanidium hydrochloride, and 50% activity at a glucose concentration of 10 mg/ml in comparison to 68% activity, 75% activity, 36% activity, and 19% activity for the control enzyme, respectively. The observations collectively aimed at the operation of an alternative secretory pathway, distinct from the target of brefeldin A, which bypassed the Golgi apparatus, but still was able to deliver the cargo to the vacuoles for secretion. This can be utilized in selectively enhancing the yield and stability of glycosidases for a successful industrial recipe.

Early Growth, Pigmentation, Protein Content, and Phenylalanine Ammonia-lyase Activity of Red Curled Lettuces Grown under Different Lighting Conditions

  • Heo, Jeong-Wook;Kang, Dong-Hyeon;Bang, Hea-Son;Hong, Seung-Gil;Chun, Chang-Hoo;Kang, Kee-Kyung
    • Horticultural Science & Technology
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    • v.30 no.1
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    • pp.6-12
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    • 2012
  • Early growth, pigmentation, protein content, and phenylalanine ammonia-lyase (PAL) activity of red curly lettuces (Lactuca sativa L.) grown under different lighting conditions were investigated. Fluorescent lamps (control), blue, red, and blue plus red light-emitting diodes (LEDs) were used as light sources for 10 days. An equal proportion (1:1) of blue and red LEDs was used in the mixed radiation condition. Compared with the control, monochromic red or blue lighting increased fresh and dry weights of 'Ttuksum' and 'Jaju' lettuces. Anthocyanin synthesis was also significantly promoted by the mixed radiation of blue and red LEDs. The mixed radiation also increased the protein content and PAL enzyme activity of 'Ttuksum' leaves by about 200% compared to other treatments. Anthocyanin content was the highest in lettuces subjected to the mixture radiation of blue and red light treatment, while anthocyanin synthesis was inhibited by monochromic red light. The results of the present study indicate that growth and pigment synthesis in lettuces are significantly enhanced by exposure to mixed radiation from blue and red LEDs.

Effect of Dietary Lipid Sources on Growth, Enzyme Activities and Immuno-hematological Parameters in Catla catla Fingerlings

  • Priya, K.;Pal, A.K.;Sahu, N.P.;Mukherjee, S.C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.11
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    • pp.1609-1616
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    • 2005
  • Ninety advanced Catla catla fingerlings (av. wt. 16 g) were randomly distributed in six treatment groups with three replicates each for an experimental period of 60 days to study the effect of dietary lipid source on growth, enzyme activities and immuno-hematological parameters. Six isoprotein (40.0-41.9%) and isocaloric (4,260 kcal $kg^{-1}$) semi-purified diets were prepared with varying levels of soybean oil (SBO) and cod liver oil (CLO) within a total of 8% lipid viz., $D_1$ (Control), $D_2$ (8% SBO), $D_3$ (6% SBO and 2% CLO), $D_4$ (4% SBO and 4% CLO), $D_5$ (2% SBO and 6% CLO) and $D_6$ (8% CLO). Highest SGR was noted in $D_5$ (0.73${\pm}$0.03) group, which was similar with $D_3$ (0.71${\pm}$0.02) and $D_4$ (0.69${\pm}$0.01) groups. Activity of intestinal lipase, hepatic glucose-6-phosphate dehydrogenase (G6PDH) and aspartate amino transferase (AST) of the lipid treatment groups were significantly higher (p<0.05) than the control group. The respiratory burst activity of the phagocytes (Nitroblue tetrazolium (NBT)) was highest in $D_2$ (1.95${\pm}$0.21) followed by $D_3$ (1.19${\pm}$0.15) group, which were significantly (p<0.05) higher than the other groups. Globulin level was significantly higher in $D_3$ (1.29${\pm}$0.08) than in the other groups expect $D_4$. Hemoglobin content and total erythrocyte count did not show any significant difference. From this study, it is concluded that a diet containing 6% soybean oil and 2% cod liver oil ($D_3$) yields higher growth and immune response in Catla catla fingerlings and would be cost effective.

Introduction of Shiva Gene into tobacco and Potato Using Tissue-Specific Tomato PAL Promoter (조직특이성 promoter를 이용한 Shiva 유전자의 식물체내 도입)

  • 이정윤;이신우;박권우
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.2
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    • pp.109-113
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    • 1998
  • In this study we tried to transform an antimicrobial peptide gene (Shiva) under the promoter of tomato phenylalanine ammonia-lyase (tPAL5) into tobacco and potato plants. Antimicrobial peptide gene was isolated originally from giant silk moth (Hyalophora cecropia) and modified ie nucleotide sequence to increase antimicrobial activity. Transgenic tobacco plants were regenerated and their seeds were tested on the media containing kanamycin (500 mg/L). The results of PCR amplification and genomic Southern blot hybridization confirmed the integration of construct (tPAL5 promoter-Shiva-NOS-GUS-NOS) into chromosome. We observed that one of the transgenic tobacco plants showed chromosome rearrangement when integrated. In case of potato transformation, the efficiency of regeneration was maximized at the medium containing Zeatin 2mg/L, NAA 0.01mg/L, GA$_3$ 0.1mg/L. We also observed the high expression of GUS (${\beta}$-glucuronidase) enzyme which was located next to the terminator sequence of nopaline synthase gene (NOS) in the vascular tissue of stem, leaves of transgenic potatoes. This result suggested that a short sequence of Shiva gene (120 bp) and NOS terminator sequence might be served as a leader sequence of transcript when translated.

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Regional Difference of ROS Generation, lipid Peroxidation, and Antioxidant Enzyme Activity In Rat Brain and Their Dietary Modulation

  • Baek, Bong-Sook;Kwon, Hyun-Joo;Lee, Kyoung-Hee;Yoo, Mi-Ae;Kim, Kyu-Won;Yuji-Ikeno;Yu, Byung-Pal;Chung, Hae-Young
    • Archives of Pharmacal Research
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    • v.22 no.4
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    • pp.361-366
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    • 1999
  • One of the potential causes of age-related neuronal damage can be reactive oxygen species (ROS), as the brain is particularly sensitive to oxidative damage. In the present study, we investigated the effects of aging and dietary restriction (DR) on ROS generation, lipid peroxidation, and antioxidant enzymes in cerebrum, hippocampus, and cerebellum of 6-, 12-, 18-, and 24-month-old rats. ROS generation significantly increased with age in cerebrum of ad libitum (AL) rats. However, no significant age-difference was observed in hippocampus and cerebellum. DR significantly decreased ROS generation in cerebrum and cerebellum at 24-months. On the other hand, the increased lipid peroxidation of AL rats during aging was significantly reduced by DR in all regions. Our results further showed that catalase activity decreased with age in cerebellum of AL rats, which was reversed by DR, although SOD activity had little change by aging and DR in all regions. In a similar way, glutathione (GSH) peroxidase activity increased with age in cerebrum of AL rats, while DR suppressed it at 24-months. These data further support the evidence that the vulnerability to oxidative stress in the brain is region-specific.

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Antifungal Activities of Crude Extractum from Camellia semiserrata Chi (Nanshancha) Seed Cake Against Colletotrichum musae, Colletotrichum gloeosporioides and Penicillium italicum in vitro and in vivo Fruit Test

  • Meng, Xiangchun;Li, Jun;Bi, Fangcheng;Zhu, Lixue;Ma, Zhiyu
    • The Plant Pathology Journal
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    • v.31 no.4
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    • pp.414-420
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    • 2015
  • Antifungal activities of crude extractum of Nanshancha Seed Cake (NSC), to inactivate postharvest pathogens were investigated. Highest inhibitory rate was found against C. musae, C. gloeosporioides and C. papaya P.Henn, which was much stronger than that by tea saponin. Compared to tea saponin, effects of NSC extractum was relatively weak and similar on C. gloeosporioides Penzig and P. italicum. In an in vivo study, best controlling effects by NSC extractum was found with banana anthracnose disease development, which showed no inhibitory effects by tea saponin. NSC extractum controlled in vitro C. musae growth through directly inhibiting germination rate and germ tube elongation, and causing distortation, rupture and indentation of C. musae mycelium. In banana fruit subject to C. musae inoculation, higher PAL, POD, GLU and CHT activity was observed in banana fruit treated with crude NSC extractum than that of water control fruits. Current study proved the best controlling effects of crude NSC extractum in C. musae in vitro and in vivo development, which through direct inhibition of C. musae growth and increasing defense system of the banana fruit.

Inhibitory Effect of Chitosan and Phosphate Cross-linked Chitosan against Cucumber Mosaic Virus and Pepper Mild Mottle Virus

  • Gangireddygari, Venkata Subba Reddy;Chung, Bong Nam;Cho, In-Sook;Yoon, Ju-Yeon
    • The Plant Pathology Journal
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    • v.37 no.6
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    • pp.632-640
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    • 2021
  • Cucumber mosaic virus (CMV) and Pepper mild mottle virus (PMMoV) causes severe economic loss in crop productivity of both agriculture and horticulture crops in Korea. The previous surveys showed that naturally available biopolymer material - chitosan (CS), which is from shrimp cells, reduced CMV accumulation on pepper. To improve the antiviral activity of CS, it was synthesized to form phosphate cross-linked chitosan (PCS) and compared with the original CS. Initially, the activity of CS and PCS (0.01%, 0.05%, and 0.1% concentration) compound against PMMoV infection and replication was tested using a half-leaf assay on Nicotiana glutinosa leaves. The total number of local lesions represented on a leaf of N. glutinosa were counted and analyzed with phosphate buffer treated leaves as a negative control. The leaves treated with a 0.1% concentration of CS or PCS compounds exhibited an inhibition effect by 40-75% compared with the control leaves. The same treatment significantly reduced about 40% CMV accumulation measured by double antibody sandwich enzyme-linked immunosorbent assay and increased the relative expression levels of the NPR1, PR-1, cysteine protease inhibitor gene, LOX, PAL, SRC2, CRF3 and ERF4 genes analyzed by quantitative reverse transcriptase-polymerase chain reaction, in chili pepper plants.

Effects of Korean Ginseng on the Visceral Alkaline Phosphatase Activity and Blood Inorganic Phosphorus Level (인삼이 장기의 Alkaline Phosphatase활성과 혈중 무기 인량에 미치는 영향)

  • Jung, Noh-Pal
    • The Korean Journal of Physiology
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    • v.7 no.1
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    • pp.1-11
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    • 1973
  • To study the effects of Korean ginseng on mice visceral alkaline phosphatase activity and inorganic phosphate phosphorus level in the blood, the experimental groups of male and female mice were injected subcutaneously with 0.01 ml or 0.02 ml of Tyrode solution per 10 g of body weight daily, and 0. 1 mg or 0.2 mg of alcohol extract of ginseng which was diluted with Tyrode solution. After groups of mice were treated for 7 days or 14 days, the alkaline phosphatase activity in the jejunum, kidney and liver homogenate and serum were determined with sodium ${\beta}-glycerophophosphate$ as substrate, and quantified the content of inorganic phosphate phosphorus in the blood of above mice. 1. Alkaline phosphatase activity in the jejunum, kidney, liver and serum of male mice for the 7 day treatment with ginseng extract (0.1 mg/10 g) was increased by 15.28%, 12.86%, lg.05% and 11.70% respectively, compared with Tyrode solution group. 2. Alkaline phosphatase activity in the jejunum, kidney, liver and serum of female mice for the 7 day treatment with ginseng extract (0.2 mg/10 g) was increased 3.46%, 6.94%, 20.37% and 4.05 % respectively. 3. Alkaline phosphatase activity in the jejunum, kidney, liver and serum of male mice for the 14 day treatment with ginseng extract(0.1 mg/10 g) was increased·15.92%, 19.76f, 10.16% and -1.63 % respectively. 4. Alkaline phosphatase activity in the jejunum, kidney, liver and serum of female mice for the 14 day treatment with ginseng extract(0.2 mg/10 g) was increased 18.89%, 24.55%, 16.97% and 27.59% respectively. 5. Increasing activity of the enzyme in the liver of both male and female mice for the 7 day treatment was declined to some extent at the 14 day treatment, on the other hand, increasing activity of the enzyme in the jejunum, kidney and serum of mice for the 7 day treatment was more promoted at the 14 day treatment. 6. The 7 day and 14 day treatment with ginseng extract increased 20.20% and 20.96% of inorganic phosphate phosphorus in male blood, 22.38% and 17.57f in female blood respectively. In accordance with the results mutual relationships of ginseng, internal secretion, nucleic acid and alkaline phosphatase activity were disscussed.

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Effects of Pseudomonas aureofaciens 63-28 on Defense Responses in Soybean Plants Infected by Rhizoctonia solani

  • Jung, Woo-Jin;Park, Ro-Dong;Mabood, Fazli;Souleimanov, Alfred;Smith, Donald L.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.4
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    • pp.379-386
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    • 2011
  • The objective of this work was to investigate the ability of the plant growth-promoting rhizobacterium Pseudomonas aureofaciens 63-28 to induce plant defense systems, including defense-related enzyme levels and expression of defense-related isoenzymes, and isoflavone production, leading to improved resistance to the phytopathogen Rhizoctonia solani AG-4 in soybean seedlings. Seven-day-old soybean seedlings were inoculated with P. aureofaciens 63-28, R. solani AG-4, or P. aureofaciens 63-28 plus R. solani AG-4 (P+R), or not inoculated (control). After 7 days of incubation, roots treated with R. solani AG-4 had obvious damping-off symptoms, but P+R-treated soybean plants had less disease development, indicating suppression of R. solani AG-4 in soybean seedlings. Superoxide dismutase (SOD) and catalase (CAT) activities of R. solani AG-4-treated roots increased by 24.6% and 54.0%, respectively, compared with control roots. Ascorbate peroxidase (APX) and phenylalanine ammonia lyase (PAL) activities of R. solani AG-4-treated roots were increased by 75.1% and 23.6%, respectively. Polyphenol oxidase (PPO) activity in soybean roots challenged with P. aureofaciens 63-28 and P+R increased by 25.0% and 11.6%, respectively. Mn-SOD (S1 band on gel) and Fe-SOD (S2) were strongly induced in P+R-treated roots, whereas one CAT (C1) and one APX (A3) were strongly induced in R. solani AG-4- treated roots. The total isoflavone concentration in P+Rtreated shoots was 27.2% greater than the control treatment. The isoflavone yield of R. solani AG-4-treated shoots was 60.9% less than the control.

Biocontrol of Damping-Off(Rhizoctonia solani) in Cucumber by Trichoderma asperellum T-5 (Trichoderma asperellum T-5를 이용한 오이 모잘록병(Rhizoctonia solani)의 생물학적 제어)

  • Ryu, Ji-Yeon;Jin, Rong-De;Kim, Yong-Woong;Lee, Hyang-Burm;Kim, Kil-Yong
    • Korean Journal of Soil Science and Fertilizer
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    • v.39 no.4
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    • pp.185-194
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    • 2006
  • A fungal strain of Trichoderma having strong chitinolytic activity was isolated from field soil enriched with crabshell for several years. Based on 5.8S rRNA, partial 18S, 28S rRNA genes, ITS1, ITS2 sequence analysis and morphological characteristics, the fungus was identified as Trichoderma asperellum and named as Trichoderma asperellum T-5 (TaT-5). The fungus released lytic enzymes such as chitinase and ${\beta}$-1, 3-glucanse, and produced six antifungal substances in chitin broth medium. To demonstrate the protective effect of TaT-5 against damping-off in cucumber plant caused by Rhizoctonia solani, TaT-5 culture broth (TA), chitin medium (CM) and distilled water (DW) were applied to each pot at 10 days after sowing, respectively. Then, the homogenized hyphae of R. solani were infected to each pot at 1 week after TaT-5 inoculation. During experimental period, fresh weight of shoot and root in cucumber plant more increased at TA treatment compared to other treatments. PR-proteins (${\beta}$-1, 3-glucanase and chitinase) activities in cucumber leaves markedly increased at CM and DW treatments, but the activity slightly increased and then decreased at TA treatment at 3 days after infection of R. solani. The activity of PR-proteins activities in cucumber roots at all treatments decreased with time where the degree of decrement was more alleviated at TA treatment than CM and DW. These results suggest that the lytic enzymes (chitinase and ${\beta}$-1, 3-glucanse) and antifungal substances produced by TaT-5 can reduce the pathogenic attack by R. solani in cucumber plants.