Biological degradation of anthroquinone and azo dyes by a novel laccase from Lentinus sp.

This study identifies a new fungal strain, Lentinus sp., that can produce extracellular forms of laccases with an activity of approximately 58 300 U/L. A purified laccase (designated lcc3) was identified by LC-ESI MS/MS as an N-linkage glycosylated protein. The isolated lcc3 cDNA is composed of 1563 bp encoding for a polypeptide of 521 amino acid residues with 4 putative Cu binding regions. Kinetic analyses revealed that the specific activity, k(cat), K(m), and k(cat)/K(m) of lcc3 at pH 2.5 and 70 °C with 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) used as a substrate was 2047 U mg(-1), 2017 s(-1), 8.4 µM, and 240 s(-1) µM(-1), respectively. Lcc3 is stable at pH 6.0-10.0 and has a midpoint temperature (T(m)) of 77.1 °C. We observed 97% decolorization efficiency on Acid Blue 80, 88% on RBBR, and 61% on Acid Red 37 by lcc3. Structural modeling analysis showed that five, four, and three hydrogen bonds can be formed between Acid Blue 80 and Arg(178), Arg(182), or Asn(358); between RBBR and His(132), Ser(134), or Asp(482); and between Acid Red 37 and Arg(178), respectively. Notably, Lentinus lcc3 efficiently reversed the toxicity of anthraquinone and azo dyes on rice seed germination and decolorized industrial textile effluent, suggesting the enzyme may be valuable for bioremediation.

Antifungal activity of essential oil and its constituents from Calocedrus macrolepis var. formosana Florin leaf against plant pathogenic fungi.

Resistance to conventional fungicides causes the poor disease control of agriculture. Natural products from plants have great potential as novel fungicide sources for controlling pathogenic fungi. In this study antipathogenic activity of the leaf essential oil and its constituents from Calocedrus macrolepis var. formosana Florin were evaluated in vitro against six plant pathogenic fungi. Chemical analysis of leaf oil by GC/MS allowed identification of alpha-pinene (44.2%), limonene (21.6%), beta-myrcene (8.9%), beta-caryophyllene (8.2%), caryophyllene oxide (2.4%), alpha-cadinol (1.6%), beta-pinene (1.2%), and T-muurolol (1.1%) as main components. Sesquiterpenoid components of the oil were more effective than monoterpenoid components of the oil. In particular, T-muurolol and alpha-cadinol strongly inhibited the growth of Rhizoctonia solani and Fusarium oxysporum, with the IC(50) values < 50 microg ml(-1). These compounds also efficiently inhibited the mycelial growths of Colletotrichum gloeosporioides, P. funerea, Ganoderma australe and F. solani. These results showed that T-muurolol and alpha-cadinol possess antifungal activities against a broad spectrum of tested plant pathogenic fungi and could be used as potential antifungal agents for the control of fungal diseases in plants.

Study on the antiinflammatory activity of methanol extract from seagrass Zostera japonica.

Methanolic extracts from the seagrass Zostera japonica were extracted successively using n-hexane (n-C(6)H(14)), dichloromethane (CH(2)Cl(2)), ethyl acetate (EtOAc), and water to give the n-C(6)H(14) (16.8%), CH(2)Cl(2) (40.6%), EtOAc (34.1%), and H(2)O (8.5%) soluble fractions, respectively. We have demonstrated that the hexane fraction has the highest capacity to inhibit proIL-1beta expression as compared to other fractions in lipopolysaccharides (LPS)-stimulated J774A.1 murine macrophages. Further analysis of the composition and antiinflammatory activity of the subfraction H5 from hexane fraction showed that it had the best antiinflammatory capacity and that it's major constituents were fatty acids, including palmitic acid methyl ester (21.5%), palmitic acid (24.02%), linoleic acid methyl ester (13.09%), oleic acid methyl ester (8.41%), and linoleic acid (7.93%), respectively. H5 inhibited LPS-induced TNFalpha, IL-1beta, and IL-6 in a dose-dependent manner, suggesting that H5 is bioactive in antiinflammation in vitro. This study is the first to report the antiinflammatory activity of extracts obtained from the seagrass Z. japonica.

Alkali-soluble polysaccharides of Rhizoclonium riparium alga induce IL-1 gene expression via protein kinase signaling pathways.

Fortification of aquaculture foodstuff with various algae may improve the resistance of certain fish or shrimp to diseases and, as a routine procedure, has become ever more popular and, seemingly, important. Herein, we isolated certain alkali-soluble polysaccharides from a Rhizoclonium riparium alga (RASP), polysaccharides that can be separated into two different groups on the basis of the polysaccharide's molecular weight. Using gas chromatography-mass spectometry analysis, we found that the major monosaccharides constituting the higher molecular-weight group of RASP were galactose (41.99%), glucose (34.53%), xylose (20.24%), and mannose (3.24%). Using a murine-derived macrophage cell line J774A.1, we found that polysaccharide constituents of the higher molecular-weight group of RASP were able to induce interleukin-1beta (IL-1) gene expression via protein kinase-mediated signal transduction pathways. In essence, we found that c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (p38), but not extracellular signal-regulated kinase (ERK), play an important role in the regulation of IL-1 gene expression in RASP-stimulated J774A.1 cells. To the best of our knowledge, this is the first occasion that polysaccharides from R. riparium have been demonstrated to exert immunomodulation properties by the induction of IL-1 within macrophages. Our current results provide support for the possible use of R. riparium as an additive to various food/foodstuff, to modulate the immune response of humans or certain animals.

Bioactivity assay of extracts from Calocedrus macrolepis var. formosana bark.

Alcoholic extracts from bark of Calocedrus macrolepis var. formosana Florin (Cupressaceae) were extracted successively using n-hexane, dichloromethane, ethyl acetate, 1-butanol and water, which gave 34.8%, 34.1%, 24.1%, 3.3% and 3.7% soluble fractions, respectively. Antioxidation activity of these fractions by DPPH assay and dissimilar IC50 values of the DPPH showed that ethyl acetate fraction had the best antioxidant activity; its IC50 was 2.6 microg/ml. Analyses of the composition and anti-inflammatory activity of the subfractions from n-C6H14 fraction showed that the T3 and H5ppt had the best anti-inflammatory activity in LPS-stimulated murine macrophage J774A. 1 cells, respectively; moreover, their major constituent was sugiol (T3 37.1%, H5ppt 81.1%), which at dosages of 10 microg/ml inhibited proIL-1beta protein production completely. Furthermore, the T1 also exhibited anti-inflammatory activity, and its major constituent was ferruginol (above 85.6%).

Essential Oil Compositions and Antimicrobial Activities of Various Parts of Litsea cubeba from Taiwan.

The essential oils of leaves, fruits, flowers, stems and twigs of Litsea cubeba were extracted by hydrodistillation. A total of 53, 50, 76, 94 and 90 compounds were identified from the leaf, fruit, flower, stem and twig oils, respectively, and their yields were 13.9 ± 0.09, 4.0 ± 0.03, 10.4 ± 0.05, 0.09 ± 0.01 and 0.4 ± 0.02 mL/100 g of the oven-dried (o.d.) materials, respectively. The main component in the leaf, flower and twig oils was 1,8-cineole, whereas in the fruit oil it was citral, and in the stem oil limonene, citronellal, and citronellol. When tested for their antibacterial activities using the paper disc diffusion method, oils from all parts showed excellent activities, particularly the fruit oil. When the oils were infused onto filter paper and tested for their antimicrobial paper capability according to the JIS L 1902 method, the fruit oil exhibited excellent antimicrobial activities. Citral was deemed the main cause of the antimicrobial activity. With the multiplicity of contagious diseases and their prevalence in hospitals, these essential oils present a potentially good choice as antibacterial agents. We think that the essential oils of this species are capable of multipurpose applications.

Composition of the Leaf Essential Oil of Phoebe formosana from Taiwan and its in vitro Cytotoxic, Antibacterial, and Antifungal Activities.

The essential oil from the leaves of Phoebe formosana from Taiwan was isolated using hydrodistillation in a Clevenger-type apparatus, and characterized by GC-FID and GC-MS. Seventy-one compounds were identified, representing 100% of the oil. The main components identified were a-humulene (16.8%), τ-cadinol (8.9%), α-pinene (8.4%), α-cadinol (8.1%), ß-caryophyllene (8.0%), ß-phellandrene (6.0%), and ß-eudesmol (5.8%). The oil exhibited cytotoxic activity against human lung, liver and oral cancer cells. The active compounds were ß-caryophyllene, α-humulene, τ-cadinol, ß-eudesmol, and α-cadinol. The antibacterial activity of the oil was tested by the disc diffusion and micro-broth dilution methods against eight bacterial species. The oil exhibited moderate growth suppression against Gram-positive bacteria with inhibition zones of 28 to 36 mm, and MIC values of 250 to 375 µg/mL. The active antibacterial compounds were determined to be τ-cadinol, ß-eudesmol, and α-cadinol. The leaf oil displayed excellent antifungal activity with the active compounds determined as α-cadinol, ß-eudesmol, τ-cadinol, α-humulene, and ß-caryophyllene.

Composition, in vitro Anti-inflammatory, Antioxidant and Antimicrobial Activities of the Leaf Essential Oil of Machilus konishii from Taiwan.

The chemical composition and in vitro anti-inflammatory, antioxidant and antimicrobial activities of the leaf essential oil of Machils konishii has been investigated. The essential oil was isolated using hydrodistillation in a Clevenger-type apparatus, and characterized by GC-FID and GC-MS. Sixty-six compounds were identified, representing 100% of the oil. The main components identified were a-pinene (33.9%), α-pinene (13.9%), and thymol (12.0%); The leaf oil was able to reduce nitric oxide production by lipopolysaccharide-activated murine macrophages RAW 264.7 without reducing the cell viability. In addition, the leaf oil showed strong antioxidant and antimicrobial activities. The major ingredient of the oil that was responsible for the anti-inflammatory, antioxidant and antimicrobial activities was thymol.

Composition, in vitro Anti-inflammatory, Antioxidant and Antimicrobial Activities of Essential Oils from Leaf and Twig Parts of Cupressus cashmeriana.

The chemical composition and in vitro anti-inflammatory, antioxidant and antimicrobial activities of the essential oils isolated from the leaf and twig of Cupressus cashmeriana have been investigated. The essential oils were isolated using hydrodistillation in a Clevenger-type apparatus, and characterized by GC-FID and GC-MS, respectively. The leaf oil mainly consisted of alpha-pinene (21.8%), epizonarene (8.0%), sabinene (7.9%), limonene (7.6%), gamma-terpinene (7.0%), and allo-aromadendrene (7.0%); the twig oil was mostly carvacrol methyl ether (35.4%), manool (16.1%), carvacrol (14.2%), and (2Z,6E)-farnesol (6.9%). Twig oil, but not leaf oil, was able to reduce nitric oxide production by lipopolysaccharide-activated murine macrophages RAW 264.7 without reducing the cell viability. In addition, twig oil showed better antioxidant and antimicrobial activities than leaf oil. The major ingredient of the twig oil that was responsible for the anti-inflammatory, antioxidant and antimicrobial activities was carvacrol.

Composition, in vitro Cytotoxic, and Antimicrobial Activities of the Flower Essential Oil of Diospyros discolor from Taiwan.

This study investigated the chemical composition, in vitro cytotoxic, and antimicrobial activities of the essential oil isolated from the flower of Diospyros discolor from Taiwan. The essential oil was isolated using hydrodistillation in a Clevenger-type apparatus, and characterized by GC-FID and GC-MS. Twenty-eight compounds were identified, representing 100% of the oil. The main components identified were (2Z,6E)-farnesol (35.0%), α-cadinol (10.9%), (E)-nerolidol (6.9%), α-humulene (6.0%), τ-cadinol (5.6%), ß-caryophyllene (5.3%), and τ-muurolol (5.3%). The oil exhibited cytotoxic activity against human colon, liver, and lung cancer cells. The active compounds were ß-caryophyllene, α-humulene, τ-cadinol, τ-muurolol, α-cadinol, and (2Z,6E)-farnesol. The antimicrobial activity of the oil was tested by the disc diffusion and micro-broth dilution methods against ten microbial species. The oil exhibited moderate to strong growth suppression against Bacillus cereus, Staphylococcus aureus, S. epidermidis, Escherichia coli, Enterobacter aerogenes, and Candida albicans, with inhibition zones of 40 to 52 mm and MIC values of 31.25-62.5 µg/mL, respectively. For the antimicrobial activities of the oil, the active compounds were determined to be α-cadinol, τ-cadinol, τ-muurolol and (2Z,6E)-farnesol.

Composition, in vitro Cytotoxicity, and Anti-mildew Activities of the Leaf Essential Oil of Machilus thunbergii from Taiwan.

This study investigated the chemical composition, in-vitro cytotoxicity, and anti-mildew fungal activities of the essential oil isolated from the leaf of Machilus thunbergii from Taiwan. The essential oil was isolated using hydrodistillation in a Clevenger-type apparatus, and characterized by GC-FID and GC-MS. Eighty-three compounds were identified, representing 99.8% of the oil. The main components identified were n-decanal (26.6%), ß-caryophyllene (15.8%), α-humulene (10.8%), and ß-eudesmol (10.5%). The oil exhibited cytotoxic activity against human oral, liver, lung, colon, melanoma, and leukemic cancer cells. The active source compounds were ß-caryophyllene, a-humulene, and ß-eudesmol. The anti-mildew activity of the leaf oil was also evaluated. Results showed that the leaf oil had excellent anti-mildew activity. For the anti-mildew activity of the leaf oil, the active source compound was determined to be ß-eudesmol.

Chemical composition and anti-mildew activities of essential oils from different parts of Michelia compressa var.formosana.

In this study, anti-mildew activities of the essential oils of the leaves, wood, twigs, flowers, and their constituents from Michelia comnpressa var. formosana were evaluated in vitro against 7 mildew fungi. The main compounds responsible for the anti-mildew activities were isolated and identified. The essential oils from the fresh leaves, wood, twigs, and flowers of M. compressa var. formosana were isolated using hydrodistillation in a Clevenger-type apparatus, and characterized by GC-FID and GC-MS. The essential oil yield from the flowers was the highest, and the oils obtained from the 4 parts of the tree were composed mainly of sesquiterpenoids. The leaf oil consisted primarily of α-cadinol (18.9%) and germacrene D (18.5%); the twig oil's main constituent was α-cadinol (19.1%); the wood oil's major components were α-cadinol (25.7%) and ß-eudesmol (20.2%); and the flower oil contained α-cadinol (11.4%). Comparing the anti-mildew activities of the oils suggested that wood oil was the most effective. Further fractionation of the wood oil produced α-cadinol, ß-eudesmol, τ-cadinol, and elemol. The 4 compounds exhibit very strong anti-mildew activities, and these in descending order are α-cadinol, elemol, ß-eudesmol, and τ-cadinol. For the anti-mildew activities of the wood oil, the active source compounds were determined to be α-cadinol, elemol, ß-eudesmol, and τ-cadinol.

Composition, in-vitro anticancer, and antimicrobial activities of the leaf essential oil of Machilus mushaensis from Taiwan.

This study investigated the chemical composition, in-vitro anticancer, and antimicrobial activities of the essential oil isolated from the leaf of Machilus mushaensis from Taiwan. The essential oil was isolated using hydrodistillation in a Clevenger-type apparatus, and characterized by GC-FID and GC-MS. Forty compounds were identified, representing 100% of the oil. The main components identified were n-decanal (61.0%), and alpha-cadinol (20.8%). The oil exhibited cytotoxic activity against human oral, liver, lung, colon, melanoma, and leukemic cancer cells. The antimicrobial activity of the oil was tested by the disc diffusion and micro-broth dilution methods against ten microbial species. The oil exhibited moderate growth suppression against Gram-positive bacteria and yeast with inhibition zones of 25-29 mm to MIC values of 375-500 microg/mL, respectively. alpha-Cadinol was found to show promising anticancer and antimicrobial activities.

Composition and in-vitro cytotoxic activities of the leaf essential oil of Beilschmiedia erythrophloia from Taiwan.

This study investigated the chemical composition and in-vitro cytotoxic activities of the essential oil isolated from the leaf of Beilschmiedia erythrophloia. The essential oil was isolated using hydrodistillation in a Clevenger-type apparatus, and characterized by GC-FID and GC-MS. Fifty-five compounds were identified, representing 100% of the oil. The main components identified were beta-caryophyllene (22.6%), alpha-humulene (21.9%), terpinen-4-ol (5.3%), cis-beta-ocimene (5.1%), sabinene (5.0%) and limonene (4.5%). The anticancer activities of oil were evaluated. The results showed that the oil exhibited cytotoxic activity against human oral, liver, lung, colon, melanoma, and leukemic cancer cells.

Composition and in vitro anticancer activities of the leaf essential oil of Neolitsea variabillima from Taiwan.

This study investigated the chemical composition and in vitro anticancer activities of the essential oil isolated from the leaf of Neolitsea variabillima. The essential oil was isolated using hydrodistillation in a Clevenger-type apparatus, and characterized by GC-FID and GC-MS. Sixty-seven compounds were identified, representing 100% of the oil. The main components identified were trans-beta-ocimene (13.4%), alpha-cadinol (10.5%), terpinen-4-ol (9.3%), tau-cadinol (9.2%), beta-caryophyllene (8.8%), and sabinene (6.7%). The anticancer activities of oil were evaluated. The results showed that the oil exhibited cytotoxic activity against human oral, liver, lung, colon, melanoma, and leukemic cancer cells. The presence of beta-caryophyllene, tau-cadinol, and alpha-cadinol significantly contributed to the anticancer activities of N. variabillima leaf oil.

The composition, anti-mildew and anti-wood-decay fungal activities of the leaf and fruit oils of Juniperus formosana from Taiwan.

In this study, anti-mildew and anti-wood-decay fungal activities of the leaf and fruits essential oil and its constituents from Juniperus formosana were evaluated in vitro against seven mildew fungi and four wood decay fungi, respectively. The main compounds responsible for the anti-mildew and anti-wood-decay fungal activities were also identified. The essential oil from the fresh leaves and fruits of J. formosana were isolated using hydrodistillation in a Clevenger-type apparatus, and characterized by GC-FID and GC-MS, respectively. The leaf oil mainly consisted of alpha-pinene (41.0%), limonene (11.5%), alpha-cadinol (11.0%), elemol (6.3%), and beta-myrcene (5.8%); the fruit oil was mostly alpha-pinene (40.9%), beta-myrcene (32.4%), alpha-thujene (5.9%) and limonene (5.9%). Comparing the anti-mildew and anti-wood-decay fungal activities of the oils suggested that the leaf oil was the most effective. For the anti-mildew and anti-wood-decay fungal activities of the leaf oil, the active source compounds were determined to be alpha-cadinol and elemol.

Composition, antioxidant and antimicrobial activities of the leaf essential oil of Machilus japonica from Taiwan.

The chemical composition, and antioxidant and antimicrobial activities of the essential oil isolated from the leaf of Machilus japonica from Taiwan have been investigated. The essential oil from the fresh leaves was isolated using hydrodistillation in a Clevenger-type apparatus, and characterized by GC-FID and GC-MS. A total of 97 compounds were identified, representing 100% of the oil. The main components identified were alpha-phellandrene (14.5%), alpha-pinene (12.8%), thymol (12.6%), beta-pinene (8.3%), alpha-terpineol (6.5%) and carvacrol (6.0%). The antioxidant activity of the oil was tested by the DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical scavenging capability test. The results showed that the IC50 was 51.8 microg/mL. The antimicrobial activity of the oil was tested by the disc diffusion and micro-broth dilution methods against ten microbial species. The oil exhibited strong growth suppression against Gram-positive bacteria and yeast, with inhibition zones of 48-54 mm and MIC values of 16.12-32.25 microg/mL, respectively. For the antioxidant and antimicrobial activities of the oil, the active source compounds were determined to be thymol and carvacrol.

Composition, anticancer, and antimicrobial activities in vitro of the heartwood essential oil of Cunninghamia lanceolata var. konishii from Taiwan.

This study investigated the chemical composition, anticancer, and antimicrobial activities in vitro of the essential oil isolated from the heartwood of Cunninghamia lanceolata var. konishii from Taiwan. The essential oil was isolated using hydrodistillation in a Clevenger-type apparatus, and characterized by GC-FID and GC-MS. Thirty-seven compounds were identified, representing 100% of the oil. The main components identified were cedrol (58.3%), alpha-cedrene (11.8%), alpha-terpineol (4.2%) and beta-cedrene (3.5%). The oil exhibited cytotoxic activity against human lung, liver and oral cancer cells. The active source compound was cedrol. The antimicrobial activity of the oil was tested by the disc diffusion and micro-broth dilution methods against ten microbial species. The oil exhibited strong growth suppression against Gram-positive bacteria and yeast with inhibition zones of 42-50 mm to MIC values of 31.25-62.5 microg/mL, respectively. For the antimicrobial activities of the oil, the active compound was determined to be cedrol.

Composition, antioxidant, antimicrobial and anti-wood-decay fungal activities of the twig essential oil of Taiwania cryptomerioides from Taiwan.

This study investigated the chemical composition, antioxidant, antimicrobial and anti-wood-decay fungal activities of the essential oil isolated from the twigs of Taiwania cryptomerioides from Taiwan. The essential oil was isolated using hydrodistillation in a Clevenger-type apparatus, and characterized by GC-FID and GC-MS. A total of 35 compounds were identified, representing 100% of the oil. The main components identified were alpha-cadinol (45.9%), ferruginol (18.9%) and beta-eudesmol (10.8%). The antioxidant activity of the oil was tested by the DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical scavenging capability test. The results showed an IC50 of 90.8 +/- 0.2 microg/mL. The active source compound was ferruginol. The antimicrobial activity of the oil was tested by the disc diffusion and micro-broth dilution methods against ten microbial species. The oil exhibited strong growth suppression against Gram-positive bacteria and yeast with inhibition zones of 45-52 mm and MIC values of 31.25-62.5 microg/mL, respectively. The anti-wood-decay fungal activity of the oil was also evaluated. The oil demonstrated excellent activity against four wood-decay-fungal species. For the antimicrobial and anti-wood-decay fungal activities of the oil, the active source compounds were determined to be alpha-cadinol, beta-eudesmol and ferruginol.

Composition and antipathogenic activities of the twig essential oil of Chamaecyparis formosensis from Taiwan.

In this study, antipathogenic activities of the twig essential oil and its constituents from Chamaecyparis formosensis Matsum were evaluated in vitro against six plant pathogenic fungi. The essential oil from the fresh twigs was isolated using hydrodistillation in a Clevenger-type apparatus, and characterized by GC-FID and GC-MS. Twenty-five compounds were identified, representing 98.9% of the oil. The main components were beta-eudesmol (25.1%), tau-muurolol (21.6%), elemol (15.0%), totarol (14.9%), and alpha-cadinol (12.4%). The twig oil (500 mcirog/mL) showed growth inhibitory activity against the phytopathogenic fungi, Fusarium oxysporum, Pestalotiopsis funereal, and Ganoderma austral, with antifungal indices of 92.7%, 71.1%, and 87.7%, respectively. In addition, the oil suppressed totally the growth of Rhizoctonia solani, Colletotrichum gloeosporioides, and Fusarium solani. In order to ascertain the source compounds of these antipathogenic activities, the main components were individually evaluated. Tau-Muurolol and alpha-cadinol exhibited excellent activity against F. oxysporum, R. solani, C. gloeosporioides, and F. solani, with IC50 < 50 microg/mL. These compounds also efficiently inhibited the mycelial growths of P. funereal and G. austral. Thus, alpha-cadinol and tau-muurolol could be considered as potential natural fungicides for controlling fungal pathogens and worth.