• Title/Summary/Keyword: fungal enzyme

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Effect of Producing Different Phenazines on Bacterial Fitness and Biological Control in Pseudomonas chlororaphis 30-84

  • Yu, Jun Myoung;Wang, Dongping;Pierson, Leland S. III;Pierson, Elizabeth A.
    • The Plant Pathology Journal
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    • v.34 no.1
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    • pp.44-58
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    • 2018
  • Pseudomonas chlororaphis 30-84 is a biological control agent selected for its ability to suppress diseases caused by fungal pathogens. P. chlororaphis 30-84 produces three phenazines: phenazine-1-carboxylic acid (PCA), 2-hydroxy-phenazine-1-carboxylic acid (2OHPCA) and a small amount of 2-hydroxy-phenazine (2OHPHZ), and these are required for fungal pathogen inhibition and wheat rhizosphere competence. The two, 2-hydroxy derivatives are produced from PCA via the activity of a phenazine-modifying enzyme encoded by phzO. In addition to the seven biosynthetic genes responsible for the production of PCA, many other Pseudomonas strains possess one or more modifying genes, which encode enzymes that act independently or together to convert PCA into other phenazine derivatives. In order to understand the fitness effects of producing different phenazines, we constructed isogenic derivatives of P. chlororaphis 30-84 that differed only in the type of phenazines produced. Altering the type of phenazines produced by P. chlororaphis 30-84 enhanced the spectrum of fungal pathogens inhibited and altered the degree of take-all disease suppression. These strains also differed in their ability to promote extracellular DNA release, which may contribute to the observed differences in the amount of biofilm produced. All derivatives were equally important for survival over repeated plant/harvest cycles, indicating that the type of phenazines produced is less important for persistence in the wheat rhizosphere than whether or not cells produce phenazines. These findings provide a better understanding of the effects of different phenazines on functions important for biological control activity with implications for applications that rely on introduced or native phenazine producing populations.

The Role of Fungal Laccase in Biodegradation of Lignin

  • Andrzej Leonowicz;Jolanta Luterek;Maria W.Wasilewska;Anna Matuszewska;M.Hofrichter;D.Ziegenhagen;Jerzy Rogalski;Cho, Nam-Seok
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.31 no.5
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    • pp.1-11
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    • 1999
  • Wood components, cellulose and lignin, are degraded simultaneously and the general outline for the complementary character of carbohydrates and lignin decomposition as well as the existence of enzymatic systems combining these processes is still valid. The degradatiion of free cellulose or hemicellulose into monosaccharides has long been known to be relatively simple, but the mechanism of lignin degradatiion wasn ot solved very clearly yet. Anyway the biodegradation of woold constituents is understood at present as an enzymatic process. Kigninolytic activity has been correlated with lignin and manganese peroxidases. At present the attention is paid to laccase. Laccase oxidizes lignin molecule to phenoxy radicals and quinones . This oxidation can lead to the cleavageo f C-C or C-O bonds in the lignin phenyl-propane subunits, resulting either in degradation of both side chains and aromatic rings, or in demethylation processes. The role of laccase lies in the "activation" of some low molecular weight mediators and radicals produced by fungal cultures. Such activated factors produced also in cooperation with other enzymes are probably exported to the wood environment where they work in degradation processes as the ' enzyme messengers." It is worth mentioning that only fungi possessing laccase show demethylating activity. Thus demethylation, the process important for ligninolysis, is probably caused exclusively by laccase. Under natural conditions laccase seems to work with other fungal enzymes , mediators and mediating radicals. It has shown the possibility of direct Bjrkman lignin depolymerization by cooperative activity of laccase and glucose oxidase.

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A Correlation Between Fibrinolytic Activity and Microflora in Korean Fermented Soybean Products (된장의 혈전용해효소 활성과 미생물 분포와의 상관관계)

  • 김동호;송현파;김기연;김정옥;변명우
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.1
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    • pp.41-46
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    • 2004
  • A correlation between fibrinolytic activity and microflora in Korean traditional soybean fermented food was investigated. The fibrinolytic activities of traditional soybean pastes and commercially processed samples were 2.42$\pm$1.01 unit/g and 1.58$\pm$0.98 unit/g, respectively. The cell density of Bacillus in traditional soybean pastes was about 10$^{7}$ CFU/g and its commercially processed one was 10$^{6}$ CFU/g. Acid producing bacteria, fungi and yeast group were higher in commercially processed one. The correlations of fibrinolytic activity and microflora in traditional and commercial Doenjang were positively correlated in Bacillus ($R^2$≒ 0.69), negatively correlated in fungal group ($R^2$≒0.40), and there were no significant correlations in acid forming bacteria and yeast group ($R^2$<0.16). Fibrinolytic activities in Meju and Koji were 6.54$\pm$1.97 unit/g and 1.46$\pm$0.43 unit/g respectively, and were positively correlated with Bacillus. Yeast and acid forming bacteria were grown by 5∼6 decimal induction during fermentation period of Doenjang, but Bacillus, fungal cells and fibrinolytic activity were nearly stable. Results indicate that fibrinolytic activity of Doenjang depends on enzyme induction in Meju or Koji processing by Bacillus, Doenjang fermentation process.

Identification, Enzymatic Activity, and Decay Ability of Basidiomycetous Fungi Isolated from the Decayed Bark of Mongolian Oak (Quercus mongolica Fisch. ex Ledeb.)

  • Nguyen, Manh Ha;Kim, Dae Ho;Park, Ji Hyun;Park, Young Ui;Lee, Moo Yeul;Choi, Myeong Hee;Lee, Dong Ho;Lee, Jong Kyu
    • Journal of Forest and Environmental Science
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    • v.37 no.1
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    • pp.52-61
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    • 2021
  • Decay fungi can decompose plant debris to recycle carbon in the ecosystem. Still, they can also be fungal pathogens, which can damage living trees and/or wood material and cause a large amount of timber loss. We isolated and identified basidiomycetous fungi from the decayed bark of Mongolian oak wrapped with sticky roll traps. The degrading enzyme activities were then tested for all fungal isolates. The decay ability of selected isolates was assessed based on the weight loss of wood discs after inoculating with culture suspension of decay fungi under the different humidity levels. A total of 46 basidiomycetous fungal isolates belonged to 12 species, and 10 genera were obtained from Jong Myo (16 isolates), Chang Kyung palace (7 isolates), Cheong Gye (10 isolates), and Gun Po (13 isolates). Gymnopus luxurians was the most dominant fungus in the present study, and this species distributed in all survey sites with 9 isolates in Jong Myo, followed by 3 isolates in Chang Kyung palace, while Cheong Gye and Gun Po had only 1 isolate each. Among 46 isolates, 44 isolates secreted at least one enzyme, while 25 isolates produced both cellulase and phenol oxidase enzymes, and 2 isolates produced neither. The assessment of decay ability by artificial inoculation indicated that the weight loss of wood discs was significantly influenced by humidity conditions when inoculated with bark decay fungi. The percent weight losses by G. luxurians inoculation in RH of 90-100% and RH of 65-75% were 4.61% and 2.45%, respectively. The weight loss caused by Abortiporus biennis were 6.67% and 0.46% in RH of 90-100% and RH of 45-55%, respectively. The humidity reduction approach should be applied for further studies to control the growth and spread of bark decay fungi on the trunks wrapped with sticky roll traps.

Occurrence of an invertase producing strain of Aspergillus niger LP5 isolated from longan pollen and its application in longan syrup production to feed honey bees (Apis mellifera L.)

  • Danmek, Khanchai;Ruenwai, Rawisara;Sorachakula, Choke;Jung, Chuleui;Chuttong, Bajaree
    • Journal of Ecology and Environment
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    • v.46 no.2
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    • pp.136-143
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    • 2022
  • Background: In northern Thailand, the longan flower is the principal nectar source for honey production. Microorganisms play a critical function in the agricultural ecology. The morphological characteristics of fungal species found in longan pollen were studied. Aspergillus spp. were found to be invertase-producing strains and were employed in the longan syrup production process. The purpose of this study was to evaluate the effects of invertase-added longan syrup on the adult honey bee population numbers that were fed by this syrup for 16 weeks. Results: Different fungal species were found in longan pollen samples. Aspergillus was the main genus, with three predominant sections: Nigri, Flavi, and Terrei. Other isolated species were Trichoderma spp., Rhizopus spp., Neurospora spp., Chaetomium spp., Fusarium spp. and Penicillium spp. However, Aspergillus spp. is the only fungal species that produces the enzyme invertase. The invertase-producing strains belonging to the Aspergillus section Nigri were found to be A. niger LP5 with an optimum activity at pH 6.0 and 60℃. When A. niger LP5 invertase was used for longan syrup processing, the highest levels of glucose (3.45%) and fructose (2.08%) were found in invertase added longan syrup (C), while fresh (A) and boiled longan syrup (B) had lower contents of both sugars. The sucrose content was detected in (A) at 4.25%, while (B) and (C) were at 4.02% and 3.08%, respectively. An appropriate amount of sugar to feed and maintain the honey bee population was considered. The data showed no statistically significant differences between the two selected forms of longan syrup compared to the sugar syrup examined by the adult honey bee population. Conclusions: The main species of isolated fungi from longan pollen were Aspergillus spp. The discovery of an invertase-producing strain of A. niger LP5 has enabled its application for enzyme utilization in the invert sugar preparation process. The adult worker bee populations fed by longan syrup from both boiled and invertase-added sources showed an increasing trend. Artificial syrup made from longan fruit to feed honey bees when natural food sources are limited can be applied.

Cloning and Expression of Isocitrate Lyase, a Key Enzyme of the Glyoxylate Cycle, of Candida albicans for Development of Antifungal Drugs

  • SHIN DONG-SUN;KIM SANGHEE;YANG HYEONG-CHEOL;OH KI-BONG
    • Journal of Microbiology and Biotechnology
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    • v.15 no.3
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    • pp.652-655
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    • 2005
  • This paper describes the development of an enzymatic assay system for the identification of inhibitors of isocitrate lyase (ICL), one of the key enzymes of the glyoxylate cycle that is considered as a new target for antifungal drugs. A 1.6 kb DNA fragment encoding the isocitrate lyase from Candida albicans ATCC10231 was amplified by PCR, cloned into a vector providing His-Patch-thioredoxin-tag at the N-terminus, expressed in Escherichia coli, and purified by metal chelate affinity chromatography. The molecular mass of the purified ICL was approximately 62 kDa, as determined by SDS-PAGE, and the enzyme activity was directly proportional to incubation time and enzyme concentration. The effects of itaconate-related compounds on ICL activity were also investigated. Among them, itaconic acid, 3-nitropropionate, and oxalate had strong inhibitory activities with $IC_{50}$ values of 5.8, 5.4 and $8.6\;{mu}g/ml$, respectively. These inhibitors also exhibited antifungal activity on YPD agar media containing acetate as a sole carbon source, albeit at high concentration. The results indicate that the C. albicans ICL may be a regulatory enzyme playing a crucial role in fungal growth and is a prime target for antifungal agents.

Isolation and Identification of Cellulomonas fimi, Characteristics of its Cellulase and Conversion of the Sawdust into Ethanol (Cellulomonas fimi의 분리(分離) 및 동정(同定), cellulase 특성(特性)과 톱밥의 Ethanol 전환(轉換))

  • Lee, Chan-Yong;Lee, Ke-Ho
    • Applied Biological Chemistry
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    • v.28 no.2
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    • pp.98-105
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    • 1985
  • In the sheep and cattle's rumen, facultative anaerobic cellulolytic bacteria were isolated by using Hungate's roll tube technique. In the 21 isolated species, one was screened by its strong cellulolytic activity and identified as Cellulomonas fimi C-14 by investigate morphological, cultural, physiological characteristics and electron microgram. Optimum conditions of the cell growth and enzyme production were pH 6.5 an $30^{\circ}C$, Thiamine and biotin support a good growth of C. fimi C-14. In the enzyme activities, Crystalline cellulose hydrolyzing activity, CMCase activity and ${\beta}-glucosidase$ activity were 20.6, 226.6 and 0.56$(unit{\times}10^3/ml)$ at pH 6.0, $40^{\circ}C$. By addition of fungal cellulase, enzyme activity was increased. Simultaneous Saccharification Fermentation is better than two step fermentation in ethanol yield with Saccharomyces cerevisiae DY2.

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Optimization of Tannase Production by Aspergillus niger in Solid-State Packed-Bed Bioreactor

  • Rodriguez-Duran, Luis V.;Contreras-Esquivel, Juan C.;Rodriguez, Raul;Prado-Barragan, L. Arely;Aguilar, Cristobal N.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.9
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    • pp.960-967
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    • 2011
  • Tannin acyl hydrolase, also known as tannase, is an enzyme with important applications in the food, feed, pharmaceutical, and chemical industries. However, despite a growing interest in the catalytic properties of tannase, its practical use is very limited owing to high production costs. Several studies have already demonstrated the advantages of solid-state fermentation (SSF) for the production of fungal tannase, yet the optimal conditions for enzyme production strongly depend on the microbial strain utilized. Therefore, the aim of this study was to improve the tannase production by a locally isolated A. niger strain in an SSF system. The SSF was carried out in packed-bed bioreactors using polyurethane foam as an inert support impregnated with defined culture media. The process parameters influencing the enzyme production were identified using a Plackett-Burman design, where the substrate concentration, initial pH, and incubation temperature were determined as the most significant. These parameters were then further optimized using a Box-Behnken design. The maximum tannase production was obtained with a high tannic acid concentration (50 g/l), relatively low incubation temperature ($30^{\circ}C$), and unique low initial pH (4.0). The statistical strategy aided in increasing the enzyme activity nearly 1.97-fold, from 4,030 to 7,955 U/l. Consequently, these findings can lead to the development of a fermentation system that is able to produce large amounts of tannase in economical, compact, and scalable reactors.

Combined Effects of Vital Gluten, Gum, Emulsifier, and Enzyme on the Properties of Rice Bread (활성글루텐, 검, 유화제 및 효소제의 복합첨가에 따른 쌀빵의 품질특성)

  • Kim, Kyung-Eun;Lee, Young-Tack
    • Food Engineering Progress
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    • v.13 no.4
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    • pp.320-325
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    • 2009
  • The effects of adding additives such as vital gluten, gum, emulsifier, and enzyme to rice flour on baking quality were examined. The effects of different gums on the pasting and dough properties of rice flour containing vital gluten were studied using a Rapid Visco Analyzer (RVA) and a Brabender farinograph. The RVA peak, breakdown, and final viscosities decreased with the addition of gums, while setback viscosity increased. The farinogram showed that rice flour supplemented with gums such as tara gum, guar gum, and locust bean gum (LBG) increased water absorption and dough stability, yielding strengthened dough similar to wheat flour dough. The addition of guar or tara gum/sodium stearoyl lactylate (SSL)/fungal $\alpha$-amylase (AMYL) or glucose oxidase (GO) blend improved the volume and reduced the crumb firmness of rice bread prepared from rice flour containing 14% vital gluten. Therefore, the combined addition of gum, emulsifier and enzyme into rice flour significantly improved the rice bread quality, allowing the decrease of the vital gluten level in rice bread formula.

Effect of Additives in Making Frozen Rice Dough on the Quality of Rice Bread (쌀가루 냉동반죽 제조 시 첨가제가 쌀빵의 품질에 미치는 영향)

  • Kim, Kyung-Eun;Lee, Young-Tack
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.10
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    • pp.1438-1443
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    • 2009
  • The effects of adding additives such as gum, emulsifier, and enzyme both individually and as mixtures to frozen rice bread dough on baking quality were examined. Rice flours containing 17% vital gluten, and gum/emulsifier/enzyme blends were mixed and stored in a freezer at $-20^{\circ}C$ for 4 weeks. The rice doughs were removed from the freezer, thawed, and then followed the rice baking procedure. The dough freezing and frozen storage resulted in decreased volume of rice bread. The addition of guar gum/sodium stearoyl lactylate (SSL)/fungal $\alpha$-amylase blend improved volume of the rice bread obtained from rice dough during frozen storage. An increase in firmness of crumb was observed in rice breads during 3 days of storage at $25^{\circ}C$. Compared to the control dough without additives, addition of guar gum/ SSL blend or guar gum/ SSL/ fungal $\alpha$-amylase blend into frozen dough significantly reduced the crumb firmness of rice bread, indicating a significant antistaling effect.