• Title/Summary/Keyword: fungal enzyme

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Towards a Miniaturized Culture Screening for Cellulolytic Fungi and Their Agricultural Lignocellulosic Degradation

  • Arnthong, Jantima;Siamphan, Chatuphon;Chuaseeharonnachai, Charuwan;Boonyuen, Nattawut;Suwannarangsee, Surisa
    • Journal of Microbiology and Biotechnology
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    • v.30 no.11
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    • pp.1670-1679
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    • 2020
  • The substantial use of fungal enzymes to degrade lignocellulosic plant biomass has widely been attributed to the extensive requirement of powerful enzyme-producing fungal strains. In this study, a two-step screening procedure for finding cellulolytic fungi, involving a miniaturized culture method with shake-flask fermentation, was proposed and demonstrated. We isolated 297 fungal strains from several cellulose-containing samples found in two different locations in Thailand. By using this screening strategy, we then selected 9 fungal strains based on their potential for cellulase production. Through sequence-based identification of these fungal isolates, 4 species in 4 genera were identified: Aspergillus terreus (3 strains: AG466, AG438 and AG499), Penicillium oxalicum (4 strains: AG452, AG496, AG498 and AG559), Talaromyces siamensis (1 strain: AG548) and Trichoderma afroharzianum (1 strain: AG500). After examining their lignocellulose degradation capacity, our data showed that P. oxalicum AG452 exhibited the highest glucose yield after saccharification of pretreated sugarcane trash, cassava pulp and coffee silverskin. In addition, Ta. siamensis AG548 produced the highest glucose yield after hydrolysis of pretreated sugarcane bagasse. Our study demonstrated that the proposed two-step screening strategy can be further applied for discovering potential cellulolytic fungi isolated from various environmental samples. Meanwhile, the fungal strains isolated in this study will prove useful in the bioconversion of agricultural lignocellulosic residues into valuable biotechnological products.

The Production and Enzymatic Properties of Extracellular Chitinase from Pseudomonas stutzeri YPL-1, as a Biocontrol Agent

  • Lim, Ho-Seong;Kim, Sang-Dal
    • Journal of Microbiology and Biotechnology
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    • v.4 no.2
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    • pp.134-140
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    • 1994
  • An antagonistic bacterium Pseudomonas stutzeri YPL-1 liberated extracellular chitinase and $\beta$-1,3-glucanase which are key enzymes in the decomposition of fungal hyphal walls. The lytic enzymes caused abnormal swelling and retreating at the hyphal tips of plant pathogenic fungus Fusarium solani in a dual culture. Scanning electron microscopy revealed the hyphal degradation of F. solani in the regions interacting with P. stutzeri YPL-1. The production of chitinase and properties of a crude preparation of the enzyme from P. stutzeri YPL-1 were investigated. Peak of the chitinase activity was detected after 4 hr of cultivation. The enzyme had optimum temperature and pH of 50$^{\circ}C$ and pH 5.3, respectively. The enzyme was stable in the pH range of 3.5 to 6.0 up to 50$^{\circ}C$. The enzyme was significantly inhibited by metal compounds such as $HgCl_2$, but was stimulated by $CoCl_2$. P. stutzeri YPL-1 produced high levels of the enzyme after 84 hr of incubation. Among the tested carbon sources, chitin was the most effective for the enzyme production, at the concentration level of 3%. As a source of nitrogen, peptone was the best for the enzyme production, at the concentration level of 4%. The maximum amount of enzyme was produced by cultivating the bacterium at a medium of initial pH 6.8.

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Isolation and Identification of the Fungi Producing a Soybean Milk Clotting Enzyme (두유 응고효소 생산 곰팡이의 분리 및 동정)

  • Lee, Chul-Woo;Kang, Chang-Hoon;Ha, Duk-Mo
    • Microbiology and Biotechnology Letters
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    • v.19 no.2
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    • pp.109-115
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    • 1991
  • Twenty-five fungal strains producing an extracellular soybean milk clotting enzyme were isolated from 146 soil samples, and identified as 11 species belonging to six genera of Aspergillus oryzae (5 strains), Aspergillus flavus (2 strains), Aspergillus parasiticus (1 strain), Aspergillus tamarii (2 strains), Aspergillus niger (4 strains), Aspergillus fumigatus (2 strains), Mucor hiemalis (2 strains), Wallemia sebi (4 strains), Scopulariopsis condida (1 strain), Fusarium redolens(1 strain) and Verticillum lecanii (1 strain). Among them, Aspergillus oryzae 020 and Aspergillus tamarii 287 showed relatively high soybean milk clotting activity. The coagulabilities of the enzyme from representative strains of those species decreased as the pH of soybean milk increased from 6.0 to 7.0 The optimum temperature for soybean milk clotting enzymes of those strains were 65$^{\circ}C$.

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Qualitative Evaluation of Ligninolytic Enzymes in Xylariaceous Fungi

  • Lee, Yang-Soo
    • Journal of Microbiology and Biotechnology
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    • v.10 no.4
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    • pp.462-469
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    • 2000
  • Sixty-one strains representing the main genera of wood-decaying xylariaceous fungi (mainly in Daldinia, Hypoxylon, Kretzschmaria, Rosellinia, Penzigia, and Xylaria) were tested for their ability to produce ligninolytic enzymes. The phenol oxidase activity and fungal growth of the xylariaceous fungi on gallic aicid and tannic acid media showed a variation in their ability to degrade lignocellulose. A number of species showed equal 개 betterligninolytic enzyme activities than Coriolus versicolor, a known basidiomycete wood-degrader. A large variation of the enzyme activity was observed by individual strains as well as a substantial variation between the isolates of the same species. The most frequent ligninolytic enzymes were peroxidase and general oxidase. With 19% of the strains tested, peroxidase showed the strongest ligninolytic enzyme activity, while tyrosinase activity was detected only in 7% of the strains. All strains of Kretzschmaria and Rosellinia tested was positive for laccase. Xylariaceous fungi were able to degrade the macromolecule, lignin, using each specific ligninolytic enzyme in the specfic lignin degradation pathway.

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Optimization of Fungal Enzyme Production by Trichoderma harzianum KUC1716 through Surfactant-Induced Morphological Changes

  • Lee, Hanbyul;Lee, Young Min;Heo, Young Mok;Hong, Joo-Hyun;Jang, Seokyoon;Ahn, Byoung Jun;Lee, Sung-Suk;Kim, Jae-Jin
    • Mycobiology
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    • v.45 no.1
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    • pp.48-51
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    • 2017
  • The morphological optimization of Trichoderma harzianum was carried out using several surfactants to achieve increased cellulase production. Addition of the surfactants to the culture medium successfully modified the fungal morphology from an aggregated form to a dispersed form. Optimization of the fungal morphology increased cellulase activity up to 177%. The morphologically optimized conditions enhanced the accessibility of the fungus to substrates and thus promoted cellulase production.

Decolorization of Three Acid Dyes by Enzymes from Fungal Strains

  • PARK , CHUL-HWAN;LEE, YU-RI;KIM, TAK-HYUN;LEE, BYUNG-HWAN;LEE, JIN-WON;KIM, SANG-YONG
    • Journal of Microbiology and Biotechnology
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    • v.14 no.6
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    • pp.1190-1195
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    • 2004
  • In recent years, there has been an intensive research on decolorization of dye and textile wastewater by various fungal strains. In this study, the decolorization ability of three commercial dyes, acid yellow 99, acid blue 350, and acid red 114, were investigated using 10 fungal strains. Among the fungal strains tested, Trametes versicolor KCTC 16781 completely decolorized all dyes in both solid and liquid experiments, and was also able to decolorize the mixture of those three dyes in liquid experiments. The secretion of the ligninolytic enzymes into the extracellular medium during decolorization by T versicolor KCTC 16781 was also studied. No lignin peroxidase activity was detected, and manganese peroxidase and laccase activities were investigated.

PTP1B Inhibitory Secondary Metabolites from Marine-Derived Fungal Strains Penicillium spp. and Eurotium sp.

  • Sohn, Jae Hak;Lee, Yu-Ri;Lee, Dong-Sung;Kim, Youn-Chul;Oh, Hyuncheol
    • Journal of Microbiology and Biotechnology
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    • v.23 no.9
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    • pp.1206-1211
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    • 2013
  • The selective inhibition of PTP1B has been widely recognized as a potential drug target for the treatment of type 2 diabetes and obesity. In the course of screening for PTP1B inhibitory fungal metabolites, the organic extracts of several fungal species isolated from marine environments were found to exhibit significant inhibitory effects, and the bioassay-guided investigation of these extracts resulted in the isolation of fructigenine A (1), cyclopenol (2), echinulin (3), flavoglaucin (4), and viridicatol (5). The structures of these compounds were determined mainly by analysis of NMR and MS data. These compounds inhibited PTP1B activity with 50% inhibitory concentration values of 10.7, 30.0, 29.4, 13.4, and 64.0 ${\mu}M$, respectively. Furthermore, the kinetic analysis of PTP1B inhibition by compounds 1 and 5 suggested that compound 1 inhibited PTP1B activity in a noncompetitive manner, whereas compound 5 inhibited PTP1B activity in a competitive manner.

Influence of Allyl Isothiocyanate on the Soil Microbial Community Structure and Composition during Pepper Cultivation

  • Gao, Jingxia;Pei, Hongxia;Xie, Hua
    • Journal of Microbiology and Biotechnology
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    • v.31 no.7
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    • pp.978-989
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    • 2021
  • Allyl isothiocyanate (AITC), as a fumigant, plays an important role in soil control of nematodes, soil-borne pathogens, and weeds, but its effects on soil microorganisms are unclear. In this study, the effects of AITC on microbial diversity and community composition of Capsicum annuum L. soil were investigated through Illumina high-throughput sequencing. The results showed that microbial diversity and community structure were significantly influenced by AITC. AITC reduced the diversity of soil bacteria, stimulated the diversity of the soil fungal community, and significantly changed the structure of fungal community. AITC decreased the relative abundance of dominant bacteria Planctomycetes, Acinetobacter, Pseudodeganella, and RB41, but increased that of Lysobacter, Sphingomonas, Pseudomonas, Luteimonas, Pseudoxanthomonas, and Bacillus at the genera level, while for fungi, Trichoderma, Neurospora, and Lasiodiplodia decreased significantly and Aspergillus, Cladosporium, Fusarium, Penicillium, and Saccharomyces were higher than the control. The correlation analysis suggested cellulase had a significant correlation with fungal operational taxonomic units and there was a significant correlation between cellulase and fungal diversity, while catalase, cellulose, sucrase, and urease were the major contributors in the shift of the community structure. Our results will provide useful information for the use of AITC in the assessment of environmental and ecological security.

Optimum Conditions for Glycoside Conversion to Aglycone by ${\gamma}-Galactosidase$ (유당분해효소에 의한 Glycoside의 Aglycone으로 전환을 위한 최적 조건 확립)

  • Kim, Nam-Chul;Jeon, Byung-Ju;Ahn, Joung-Jwa;Kwak, Hae-Soo
    • Food Science of Animal Resources
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    • v.27 no.3
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    • pp.329-336
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    • 2007
  • This study was designed to find the optimum conditions for converting isoflavone glycoside to aglycone by ${\beta}-galactosidase$. Three different forms of the enzyme were tested and the optimum enzyme concentration, incubation temperature, pH, and incubation time were determined. Before treatment with enzyme, isoflavone contained 89.4% glycoside including daidzin, glycitin and genistin, and only 10.6% aglycone including daidzein, glycitein and genistein. Among the enzymes tested, the highest rate of isoflavone hydrolysis to aglycone, 35%, was observed when 3 unit/g Fungal Lactase (Amano Enzyme) was used. Higher incubation temperatures resulted in a higher rate of hydrolysis along with a greater loss of isoflavone mass. Therefore, body temperature $(37^{\circ}C)$ may be adequate for isoflavone conversion, with 44.9% hydrolysis and less than 10% loss of mass. As expected, a higher amount of aglycone was produced at pH 7 compared with other pH values. During 5hr of incubation, the conversion of glycoside to aglycone increased dramatically from 0 to 1hr, and plateaued thereafter. In addition, commercial soy-based milk was hydrolyzed more effectively with ${\beta}-galactosidase$ when incubated for 5hr. Based on the above results, the optimum conditions for isoflavone hydrolysis by ${\beta}-galactosidase$ were for 3 hr at $37^{\circ}C$, pH 7 with 3 unit/g Fungal Lactase (Amano Lactase), yielding an average total amount of aglycone ranging from 40 to 47%.

Effects of Cordyceps militaris Mycelia on Fibrolytic Enzyme Activities and Microbial Populations In vitro

  • Yeo, Joon-Mo;Lee, Shin-Ja;Shin, Sung-Hwan;Lee, Sung-Hoon;Ha, Jong-Kyu;Kim, Wan-Young;Lee, Sung-Sill
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.3
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    • pp.364-368
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    • 2011
  • An experiment was conducted to examine the effects of Cordyceps militaris mycelia on microbial populations and fibrolytic enzyme activities in vitro. C. militaris mycelia was added to buffered rumen fluid with final concentrations of 0.00, 0.10, 0.15, 0.20, 0.25 and 0.30 g/L and incubation times were for 3, 6, 9, 12, 24, 36, 48 and 72 h. At all incubation times, the supplementation of C. militaris mycelia linearly increased the number of total viable and celluloytic bacteria; maximum responses were seen with 0.25 g/L supplementation of C. militaris mycelia. The addition of C. militaris mycelia above the level of 0.20 g/L significantly (p<0.01) increased the number of total and cellulolytic bacteria compared with the control. On the other hand, the response of fungal counts to the supplementation of C. militaris mycelia showed a linear decrease; the lowest response was seen with 0.30 g/L supplementation of C. militaris mycelia. It would seem that C. militaris mycelia possess a strong negative effect on rumen fungi since the lowest level of C. militaris mycelia supplementation markedly decreased fungal counts. Carboxylmethyl cellulase activities were linearly increased by the addition of C. militaris mycelia except at 3 and 9 h incubation times. At all incubation times, the supplementation of C. militaris mycelia linearly increased the activities of xylanase and avicelase. In conclusion, the supplementation of C. militaris mycelia to the culture of mixed rumen microorganisms showed a positive effect on cellulolytic bacteria and cellulolytic enzyme activities but a negative effect on fungi.