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Improvement of efficiency of brown coal biosolubilization by novel recombinant Fusarium oxysporum laccase.

ABSTRACT: Clean coal technologies (e.g. coal biosolubilization) are of essential value, especially in Europe, where coal is the national wealth and other energy sources like crude oil are not available. Fusarium oxysporum LOCK 1134, the strain isolated from brown coal, efficiently biosolubilizes lignite. The obtained liquefied products contain 50% less sulfur and over 99% less mercury than the crude coal. Moreover, the liquefied coal can be modified further by laccase. In this study F. oxysporum laccase was expressed in Pichia pastoris for the first time and was assessed as an additional agent for coal degradation. The novel laccase contributes to humic and fulvic acids release from liquefied coal due to introduction of oxygen into coal structure. The effect is increased when a natural redox mediator, sinapic acid, is present in the reaction mixture-up to 30% and 80% respectively. Humic acids obtained by biological process are environmentally friendly fertilizers that may have stimulating effects on crop growth.

SUBMITTER: Kwiatos N

PROVIDER: S-EPMC6104410 | biostudies-literature | 2018 Aug

REPOSITORIES: biostudies-literature

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Improvement of efficiency of brown coal biosolubilization by novel recombinant Fusarium oxysporum laccase.

Kwiatos Natalia N Jędrzejczak-Krzepkowska Marzena M Strzelecki Bartosz B Bielecki Stanisław S

AMB Express 20180822 1

Clean coal technologies (e.g. coal biosolubilization) are of essential value, especially in Europe, where coal is the national wealth and other energy sources like crude oil are not available. Fusarium oxysporum LOCK 1134, the strain isolated from brown coal, efficiently biosolubilizes lignite. The obtained liquefied products contain 50% less sulfur and over 99% less mercury than the crude coal. Moreover, the liquefied coal can be modified further by laccase. In this study F. oxysporum laccase w ...[more]

PMID: 30136100

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Project description:Laccase is a copper-containing polyphenol oxidase with a wide range of substrates, possessing a good application prospect in wastewater treatment and dye degradation. The purpose of this research is to study the degradation of various industrial dyes by recombinant laccase rlac1338 and the mutant enzyme lac2-9 with the highest enzyme activity after modification by error-prone PCR. Four enzyme activities improved mutant enzymes were obtained through preliminary screening and rescreening, of which lac2-9 has the highest enzyme activity. There are four mutation sites, including V281A, V281A, P309L, S318G, and D232V. The results showed that the expression of the optimized mutant enzyme also increased by 22 ± 2% compared to the unoptimized enzyme and the optimal reaction temperature of the mutant enzyme lac2-9 was 5°C higher than that of the rlac1338, and the optimal pH increased by 0.5 units. The thermal stability and pH stability of mutant enzyme lac2-9 were also improved. With ABTS as the substrate, the kcat/Km of rlac1338 and mutant strain lac2-9 are the largest than other substrates, 0.1638 and 0.618 s-1M-1, respectively, indicating that ABTS is the most suitable substrate for the recombinant enzyme and mutant enzyme. In addition, the Km of the mutant strain lac2-9 (76 μM) was significantly lower, but the kcat/Km (0.618 s-1M-1) was significantly higher, and the specific enzyme activity (79.8 U/mg) increased by 3.5 times compared with the recombinant laccase (22.8 U/mg). The dye degradation results showed that the use of rlac1338 and lac2-9 alone had no degradation effect on the industrial dyes [indigo, amaranth, bromophenol blue, acid violet 7, Congo red, coomassie brilliant blue (G250)], however, adding small molecular mediators Ca2+ and ABTS at the same time can significantly improve the degradation ability. Compared to the rlac1338, the degradation rates with the simultaneous addition of Ca2+ and ABTS of mutant enzyme lac2-9 for acid violet 7, bromophenol blue and coomassie brilliant blue significantly improved by 8.3; 3.4 and 3.4 times. Therefore, the results indicated that the error-prone PCR was a feasible method to improve the degradation activity of laccase for environmental pollutants, which provided a basis for the application of laccase on dye degradation and other environmental pollutants.

Project description:Mycoviruses that induce hypovirulence in phytopathogenic fungi are interesting because their potential use as biological control agents of the plant diseases caused by their fungal hosts. The recently identified chrysovirus Fusarium oxysporum f. sp. dianthi virus 1 (FodV1) has been associated to the induction of hypovirulence in Fusarium oxysporum f. sp. dianthi, the forma specialis of F. oxysporum that causes vascular wilt in carnation (Dianthus caryophyllus). In this work, we have used confocal laser scanner microscopy and two isogenic GFP-labeled strains of F. oxysporum f. sp. dianthi infected (V+) and not infected (V-) with the Fusarium oxysporum f. sp. dianthi virus 1, respectively, to analyze the effect of mycovirus FodV1 on the plant colonization pattern of its fungal host. Results demonstrate that FodV1-viral infection affects the speed and spatial distribution of fungal colonization into the plant. Initial stages of external root colonization were similar for both strains, but the virus-free strain colonized the internal plant tissues faster than the virus-infected strain. In addition, other differences related to the specific zone colonized and the density of colonization were observed between both F. oxysporum f. sp. dianthi strains. The hyphae of both V- and V+ strains progressed up through the xylem vessels but differences in the number of vessels colonized and of hyphae inside them were found. Moreover, as colonization progressed, V- and V+ hyphae propagated horizontally reaching the central medulla but, while the virus-free strain V- densely colonized the interior of the medulla cells, the virus-infected strain V+ appeared mainly in the intercellular spaces and with a lower density of colonization. Finally, the incidence of FodV1-viral infections in a collection of 221 isolates sampled between 2008 and 2012 in the geographic area where the originally infected isolate was obtained has been also analyzed. The very low (<2%) incidence of viral infections is discussed here. To the best of our knowledge, this work provides the first microscopic evidence about the effect of a hypovirulence-inducing mycovirus on the pattern of plant colonization by its fungal host.

Dataset Information

Improvement of efficiency of brown coal biosolubilization by novel recombinant Fusarium oxysporum laccase.

Publications

Improvement of efficiency of brown coal biosolubilization by novel recombinant Fusarium oxysporum laccase.

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