Expression of a thermotolerant laccase from Pycnoporus sanguineus in Trichoderma reesei and its application in the degradation of bisphenol A

Zhao, Jie; Zeng, Shengquan; Xia, Ying; Xia, Liming

doi:10.1016/j.jbiosc.2017.11.010

Cited by 28 publications

(19 citation statements)

References 53 publications

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“…After the discovery of transformation techniques [37,38], various proteins of fungal and non-fungal origin have been successfully expressed in T. reesei [39,40]; the production of fungal proteins, in particular, was more efficient than that of mammalian proteins. For instance, fungal proteins such as laccase, lipase, xylanase, and glucose oxidase were expressed using the cbh1 promoter at levels of 100 mg/L and several grams per liter in shake-flask and fermentation cultures, respectively [26,[41][42][43]. One of the most advantageous properties of T. reesei is that it does not hyperglycosylate proteins.…”

Section: Discussionmentioning

confidence: 99%

“…Another study yielded 28.6-fold higher laccase production by expressing a laccase gene from Pleurotus ostreatus in the T. reesei strain Tu6 [25]. Recently, a thermotolerant laccase from Pycnoporus sanguineus was expressed in T. reesei with a yield of 17.7 U/mL [26]. These studies on the expression of ligninolytic enzymes in T. reesei suggest that T. reesei can serve as a compatible host for effective ligninolytic enzymes production.…”

Section: Introductionmentioning

confidence: 99%

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Improvement of Saccharification and Delignification Efficiency of Trichoderma reesei Rut-C30 by Genetic Bioengineering

et al. 2020

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Trichoderma reesei produces various saccharification enzymes required for biomass degradation. However, the lack of an effective lignin-degrading enzyme system reduces the species’ efficiency in producing fermentable sugars and increases the pre-treatment costs for biofuel production. In this study, we heterologously expressed the Ganoderma lucidum RMK1 versatile peroxidase gene (vp1) in the Rut-C30 strain of T. reesei. The expression of purified 6×His-tag–containing recombinant G. lucidum-derived protein (rVP1) was confirmed through western blot, which exhibited a single band with a relative molecular weight of 39 kDa. In saccharification and delignification studies using rice straw, the transformant (tVP7, T. reesei Rut-C30 expressing G. lucidum-derived rVP1) showed significant improvement in the yield of total reducing sugar and delignification, compared with that of the parent T. reesei Rut-C30 strain. Scanning electron microscopy (SEM) of tVP7-treated paddy straw showed extensive degradation of several layers of its surface compared with the parent strain due to the presence of G. lucidum-derived rVP1. Our results suggest that the expression of ligninolytic enzymes in cellulase hyperproducing systems helps to integrate the pre-treatment and saccharification steps that may ultimately reduce the costs of bioethanol production.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Improvement of Saccharification and Delignification Efficiency of Trichoderma reesei Rut-C30 by Genetic Bioengineering

et al. 2020

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“…Most fungal laccases have their optimal activity at higher temperatures. For example, laccase 32 , Pycnoporus sanguineus at 65 °C 33 and Melanocarpus albomyces (ascomycetes) at 60-70 °C 34 have their optimal activity. The thermostability of the enzymes is comparable to other laccases expressed in S. cerevisiae (T 50 around 70 °C): P. cinabarinus laccase, PM1 laccase or T. versicolor laccase 22,35,36 (Fig.…”

Section: Expression and Engineering Of Laccasementioning

confidence: 99%

Evolved Fusarium oxysporum laccase expressed in Saccharomyces cerevisiae

Kwiatos

Jędrzejczak-Krzepkowska

Krzemińska

et al. 2020

Sci Rep

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Fusarium oxysporum laccase was functionally expressed in Saccharomyces cerevisiae and engineered towards higher expression levels and higher reactivity towards 2,6-dimethoxyphenol, that could be used as a mediator for lignin modification. A combination of classical culture optimization and protein engineering led to around 30 times higher activity in the culture supernatant. The winner mutant exhibited three times lower Km, four times higher kcat and ten times higher catalytic efficiency than the parental enzyme. The strategy for laccase engineering was composed of a combination of random methods with a rational approach based on QM/MM MD studies of the enzyme complex with 2,6-dimethoxyphenol. Laccase mediator system with 2,6-dimethoxyphenol caused fulvic acids release from biosolubilized coal. Laccases are oxidoreductases that catalyze the 4-electron reduction of O 2 to water with simultaneous oxidation of organic substrates. Laccases are able to catalyze oxidation of a substrate of interest directly or indirectly by the formation of a radical, which then may take part in a non-enzymatic event that effects in the oxidation of a substrate of interest. Laccases oxidize phenolic substrates but are also able to oxidize non-phenolic or bigger substrates by Laccase Mediator System (LMS), where a small phenolic compound acts as a mediator 1. Laccases contain 4 copper atoms buried in their 3D structure, which are located in two separate Cu centers. T1 copper ion is a mononuclear center, whereas T2 copper ion and two T3 copper ions are positioned in T2/T3 copper center. The substrate is oxidized closed to T1 copper ion and then the electrons are transferred to the tri-nuclear center where O 2 is reduced to water. The coppers are coordinated by nearby located residues: the T1 copper ion by one Cys and two His residues, the T2 copper by two His residues and a solvent molecule and T3 copper to three His residues 2,3. Biosolubilization of brown coal is a clean coal technology that aims at the conversion of the lignite to its cleaner form or to change its structure to gain new features 4,5. Such solubilized material could be used as a source of value-added products 4. The liquid form of coal-microorganisms, such as Fusarium oxysporum reported before as an excellent coal solubilizer 6 , uses their metabolites, alkaline substances, biosurfactants and enzymes to turn the solid polymer to black liquid. Laccases, from bacteria and fungi, for example from species Pleurotus 7 or Streptomyces 8 , are known to take part in the degradation of lignin and lignite. These two polymers are similar in structure, thus, the mechanism of their degradation is expected to be the same. Lignin comprises only 10-20% of phenolic subunits. In theory, laccase could act on those subunits present on the lignin surface and modify the polymer. However, the possibility of lignin subunits entering the laccase active site is very limited. Moreover, it is not known to what extent lignin could be modified in this way 9. According to literature data, the treatme...

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“…Laccases (EC 1.10.3.2) are polyphenol copper blue proteases containing four copper ions (except Phlebia radiata laccase, which has two copper ions) and exist in plants, fungi, insects, and a small number of bacteria [1][2][3][4][5]. Since Yoshida [6] discovered laccase 131 in lacquer sap in 1883, laccase has become one of the hot research topics in the chemical industry, biological sciences, and environmental science [7][8][9].…”

Section: Introductionmentioning

confidence: 99%

“…In nature, white-rot fungi have been widely studied as one of the most effective producers of fungal laccase; So far, the heterologous expression of most laccases has been successful, for example, laccase genes have been cloned from Fome lignosus [17], Pycnoporus cinnabarinus [18], and various Trametes strains [19][20][21]. The difference between this study and the previous work mainly includes the following two points: 1) In previous studies, most recombinant laccases were used to degrade phenolic compounds, dye decolorization and degradation, etc., such as the study of Zhao [5], Nagai [22], Baldrian [23], etc. While in this study, recombinant laccase was used to degrade lignin, and the its ability was determined from qualitative and quantitative methods, which were not available in previous studies; 2) in the previous research on the degradation of lignin, the most studied was woody lignin, such as the research of Wei [24], Liu [25], etc., however, in this study, herbal lignin was used as the research object, to explore the effect of recombinant laccase on the degradability of herbal lignin.…”

Section: Introductionmentioning

confidence: 99%

Expression of Pleurotus ostreatus Laccase Gene in Pichia pastoris and Its Degradation of Corn Stover Lignin

2020

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Pleurotus ostreatus is a species of white-rot fungi that effectively degrades lignin. In this study, we aimed to efficiently express the lac-2 gene of Pleurotus ostreatus in the Pichia pastoris X33 yeast strain. The enzymatic properties of recombinant yeast were determined, and its ability to degrade corn stover lignin was determined. The results showed the optimum pH values of recombinant laccase for 2,2'-Azinobis-3-ethylbenzothiazoline-6-sulfonic acid, 2,6-dimethoxyphenol, and 2-methoxyphenol were 3.0, 3.0, and 3.5, respectively. The optimum reaction temperature was 50 • C, and it had good thermal stability and acid and alkali resistance. The degradation rate of lignin in corn stover by recombinant laccase was 18.36%, and the native Pleurotus ostreatus degradation rate was 14.05%, the difference between them is significant (p < 0.05). This experiment lays a foundation for the study of the degradation mechanism of lignin by laccase.

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Expression of a thermotolerant laccase from Pycnoporus sanguineus in Trichoderma reesei and its application in the degradation of bisphenol A

Cited by 28 publications

References 53 publications

Improvement of Saccharification and Delignification Efficiency of Trichoderma reesei Rut-C30 by Genetic Bioengineering

Improvement of Saccharification and Delignification Efficiency of Trichoderma reesei Rut-C30 by Genetic Bioengineering

Evolved Fusarium oxysporum laccase expressed in Saccharomyces cerevisiae

Expression of Pleurotus ostreatus Laccase Gene in Pichia pastoris and Its Degradation of Corn Stover Lignin

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