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27 pages, 2689 KiB  
Review
Novel Approach of Tackling Wax Deposition Problems in Pipeline Using Enzymatic Degradation Process: Challenges and Potential Solutions
by Shazleen Saadon, Raja Noor Zaliha Raja Abd Rahman, Nor Hafizah Ahmad Kamarudin, Sara Shahruddin, Siti Rohaida Mohd Shafian, Norhidayah Ahmad Wazir and Mohd Shukuri Mohamad Ali
Processes 2024, 12(10), 2074; https://doi.org/10.3390/pr12102074 - 25 Sep 2024
Viewed by 365
Abstract
Anthropogenic activities have led to hydrocarbon spills, and while traditional bioremediation methods are costly and time-consuming, recent research has focused on engineered enzymes for managing pollutant. The potential of enzymes for resolving wax flow problems in the petroleum industry remains unexplored. This paper [...] Read more.
Anthropogenic activities have led to hydrocarbon spills, and while traditional bioremediation methods are costly and time-consuming, recent research has focused on engineered enzymes for managing pollutant. The potential of enzymes for resolving wax flow problems in the petroleum industry remains unexplored. This paper offers a comprehensive review of the current state of research activities related to the bioremediation of petroleum-polluted sites and the biodegradation of specific petroleum hydrocarbons. The assayed enzymes that took part in the degradation were discussed in detail. Lipase, laccase, alkane hydroxylase, alcohol dehydrogenase, esterase, AlkB homologs and cytochrome P450 monooxygenase are among the enzymes responsible for the degradation of more than 50% of the hydrocarbons in contaminated soil and wastewater and found to be active on carbon C8 to C40. The possible biodegradation mechanism of petroleum hydrocarbons was also elucidated. The enzymes’ primary metabolic pathways include terminal, subterminal, and ω-oxidation. Next, given the successful evidence of the hydrocarbon treatment efficiency, the authors analyzed the opportunity for the enzymatic degradation approach if it were to be applied to a different scenario: managing wax deposition in petroleum-production lines. With properties such as high transformation efficiency and high specificity, enzymes can be utilized for the treatment of viscous heavy oil for transportability, evidenced by the 20 to 99% removal of hydrocarbons. The challenges associated with the new approach are also discussed. The production cost of enzymes, the characteristics of hydrocarbons and the operating conditions of the production line may affect the biocatalysis reaction to some extent. However, the challenges can be overcome by the usage of extremophilic enzymes. The combination of technological advancement and deployment strategies such as the immobilization of a consortium of highly thermophilic and halotolerant enzymes is suggested. Recovering and reusing enzymes offers an excellent strategy to improve the economics of the technology. This paper provides insights into the opportunity for the enzymatic degradation approach to be expanded for wax deposition problems in pipelines. Full article
(This article belongs to the Special Issue Application of Enzymes in Sustainable Biocatalysis)
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16 pages, 2437 KiB  
Article
Highly Efficient Degradation of 2-Methylisoborneol by Laccase Assisted by a Micro-Electric Field
by Ling Xu, Beidian Li, Tingting Liu, Anzhou Ma, Guoqiang Zhuang, Jingya Qian, Yi Cui, Shuhao Huo, Jiexiang Xia and Feng Wang
Catalysts 2024, 14(9), 649; https://doi.org/10.3390/catal14090649 - 23 Sep 2024
Viewed by 377
Abstract
Taste and odor (T&O) compounds have emerged as crucial parameters for assessing water quality. Therefore, identifying effective methodologies for the removal of these compounds is imperative. In this study, an effective approach utilizing laccase assisted by a micro-electric field was developed for the [...] Read more.
Taste and odor (T&O) compounds have emerged as crucial parameters for assessing water quality. Therefore, identifying effective methodologies for the removal of these compounds is imperative. In this study, an effective approach utilizing laccase assisted by a micro-electric field was developed for the degradation of 2-methylisoborneol (2-MIB). For this purpose, the optimal conditions for the laccase-catalyzed degradation of 2-MIB were determined, and they were pH 4.0, 25 °C, 150 rpm, 0.1 U/mL of laccase, and 200 ng/L of 2-MIB. Under these specified conditions, the degradation efficiency of 2-MIB was approximately 78% after a 4 h reaction period. Subsequently, the introduction of an electric field yielded a synergistic effect with the enzyme for 2-MIB degradation. At an electric current intensity of 0.04 A over a 4 h duration, the degradation efficiency increased to 90.78%. An analysis using SPME-GC/MS provided information on the degradation intermediates of 2-MIB resulting from laccase-catalyzed degradation, electrocatalytic degradation, and micro-electric-assisted laccase degradation. The potential degradation pathways of 2-MIB illustrated that these three methods result in common degradation products, such as capric aldehyde, nonylaldehyde, and 2-ethylhexanol, and their final products include 3-pentanone, acetone, and 2-butanone. This study provides an enzyme–electrochemical method for the efficient and rapid degradation and removal of 2-MIB. The strategy of laccase catalysis assisted by a micro-electric field has good potential for the removal of pollutants from the natural environment. Full article
(This article belongs to the Special Issue Design and Application of Combined Catalysis)
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16 pages, 2389 KiB  
Article
Enhancing Laccase Production by Trametes hirsuta GMA-01 Using Response Surface Methodology and Orange Waste: A Novel Breakthrough in Sugarcane Bagasse Saccharification and Synthetic Dye Decolorization
by Guilherme Guimarães Ortolan, Alex Graça Contato, Guilherme Mauro Aranha, Jose Carlos Santos Salgado, Robson Carlos Alnoch and Maria de Lourdes Teixeira de Moraes Polizeli
Reactions 2024, 5(3), 635-650; https://doi.org/10.3390/reactions5030032 - 19 Sep 2024
Viewed by 587
Abstract
Trametes hirsuta GMA-01 was cultivated in a culture medium supplemented with orange waste, starch, wheat bran, yeast extract, and salts. The fungus produced several holoenzymes, but the laccase levels were surprisingly high. Given the highlighted applicability of laccases in various biotechnological areas with [...] Read more.
Trametes hirsuta GMA-01 was cultivated in a culture medium supplemented with orange waste, starch, wheat bran, yeast extract, and salts. The fungus produced several holoenzymes, but the laccase levels were surprisingly high. Given the highlighted applicability of laccases in various biotechnological areas with minimal environmental impact, we provided a strategy to increase its production using response surface methodology. The immobilization of laccase into ionic supports (CM-cellulose, DEAE-agarose, DEAE-cellulose, DEAE-Sephacel, MANAE-agarose, MANAE-cellulose, and PEI-agarose) was found to be efficient and recuperative, showcasing the technical prowess of research. The crude extract laccase (CE) and CM-cellulose-immobilized crude extract (ICE) showed optimum activity in acidic conditions (pH 3.0) and at 70 °C for the CE and 60 °C for the ICE. The ICE significantly increased thermostability at 60 °C for the crude extract, which retained 21.6% residual activity after 240 min. The CE and ICE were successfully applied to sugarcane bagasse hydrolysis, showing 13.83 ± 0.02 µmol mL−1 reducing sugars after 48 h. Furthermore, the CE was tested for dye decolorization, achieving 96.6%, 71.9%, and 70.8% decolorization for bromocresol green, bromophenol blue, and orcein, respectively (0.05% (w/v) concentration). The properties and versatility of T. hirsuta GMA-01 laccase in different biotechnological purposes are interesting and notable, opening several potential applications and providing valuable insights into the future of biotechnological development. Full article
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13 pages, 17756 KiB  
Article
PlLAC15 Facilitates Syringyl Lignin Deposition to Enhance Stem Strength in Herbaceous Peony
by Yuehan Yin, Shiqi Zuo, Minghao Zhao, Jun Tao, Daqiu Zhao and Yuhan Tang
Agriculture 2024, 14(9), 1609; https://doi.org/10.3390/agriculture14091609 - 14 Sep 2024
Viewed by 364
Abstract
Stems are prone to bending or lodging due to inadequate stem strength, which seriously reduces the cut-flower ornamental quality of herbaceous peony (Paeonia lactiflora Pall.). Plant LACCASE (LAC), a copper-containing polyphenol oxidase, has been shown to participate in the polymerization process of [...] Read more.
Stems are prone to bending or lodging due to inadequate stem strength, which seriously reduces the cut-flower ornamental quality of herbaceous peony (Paeonia lactiflora Pall.). Plant LACCASE (LAC), a copper-containing polyphenol oxidase, has been shown to participate in the polymerization process of monolignols; however, the role of LAC in regulating the stem strength of P. lactiflora remains unclear. Here, the full-length cDNA of PlLAC15, which demonstrated a positive association with stem strength, was isolated. It consisted of 1790 nucleotides, encoding 565 amino acids that had four typical laccase copper ion-binding domains. Moreover, PlLAC15 was highly expressed in the stem, and its expression level gradually significantly increased during stem development. Furthermore, PlLAC15 was found to be localized specifically to the cell wall, and its recombinant protein exhibited laccase activity. Additionally, the role of PlLAC15 in regulating the stem strength of P. lactiflora was confirmed by transgenic studies. When PlLAC15 was overexpressed in tobacco, stem strength increased by more than 50%, S-lignin was significantly deposited, and the lignification degree of stem xylem fiber cells increased. These results suggested that PlLAC15 facilitated S-lignin deposition to enhance stem strength in P. lactiflora, which would provide precious information that benefits future exploration of stem bending or lodging resistance in plants. Full article
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15 pages, 5040 KiB  
Article
Transcriptome Analysis Reveals the Effect of Oyster Mushroom Spherical Virus Infection in Pleurotus ostreatus
by Yifan Wang, Junjie Yan, Guoyue Song, Zhizhong Song, Matthew Shi, Haijing Hu, Lunhe You, Lu Zhang, Jianrui Wang, Yu Liu, Xianhao Cheng and Xiaoyan Zhang
Int. J. Mol. Sci. 2024, 25(17), 9749; https://doi.org/10.3390/ijms25179749 - 9 Sep 2024
Viewed by 425
Abstract
Oyster mushroom spherical virus (OMSV) is a mycovirus that inhibits mycelial growth, induces malformation symptoms, and decreases the yield of fruiting bodies in Pleurotus ostreatus. However, the pathogenic mechanism of OMSV infection in P. ostreatus is poorly understood. In this study, RNA [...] Read more.
Oyster mushroom spherical virus (OMSV) is a mycovirus that inhibits mycelial growth, induces malformation symptoms, and decreases the yield of fruiting bodies in Pleurotus ostreatus. However, the pathogenic mechanism of OMSV infection in P. ostreatus is poorly understood. In this study, RNA sequencing (RNA-seq) was conducted, identifying 354 differentially expressed genes (DEGs) in the mycelium of P. ostreatus during OMSV infection. Verifying the RNA-seq data through quantitative real-time polymerase chain reaction on 15 DEGs confirmed the consistency of gene expression trends. Both Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses highlighted the pivotal role of primary metabolic pathways in OMSV infection. Additionally, significant changes were noted in the gene expression levels of carbohydrate-active enzymes (CAZymes), which are crucial for providing the carbohydrates needed for fungal growth, development, and reproduction by degrading renewable lignocellulose. The activities of carboxymethyl cellulase, laccase, and amylase decreased, whereas chitinase activity increased, suggesting a potential mechanism by which OMSV influenced mycelial growth through modulating CAZyme activities. Therefore, this study provided insights into the pathogenic mechanisms triggered by OMSV in P. ostreatus. Full article
(This article belongs to the Special Issue Advances in Plant Virus Diseases and Virus-Induced Resistance)
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24 pages, 8731 KiB  
Article
Involvement of MicroRNAs in the Hypersensitive Response of Capsicum Plants to the Capsicum Chlorosis Virus at Elevated Temperatures
by Wei-An Tsai, Christopher A. Brosnan, Neena Mitter and Ralf G. Dietzgen
Pathogens 2024, 13(9), 745; https://doi.org/10.3390/pathogens13090745 - 31 Aug 2024
Viewed by 348
Abstract
The orthotospovirus capsicum chlorosis virus (CaCV) is an important pathogen affecting capsicum plants. Elevated temperatures may affect disease progression and pose a potential challenge to capsicum production. To date, CaCV-resistant capsicum breeding lines have been established; however, the impact of an elevated temperature [...] Read more.
The orthotospovirus capsicum chlorosis virus (CaCV) is an important pathogen affecting capsicum plants. Elevated temperatures may affect disease progression and pose a potential challenge to capsicum production. To date, CaCV-resistant capsicum breeding lines have been established; however, the impact of an elevated temperature of 35 °C on this genetic resistance remains unexplored. Thus, this study aimed to investigate how high temperature (HT) influences the response of CaCV-resistant capsicum to the virus. Phenotypic analysis revealed a compromised resistance in capsicum plants grown at HT, with systemic necrotic spots appearing in 8 out of 14 CaCV-infected plants. Molecular analysis through next-generation sequencing identified 105 known and 83 novel microRNAs (miRNAs) in CaCV-resistant capsicum plants. Gene ontology revealed that phenylpropanoid and lignin metabolic processes, regulated by Can-miR408a and Can- miR397, are likely involved in elevated-temperature-mediated resistance-breaking responses. Additionally, real-time PCR validated an upregulation of Can-miR408a and Can-miR397 by CaCV infection at HT; however, only the Laccase 4 transcript, targeted by Can-miR397, showed a tendency of negative correlation with this miRNA. Overall, this study provides the first molecular insights into how elevated temperature affects CaCV resistance in capsicum plants and reveals the potential role of miRNA in temperature-sensitive tospovirus resistance. Full article
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21 pages, 9180 KiB  
Article
Study on Enzyme Activity and Metabolomics during Culture of Liquid Spawn of Floccularia luteovirens
by Yanqing Ni, Qiuhong Liao, Siyuan Gou, Tongjia Shi, Wensheng Li, Rencai Feng, Zhiqiang Zhao and Xu Zhao
J. Fungi 2024, 10(9), 618; https://doi.org/10.3390/jof10090618 - 29 Aug 2024
Viewed by 604
Abstract
To comprehensively investigate the physiological characteristics and metabolic processes of the mycelium of Floccularia luteovirens (F. luteovirens), a wild edible fungus unique to the plateau region, we conducted an in-depth analysis of the mycelium enzyme activity and metabolites during different culture [...] Read more.
To comprehensively investigate the physiological characteristics and metabolic processes of the mycelium of Floccularia luteovirens (F. luteovirens), a wild edible fungus unique to the plateau region, we conducted an in-depth analysis of the mycelium enzyme activity and metabolites during different culture periods. The activity of seven enzymes all followed a trend of initially increasing and then decreasing. The intra- and extracellular activity peaks of three hydrolases—amylase, protease, and cellulase—all occurred on the 20th day, except for the extracellular amylase, which peaked on the 15th day. In contrast, the peak activity of laccase occurred on the 10th day. Moreover, three types of oxidoreductases in the mycelium (catalase (CAT), superoxide dismutase (SOD), and 2,3,5-triphenyltetrazolium chloride (TTC)-dehydrogenase (TTC-DH)) also exhibited significant changes in activity. CAT and SOD activity reached their maximum on the 20th day, whereas TTC-DH showed high activity on both the 10th and 20th days. Through a comprehensive assessment of the evolving trends of these physiological parameters, we determined that the optimal cultivation cycle for F. luteovirens liquid spawn is 20 days. An untargeted metabolomic analysis revealed that 3569 metabolites were detected in the F. luteovirens mycelium, including a variety of secondary metabolites and functional components, with terpenoids being particularly abundant, accounting for 148 types. By comparing three different culture stages (10 days, 20 days, and 30 days), 299, 291, and 381 metabolites, respectively, showed different accumulation patterns in the comparison groups of 10d vs. 20d, 20d vs. 30d, and 10d vs. 30d. These differential metabolites were primarily concentrated in carboxylic acids and their derivatives, fatty acyl groups, organic oxygen compounds, and lipid compounds. In addition, there were several amino acids whose abundance continued to grow during culturing. The metabolism of amino acids greatly affects mycelium growth and development. This research delineates the interplay between mycelium growth and metabolism, offering empirical support for a cultivation strategy for liquid F. luteovirens, and an exploration of its metabolites for potential applications. Full article
(This article belongs to the Section Fungal Cell Biology, Metabolism and Physiology)
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6 pages, 1218 KiB  
Proceeding Paper
Screening and Isolation of Microbes for Promoting Humification of Agricultural Organic Wastes
by Kai-Siang Chen, Sheng-Chin Hu, Pin-Chen Yeh, Jia-Xiang You and Rey-May Liou
Eng. Proc. 2024, 74(1), 24; https://doi.org/10.3390/engproc2024074024 - 29 Aug 2024
Viewed by 206
Abstract
Agroecosystems play a crucial role in carbon sequestration and reducing atmospheric CO2 emissions by storing organic carbon in soil. Soil fertility and productivity are essential for global crop demands and depend on soil organic matter, particularly humic substances (HS). HS is crucial [...] Read more.
Agroecosystems play a crucial role in carbon sequestration and reducing atmospheric CO2 emissions by storing organic carbon in soil. Soil fertility and productivity are essential for global crop demands and depend on soil organic matter, particularly humic substances (HS). HS is crucial for soil health and carbon sequestration as it involves the carbon cycle, supplies nutrients for plants, and reduces the emission of pollutants through microbial processes for the enhancement of CO2 sequestration. Humification is a natural process of organic matter stabilization, playing a crucial role in maintaining soil organic content and carbon storage. Laccase is used to polymerize monomeric compounds such as phenols and their derivatives into highly polymerized HS. We screened five cellulose-degrading isolates, among which three strains demonstrate lignin-degrading capabilities. WSC-7 exhibited the highest laccase activity and showed a high similarity [99.51%] to Arthrobacter defluvii based on 16S rDNA analysis. Strain LiPK-078-5 in nutrient broth medium and WSC-7 in Difco Sporulation Medium exhibited optimal catalytic activity for catechol, indicating their efficiency for aromatic polymerization of soluble organic carbon. The addition of rice husk biochar with strains LiPK-078-5, WSC-6, and WSC-7 increased organic carbon content effectively. The synthesis of humic substances in soil through microbial processes increases soil carbon sequestration and reduces greenhouse gas emissions in the environment. Full article
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16 pages, 2011 KiB  
Article
A Treatment for Rice Straw and Its Use for Mealworm (Tenebrio molitor L.) Feeding: Effect on Insect Performance and Diet Digestibility
by Jorge Saura-Martínez, Ana Montalbán, Jesús Manzano-Nicolás, Amaury Taboada-Rodríguez, Fuensanta Hernández and Fulgencio Marín-Iniesta
Insects 2024, 15(8), 631; https://doi.org/10.3390/insects15080631 - 22 Aug 2024
Viewed by 552
Abstract
The development of reuse processes for plant by-products for both animal and human food offers numerous possibilities for quality-of-life improvements that align with a circular economy model. For this reason, we divided this study into two experiments. First, we designed a combined treatment [...] Read more.
The development of reuse processes for plant by-products for both animal and human food offers numerous possibilities for quality-of-life improvements that align with a circular economy model. For this reason, we divided this study into two experiments. First, we designed a combined treatment consisting of laccase, ultrasound, and ascorbic acid to hydrolyze rice straw plant fibers and used the resulting feed as the basis for T. molitor diets. Second, we formulated diets with different inclusion levels (0%, 25%, 50%, 75%, and 100%) of rice straw and treated rice straw to assess their impact on larvae growth and diet digestibility. For each treatment, six replicates were employed: four for the growth–performance–digestibility trial and two for complementary uric acid determination tests. The combined laccase enzyme, ultrasound, and ascorbic acid treatment hydrolyzed 13.2% of the vegetable fibers. The diets containing treated rice straw resulted in higher larvae weight and a better feed conversion ratio; however, reaching 100% by-product inclusion values led to similar results between both diets. In conclusion, these treatments improve the potential of low-nutritional-value vegetable by-products as part of a T. molitor diet, opening the possibility of new methodologies for the use of recalcitrant vegetable by-products for insect rearing. Full article
(This article belongs to the Collection Science of Insect Rearing Systems)
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16 pages, 7015 KiB  
Article
Degradation Characteristics of Nicosulfuron in Water and Soil by MnO2 Nano-Immobilized Laccase
by Wanlei Yue, Xin Wang, Jiale Zhang, Jia Bao and Mengqin Yao
Toxics 2024, 12(8), 619; https://doi.org/10.3390/toxics12080619 - 21 Aug 2024
Viewed by 523
Abstract
As a typical sulfonylurea herbicide, nicosulfuron is mainly used to control grass weeds and some broadleaf weeds in corn fields. However, as the amount of use continues to increase, it accumulates in the environment and eventually becomes harmful to the ecosystem. In the [...] Read more.
As a typical sulfonylurea herbicide, nicosulfuron is mainly used to control grass weeds and some broadleaf weeds in corn fields. However, as the amount of use continues to increase, it accumulates in the environment and eventually becomes harmful to the ecosystem. In the present study, a new metallic nanomaterial, δ-MnO2, was prepared, which not only has a similar catalytic mechanism as laccase but also has a significant effect on pesticide degradation. Therefore, the bicatalytic property of MnO2 can be utilized to improve the remediation of nicosulfuron contamination. Firstly, MnO2 nanomaterials were prepared by controlling the hydrothermal reaction conditions, and immobilized laccase was prepared by the adsorption method. Next, we investigate the effects of different influencing factors on the effect of immobilized laccase, MnO2, and free laccase on the degradation of nicosulfuron in water and soil. In addition, we also analyze the metabolic pathway of nicosulfuron degradation in immobilized laccase and the bicatalytic mechanism of MnO2. The results demonstrated that the degradation rate of nicosulfuron in water by immobilized laccase was 88.7%, and the optimal conditions were 50 mg/L, 25 h, 50 °C, and pH 5. For nicosulfuron in soil, the optimal conditions for the degradation by immobilized laccase were found to be 151.1 mg/kg, 46 °C, and pH 5.9; under these conditions, a degradation rate of 90.1% was attained. The findings of this study provide a theoretical reference for the immobilized laccase treatment of sulfonylurea herbicide contamination in water and soil. Full article
(This article belongs to the Section Toxicity Reduction and Environmental Remediation)
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14 pages, 1904 KiB  
Article
Aflatoxin B1 Control by Various Pseudomonas Isolates
by Dóra Anna Papp, Sándor Kocsubé, Zoltán Farkas, András Szekeres, Csaba Vágvölgyi, Zsuzsanna Hamari and Mónika Varga
Toxins 2024, 16(8), 367; https://doi.org/10.3390/toxins16080367 - 20 Aug 2024
Viewed by 743
Abstract
The climate-change-coupled fungal burden in crop management and the need to reduce chemical pesticide usage highlight the importance of finding sustainable ways to control Aspergillus flavus. This study examines the effectiveness of 50 Pseudomonas isolates obtained from corn rhizospheres against A. flavus [...] Read more.
The climate-change-coupled fungal burden in crop management and the need to reduce chemical pesticide usage highlight the importance of finding sustainable ways to control Aspergillus flavus. This study examines the effectiveness of 50 Pseudomonas isolates obtained from corn rhizospheres against A. flavus in both solid and liquid co-cultures. The presence and quantity of aflatoxin B1 (AFB1) and AFB1-related compounds were determined using high-performance liquid chromatography–high resolution mass spectrometry analysis. Various enzymatic- or non-enzymatic mechanisms are proposed to interpret the decrease in AFB1 production, accompanied by the accumulation of biosynthetic intermediates (11-hydroxy-O-methylsterigmatocystin, aspertoxin, 11-hydroxyaspertoxin) or degradation products (the compounds C16H10O6, C16H14O5, C18H16O7, and C19H16O8). Our finding implies the upregulation or enhanced activity of fungal oxidoreductases and laccases in response to bacterial bioactive compound(s). Furthermore, non-enzymatic reactions resulted in the formation of additional degradation products due to acid accumulation in the fermented broth. Three isolates completely inhibited AFB1 or any AFB1-related compounds without significantly affecting fungal growth. These bacterial isolates supposedly block the entire pathway for AFB1 production in the fungus during interaction. Apart from identifying effective Pseudomonas isolates as potential biocontrol agents, this work lays the foundation for exploring new bacterial bioactive compounds. Full article
(This article belongs to the Section Mycotoxins)
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18 pages, 2701 KiB  
Article
Fungal Laccases and Fumonisin Decontamination in Co-Products of Bioethanol from Maize
by Marianela Bossa, Noelia Edith Monesterolo, María del Pilar Monge, Paloma Rhein, Sofía Noemí Chulze, María Silvina Alaniz-Zanon and María Laura Chiotta
Toxins 2024, 16(8), 350; https://doi.org/10.3390/toxins16080350 - 10 Aug 2024
Viewed by 843
Abstract
Maize (Zea mays L.) may be infected by Fusarium verticillioides and F. proliferatum, and consequently contaminated with fumonisins (FBs), as well as the co-products of bioethanol intended for animal feed. Laccase enzymes have a wide industrial application such as mycotoxin degradation. [...] Read more.
Maize (Zea mays L.) may be infected by Fusarium verticillioides and F. proliferatum, and consequently contaminated with fumonisins (FBs), as well as the co-products of bioethanol intended for animal feed. Laccase enzymes have a wide industrial application such as mycotoxin degradation. The aims were to isolate and identify fungal laccase-producing strains, to evaluate laccase production, to determine the enzymatic stability under fermentation conditions, and to analyse the effectiveness in vitro of enzymatic extracts (EEs) containing laccases in degrading FB1. Strains belonging to Funalia trogii, Phellinus tuberculosus, Pleurotus ostreatus, Pycnoporus sanguineus and Trametes gallica species showed laccase activity. Different isoforms of laccases were detected depending on the evaluated species. For the FB1 decontamination assays, four enzymatic activities (5, 10, 15 and 20 U/mL) were tested, in the absence and presence of vanillic acid (VA) and 2,2,6,6-tetramethylpiperidine-N-oxyl (TEMPO) as redox mediators (1 and 10 mM). Trametes gallica B4-IMICO-RC EE was the most effective strain in buffer, achieving a 60% of FB1 reduction. Laccases included in EEs remained stable at different alcoholic degrees in maize steep liquor (MSL), but no significant FB1 reduction was observed under the conditions evaluated using MSL. This study demonstrate that although laccases could be good candidates for the development of a strategy to reduce FB1, further studies are necessary to optimise this process in MSL. Full article
(This article belongs to the Section Mycotoxins)
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15 pages, 6204 KiB  
Article
Identification of Laccase Genes in Athelia bombacina and Their Interactions with the Host
by Xiaonan Sun, Weiwei Yan, Xinnan Zhang, Wenhui Wang and Xiaohui Jia
Horticulturae 2024, 10(8), 842; https://doi.org/10.3390/horticulturae10080842 - 9 Aug 2024
Viewed by 529
Abstract
Laccase (LAC), a copper-containing polyphenol oxidase, is an important pathogenic factor of pathogenic fungi, and has been identified as an important virulence factor in numerous pathogenic fungi. LAC is encoded by a gene family and belongs to the class of multicopper oxidases. The [...] Read more.
Laccase (LAC), a copper-containing polyphenol oxidase, is an important pathogenic factor of pathogenic fungi, and has been identified as an important virulence factor in numerous pathogenic fungi. LAC is encoded by a gene family and belongs to the class of multicopper oxidases. The study aimed to identify the LAC genes in Athelia bombacina (Link) Pers, and their interactions with the host. The expression levels of the LAC genes were quantified using RT-qPCR. The LAC activity, level of malondialdehyde (MDA) and activities of protective enzymes in ‘Huangguan’ pears during the interaction were measured. The AbLac4 gene deletion mutant strain was constructed. Six LAC genes were identified in A. bombacina, distributed across three chromosomes. Interspecies collinearity analysis suggested that LAC genes could serve as crucial pathogenic factors in A. bombacina. The LAC gene family can be classified into three distinct subgroups. Among the subgroups, variations were observed in their characteristic sequences and conserved motifs. However, the LAC genes within the same subgroup exhibited a high degree of conservation. The genes showed diverse expression profiles, with their promoters harboring multiple stress-responsive elements. Signal peptide prediction showed that all LAC proteins, with the exception of the AbLac3 protein, possessed signal peptides, indicating that they are secretory proteins. The subcellular localization analysis showed that all LAC proteins may be localized extracellularly. RT-qPCR revealed differential expression patterns among LAC genes; specifically, AbLac1 and AbLac4 exhibited distinct expression dynamics during the infection process. The LAC activity first increased and then decreased, with the highest increase rate occurring in the early stage of culture. The MDA content and catalase (CAT) activity at the inoculated site were found to be significantly higher than the uninoculated control. In addition, the deletion of AbLac4 gene reduced the growth rate and pathogenic ability of A. bombacina. This investigation found that AbLac1 and AbLac4 may play pivotal roles in mediating host interactions, and the fruit may combat pathogen infection through increasing the activities of CAT, phenylalanine ammonia lyase and peroxidase. This study provides valuable new insights into the pathogenic mechanisms of A. bombacina, significantly contributing to the field. Full article
(This article belongs to the Special Issue Microbial Interaction with Horticulture Plant Growth and Development)
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20 pages, 5565 KiB  
Article
Biocatalytic Screening of the Oxidative Potential of Fungi Cultivated on Plant-Based Resources
by Alina Kinner, Stephan Lütz and Katrin Rosenthal
AppliedChem 2024, 4(3), 282-301; https://doi.org/10.3390/appliedchem4030018 - 8 Aug 2024
Viewed by 711
Abstract
The environmental impacts of the postindustrial era, which rely on fossil fuels, have compelled a reconsideration of the future of energy and chemical industries. Fungi are a valuable resource for improving a circular economy through the enhanced valorization of biomass and plant waste. [...] Read more.
The environmental impacts of the postindustrial era, which rely on fossil fuels, have compelled a reconsideration of the future of energy and chemical industries. Fungi are a valuable resource for improving a circular economy through the enhanced valorization of biomass and plant waste. They harbor a great diversity of oxidative enzymes, especially in their secretome. Enzymatic breakdown of the plant cell wall complex and lignocellulosic biomass yields sugars for fermentation and biofuel production, as well as aromatic compounds from lignin that can serve as raw materials for the chemical industry. To harness the biocatalytic potential, it is essential to identify and explore wild-type fungi and their secretomes. This study successfully combined genome mining and activity screening to uncover the oxidative potential of a collection of underexploited ascomycetes and basidiomycetes. The heme peroxidase and laccase activities of four promising candidates, Bipolaris victoriae, Colletotrichum sublineola, Neofusicoccum parvum and Moesziomyces antarcticus, were investigated to gain a deeper insight into their enzyme secretion. Furthermore, a plant-based medium screening with the phytopathogen C. sublineola revealed that soybean meal is a beneficial component to trigger the production and secretion of enzymes that catalyze H2O2-dependent oxidations. These results demonstrate that understanding fungal secretomes and their enzymatic potential opens exciting avenues for sustainable biotechnological applications across various industries. Full article
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17 pages, 3367 KiB  
Article
Blue and Yellow Laccases from Alternaria sp. Strain HU: Characterization and Immobilization on Magnetic Nanoparticles
by Ingrida Radveikienė, Regina Vidžiūnaitė, Rolandas Meškys and Vida Časaitė
J. Fungi 2024, 10(8), 559; https://doi.org/10.3390/jof10080559 - 8 Aug 2024
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Abstract
Laccases are important and valuable enzymes with a great potential for biotechnological applications. In this study, two novel laccases, LacHU1 and LacHU2, from Alternaria sp. HU have been purified and characterized. The molecular mass of each isoenzyme was ~66 kDa. LacHU1 laccases was [...] Read more.
Laccases are important and valuable enzymes with a great potential for biotechnological applications. In this study, two novel laccases, LacHU1 and LacHU2, from Alternaria sp. HU have been purified and characterized. The molecular mass of each isoenzyme was ~66 kDa. LacHU1 laccases was yellow and had no typical blue oxidase spectra and LacHU2 had a blue color and characteristic absorption spectra. The catalytic efficiency of LacHU1 for most substrates was higher than that of LacHU2 laccase. Both isoenzymes effectively oxidize flavonoids. Alternaria sp. laccases were successfully immobilized on magnetic nanoparticles. The thermostability of immobilized laccases increased and optimal pH shifted to more alkaline compared to the free laccases. Potential applications of laccases from Alternaria sp. HU are in the oxidation of flavonoids in cotton or in water treatment processes. Full article
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