Identification of lipolytic enzymes isolated from bacteria indigenous to Eucalyptus wood species for application in the pulping industry

Ramnath, Lucretia; Sitholé, Bruce; Govinden, Roshini

doi:10.1016/j.btre.2017.07.004

Cited by 90 publications

(56 citation statements)

References 86 publications

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“…Bioassays confirmed the ability of C. truncatum and C. brevisporum to produce and secrete cutinase enzyme in lipid substrate media. Growth characteristics for the identification of secreted lipase activity using Tween‐20, Rhodamine, and tributyrin agar plates were congruent with the results obtained from other studies (Lanka & Latha, ; Ramnath, Sithole, & Govinden, ; Wadia & Jain, ). As both Tween‐20 and Tributyrin can test positive with esterases, the positive fluorescence with Rhodamine B was used as the determining factor for the presence of lipase activity (Lanka & Latha, ; Tomulescu et al, ).…”

Section: Discussionsupporting

confidence: 88%

Section: Lipid Substrate Bioassayssupporting

confidence: 88%

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Diversity, structure, and synteny of the cutinase gene of Colletotrichum species

Villafana

Rampersad

2020

Ecology and Evolution

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Colletotrichum species complexes are among the top 10 economically important fungal plant pathogens worldwide because they can infect climacteric and nonclimacteric fruit at the pre and/or postharvest stages. C. truncatum is the major pathogen responsible for anthracnose of green and red bell pepper fruit worldwide. C. brevisporum was recently reported to be a minor pathogen of red bell pepper fruit in Trinidad, but has recently been reported as pathogenic to other host species in other countries. The ability of these phytopathogens to produce and secrete cutinase is required for dismantling the cuticle of the host plant and, therefore, crucial to the necrotrophic phase of their infection strategy. In vitro bioassays using different lipid substrates confirmed the ability of C. truncatum and C. brevisporum isolates from green and red bell peppers to secrete cutinase. The diversity, structure and organization and synteny of the cutinase gene were determined among different Colletotrichum species. Cluster analysis indicated a low level of nucleotide variation among C. truncatum sequences. Nucleotide sequences of C. brevisporum were more related to C. truncatum cutinase nucleotide sequences than to C. gloeosporioides. Cluster patterns coincided with haplotype and there was evidence of significant positive selection with no recombination signatures. The structure of the cutinase gene included two exons with one intervening intron and, therefore, one splice variant. Although amino acid sequences were highly conserved among C. truncatum isolates, diversity “hot spots” were revealed when the 66‐amino acid coding region of 200 fungal species was compared. Twenty cutinase orthologues were detected among different fungal species, whose common ancestor is Pezizomycotina and it is purported that these orthologues arose through a single gene duplication event prior to speciation. The cutinase domain was retained both in structure and arrangement among 34 different Colletotrichum species. The order of aligned genomic blocks between species and the arrangement of flanking protein domains were also conserved and shared for those domains immediately located at the N‐ and C‐terminus of the cutinase domain. Among these were an RNA recognition motif, translation elongation factor, signal peptide, pentatricopeptide repeat, and Hsp70 family of chaperone proteins, all of which support the expression of the cutinase gene. The findings of this study are important to understanding the evolution of the cutinase gene in C. truncatum as a key component of the biotrophic–necrotrophic switch which may be useful in developing gene‐targeting strategies to decrease the pathogenic potential of Colletotrichum species.

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Section: Discussionsupporting

confidence: 88%

Section: Lipid Substrate Bioassayssupporting

confidence: 88%

Diversity, structure, and synteny of the cutinase gene of Colletotrichum species

Villafana

Rampersad

2020

Ecology and Evolution

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“…Microbial lipases have their production greatly influenced by medium composition being the carbon source a major factor; since lipases are inducible and mostly extracellular, they are generally produced in the presence of a lipid source , therefore the use of olive oil as an inductor relays in that its major components are triacylglycerols; additionally, lipases are significantly influenced by physicochemical factors such as temperature and pH .…”

Section: Resultsmentioning

confidence: 99%

“…) the best results were obtained with sonication with 211 U/L. Centering the analysis on this method, diverse reports indicate that the use of sonication is an efficient method for cell lysis for organisms with resistant cell membranes and has been used by several authors, with some mentions that physical methods are the most common at industrial level . The discussion about sonication presented by Klimek et al .…”

Section: Resultsmentioning

confidence: 99%

“…8) the best results were obtained with sonication with 211 U/L. Centering the analysis on this method, diverse reports indicate that the use of sonication is an efficient method for cell lysis for organisms with resistant cell membranes and has been used by several authors, with some mentions that physical methods are the most common at industrial level [29,34]. The discussion about sonication presented by Klimek et al (2011) [35] refers that the use of it depends on factors such as the percentage of chitin in the cell walls which can lead to the variation in effectiveness from the same lysis method between organisms; however, said methods do not leave residues and by not having a direct interaction with the enzyme, like some detergents may have for the inherent presence of charges that could affect the integrity or enzymatic yield of the enzyme of interest, comprise in our case a preferable technique.…”

Section: Intracellular Lysismentioning

confidence: 99%

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All‐around management of a fungal isolate obtained from cheese spoilage as an environmental source: Direct approach from an undergrad student to a biotechnological characterization

Lechuga

Herrera

Árevalo-Niño

2019

Biochem Molecular Bio Educ

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In this work, we present the results of an undergrad student from the perspective of its first approach as a principal researcher in a project. In order to gain practical experience, one of the options for students that have interest in pursuing a postgraduate program corresponds to a research stay in a laboratory of their selected field conducting a project for a period of 6 months. In this particular project, a fungal sample was isolated from Parmesan cheese spoilage and its enzymatic activity evaluated. Using simple and standardized protocols, the student was capable of identifying a possible biotechnological application for the isolate by detecting and categorizing the lipolytic activity. Through microculture characterization in potato dextrose agar (PDA) the genus of the sample was determined as Penicillium and confirmed by molecular analysis of the ITS1 and ITS4 regions. In order to examine comprehensively the potential of the new isolate, the extracellular and intracellular enzymatic activities were analyzed as well as four methods of cell rupture. From these results, sonication was determined as the best technique with 211 U/L as a maximum lipolytic value. To finalize the evaluation of the sample, the student determined the optimal pH and temperature as well as the thermotolerance of the crude extract obtaining a residual activity of 13% after 60 minutes of incubation at 45 °C. Upon conclusion of the research we could recognize that through a direct characterization of a fungal isolate using techniques that are widely known, the student was capable of determining and value one of the most interesting capabilities fungi has to offer; enzymatic activity, and that the knowledge obtained from established protocols enables and encourages the students to correlate the source from where they were obtained with potential biotechnological applications. © 2019 International Union of Biochemistry and Molecular Biology, 47(6):681–688, 2019.

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Enzymatic Intervention as an Ecofriendly Approach in Industries

Gupta¹,

Dwivedi²,

Yadav³

et al. 2022

Microbial Biotechnology

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Identification of lipolytic enzymes isolated from bacteria indigenous to Eucalyptus wood species for application in the pulping industry

Cited by 90 publications

References 86 publications

Diversity, structure, and synteny of the cutinase gene of Colletotrichum species

Diversity, structure, and synteny of the cutinase gene of Colletotrichum species

All‐around management of a fungal isolate obtained from cheese spoilage as an environmental source: Direct approach from an undergrad student to a biotechnological characterization

Enzymatic Intervention as an Ecofriendly Approach in Industries

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