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Metarhizium anisopliae overexpression

Min-Nan Tseng, Chia-Ling Chung, Shean-Shong Tzean
The entomopathogenic fungus Metarhizium anisopliae MA05-169 is a transformant strain that has been metabolically engineered to express dihydroxynaphthalene-melanin biosynthesis genes. In contrast to the wild type strain, the transformant displays a greater resistance to environmental stress and a higher virulence toward target insect host. However, the underlying mechanisms for these characteristics remain unclear; hence experiments were initiated to explore the possible mechanism(s) through physiological and molecular approaches...
2014: PloS One
Yingying Li, Kehe Fu, Shigang Gao, Qiong Wu, Lili Fan, Yaqian Li, Jie Chen
The chit42 gene cloned from Metarhizium anisopliae lacks chitin-binding domain (chBD), which plays important roles in binding insoluble chitin. Five kinds of hybrid chitinase Trichoderma transformants were constructed in this study, where the chit42 gene was fused to chBDs derived from plant, bacterial, and insect sources. The transformant Mc4 harboring chBDs from bitter melon (Momordica charantia) displayed the highest chitinase activity among all chBDs. The chitinase activities of Mc4, chit42 Trichoderma transformant Mchit3, and wild-type strain T30 were 44...
2013: Journal of Environmental Science and Health. Part. B, Pesticides, Food Contaminants, and Agricultural Wastes
Federica Volontè, Ines Pisanelli, Paola D'Arrigo, Fiorenza Viani, Gianluca Molla, Stefano Servi, Loredano Pollegioni
In this work, a reliable protocol was designed to rapidly express and purify a microbial chymotrypsin(ogen) as a useful alternative to using animal proteases. The cDNA encoding for chymotrypsinogen from the deuteromycete Metarhizium anisopliae (chy1) was overexpressed in an Origami2(DE3) E. coli strain deficient in thioredoxin reductase and glutathione reductase activities, thus possibly allowing disulfide exchange. By using a quick purification protocol, in which the hexahistidine tag was added at the C-terminal end of the protease, the recombinant CHY1 protein could be purified in a single step on an Ni-NTA column as a mixture of 19...
December 10, 2011: Enzyme and Microbial Technology
Sibao Wang, Weiguo Fang, Chengshu Wang, Raymond J St Leger
An enduring theme in pathogenic microbiology is poor understanding of the mechanisms of host specificity. Metarhizium is a cosmopolitan genus of invertebrate pathogens that contains generalist species with broad host ranges such as M. robertsii (formerly known as M. anisopliae var. anisopliae) as well as specialists such as the acridid-specific grasshopper pathogen M. acridum. During growth on caterpillar (Manduca sexta) cuticle, M. robertsii up-regulates a gene (Mest1) that is absent in M. acridum and most other fungi...
June 2011: PLoS Pathogens
Claudia Erika Morales Hernandez, Israel Enrique Padilla Guerrero, Gloria Angelica Gonzalez Hernandez, Eduardo Salazar Solis, Juan Carlos Torres Guzman
Catalases and peroxidases are the most important enzymes that degrade hydrogen peroxide into water and oxygen. These enzymes and superoxide dismutase are the first lines of cell defense against reactive oxygen species. Metarhizium anisopliae displays an increase in catalase-peroxidase activity during germination and growth. To determine the importance of catalase during the invasion process of M. anisopliae, we isolated the cat1 gene. cat1 cDNA expression in Escherichia coli and the subsequent purification of the protein confirmed that the cat1 gene codes for a monofunctional catalase...
July 2010: Applied Microbiology and Biotechnology
Weiguo Fang, Jin Feng, Yanhua Fan, Yongjun Zhang, Michael J Bidochka, Raymond J St Leger, Yan Pei
Entomopathogenic fungi, such as Beauveria bassiana and Metarhizium anisopliae are being developed as alternatives to chemical insecticides. They infect insects by direct penetration of the cuticle using a combination of physical pressure and extracellular hydrolytic enzymes such as proteases and chitinases. Previously we found that overexpression of a subtilisin-like protease (Pr1A) or a chitinase (Bbchit1) resulted in increased virulence of M. anisopliae and B. bassiana, respectively. In this study, we found that a mixture of the B...
October 2009: Journal of Invertebrate Pathology
Juliano Tomazzoni Boldo, Angela Junges, Karina Bohrer do Amaral, Charley Christian Staats, Marilene Henning Vainstein, Augusto Schrank
Chitinases have been implicated in fungal cell wall remodeling and play a role in exogenous chitin degradation for nutrition and competition. Due to the diversity of these enzymes, assigning particular functions to each chitinase is still ongoing. The entomopathogenic fungus Metarhizium anisopliae produces several chitinases, and here, we evaluate whether endochitinase CHI2 is involved in the pathogenicity of this fungus. We constructed strains either overexpressing or lacking the CHI2 chitinase. These constructs were validated by Southern, Northern and Western blot analysis, and chitinase production...
October 2009: Current Genetics
Berit Schuhmann, Volkhard Seitz, Andreas Vilcinskas, Lars Podsiadlowski
A novel defensin-like peptide was identified in the greater wax moth, Galleria mellonella. It was discovered in a haemocyte cDNA bank enriched with transcripts upregulated after immune challenge via subtractive hybridisation and suppressive PCR. The deduced amino acid sequence of the defensin-like peptide exhibits similarities to the antifungal peptides drosomycin from Drosophila melanogaster and heliomicin from Heliothis virescens. Therefore, it has been termed gallerimycin. Upregulation of gallerimycin after stimulation of the immune system by LPS-injection was demonstrated by quantitative real-time PCR...
July 2003: Archives of Insect Biochemistry and Physiology
S E Screen, G Hu, R J St Leger
Extracellular chitinase activity has been implicated in the pathogenesis of several fungal infections. Following induction with chitin, the insect pathogens Metarhizium anisopliae sf. acridum ARSEF strain 324 and Metarhizium anisopliae sf. anisopliae ARSEF strain 2575 secrete 44-kDa basic and acidic isoforms of endochitinase, respectively. The gene from strain 324 (Chit1) was cloned and inserted into the genome of strain 2575 under the control of Aspergillus regulatory elements such that transgenic 2575 (2575-Chit(+)) expressed CHIT1 in a noninducing medium (i...
November 2001: Journal of Invertebrate Pathology
R St Leger, L Joshi, M J Bidochka, D W Roberts
Mycoinsecticides are being used for the control of many insect pests as an environmentally acceptable alternative to chemical insecticides. A key aim of much recent work has been to increase the speed of kill and so improve commercial efficacy of these biocontrol agents. This might he achieved by adding insecticidal genes to the fungus, an approach considered to have enormous potential for the improvement of biological pesticides. We report here the development of a genetically improved entomopathogenic fungus...
June 25, 1996: Proceedings of the National Academy of Sciences of the United States of America
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