Project description:Aspergillus terreus is a filamentous ascomycota, which is prominent for its production of lovastatin, an antihypercholesterolemic drug. The commercial importance of lovastatin with annual sales of billions of dollars made us to focus on lovastatin biosynthetic cluster proteins. The analysis of these lovastatin biosynthetic cluster proteins with different perspectives such as physicochemical property, structure based analysis and functional studies were done to find out the role and function of every protein involved in the lovastatin biosynthesis pathway. Several computational tools are used to predict the physicochemical properties, secondary structural features, topology, patterns, domains and cellular location. There are 8 unidentified proteins in lovastatin biosynthetic cluster, in which 6 proteins have homologous partners, and annotation transfer is done based on the closely related homologous genes, and their structures are also modeled. The two other proteins that do not have homologous partners are predicted as PQ loop repeat protein that may be involved in glycosylation machinery and as thiolase-acyl activity by the integrated functional analysis approach.
Project description:Cultivation of Aspergillus terreus ATCC 20542 in a stirred tank bioreactor was performed to induce the biosynthesis of secondary metabolites and provide the bioprocess-related insights into the metabolic capabilities of the investigated strain. The activation of biosynthetic routes was attempted by the diversification of process conditions and growth media. Several strategies were tested, including the addition of rapeseed oil or inulin, changing the concentration of nitrogen source, reduction of chlorine supply, cultivation under saline conditions, and using various aeration schemes. Fifteen secondary metabolites were identified in the course of the study by using ultra-high performance liquid chromatography coupled with mass spectrometry, namely mevinolinic acid, 4a,5-dihydromevinolinic acid, 3?-hydroxy-3,5-dihydromonacolin L acid, terrein, aspulvinone E, dihydroisoflavipucine, (+)-geodin, (+)-bisdechlorogeodin, (+)-erdin, asterric acid, butyrolactone I, desmethylsulochrin, questin, sulochrin, and demethylasterric acid. The study also presents the collection of mass spectra that can serve as a resource for future experiments. The growth in a salt-rich environment turned out to be strongly inhibitory for secondary metabolism and the formation of dense and compact pellets was observed. Generally, the addition of inulin, reducing the oxygen supply, and increasing the content of nitrogen source did not enhance the production of examined molecules. The most successful strategy involved the addition of rapeseed oil to the chlorine-deficient medium. Under these conditions, the highest levels of butyrolactone I, asterric acid, and mevinolinic acid were achieved and the presence of desmethylsulochrin and (+)-bisdechlorogeodin was detected in the broth. The constant and relatively high aeration rate in the idiophase was shown to be beneficial for terrein and (+)-geodin biosynthesis.
Project description:In the present study, the complete genome of a filamentous fungus Aspergillus terreus ATCC 20542 was sequenced, assembled, and annotated. This strain is mainly recognized for being a model wild-type lovastatin producer and a parental strain of high-yielding industrial mutants. It is also a microorganism with a rich repertoire of secondary metabolites that has been a subject of numerous bioprocess-related studies. In terms of continuity, the genomic sequence provided in this work is of the highest quality among all the publicly available genomes of A. terreus strains. The comparative analysis revealed considerable diversity with regard to the catalog of biosynthetic gene clusters found in A. terreus. Even though the cluster of lovastatin biosynthesis was found to be well-conserved at the species level, several unique genes putatively associated with metabolic functions were detected in A. terreus ATCC 20542 that were not detected in other investigated genomes. The analysis was conducted also in the context of the primary metabolic pathways (sugar catabolism, biomass degradation potential, organic acid production), where the visible differences in gene copy numbers were detected. However, the species-level genomic diversity of A. terreus was more evident for secondary metabolism than for the well-conserved primary metabolic pathways. The newly sequenced genome of A. terreus ATCC 20542 was found to harbor several unique sequences, which can be regarded as interesting subjects for future experimental efforts on A. terreus metabolism and fungal biosynthetic capabilities. KEY POINTS: • The high-quality genome of Aspergillus terreus ATCC 20542 has been assembled and annotated. • Comparative analysis with other sequenced Aspergillus terreus strains has revealed considerable diversity in biosynthetic gene repertoire, especially related to secondary metabolism. • The unique genomic features of A. terreus ATCC 20542 are discussed.
Project description:Aspergillus terreus is well-known for lovastatin and itaconic acid production with biomedical and commercial importance. The mechanisms of metabolite formation have been extensively studied to improve their yield through genetic engineering. However, the combined repertoire of carbohydrate-active enzymes (CAZymes), cytochrome P450s (CYP) enzymes, and secondary metabolites (SMs) in the different A. terreus strains has not been well studied yet, especially with respect to the presence of biosynthetic gene clusters (BGCs). Here we present a 30 Mb whole genome sequence of A. terreus ATCC 20541 in which we predicted 10,410 protein-coding genes. We compared the CAZymes, CYPs enzyme, and SMs across eleven A. terreus strains, and the results indicate that all strains have rich pectin degradation enzyme and CYP52 families. The lovastatin BGC of lovI was linked with lovF in A. terreus ATCC 20541, and the phenomenon was not found in the other strains. A. terreus ATCC 20541 lacked a non-ribosomal peptide synthetase (AnaPS) participating in acetylaszonalenin production, which was a conserved protein in the ten other strains. Our results present a comprehensive analysis of CAZymes, CYPs enzyme, and SM diversities in A. terreus strains and will facilitate further research in the function of BGCs associated with valuable SMs.
Project description:S. aureus ATCC 25923 is performance standard for antimicrobial susceptibility testing. S. aureus ATCC 33591 showed resistance against erytrhromycin, penicillin, and streptomycin. We used microarray to compare RNA expression between sensitive and resistant strain of S. aureus as a preliminary research for MRSA inhibition.