RESUMO
Filamentous fungi are important industrial microorganisms that play important roles in the production of bio-based products such as organic acids, proteins and secondary metabolites. The development of metabolic engineering and its enabling techniques have greatly promoted the design, construction and application of filamentous fungal cell factories. This article systematically reviews the development of filamentous fungal cell factories constructed through metabolic engineering, and discusses the challenges and future perspectives for systems metabolic engineering of filamentous fungi.
Assuntos
Fungos/genética , Engenharia MetabólicaRESUMO
The lignocellulolytic filamentous fungus Neurospora crassa is able to assimilate various mono- and oligo-saccharides. However, more than half of predicted sugar transporters in the genome are still waiting for functional elucidation. In this study, system analysis of substrate spectra of predicted sugar transporters in N. crassa was performed at genome-wide level. NCU01868 and NCU08152 have the capability of uptaking various hexose, which are named as NcHXT-1 and NcHXT-2 respectively. Their transport activities for glucose were further confirmed by fluorescence resonance energy transfer analysis. Over-expression of either NcHXT-1 or NcHXT-2 in the null-hexose-transporter yeast EBY.VW4000 restored the growth and ethanol fermentation under submerged fermentation with glucose, galactose, or mannose as the sole carbon source. NcHXT-1/-2 homologues were found in a variety of cellulolytic fungi. Functional identification of two filamentous fungal-conserved hexose transporters NcHXT-1/-2 via genome scanning would represent novel targets for ongoing efforts in engineering cellulolytic fungi and hexose fermentation in yeast.
RESUMO
Filamentous fungi are widely used for large-scale production of cellulases. Morphological characteristics of mycelia under submerged condition are closely correlated with cellulases productivity. In order to find out the critical genes involved in the mycelial morphology development and cellulases production in liquid fermentation, 95 Neurospora crassa morphological mutants (named as SZY1-95) were screened for cellulases production. Compared with the wild type, cellulases production in four mutants SZY32, SZY35, SZY39 and SZY43 were significantly decreased, whereas mutants SZY63, SZY69, SZY87 and SZY11 produced much more cellulases than that of the wild type strain. Meanwhile, endo-beta-1,4-glucanase activity, beta-glucosidase activity, viscosity of broth and dry weight of these mutants were measured. The mycelial morphology of the mutants was also studied by microscope. Particularly, pellets were formed in mutant SZY11 and SZY43, whose viscosities were 25% and 50% of the wild type strain, respectively. Mutant SZY87 appeared long hyphae, and the viscosity of its broth was at least 2 folds of the wild type strain. These results indicate that a single gene deletion could influence the mycelial morphology in liquid fermentation, and increased the cellulases production. The low-viscosity related genes identified in our study will be the potential candidates for genetic modification of filamentous fungi.
Assuntos
Celulases , Fermentação , Deleção de Genes , Microbiologia Industrial , Neurospora crassa , Genética , MetabolismoRESUMO
Industrial biotechnology provides practical solutions to the challenges in the areas of resources, energy and environment. Based on the 7th China Summit Forum on Industrial Biotechnology Development, this special issue reports the latest advances in the fields of bioinformatics, microbial cell factories, fermentation engineering, industrial enzymes and high throughput screening methods.
Assuntos
Biotecnologia , China , Enzimas , Química , Fermentação , Ensaios de Triagem em Larga Escala , Microbiologia IndustrialRESUMO
Inefficient degradation of lignocellulose is one of the main barriers for the utilization of renewable plant biomass for biofuel production. The bottleneck of the biorefinery process is the generation of fermentable sugars from complicated biomass polymers. In nature, the main microbes of lignocelluloses deconstruction are fungi. Therefore, elucidating the mechanism of lignocelluloses degradation by fungi is of critical importance for the commercialization of lignocellulosic biofuels. This review focuses on the progress in lignocelluloses degradation pathways in fungi, especially on the advances made by functional genomics studies.