Flow cytometric sorting of the filamentous fungus Trichoderma reesei for improved strains

Throndset, William; Kim, Steve; Bower, B.; Lantz, S. E.; Kelemen, Bradley R.; Pepsin, Michael; Chow, Nicole; Mitchinson, Colin; Ward, Mick

doi:10.1016/j.enzmictec.2010.09.003

Cited by 24 publications

(9 citation statements)

References 33 publications

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“…Thus, high-throughput screening methods are highly required to ensure the success of genome engineering. In fact, the fluorescence-activated cell sorting (FACS) assisted the intracellular protein production has been extensively adopted in filamentous fungi, but such strategy is difficult to screen out the beneficial mutants with enhanced protein secretion capacity (Throndset et al, 2010). To solve this problem, displaying the fluorescence protein on the cell surface, coupled by FACS, allows screening of the cellulose hypersecretors from T. reesei (Gao et al, 2018).…”

Section: Discussionmentioning

confidence: 99%

Genetic Engineering of Filamentous Fungi for Efficient Protein Expression and Secretion

Wang

Zhong

Xiao

2020

Front. Bioeng. Biotechnol.

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Filamentous fungi are considered as unique cell factories for protein production due to the high efficiency of protein secretion and superior capability of post-translational modifications. In this review, we firstly introduce the secretory pathway in filamentous fungi. We next summarize the current state-of-the-art works regarding how various genetic engineering strategies are applied for enhancing protein expression and secretion in filamentous fungi. Finally, in a future perspective, we discuss the great potential of genome engineering for further improving protein expression and secretion in filamentous fungi.

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Section: Discussionmentioning

confidence: 99%

Genetic Engineering of Filamentous Fungi for Efficient Protein Expression and Secretion

Wang

Zhong

Xiao

2020

Front. Bioeng. Biotechnol.

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“…However, some studies reported the use of flow cytometry for the improvement of non-oleaginous fungal strains. [98][99][100][101][102] Successful isolation of fluorescent dye stained and/or fluorescent protein/tag expressed fungal spores and conidia was demonstrated in different non-oleaginous fungal strains including, Aspergillus fumigatus, Coniothyrium minitans, Trichoderma reesei, Trichoderma viridae, Penicillium expansum and Phoma herbarum. Green fluorescent protein (GFP) was the most frequently used fluorescent protein for the improvement of non-oleaginous fungal strains.…”

Section: Applications Of Flow Cytometry In Oleaginous Microorganismsmentioning

confidence: 99%

Role of flow cytometry for the improvement of bioprocessing of oleaginous microorganisms

Sonowal

Chikkaputtaiah

Velmurugan

2019

J of Chemical Tech & Biotech

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Oleaginous microorganisms, such as yeast, fungi, microalgae and bacteria, represent a key segment of second generation feed‐stock materials and are considered to synthesize a wide range of industrially important chemical compounds. Oleaginous microorganisms possess a broad varieties of chemical compounds such as carotenoids, pigments, carbohydrates, chlorophyll, and storage‐material lipids. Oleaginous microorganisms have been recognized as promising sources for the synthesis of unsaturated, especially polyunsaturated fatty acids (PUFA). So far, a variety of high‐throughput screening methodologies (HTMs) have been employed for the development of bioprocessing of oleaginous microorganisms for sustainable production of industrially valuable compounds. Of HTMs, flow cytometry (FC) and sorters (FACS) have received substantial interest as better HTMs because of their ability to screen large numbers of cells within seconds, and interrogate and isolate living cells at single‐cell level. Forward and side scattering signals of FC are used to determine the physiological state of the cell while different channels available in the FC facilitate the detection of signals produced from fluorophores. Simultaneous measurement of physiological characteristics along with specific compound accumulation at single‐cell level enables the possibility of separating a particular phenotype with specific properties from a population. Different microbial strain development strategies in combination with FACS produced improved phenotypes with desired properties. This review first summarizes the FACS methodologies suitable for oleaginous microorganisms and the significant progress that has been achieved in oleaginous microorganisms using FACS, and highlights the important, advanced and future prospects of FACS methodologies that are suitable for the development of bioprocessing in oleaginous microorganisms. © 2018 Society of Chemical Industry

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“…This organism can produce 100 g/L of extracellular protein of which 60% is the major cellulase Cel7a (CBH I) and 20% is Cel6a (CBH II) (Shuster & Schmoll, 2010). Genencor scientists produced a strain doubled in ability to hydrolyze pretreated corn stover and sugar cane bagasse (Throndset et al, 2010). A cellulase/hemicellulase system, more than twice as active as that from T. reesei, was obtained from the extreme thermophilic bacterium Caldicellulosiruptor bescil (Kanafusa-Shinkai et al, 2013).…”

Section: Additional Applications Of Microbial Enzymesmentioning

confidence: 99%

Enzymes of industrial interest

Demain

Sánchez

2017

Mexjbiotechnol

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For many years, industrial enzymes have played an important role in the benefit of our society due to their many useful properties and a wide range of applications. They are key elements in the progress of many industries including foods, beverages, pharmaceuticals, diagnostics, therapy, personal care, animal feed, detergents, pulp and paper, textiles, leather, chemicals and biofuels. During recent decades, microbial enzymes have replaced many plant and animal enzymes. This is because microbial enzymes are widely available and produced economically in short fermentations and inexpensive media. Screening is simple, and strain improvement for increased production has been very successful. The advances in recombinant DNA technology have had a major effect on production levels of enzymes and represent a way to overproduce industrially important microbial, plant and animal enzymes. It has been calculated that 50-60% of the world enzyme market is supplied with recombinant enzymes. Molecular methods, including genomics and metagenomics, are being used for the discovery of new enzymes from microbes. Also, directed evolution has allowed the design of enzyme specificities and better performance.

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Flow cytometric sorting of the filamentous fungus Trichoderma reesei for improved strains

Cited by 24 publications

References 33 publications

Genetic Engineering of Filamentous Fungi for Efficient Protein Expression and Secretion

Genetic Engineering of Filamentous Fungi for Efficient Protein Expression and Secretion

Role of flow cytometry for the improvement of bioprocessing of oleaginous microorganisms

Enzymes of industrial interest

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