Fluorescent proteins in microbial biotechnology—new proteins and new applications

Vizcaino-Caston, Isaac; Wyre, Chris; Overton, Tim W.

doi:10.1007/s10529-011-0767-5

Cited by 18 publications

(6 citation statements)

References 87 publications

(86 reference statements)

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“…When fungal secretory enzymes were fused with GFP, intense fluorescence was observed near (or at) the tip, cell wall, and septa regions of Aspergillus hyphae [1]. Detection of GFP requires only irradiation with blue light and can be rapidly observed and analyzed [34]. The results of fluorescence measurements indicated that the gfp gene was successfully expressed in A. oryzae, which demonstrated the applicability of the newly constructed system.…”

Section: Discussionmentioning

confidence: 87%

Construction of a Shuttle Vector for Heterologous Expression of a Novel Fungal ��-Amylase Gene in Aspergillus oryzae

Yin¹,

YouZhi²,

Yin³

et al. 2015

Journal of Microbiology and Biotechnology

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The filamentous fungus Aspergillus oryzae is a well-known expression host used to express homologous and heterologous proteins in a number of industrial applications. To facilitate higher yields of proteins of interest, we constructed the pAsOP vector to express heterologous proteins in A. oryzae. pAsOP carries a selectable marker, pyrG, derived from Aspergillus nidulans, and a strong promoter and a terminator of the amyB gene derived from A. oryzae. pAsOP transformed A. oryzae efficiently via the PEG-CaCl2-mediated transformation method. As proof of concept, green fluorescent protein (GFP) was successfully expressed in A. oryzae transformed by pAsOP-GFP. Additionally, we identified a novel fungal α-amylase (PcAmy) gene from Penicillium sp. and cloned the gene into the vector. After transformation by pAsOPPcAmy, the α-amylase PcAmy from Penicillium sp. was successfully expressed in a heterologous host system for the first time. The α-amylase activity in the A. oryzae transformant was increased by 62.3% compared with the untransformed A. oryzae control. The PcAmy protein produced in the system had an optimum pH of 5.0 and optimum temperature of 30°C. As a cold-adapted enzyme, PcAmy shows potential value in industrial applications because of its high catalytic activity at low temperature. Furthermore, the expression vector reported in this study provides promising utility for further scientific research and biotechnological applications.

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Section: Discussionmentioning

confidence: 87%

Construction of a Shuttle Vector for Heterologous Expression of a Novel Fungal ��-Amylase Gene in Aspergillus oryzae

Yin¹,

YouZhi²,

Yin³

et al. 2015

Journal of Microbiology and Biotechnology

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“…Only by doing this step, could we have confidence to do the next following purification step. The bright green fluorescence was observed under excitation at 488 nm and emission at 525 nm[ Fig.2 2 ), it can be seen that almost all the yeast cells have expressed the fusion protein HGFI-GFP in vivo after the methanol induction for 96 h. As one of the most widely used reporter proteins，GFP is usually fused to the N-terminus or C-terminus of recombinant genes of interest, recombinant fusions possess many advantages, including its fluorescence, stability and simple detection in living cells [20] . Moreover, they were expressed successfully in a wide variety of organisms, such as E. coli, P. pastoris, A. niger and mammalian cell CHO [21][22][23] .…”

Section: Fluorescent Detection Of Hgfi-gfp In P Pastorismentioning

confidence: 99%

A visualized fusion protein based on self-assembly hydrophobin HGFI

Zhao

Liu

Song

et al. 2015

Chem. Res. Chin. Univ.

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Hydrophobins are a type of small amphipathic proteins with a unique self-assembly property, which can be used to modify material surfaces and adsorb enzymes, antibodies and even cells. In this study, a fusion protein consisting of hydrophobin HGFI and green fluorescent protein(GFP) was successfully obtained from Pichia patoris (P. pastoris). Water contact angle(WCA) measurement proves that the wettability of the surfaces of different materials was changed. We further demonstrated the self-assembly ability of HGFI-GFP, which can be used to disperse the multi-walled carbon nanotubes(MWCNTs). Finally, the adsorption of HGFI-GFP onto the surface of the tissue engineering material poly(ε-caprolactone)(PCL) was evaluated by detecting the fluorescence of the fusion protein itself. The resalt demonstrates that both the basic self-assembly activity of the HGFI domain and the functional activity of the GFP domain were still remained.

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“…One approach is to use cells with fl uorescent proteins as reporters (Vizcaino-Caston et al 2012 ). They can be used: to localize cells or cell components; to report specifi c protein synthesis; and to sense physicochemical states.…”

Section: Cheese As a Solid State Fermentationmentioning

confidence: 99%

Diversity, Dynamics and Functional Role of Actinomycetes on European Smear Ripened Cheeses

Bora¹,

Dodd²,

Desmasures³

2015

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Fluorescent proteins in microbial biotechnology—new proteins and new applications

Cited by 18 publications

References 87 publications

Construction of a Shuttle Vector for Heterologous Expression of a Novel Fungal ��-Amylase Gene in Aspergillus oryzae

Construction of a Shuttle Vector for Heterologous Expression of a Novel Fungal ��-Amylase Gene in Aspergillus oryzae

A visualized fusion protein based on self-assembly hydrophobin HGFI

Diversity, Dynamics and Functional Role of Actinomycetes on European Smear Ripened Cheeses

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