Cloning, functional expression and characterization of Aspergillus sulphureus β-mannanase in Pichia pastoris

2007 ◽  
Vol 128 (3) ◽  
pp. 452-461 ◽  
Author(s):  
Xiaoling Chen ◽  
Yunhe Cao ◽  
Yuhua Ding ◽  
Wenqing Lu ◽  
Defa Li
Keyword(s):  
Pichia Pastoris ◽  
Functional Expression ◽  
Aspergillus Sulphureus

In-fusion expression and characterization of β-xylanase and β-1,3-1,4-glucanase in Pichia pastoris

Biologia ◽  
2012 ◽  
Vol 67 (4) ◽  
Author(s):  
Jiayun Qiao ◽  
Yunhe Cao
Keyword(s):  
Bacillus Subtilis ◽  
Pichia Pastoris ◽  
Fusion Protein ◽  
Fusion Expression ◽  
Chimeric Genes ◽  
Fusion Enzyme ◽  
Fusion Enzymes ◽  
Aspergillus Sulphureus

AbstractTwo chimeric genes, XynA-Bs-Glu-1 and XynA-Bs-Glu-2, encoding Aspergillus sulphureus β-xylanase (XynA, 26 kDa) and Bacillus subtilis β-1,3-1,4-glucanase (Bs-Glu, 30 kDa), were constructed via in-fusion by different linkers and expressed successfully in Pichia pastoris. The fusion protein (50 kDa) exhibited both β-xylanase and β-1,3-1,4-glucanase activities. Compared with parental enzymes, the moiety activities were decreased in fermentation supernatants. Parental XynA and Bs-Glu were superior to corresponding moieties in each fusion enzymes because of lower Kn higher kcat. Despite some variations, common optima were generally 50°C and pH 3.4 for the XynA moiety and parent, and 40°C and pH 6.4 for the Bs-Glu counterparts. Thus, the fusion enzyme XynA-Bs-Glu-1 and XynA-Bs-Glu-2 were bifunctional.

scholarly journals Characterization of a thermostable β-glucosidase from Aspergillus fumigatus Z5, and its functional expression in Pichia pastoris X33

2012 ◽  
Vol 11 (1) ◽  
pp. 25 ◽  
Author(s):  
Dongyang Liu ◽  
Ruifu Zhang ◽  
Xingming Yang ◽  
Zhenhua Zhang ◽  
Song Song ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Aspergillus Fumigatus ◽  
Functional Expression

Functional expression and characterization of an endo-1,4-β-mannosidase from Triticum aestivum in Pichia pastoris

Biologia ◽  
2020 ◽  
Vol 75 (11) ◽  
pp. 2073-2081 ◽  
Author(s):  
Shuai-Bing Zhang ◽  
Wei-Ji Zhang ◽  
Na Li ◽  
Huan-Chen Zhai ◽  
Yang-Yong Lv ◽  
...  
Keyword(s):  
Triticum Aestivum ◽  
Pichia Pastoris ◽  
Functional Expression

Functional expression, purification, and characterization of human Flt3 ligand in the Pichia pastoris system

2005 ◽  
Vol 42 (2) ◽  
pp. 246-254 ◽  
Author(s):  
Yan-Li Zhang ◽  
Song-Sen Chen ◽  
Ke-Gong Yang ◽  
Lin Su ◽  
Yan-Chun Deng ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Functional Expression ◽  
Flt3 Ligand ◽  
Purification And Characterization

scholarly journals Functional expression of the eukaryotic proton pump rhodopsin OmR2 in Escherichia coli and its photochemical characterization

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Masuzu Kikuchi ◽  
Keiichi Kojima ◽  
Shin Nakao ◽  
Susumu Yoshizawa ◽  
Shiho Kawanishi ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Proton Pump ◽  
Expression System ◽  
Spectroscopic Characterization ◽  
Functional Expression ◽  
Proton Pumping ◽  
E Coli ◽  
Oxyrrhis Marina ◽  
Domains Of Life

AbstractMicrobial rhodopsins are photoswitchable seven-transmembrane proteins that are widely distributed in three domains of life, archaea, bacteria and eukarya. Rhodopsins allow the transport of protons outwardly across the membrane and are indispensable for light-energy conversion in microorganisms. Archaeal and bacterial proton pump rhodopsins have been characterized using an Escherichia coli expression system because that enables the rapid production of large amounts of recombinant proteins, whereas no success has been reported for eukaryotic rhodopsins. Here, we report a phylogenetically distinct eukaryotic rhodopsin from the dinoflagellate Oxyrrhis marina (O. marina rhodopsin-2, OmR2) that can be expressed in E. coli cells. E. coli cells harboring the OmR2 gene showed an outward proton-pumping activity, indicating its functional expression. Spectroscopic characterization of the purified OmR2 protein revealed several features as follows: (1) an absorption maximum at 533 nm with all-trans retinal chromophore, (2) the possession of the deprotonated counterion (pKa = 3.0) of the protonated Schiff base and (3) a rapid photocycle through several distinct photointermediates. Those features are similar to those of known eukaryotic proton pump rhodopsins. Our successful characterization of OmR2 expressed in E. coli cells could build a basis for understanding and utilizing eukaryotic rhodopsins.

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