scholarly journals Generation of Xylose-Fermenting Saccharomyces Cerevisiae by Protein-Engineering

10.5772/27603 ◽  
2012 ◽  
Author(s):  
Seiya Watanabe ◽  
Keisuke Makino
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Protein Engineering

scholarly journals Conversion of Waste Agriculture Biomass to Bioethanol by Recombinant Saccharomyces cerevisiae

2010 ◽  
Vol 2 (2) ◽  
pp. 351-361
Author(s):  
A. A. Saleh ◽  
S. Hamdan ◽  
N. Annaluru ◽  
S. Watanabe ◽  
M. R. Rahman ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Enzyme Activity ◽  
Protein Engineering ◽  
Agricultural Waste ◽  
Xylitol Dehydrogenase ◽  
Ethanol Conversion ◽  
Waste Biomass ◽  
Wild Type ◽  
Coenzyme Specificity ◽  
Recombinant Yeasts

Agricultural waste biomass has already been transferred to bioethanol and used as energy related products, although many issues such as efficiency and productivity still to be overcome. In this study, the protein engineering was applied to generate enzymes with completely reversed coenzyme specificity and developed recombinant yeasts containing those engineered enzymes for construction of an efficient biomass-ethanol conversion system. Recombinant yeasts were constructed with the genes encoding a wild type xylose reductase (XR) and the protein engineered xylitol dehydrogenase (XDH) (with NADP) of Pichia stipitis.  These recombinant yeasts were characterized based on the enzyme activity and fermentation ability of xylose to ethanol. The protein engineered enzymes were expressed significantly in Saccharomyces cerevisiae as judged by the enzyme activity in vitro. Ethanol fermentation was measured in batch culture under anaerobic conditions. The significant enhancement was found in Y-ARS strain, in which NADP+-dependent XDH was expressed; 85% decrease of unfavorable xylitol excretion with 26% increased ethanol production, when compared with the reference strain expressing the wild-type XDH.  Keywords: Agricultural waste biomass; Protein engineering; Xylitol dehydrogenase; Xylose-fermentation; Eethanol production. © 2010 JSR Publications. ISSN: 2070-0237 (Print); 2070-0245 (Online). All rights reserved. DOI: 10.3329/jsr.v2i2.2882               J. Sci. Res. 2 (2), 351-361 (2010) 

scholarly journals A deeper understanding of the spontaneous derepression of the URA3 gene in MaV203 Saccharomyces cerevisiae and its implications for protein engineering and the reverse two‐hybrid system

Yeast ◽  
2019 ◽  
Vol 36 (12) ◽  
pp. 701-710
Author(s):  
David Cortens ◽  
Rebekka Hansen ◽  
Geert‐Jan Graulus ◽  
Erik Steen Redeker ◽  
Peter Adriaensens ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Protein Engineering ◽  
Hybrid System ◽  
Ura3 Gene ◽  
Two Hybrid

scholarly journals Protein engineering of Saccharomyces cerevisiae transporter Pdr5p identifies key residues that impact Fusarium mycotoxin export and resistance to inhibition

2016 ◽  
Vol 5 (6) ◽  
pp. 979-991 ◽  
Author(s):  
Amanda B. Gunter ◽  
Anne Hermans ◽  
Whynn Bosnich ◽  
Douglas A. Johnson ◽  
Linda J. Harris ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Protein Engineering ◽  
Fusarium Mycotoxin ◽  
Key Residues

scholarly journals Engineering the Enantioselectivity of Yeast Old Yellow Enzyme OYE2y in Asymmetric Reduction of (E/Z)-Citral to (R)-Citronellal

Molecules ◽  
2019 ◽  
Vol 24 (6) ◽  
pp. 1057 ◽  
Author(s):  
Xiangxian Ying ◽  
Shihua Yu ◽  
Meijuan Huang ◽  
Ran Wei ◽  
Shumin Meng ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Protein Engineering ◽  
Active Site ◽  
Active Sites ◽  
Asymmetric Reduction ◽  
Saturation Mutagenesis ◽  
Old Yellow Enzyme ◽  
Docking Analysis ◽  
Site Saturation ◽  
Low Activity

The members of the Old Yellow Enzyme (OYE) family are capable of catalyzing the asymmetric reduction of (E/Z)-citral to (R)-citronellal—a key intermediate in the synthesis of L-menthol. The applications of OYE-mediated biotransformation are usually hampered by its insufficient enantioselectivity and low activity. Here, the (R)-enantioselectivity of Old Yellow Enzyme from Saccharomyces cerevisiae CICC1060 (OYE2y) was enhanced through protein engineering. The single mutations of OYE2y revealed that the sites R330 and P76 could act as the enantioselectivity switch of OYE2y. Site-saturation mutagenesis was conducted to generate all possible replacements for the sites R330 and P76, yielding 17 and five variants with improved (R)-enantioselectivity in the (E/Z)-citral reduction, respectively. Among them, the variants R330H and P76C partly reversed the neral derived enantioselectivity from 32.66% e.e. (S) to 71.92% e.e. (R) and 37.50% e.e. (R), respectively. The docking analysis of OYE2y and its variants revealed that the substitutions R330H and P76C enabled neral to bind with a flipped orientation in the active site and thus reverse the enantioselectivity. Remarkably, the double substitutions of R330H/P76M, P76G/R330H, or P76S/R330H further improved (R)-enantioselectivity to >99% e.e. in the reduction of (E)-citral or (E/Z)-citral. The results demonstrated that it was feasible to alter the enantioselectivity of OYEs through engineering key residue distant from active sites, e.g., R330 in OYE2y.

Protein Engineering of Saccharomyces cerevisiae Oxidosqualene-Lanosterol Cyclase into Parkeol Synthase

2012 ◽  
Vol 14 (20) ◽  
pp. 5222-5225 ◽  
Author(s):  
Yuan-Ting Liu ◽  
Tain-Chang Hu ◽  
Cheng-Hsiang Chang ◽  
Wen-Shiang Shie ◽  
Tung-Kung Wu
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Protein Engineering ◽  
Oxidosqualene Lanosterol Cyclase

Insights into the biochemical and functional characterization of sortase E transpeptidase of Corynebacterium glutamicum

2019 ◽  
Vol 476 (24) ◽  
pp. 3835-3847 ◽  
Author(s):  
Aliyath Susmitha ◽  
Kesavan Madhavan Nampoothiri ◽  
Harsha Bajaj
Keyword(s):  
Protein Engineering ◽  
Cell Attachment ◽  
Functional Characterization ◽  
Protein Structures ◽  
Structural Similarity ◽  
Surface Proteins ◽  
Sortase A ◽  
Peptide Cleavage ◽  
New Findings

Most Gram-positive bacteria contain a membrane-bound transpeptidase known as sortase which covalently incorporates the surface proteins on to the cell wall. The sortase-displayed protein structures are involved in cell attachment, nutrient uptake and aerial hyphae formation. Among the six classes of sortase (A–F), sortase A of S. aureus is the well-characterized housekeeping enzyme considered as an ideal drug target and a valuable biochemical reagent for protein engineering. Similar to SrtA, class E sortase in GC rich bacteria plays a housekeeping role which is not studied extensively. However, C. glutamicum ATCC 13032, an industrially important organism known for amino acid production, carries a single putative sortase (NCgl2838) gene but neither in vitro peptide cleavage activity nor biochemical characterizations have been investigated. Here, we identified that the gene is having a sortase activity and analyzed its structural similarity with Cd-SrtF. The purified enzyme showed a greater affinity toward LAXTG substrate with a calculated KM of 12 ± 1 µM, one of the highest affinities reported for this class of enzyme. Moreover, site-directed mutation studies were carried to ascertain the structure functional relationship of Cg-SrtE and all these are new findings which will enable us to perceive exciting protein engineering applications with this class of enzyme from a non-pathogenic microbe.

Anti-Saccharomyces cerevisiae Antibodies in Inflammatory Bowel Disease: a Family Study

2001 ◽  
Vol 36 (2) ◽  
pp. 196-201 ◽  
Author(s):  
F. Seibold ◽  
O. Stich ◽  
R. Hufnagl ◽  
S. Kamil ◽  
M. Scheurlen
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Inflammatory Bowel Disease ◽  
Bowel Disease ◽  
Family Study ◽  
Inflammatory Bowel
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