Recombinant chymotrypsin inhibitor 2: expression, kinetic analysis of inhibition with .alpha.-chymotrypsin and wild-type and mutant subtilisin BPN', and protein engineering to investigate inhibitory specificity and mechanism

Biochemistry ◽  
1990 ◽  
Vol 29 (31) ◽  
pp. 7339-7347 ◽  
Author(s):  
Colin Longstaff ◽  
Alison F. Campbell ◽  
Alan R. Fersht
Keyword(s):  
Protein Engineering ◽  
Kinetic Analysis ◽  
Wild Type ◽  
Chymotrypsin Inhibitor ◽  
Inhibitory Specificity ◽  
Chymotrypsin Inhibitor 2 ◽  
Expression Kinetic

Laser Flash-Induced Kinetic Analysis of CytochromefOxidation by Wild-Type and Mutant Plastocyanin from the CyanobacteriumNostocsp. PCC 7119†

Biochemistry ◽  
2005 ◽  
Vol 44 (34) ◽  
pp. 11601-11607 ◽  
Author(s):  
Cristina Albarrán ◽  
José A. Navarro ◽  
Fernando P. Molina-Heredia ◽  
Piedad del S. Murdoch ◽  
Miguel A. De la Rosa ◽  
...  
Keyword(s):  
Kinetic Analysis ◽  
Laser Flash ◽  
Wild Type

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) 

Immunoengineering approaches for cytokine therapy

2021 ◽  
Author(s):  
Aslan Mansurov ◽  
Abigail Lauterbach ◽  
Erica Budina ◽  
Aaron T. Alpar ◽  
Jeffrey A. Hubbell ◽  
...  
Keyword(s):  
Side Effects ◽  
Protein Engineering ◽  
Autoimmune Diseases ◽  
Inflammatory Cytokines ◽  
Therapeutic Efficacy ◽  
Cytokine Therapy ◽  
Toxicity Profile ◽  
Wild Type ◽  
Systemic Side Effects ◽  
Secondary Lymphoid Organs

Since the discovery of cytokines, much effort has been put forth to achieve therapeutic translation for treatment of various diseases, including cancer and autoimmune diseases. Despite these efforts, very few cytokines have cleared regulatory approval, and those that were approved are not commonly used due to their challenging toxicity profile and/or limited therapeutic efficacy. The main limitation in translation has been that wild-type cytokines have unfavorable pharmacokinetic and pharmacodynamic profiles, either eliciting unwanted systemic side effects or insufficient residence in secondary lymphoid organs. In this review, we address protein engineering approaches that have been applied to both pro- and anti-inflammatory cytokines to enhance their therapeutic indices, and we highlight diseases in which administration of engineered cytokines is especially relevant.

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