Improved Enantioselectivity of Subtilisin Carlsberg towards Secondary Alcohols by Protein Engineering

ChemBioChem ◽  
2017 ◽  
Vol 19 (4) ◽  
pp. 338-346
Author(s):  
Robin Dorau ◽  
Tamás Görbe ◽  
Maria Svedendahl Humble
Keyword(s):  
Protein Engineering ◽  
Secondary Alcohols ◽  
Subtilisin Carlsberg

scholarly journals Novel oleate hydratases and potential biotechnological applications

2021 ◽  
Author(s):  
Peter Leon Hagedoorn ◽  
Frank Hollmann ◽  
Ulf Hanefeld
Keyword(s):  
Fatty Acids ◽  
Protein Engineering ◽  
High Throughput Screening ◽  
Unsaturated Fatty Acids ◽  
Dicarboxylic Acids ◽  
Secondary Amines ◽  
Secondary Alcohols ◽  
Product Portfolio ◽  
Conversion Rates ◽  
Wide Range

Abstract Oleate hydratase catalyses the addition of water to the CC double bond of oleic acid to produce (R)-10-hydroxystearic acid. The enzyme requires an FAD cofactor that functions to optimise the active site structure. A wide range of unsaturated fatty acids can be hydrated at the C10 and in some cases the C13 position. The substrate scope can be expanded using ‘decoy’ small carboxylic acids to convert small chain alkenes to secondary alcohols, albeit at low conversion rates. Systematic protein engineering and directed evolution to widen the substrate scope and increase the conversion rate is possible, supported by new high throughput screening assays that have been developed. Multi-enzyme cascades allow the formation of a wide range of products including keto-fatty acids, secondary alcohols, secondary amines and α,ω-dicarboxylic acids. Key points • Phylogenetically distinct oleate hydratases may exhibit mechanistic differences. • Protein engineering to improve productivity and substrate scope is possible. • Multi-enzymatic cascades greatly widen the product portfolio.

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.

Protein Engineering of Penicillin Acylase

Acta Naturae ◽  
2010 ◽  
Vol 2 (3) ◽  
pp. 47-61 ◽  
Author(s):  
V I Tishkov ◽  
S S Savin ◽  
A S Yasnaya
Keyword(s):  
Protein Engineering ◽  
Penicillin Acylase

Application of Dess-Martin oxidation in total synthesis of natural products

2020 ◽  
Vol 17 ◽  
Author(s):  
Majid M. Heravi ◽  
Tayebe Momeni ◽  
Vahideh Zadsirjan ◽  
Leila Mohammadi
Keyword(s):  
Natural Products ◽  
Total Synthesis ◽  
Selective Oxidation ◽  
Secondary Alcohols ◽  
Aldehydes And Ketones

: Dess–Martin periodinane (DMP), is a commercially available chemical, frequently being utilized as a mild oxidative agent for the selective oxidation of primary and secondary alcohols to their corresponding aldehydes and ketones, respectively. DMP shows several merits over other common oxidative agent such as chromium- and DMSO-based oxidants, thus it is habitually employed in the total synthesis of natural products. In this review, we try to underscore the applications of DMP as an effective oxidant in an appropriate step (steps) in the multistep total synthesis of natural products.

Oxidation rates of triterpenoid secondary alcohols with chromic acid

1976 ◽  
Vol 41 (3) ◽  
pp. 770-779 ◽  
Author(s):  
M. R. Nair ◽  
S. Hilgard ◽  
J. Klinot ◽  
K. Waisser ◽  
A. Vystrčil
Keyword(s):  
Chromic Acid ◽  
Secondary Alcohols ◽  
Oxidation Rates
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