A patent-based consideration of latest platforms in the art of directed evolution: a decade long untold story

Iqbal, Zarina; Sadaf, Saima

doi:10.1080/02648725.2021.2017638

Cited by 3 publications

(4 citation statements)

References 218 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Directed evolution relies on creating mutant libraries via iterative random mutagenesis (using PCR techniques, chemical mutagenesis, UV irradiation, or DNA-shuffling techniques) [ 12 , 34 , 35 ]. Systematic screening and evaluation processes are required to identify the enzymes having the desired properties [ 36 ].…”

Section: Strategies and Challenges For The Development Of Engineered ...mentioning

confidence: 99%

Bioengineered Enzymes and Precision Fermentation in the Food Industry

Boukid¹,

Ganeshan²,

Wang³

et al. 2023

IJMS

View full text Add to dashboard Cite

Enzymes have been used in the food processing industry for many years. However, the use of native enzymes is not conducive to high activity, efficiency, range of substrates, and adaptability to harsh food processing conditions. The advent of enzyme engineering approaches such as rational design, directed evolution, and semi-rational design provided much-needed impetus for tailor-made enzymes with improved or novel catalytic properties. Production of designer enzymes became further refined with the emergence of synthetic biology and gene editing techniques and a plethora of other tools such as artificial intelligence, and computational and bioinformatics analyses which have paved the way for what is referred to as precision fermentation for the production of these designer enzymes more efficiently. With all the technologies available, the bottleneck is now in the scale-up production of these enzymes. There is generally a lack of accessibility thereof of large-scale capabilities and know-how. This review is aimed at highlighting these various enzyme-engineering strategies and the associated scale-up challenges, including safety concerns surrounding genetically modified microorganisms and the use of cell-free systems to circumvent this issue. The use of solid-state fermentation (SSF) is also addressed as a potentially low-cost production system, amenable to customization and employing inexpensive feedstocks as substrate.

show abstract

Section: Strategies and Challenges For The Development Of Engineered ...mentioning

confidence: 99%

Bioengineered Enzymes and Precision Fermentation in the Food Industry

Boukid¹,

Ganeshan²,

Wang³

et al. 2023

IJMS

View full text Add to dashboard Cite

show abstract

“…In 2010, Codexis Inc. disclosed the bioreduction of (E)-methyl-2-(3-(3-(2-(7-chloroquinolyn-2-yl)vinyl)phenyl)oxopropyl)benzoate to the optically pure (S)-16 catalyzed by the KRED CDX-026 [61]. A panel of these biocatalysts which showed activity on the starting material were subjected to directed evolution; this is a consecutive methodology of random mutations and high-throughput screening used to identify the best mutants, which are used for further rounds of mutagenesis, picking out the best example from each generation, and so gradually evolving the enzyme's structure [22][23][24]. In this case, the desired target was to improve the enzymatic activity and (thermal) stability.…”

Section: Redox Enzymes: Selected Examples In the Synthesis Of Apismentioning

confidence: 99%

“…Two different solutions could be proposed; on the one hand, rational design, which requires making a logical guess about which amino acids in an enzyme must be altered to achieve the desired enzymatic improvement, and making the amino acids replacement by site-directed mutagenesis of the corresponding gene [85,86]. On the other hand, directed evolution, an already-mentioned genetic improvement (see section 2.2) [22][23][24]87], which, worth to be mentioned, granted F. Arnold the Chemistry Nobel Prize in 2018 [88]. In this particular case, for searching a TA suitable to be used for the transamination of 20, a hybrid of the two mentioned approaches was used.…”

Section: Selected Examples Of Transaminases In the Preparation Of Apismentioning

confidence: 99%

See 1 more Smart Citation

Biocatalysis for the asymmetric synthesis of Active Pharmaceutical Ingredients (APIs): this time is for real

Gonzalo

Alcántara

Marı́a³

et al. 2022

Expert Opinion on Drug Discovery

View full text Add to dashboard Cite

Introduction: Biocatalysis has emerged as a powerful and useful strategy for the synthesis of active pharmaceutical ingredients (APIs). The outstanding developments in molecular biology techniques allow nowadays the screening, large-scale production, and designing of biocatalysts, adapting them to desired reactions. Many enzymes can perform reactions both in aqueous and non-aqueous media, broadening even further the opportunities to integrate them in complex pharmaceutical multi-step syntheses.Areas covered: This paper showcases several examples of biocatalysis in the pharmaceutical industry, covering examples of different enzymes, such as lipases, oxidoreductases, and transaminases, to deliver active drugs through complex synthetic routes. Examples are critically discussed in terms of reaction conditions, motivation for using an enzyme, and how biocatalysts can be integrated in multi-step syntheses. When possible, biocatalytic routes are benchmarked with chemical reactions. Expert Opinion: The reported enzymatic examples are performed with high substrate loadings (>100 g L −1 ) and with excellent selectivity, making them inspiring strategies for present and future industrial applications. The combination of powerful molecular biology techniques with the needs of the pharmaceutical industry can be aligned, creating promising platforms for synthesis under more sustainable conditions.

show abstract

Baicalein Protects Against Iron Overload Induced Liver Dysfunction in Thalassemic Mice

Fan,

Chen,

Yang

et al. 2023

j biomed nanotechnol

View full text Add to dashboard Cite

Liver iron overload is a common and serious organ injury in β-thalassemia patients. Ferroptosis has been shown to play a crucial role in the pathological injury in iron overloaded hepatocytes. In our study, we focused on the protective effects of baicalein, a natural, active flavone extracted from an herb used in traditional Chinese medicine against RSL3-induced ferroptosis in hepatocytes and high iron diet (HID) induced liver iron-overload in a murine β654-thalassemia model. In vitro, the effects of baicalein on RSL3-induced hepatocyte ferroptosis were examined by testing ferroptosis related genes, protein, and the GSH, MDA, iron ion level by RT-PCR, Western blot and the commercial kits respectively. The anti-iron overload injury effects of baicalein were assessed in a β654-thalassemia mouse model of high iron diet induced liver injury by the same method In vitro experiments. Here, we found that baicalein could reverse cell ferroptosis in hepatocytes treated with RSL3. Importantly, changes in the iron ion content, and MDA and GSH levels in β654-thalassemia mouse livers were significantly restored by baicalein. Mechanistically, baicalein may activated the NRF2 antioxidant pathway and increased the expression of GPX4 in vivo and In vitro. We concluded that baicalein is a potential therapeutic drug for the treatment of iron overload in β-thalassemia.

show abstract

A patent-based consideration of latest platforms in the art of directed evolution: a decade long untold story

Cited by 3 publications

References 218 publications

Bioengineered Enzymes and Precision Fermentation in the Food Industry

Bioengineered Enzymes and Precision Fermentation in the Food Industry

Biocatalysis for the asymmetric synthesis of Active Pharmaceutical Ingredients (APIs): this time is for real

Baicalein Protects Against Iron Overload Induced Liver Dysfunction in Thalassemic Mice

Contact Info

Product

Resources

About