NDLI: Ligation independent cloning (LIC) as a rapid route to families of recombinant biocatalysts from sequenced prokaryotic genomes

Please wait, while we are loading the content...

Asymmetric allylation of aryl aldehydes: studies on the scope and mechanism of the palladium catalysed diethylzinc mediated umpolung using phosphoramidite ligands

Synthesis of 3′-β-carbamoylmethylcytidine (CAMC) and its derivatives as potential antitumor agents

Positive effect of natural and negatively charged cyclodextrins on the stabilization of penicillins towards β-lactamase degradation due to inclusion and external guest–host association. An NMR and MS study

New mustard-linked 2-aryl-bis-benzimidazoles with anti-proliferative activity

Ligand-mediated enantioselective addition of lithium carbazolates to aldehydes

Trisaccharide mimetics of the aminoglycoside antibiotic neomycin

Photoinduced switching of intramolecular hydrogen bond between amide NH and carboxyl oxygen

Chemical synthesis of β-O-4 type artificial lignin

Facile synthesis of chromeno[4,3-b]quinolin-6-ones from unexpected reactions of aryl isocyanides with 4-chloro-2-oxo-2H-chromene-3-carbaldehyde

A computational exploration of some transnitrosation and thiolation reactions involving CH₃SNO, CH₃ONO and CH₃NHNO

Synthesis and antimicrobial activity of N-analogous corollosporines

Evaluation of sparteine-like chiral diamines in the enantioselective lithiation–electrophilic trapping of an O-alkyl carbamate

Synthesis and properties of nanospheres copolymerised with luminescent europium(iii) chelates

Synthesis of the ABC tricyclic fragment of the pectenotoxins via stereocontrolled cyclization of a γ-hydroxyepoxide appended to the AB spiroacetal unit

Synthesis and biological activity of tetralone abscisic acid analogues

A novel heterobifunctional linker for facile access to bioconjugates

Michael addition of thiols to α-enones in ionic liquids with and without organocatalysts

Back matter

Back cover

Organic & Biomolecular Chemistry Vol - 4 Issue - 6

Organic & Biomolecular Chemistry Vol - 4 Issue - 5

Organic & Biomolecular Chemistry Vol - 4 Issue - 4

Organic & Biomolecular Chemistry Vol - 4 Issue - 3

Organic & Biomolecular Chemistry Vol -3

Organic & Biomolecular Chemistry Vol -2

Ligation independent cloning (LIC) as a rapid route to families of recombinant biocatalysts from sequenced prokaryotic genomes

Content Provider	Royal Society of Chemistry (RSC)
Author	Bonsor, Daniel Grant, Stephanie Fogg, Mark J. Solomons, Julianna Butz, Stephanie F. Fairlamb, Ian J. S. Grogan, Gideon
Copyright Year	2006
Abstract	A technique is presented for the high throughput generation of families of recombinant biocatalysts sourced from prokaryotic genomes, providing rapid access to the naturally evolved diversity of enzyme specificity for biocatalyst discovery. The method exploits a novel ligation independent cloning strategy, based on the locally engineered vector pET-YSBLIC and has been used for the rapid generation of a suite of expression plasmids containing genes encoding a family of six Baeyer–Villiger monooxygenases (BVMOs) from Mycobacterium tuberculosis H37Rv (MTb). The six resultant recombinant strains of E. coli B834 (DE3) expressing the genes were assayed for oxygenating activity in respect of the target reaction; the resolution of bicyclo[3.2.0]hept-2-en-6-one. The analysis of biotransformations catalysed by growing cells of E. coli was complicated by the production of indole in the reaction mixtures, possibly resulting from the in vivo activity of E. coli tryptophanase. Four of the recombinant strains expressing different BVMOs catalysed the oxidation of one or more of four screening substrates, well above controls that had been transformed with the re-ligated parent vector. One of the recombinant strains, E. coli B834 (DE3) pDB5, expressing the Rv3049c gene from MTb, was found to effectively resolve the target substrate, yielding a 19% yield of (1R, 5S)-(+)-bicyclo[3.2.0]hept-2-en-6-one with >95% enantiomeric excess in a 4 L fermentation reaction.
Starting Page	1252
Ending Page	1260
Page Count	9
File Format	HTM / HTML PDF
ISSN	14770520
Volume Number	4
Issue Number	7
Journal	Organic & Biomolecular Chemistry
DOI	10.1039/b517338h
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Open
Subject Keyword	DNA ligase Enzyme Euclidean vector Baeyer Escherichia coli Indole coli Tryptophanase Fermentation Mycobacterium tuberculosis Gene Horse length Prokaryote Enantiomeric excess
Content Type	Text
Resource Type	Article
Subject	Organic Chemistry Biochemistry Physical and Theoretical Chemistry

Sl.	Authority	Responsibilities	Communication Details
1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
5	Website/Portal (Helpdesk)	Queries regarding NDLI and its services	support@ndl.gov.in
6	Contents and Copyright Issues	Queries related to content curation and copyright issues	content@ndl.gov.in
7	National Digital Library of India Club (NDLI Club)	Queries related to NDLI Club formation, support, user awareness program, seminar/symposium, collaboration, social media, promotion, and outreach	clubsupport@ndl.gov.in
8	Digital Preservation Centre (DPC)	Assistance with digitizing and archiving copyright-free printed books	dpc@ndl.gov.in
9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

Escherichia coli and mycobacterium tuberculosis.

Bacillus stearothermophilus NAD+ -dependent DNA ligase Escherichia coli NAD+ -dependent DNA ligase Enterococcus faecalis NAD+ -dependent DNA ligase Helix-hairpin-helix Human DNA ligase I NAD+ -dependent DNA ligase(s) Mycobacterium tuberculosis NAD+ -dependent DNA ligase Mycobacterium tuberculosis Ly

Cloning and expression of Mycobacterium tuberculosis and Mycobacterium leprae dihydropteroate synthase in Escherichia coli.

L-tryptophan production by Escherichia coli in the presence of Iranian cane molasses

Substrate Specificity and Enantioselectivity of (2002)

Ligation, Transformation and Characterization of Rv 1926c Mycobacterium tuberculosis to Escherichia coli JM 109 For Latent Tuberculosis Immunodiagnostic

Recombinant Expressed Vector pET32a (+) S Constructed by Ligation Independent Cloning

Cloning, expression, identification and bioinformatics analysis of Rv3265c gene from Mycobacterium tuberculosis in Escherichia coli.

Cloning and expression in Escherichia coli of a recA homologue from Mycobacterium tuberculosis.

Ligation independent cloning (LIC) as a rapid route to families of recombinant biocatalysts from sequenced prokaryotic genomes

Similar Documents

Escherichia coli and mycobacterium tuberculosis.

Cloning and expression of Mycobacterium tuberculosis and Mycobacterium leprae dihydropteroate synthase in Escherichia coli.

L-tryptophan production by Escherichia coli in the presence of Iranian cane molasses

Substrate Specificity and Enantioselectivity of (2002)

Ligation, Transformation and Characterization of Rv 1926c Mycobacterium tuberculosis to Escherichia coli JM 109 For Latent Tuberculosis Immunodiagnostic

Recombinant Expressed Vector pET32a (+) S Constructed by Ligation Independent Cloning

Cloning, expression, identification and bioinformatics analysis of Rv3265c gene from Mycobacterium tuberculosis in Escherichia coli.

Cloning and expression in Escherichia coli of a recA homologue from Mycobacterium tuberculosis.

Ligation independent cloning (LIC) as a rapid route to families of recombinant biocatalysts from sequenced prokaryotic genomes