Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Methods, reagents and cells for biosynthesizing compounds

A technology of conversion and activity, applied in biochemical equipment and methods, organic chemistry, oxidoreductase, etc.

Inactive Publication Date: 2017-05-31
INVISTA TEXTILES (U K) LTD
View PDF3 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] However, no wild-type prokaryotes or eukaryotes naturally overproduce or secrete the C6 building blocks into the extracellular milieu

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Methods, reagents and cells for biosynthesizing compounds
  • Methods, reagents and cells for biosynthesizing compounds
  • Methods, reagents and cells for biosynthesizing compounds

Examples

Experimental program
Comparison scheme
Effect test

example C4-9

[0190] where C 3-8 Hydroxyalkyl is as defined herein. Example C 4-9 Hydroxyalkanoic acids include, but are not limited to, 6-hydroxycaproic acid (ie, 6-hydroxycaproic acid), 5-hydroxycaproic acid salt (ie, 5-hydroxycaproic acid), 4-hydroxycaproic acid salt (ie, 4-hydroxycaproic acid acid), 3-hydroxyhexanoate (ie, 3-hydroxyhexanoic acid), etc. Those skilled in the art will appreciate that the particular form will depend on pH (eg, neutral or ionized form, including any salt form thereof). In some embodiments, C 3-8 A hydroxyalkyl group is a group having the formula:

[0191]

[0192] In some embodiments, one or more hydroxy-substituted (C 4-9 Alkyl)-OC(=O)-(C 3-8 The alkyl) ester method is a method that produces one or more hexylhydroxy hexanoates. In some embodiments, the method includes:

[0193] a) enzymatically converting hexanoyl-CoA to hexyl hexanoate; and

[0194] b) enzymatically converting hexyl caproate to any of 6-hydroxyhexyl caproate, 6-hydroxyhexyl 6-h...

Embodiment 1

[0360] Enzymatic activity of ω-transaminase using adipate semialdehyde as substrate and forming 6-aminocaproic acid

[0361] The nucleotide sequence encoding the His tag was added to the omega-transaminases from Chromobacterium violaceum, Pseudomonas aeruginosa, Pseudomonas syringae, Sphaeroides spp. Rhodobacter, and vibrio genes (see Figure 9 ), allowing the production of ω-transaminases with a HIS tag at the N-terminus. Each resulting modified gene was cloned into a pET21a expression vector under the control of the T7 promoter, and each expression vector was transformed into a BL21[DE3] E. coli host. The resulting recombinant E. coli strain was grown in a 250 mL shake flask culture containing 50 mL LB medium and antibiotic selection pressure at 37°C, shaking at 230 rpm. Each culture was induced overnight at 16°C with 1 mM IPTG.

[0362] Pellets from each induced shake flask culture were harvested via centrifugation. Each pellet was resuspended and lysed via sonication. ...

Embodiment 2

[0368] Carboxylate reductase Enzyme activity that uses adipate as a substrate and forms adipate semialdehyde

[0369] The sequence encoding the HIS tag was added to the genes from Segniliparus rugosus and Segniliparus rotundus encoding the carboxylic acid reductases of SEQ ID NO: 4 and 6, respectively (see Figure 9 ), making it possible to generate a carboxylic acid reductase with a HIS tag at the N-terminus. Each modified gene was cloned into the pET Duet expression vector together with the sfp gene encoding the HIS-tagged phosphopantetheinyltransferase from Bacillus subtilis, both under the T7 promoter. Each expression vector was transformed into a BL21[DE3] E. coli host, and the resulting recombinant E. coli strains were grown at 37°C in 250 mL shake flask cultures containing 50 mL LB medium and antibiotic selection pressure with shaking at 230 rpm . Each culture was induced overnight at 37°C using autoinduction culture.

[0370] Pellets from each induced shake flask cu...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

This document describes biochemical pathways for producing 6- hydroxyhexanoate methyl ester and hexanoic acid hexyl ester using one or more of a fatty acid O-methyltransferase, an alcohol O-acetyltransferase and a monooxygenase, as well as recombinant hosts expressing one or more of such enzymes. 6-hydroxyhexanoate methyl esters and hexanoic acid hexyl ester can be enzymatically converted to adipic acid, adipate semialdehyde, 6-aminohexanoate, 6-hydroxyhexanoate, hexamethylenediamine, and 1,6-hexanediol.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of U.S. Provisional Application Serial Nos. 62 / 012,659, filed June 16, 2014, 62 / 012,666, filed June 16, 2014, and 62 / 012,604, filed June 16, 2014, which The entire contents are incorporated herein by reference. technical field [0003] The present invention relates to methods for the biosynthesis of methyl-6-hydroxycaproate and hexyl-caproate using one or more isolated enzymes such as fatty acid O-methyltransferase, alcohol O-acetyltransferase and monooxygenase, and recombinant host cells expressing one or more such enzymes. The present invention also relates to the enzymatic conversion of methyl 6-hydroxyhexanoate and hexyl hexanoate to 6-hydroxyhexanoate and 1,6 using one or more enzymes such as esterases, monooxygenases and demethylases - The hexanediol method. Furthermore, the present invention relates to the enzymatic conversion of 6-hydroxycaproic acid and / or 1,6-hexanediol to...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12P7/42C12P7/44C12P7/62C12P7/6436
CPCC12P7/44C12N9/1007C12P7/62C12N9/1096C12N9/0008C12N9/0077C12Y102/99006C12Y114/15001C12Y114/15003C12Y206/01018C12Y206/01019C12Y206/01029C12Y206/01048C12Y206/01082C12P7/42C12P7/6436C12P13/001C12P7/6409C12P7/24C07C59/01C08G63/06C12P7/40C07C47/02C07C47/12C07C59/325C07C223/02
Inventor A.L.博特斯A.V.E.康拉迪R.哈杜奇
Owner INVISTA TEXTILES (U K) LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com