Supercharge Your Innovation With Domain-Expert AI Agents!

Nepetalactol oxidoreductases, nepetalactol synthases, and microbes capable of producing nepetalactone

A technology of nepetalactone and lactone, which is applied in the directions of oxidoreductase, biochemical equipment and methods, enzymes, etc., and can solve the problems such as the effective chemical enzymatic method and overview of nepetalactone that are not recorded.

Pending Publication Date: 2020-10-30
ZYMERGEN INC
View PDF16 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, prior to this study, the complete pathway in any microorganism had not been outlined nor was an efficient chemoenzymatic method for the production of nepetalactone from the precursor documented.

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
  • Nepetalactol oxidoreductases, nepetalactol synthases, and microbes capable of producing nepetalactone
  • Nepetalactol oxidoreductases, nepetalactol synthases, and microbes capable of producing nepetalactone
  • Nepetalactol oxidoreductases, nepetalactol synthases, and microbes capable of producing nepetalactone

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2

[0471]Example 2 and the Sequence Listing submitted herein provide an exemplary set of nucleotide sequences encoding these enzymes. These polynucleotides comprising these sequences or variants thereof can be used to reconstitute the nepetalactone pathway in microorganisms. Due to the degeneracy of the genetic code, one or more nucleotide bases can be substituted without changing the amino acid sequence of the encoded protein. Variants of these polynucleotides include, for example, those having sequences codon optimized for expression in a particular host cell. In various embodiments, variants of these polynucleotides include those having at least 60, 65, 70, 75, 80, Sequences with 85, 90, 95, 96, 97, 98 or 99% sequence identity.

[0472] Novel polynucleotide encoding nepetalactone oxidoreductase

[0473] Examples 1 and 6 describe the identification and isolation of a polynucleotide comprising a nucleotide sequence encoding an active Nepeta lactal oxidoreductase (NOR). Nuc...

Embodiment 4

[0480] Example 4 describes the identification and isolation of a polynucleotide comprising a nucleotide sequence encoding an active nepetalactal synthase (NEPS). Nucleotide sequences encoding tested and / or putative NEPS are described in the accompanying Sequence Listing, eg, SEQ ID NO: 1506-1562.

[0481] These polynucleotides or variants thereof can be used to generate NEPS. Due to the degeneracy of the genetic code, one or more nucleotide bases can be substituted without changing the amino acid sequence of the encoded protein. Variants of NEPS polynucleotide sequences include, for example, those having sequences codon optimized for expression in a particular host cell. In various embodiments, variants of NEPS polynucleotides have at least 60, 65, 70, 75 , 80, 85, 90, 95, 96, 97, 98 or 99% sequence identity.

[0482] New Nepeta Lactal Synthase Polypeptides

[0483] The deduced amino acid sequences of these NEPS are also provided in the Sequence Listing (SEQ ID NO: 718-7...

Embodiment 1

[0561] Example 1 - Nepetalactone redox in Escherichia coli capable of converting nepetalactone to nepetalactone Cloning and expression of the proenzyme

[0562] Identify NOR candidates

[0563] Publicly available next-generation RNA sequencing data from Nepeta cataria (SRR5150709) were obtained from NCBI. Reads are extracted and composed into transcriptomes. Using the protein sequence of horse liver alcohol dehydrogenase (HLADH) as a BLAST query, alcohol dehydrogenase candidates from Nepeta sativa that might catalyze the conversion of nepetalactin to nepetalactone were identified.

[0564] Thirty-nine candidates were identified and the coding sequences were codon optimized for expression in E. coli. The codon-optimized nucleotide sequence was synthesized by Integrated DNA Technologies (IDT) using the upstream T7 promoter and ribosome binding site (RBS) and downstream T7 terminator sequence. Synthetic DNA was recovered as a plasmid (provided by IDT) containing the expr...

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

PropertyMeasurementUnit
optical densityaaaaaaaaaa
optical densityaaaaaaaaaa
Login to View More

Abstract

The present disclosure provides isolated nepetalactone oxidoreductase polypeptides (NORs), nepetalactol synthases (NEPSs), and related polynucleotides, engineered host cells, and cultures, as well asmethods for producing NORs and NEPSs, and for using them to produce nepetalactol, nepetalactone, and dihydronepetalactone. The present disclosure also provides methods for engineering cells (e.g., microbial cells) to produce nepetalactone from a fermentation substrate such as glucose, as well as engineered cells having this capability and related cultures and methods for producing nepetalactone.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of U.S. Provisional Application 62 / 609,272 filed December 21, 2017, U.S. Provisional Application 62 / 609,279, filed December 21, 2017, and U.S. Provisional Application 62 / 669,919, filed May 10, 2018 , the entire contents of these provisional applications are incorporated into this application by reference. [0003] Incorporated by reference into the sequence listing [0004] This application includes a Sequence Listing, which is filed electronically in ASCII format and is incorporated by reference in its entirety. Said ASCII copy was created on December 21, 2018, is named ZMGNP012WO_SL.txt, and is 5,630,568 bytes in size. technical field [0005] The present disclosure relates broadly to the field of nepetalactone production. In particular, the present disclosure relates to newly discovered enzymes encoding nepetalactal oxidoreductase (NOR), which catalyzes the oxidation of nepetalac...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C07D311/94
CPCC12Y103/01C07D311/94C12N1/20C12N9/0006C12N9/001C12Y101/01C12N15/52C12P5/007C12P17/06C12Y101/01324C12N15/70C12N15/81
Inventor G·瓦沃泽恩C·罗契P·韦斯特福尔W·劳S·加内桑F·麦克索利Z·赛尔贝尔
Owner ZYMERGEN INC
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

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

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com