IP Library Granted Patent US 11,001,838
Granted Patent B2
US 11,001,838 · App. 15/943,632 · Granted May 11, 2021

Platform for screening phosphomevalonate decarboxylase

Inventors: Taek Soon Lee (Berkeley, CA); Aram Kang (Richmond, CA)
Assignee: The Regents of the University of California
C12N15/1079C12N9/88C12N15/01C12Y401/01033
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Quick Facts
Patent No.
US 11,001,838
App. No.
15/943,632
Granted
May 11, 2021
Kind
B2
Abstract

The present invention provides for a method to identify a second or mutant phosphomevalonate decarboxylase (PMD) with a higher PMD activity compared to a first PMD, comprising (a) culturing a medium comprising a first host cell expressing the first PMD and a second host cell expressing the second or mutant PMD wherein the first and second host cells have their respective PMD enzymatic activities coupled to the growth rates of the host cells, and (b) identifying the second host cell that has a higher growth rate than the first host cell, thereby identifying the second or mutant PMD having a higher PMD activity.

Claims (4)

1. A method to identify a second or mutant phosphomevalonate decarboxylase (PMD) with a higher PMD activity compared to a first PMD, comprising (a) culturing a medium comprising a first host cell expressing the first PMD and a second host cell expressing the second or mutant PMD wherein the first and second host cells have their respective PMD enzymatic activities coupled to the growth rates of the host cells, wherein the host cell is microorganism which produces IPP via the MEP pathway transformed with one or more polynucleotides encoding 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR), 3-hydroxy-3-methylglutaryl-coenzyme A synthase (HMGS), mevalonate kinase (MK) and isopentenyl phosphate kinase (IPK) and treated with fosmidomycin, and (b) identifying the second host cell that has a higher growth rate than the first host cell, thereby identifying the second or mutant PMD having a higher PMD activity; wherein the host cell is a prokaryotic cell.

2. The method of claim 1 , wherein the prokaryotic cell is an Escherichia, Enterobacter, Azotobacter, Erwinia, Bacillus, Pseudomonas, Klebsielia, Proteus, Salmonella, Serratia, Shigella, Rhizobia, Vitreoscilla, Paracoccus , or Clostridia cell.

3. The method of claim 2 , wherein the prokaryotic cell is an Escherichia cell.

4. The method of claim 3 , wherein the Escherichia cell is an Escherichia coli cell.

Assignments (2)
CONFIRMATORY LICENSE Recorded Apr 16, 2019
From: UNIVERSITY OF CALIF-LAWRENC BERKELEY LAB
To: UNITED STATES DEPARTMENT OF ENERGY
Reel/Frame 048899/0538 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Apr 5, 2018
From: LEE, TAEK SOON; KANG, ARAM
To: THE REGENTS OF THE UNIVERSITY OF CALIFORNIA
Reel/Frame 045448/0347 →
Continuity (2)
Provisional Application 62480279 · Mar 31, 2017
Related Publication 20180362970A1 · Dec 20, 2018