Compositions and methods for clostridial transformation

• US 9,926,568 B2
• Filed: 06/20/2014
• Issued: 03/27/2018
• Est. Priority Date: 06/21/2013
• Status: Expired due to Fees

• Current Assignee: Danisco US, Inc. (International Flavors & Fragrances Incorporated)
• Original Assignee: Danisco US, Inc. (International Flavors & Fragrances Incorporated)
• Inventors: Scotcher, Miles C., Wells, Derek H.
• Primary Examiner(s): LYONS, MARY M
• Application Number:
  US14/899,846
• Publication Number:
  US 20160138030A1
• Time in Patent Office: 1,376 Days
• Field of Search: None
• CPC Class Codes: C12N 15/70 Vectors or expression systems specially adapted for E. coli
• CPC Class Codes: C12N 15/74 Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora
• CPC Class Codes: C12N 9/1007 Methyltransferases (general) (2.1.1.)
• CPC Class Codes: C12P 5/007 containing one or more isoprene units, i.e. terpenes carotenes C12P23/00
• CPC Class Codes: C12P 7/065 with microorganisms other than yeasts
• CPC Class Codes: C12Y 201/01 Methyltransferases (2.1.1)
• CPC Class Codes: Y02E 50/10 Biofuels, e.g. bio-diesel

Abstract

The invention provides compositions and methods for clostridial bacteria that have been engineered to produce and/or to improve efficiency of production of industrial bioproducts.

Claims

1. A method of producing a recombinant Clostridium bacterial transformant, comprising:

a) introducing a polynucleotide encoding for a DNA methyltransferase into a Escherichia bacterial host cell, wherein the polynucleotide encoding for a DNA methyltransferase has at least 90% sequence identity to SEQ ID NO;

1,

b) culturing the Escherichia bacterial host cell under conditions suitable for expression of the DNA methyltransferase, wherein the encoded DNA methyltransferase methylates a polynucleotide,

c) transferring the methylated polynucleotide from the Escherichia bacterial host cell to a Clostridium bacterial host cell, wherein the bacteria transformed using this method are selected from the group consisting of Clostridium aceticum, Clostridium ljungdahlii, Clostridium acetobutylicum, and Clostridium autoethanogenum.

2. The method of claim 1, wherein the polynucleotide encoding for a DNA methyltransferase is SEQ ID NO:

2.

3. The method of claim 1, wherein the encoded DNA methyltransferase methylates a polynucleotide at a sequence comprising CCWGG.

4. The method of claim 3, wherein the sequence comprising CCWGG is selected from the group consisting of CCAGG (SEQ ID NO:

9) and CCTGG (SEQ ID NO;

     10).