# US 11,299,706 B2 · Microbial bioconversion of curcuminoids to calebin-A

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- Number: US 11,299,706 B2
- Document ID: US11299706B2
- Status: Granted
- Kind: B2
- Application number: 16/281,120
- Filed: 2019-02-21
- Published: 2019-08-29
- Granted: 2022-04-12
- Earliest claimed: 2018-02-23
- Assignees: Sami-Sabinsa Group Limited
- Applicants: Muhammed Majeed; Kalyanam Nagabhushanam; Anju Majeed; Nooruddin Thajuddin; Sivakumar Arumugam; Krishnamurthy Kulithalai Viswanathan; Samuel Thomas Manoharan; Furqan Ali; John Adams Sebastian; Jamsheeda Moothedath; Muthuraman Gnanamani; Kirankumar Beede
- Inventors: Muhammed Majeed; Kalyanam Nagabhushanam; Anju Majeed; Nooruddin Thajuddin; Sivakumar Arumugam; Krishnamurthy Kulithalai Viswanathan; Samuel Thomas Manoharan; Furqan Ali; John Adams Sebastian; Jamsheeda Moothedath; Muthuraman Gnanamani; Kirankumar Beede
- Web page: https://patentgrep.com/patent/US11299706B2

## CPC

- C12N 1/145: Fungi isolates
- A61K 31/216: of acids having aromatic rings, e.g. benactizyne, clofibrate
- A61K 36/9066: Curcuma, e.g. common turmeric, East Indian arrowroot or mango ginger
- C12N 1/205: Bacterial isolates
- C12R 2001/38: Pseudomonas
- C12R 2001/645: Fungi {; Processes using fungi}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- C12R: INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS

## Abstract

The present invention discloses a novel endophytic fungi, Ovatospora brasiliensis MTCC 25236 for the bioconversion of curcuminoids to Calebin-A and a method for its isolation from the rhizomes of Curcuma sp. The invention also discloses a method for the bioconversion of curcuminoids to Calebin-A using an endophytic fungi Ovatospora brasiliensis MTCC 25236 and bacterial species, Acinetobacter johnsonii and Pseudomonas putida.

## Claims

### Claim 1

A method for the bioconversion of curcuminoids to Calebin-A using fungal or bacterial species, said method comprising steps of:
i. culturing the fungal or bacterial species in main two batches (flasks containing batch 1 and flasks containing batch 2) in 50 ml-500 ml volumes of suitable media and incubating at 35-37° C. with shaking at 100-120 rpm for 1 to 21 days;
ii. on the 7th day of incubation, adding varying concentrations of curcuminoids to batch 1 flasks of step i) and incubating at 37° C. with 120 rpm shaking;
iii. maintaining the batch 2 flasks of step i) without adding curcuminoids at 37° C. with 120 rpm shaking;
iv. harvesting media from batch 1 and batch 2 flasks of step ii) and step iii) after 24, 48, 72, 96, 120 hrs;
v. drying the harvested media of step iv) under vacuum and extracting using a suitable solvent;
vi. identifying the presence of Calebin-A using HPLC (high performance liquid chromatography), LC-MS (liquid chromatography-mass spectrometry) and NMR (nuclear magnetic resonance);
vii. refluxing the harvested media of step iv) with a suitable solvent followed by separating and drying the solvent layer under vacuum; and
viii. identifying the presence of Calebin-A using HPTLC, HPLC, LC-MS and NMR, wherein the fungal species is Ovatospora brasiliensis MTCC 25236, and the bacterial species is Acinetobacter johnsonii NCIMB 9871 or Pseudomonas putida NCIMB 10007.

### Claim 2 (depends on 1)

The method as in claim 1, wherein the media of step i) is selected from the group consisting of potato dextrose broth (PDB), Sabouraud Dextrose Broth, Malt Extract Broth, and Czapek Dox Broth.

### Claim 3 (depends on 1)

The method as in claim 1, wherein the solvent of step v) and vii) is selected from the group consisting of ethyl acetate, methanol, hexane, ethanol, and acetone.

### Claim 4 (depends on 1)

The method as in claim 1, wherein the solvent of step v) and vii) is ethyl acetate and methanol.