Method of production of monosaccharides
AssigneePetiva Private Limited
InventorsSaravanakumar Iyappan, Samir Kumar Roy, Saju Varghese, Aparna Devi Addala, Venkata Narayanan Karthikeyan, Kanumuru Rahul Raju, Banibrata Pandey
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Abstract
The present invention is directed towards genetic modification of native gene encoding for D-tagatose 3-epimerase and rhamnose isomerase to substantially increase the expression level of these enzymes and use of the enzymes in a process to produce rare monosaccharides such as psicose and allose. Also disclosed in the present invention is expression constructs comprising the modified genes and a host cells to express the same.
Claims13
A modified polynucleotide sequence comprising SEQ ID NO:
- 1 encoding D-tagatose 3-epimerase;
- or a modified polynucleotide sequence comprising SEQ ID NO:
- 2 encoding rhamnose isomerase.
A process of production of recombinant D-tagatose 3-epimerase or rhamnose isomerase, said process comprising the steps of:
- 1. culturing host cell transformed with an expression construct comprising SEQ ID NO:
- 1 or SEQ ID NO:
- 2 in a suitable medium in presence of IPTG or lactose for a period in the range of 2-3 hours,
- 2. isolation of expressed protein from the host cells by conventional method, and
- 3. purifying the recombinant proteins using chromatographic techniques.
A process of overproduction of rare monosaccharides from fructose, said process comprising the steps of:
- 1. culturing host cells transformed with an expression construct comprising SEQ ID NO 1:
- and SEQ ID NO:
- 2 in a separate suitable medium in presence of IPTG or lactose for a period in the range of 2-3 hours to produce D-tagatose 3-epimerase and rhamnose isomerase respectively,
- 2. isolating the expressed protein from the host cells by conventional method, and purifying the isolated protein using chromatographic techniques,
- 3. immobilizing D-tagatose 3-epimerase and rhamnose isomerase thus produced in the previous step in a suitable matrix,
- 4. contacting fructose with immobilized D-tagatose 3-epimerase for a period in the range of 5 to 10 hours to produce psicose, and
- 5. contacting D-psicose produced in the previous step with immobilized rhamnose isomerase for a period in the range of 6-12 hours to produce D-allose.
Dependencies are inferred from the claim text.