Method for biologically producing acetin compound
A method for biologically producing acetin such as monoacetin, diacetin, or triacetin according to an embodiment of the present disclosure includes reacting acetyl-CoA with glycerol in the presence of a first O-acetyl transferase to obtain the acetin. With the method, acetin which is sustainable and safe, and has more excellent quality while not causing environmental pollution may be obtained.
1. A method for production of acetin, the method comprising:
preparing a transformed microorganism including a gene encoding a first O-acetyl transferase to express the first O-acetyl transferase; and
reacting acetyl-CoA with glycerol in the presence of the first O-acetyl transferase to obtain the acetin,
wherein the first O-acetyl transferase is maltose O-acetyl transferase.
2. The method according to claim 1 , wherein an amino acid sequence of the first O-acetyl transferase has a sequence motif comprising:
aspartic acid (ASP) at a position corresponding to position 70 of the amino acid sequence of SEQ ID NO: 5;
asparagine (ASN) at a position corresponding to position 84 of the amino acid sequence of SEQ ID NO: 5;
histidine (HIS) at a position corresponding to position 114 of the amino acid sequence of SEQ ID NO: 5; and
glutamic acid (GLU) at a position corresponding to position 126 of the amino acid sequence of SEQ ID NO: 5.
3. The method according to claim 2 , wherein an amino acid sequence of the first O-acetyl transferase has a sequence motif comprising:
tyrosine (TYR) or phenylalanine (PHE) at a position corresponding to position 15 of the amino acid sequence of SEQ ID NO: 5;
arginine (ARG) or glutamine (GLN) at a position corresponding to position 26 of the amino acid sequence of SEQ ID NO: 5;
arginine (ARG) or lysine (LYS) at a position corresponding to position 30 of the amino acid sequence of SEQ ID NO: 5; and
phenylalanine (PHE) or tyrosine (TYR) at a position corresponding to position 82 of the amino acid sequence of SEQ ID NO: 5.
4. The method according to claim 1 , wherein the first O-acetyl transferase is composed of any one sequence of SEQ ID NOS: 1 to 6.
5. The method according to claim 1 , wherein the acetin comprises monoacetin and diacetin, and the method further comprises reacting the diacetin as a reaction product with acetyl-CoA in the presence of chloramphenicol-O-acetyl transferase or a second O-acetyl transferase, thus to obtain triacetin.
6. The method according to claim 5 , wherein an amino acid sequence of the second O-acetyl transferase has a sequence motif comprising:
cysteine (CYS) or leucine (LEU) at a position corresponding to position 31 of the amino acid sequence of SEQ ID NO: 7;
phenylalanine (PHE) or isoleucine (ILE) at a position corresponding to position 102 of the amino acid sequence of SEQ ID NO: 7;
phenylalanine (PHE) or tyrosine (TYR) at a position corresponding to position 143 of the amino acid sequence of SEQ ID NO: 7;
phenylalanine (PHE) or tyrosine (TYR) at a position corresponding to position 166 of the amino acid sequence of SEQ ID NO: 7;
valine (VAL), isoleucine (ILE) or leucine (LEU) at a position corresponding to position 170 of the amino acid sequence of SEQ ID NO: 7; and
histidine (HIS) at a position corresponding to position 193 of the amino acid sequence of SEQ ID NO: 7.
7. The method according to claim 5 , wherein the chloramphenicol-O-acetyl transferase is composed of the sequence of SEQ ID NO: 7, and the second O-acetyl transferase is composed of any one sequence of SEQ ID NOS: 8 to 10.
8. A method for production of acetin, the method comprising:
preparing a transformed microorganism that expresses a gene encoding a first O-acetyl transferase; and
culturing the transformed microorganism in a medium containing glycerol to produce the acetin,
wherein the first O-acetyl transferase is maltose O-acetyl transferase.
9. The method according to claim 8 , wherein an amino acid sequence of the first O-acetyl transferase has a sequence motif comprising:
aspartic acid (ASP) at a position corresponding to position 70 of the amino acid sequence of SEQ ID NO: 5;
asparagine (ASN) at a position corresponding to position 84 of the amino acid sequence of SEQ ID NO: 5;
histidine (HIS) at a position corresponding to position 114 of the amino acid sequence of SEQ ID NO: 5; and
glutamic acid (GLU) at a position corresponding to position 126 of the amino acid sequence of SEQ ID NO: 5.
10. The method according to claim 8 , wherein an amino acid sequence of the first O-acetyl transferase has a sequence motif comprising:
tyrosine (TYR) or phenylalanine (PHE) at a position corresponding to position 15 of the amino acid sequence of SEQ ID NO: 5;
arginine (ARG) or glutamine (GLN) at a position corresponding to position 26 of the amino acid sequence of SEQ ID NO: 5;
arginine (ARG) or lysine (LYS) at a position corresponding to position 30 of the amino acid sequence of SEQ ID NO: 5; and
phenylalanine (PHE) or tyrosine (TYR) at a position corresponding to position 82 of the amino acid sequence of SEQ ID NO: 5.
11. The method according to claim 8 , wherein the gene encoding the first O-acetyl transferase is composed of any one sequence of SEQ ID NOS: 16 to 21.
12. The method according to claim 8 , wherein the transformed microorganism further expresses a gene encoding chloramphenicol-O-acetyl transferase or a second O-acetyl transferase that transfers an acetyl group to diacetin.
13. The method according to claim 12 , wherein an amino acid sequence of the second O-acetyl transferase has a sequence motif comprising:
cysteine (CYS) or leucine (LEU) at a position corresponding to position 31 of the amino acid sequence of SEQ ID NO: 7;
phenylalanine (PHE) or isoleucine (ILE) at a position corresponding to position 102 of the amino acid sequence of SEQ ID NO: 7;
phenylalanine (PHE) or tyrosine (TYR) at a position corresponding to position 143 of the amino acid sequence of SEQ ID NO: 7;
phenylalanine (PHE) or tyrosine (TYR) at a position corresponding to position 166 of the amino acid sequence of SEQ ID NO: 7;
valine (VAL), isoleucine (ILE) or leucine (LEU) at a position corresponding to position 170 of the amino acid sequence of SEQ ID NO: 7; and
histidine (HIS) at a position corresponding to position 193 of the amino acid sequence of SEQ ID NO: 7.
14. The method according to claim 12 , wherein the gene encoding chloramphenicol-O-acetyl transferase is composed of the sequence of SEQ ID NO: 24, and the gene encoding the second O-acetyl transferase is composed of any one sequence of SEQ ID NOS: 25 to 27.
15. The method according to claim 8 , wherein the transformed microorganism includes a gene encoding acetylesterase, which is attenuated or deleted.
16. The method according to claim 8 , wherein the transformed microorganism further expresses genes encoding glycerol-3-phosphate dehydrogenase and DL-glycerol-3-phosphatase, and the medium includes glucose.