Synthetic catalytic mimics of esterases, lipases or desaturases

Patent metadata

Field	Value
Publication number	US-2017022487-A1
Application number	US-201515301604-A
Country	US
Kind code	A1
Filing date	Apr 1, 2015
Priority date	Apr 2, 2014
Publication date	Jan 26, 2017
Grant date	—

Abstract

Novel synthetic catalytic structures or “synzymes,” e.g., fatty acid modified polypeptides, with catalytic properties are provided. It is believed that these synthetic catalytic structures mimic some of the precise conformational changes necessary for catalytic activities seen in enzymes. The catalytic properties of these synthetic catalytic structures or synzymes can be further improved by the application of controlled external forces, e.g., electric fields, or fluidized bed.

First claim

1 . A modified polypeptide comprising a synthetic polypeptide attached to a fatty acid,
wherein the synthetic polypeptide comprises the amino acid sequence X 1 -X 2 -X 3 -X 4 -X 5 (SEQ ID NO:1), wherein X 1 , X 3 , and X 5 are independently selected from the group consisting of alanine, an alanine analog, phenylalanine, and a phenylalanine analog; and X 2 and X 4 are independently selected from the group consisting of cysteine, a cysteine analog, serine, a serine analog, histidine, and a histidine analog; wherein when X 2 is histidine or a histidine analog, X 4 is cysteine or a cysteine analog, or serine or a serine analog; wherein when X 4 is histidine or a histidine analog, X 2 is cysteine or a cysteine analog, or serine or a serine analog; wherein the synthetic polypeptide is from 6 to 30 amino acids total in length; wherein the alanine analog is selected from the group consisting of β-alanine, dehydroalanine, aminoisobutyric acid, valine and norvaline; wherein the phenylalanine analog is selected from the group consisting of methylphenylalanine, 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid, phenylglycine, ethyltyrosine, and methyltyrosine; wherein the cysteine analog is selected from the group consisting of homocysteine and penicillamine; wherein the serine analog is selected from the group consisting of methylserine, threonine, 2-amino-3-hydroxy-4-methylpentanoic acid, 3-amino-2-hydroxy-5-methylhexanoic acid, 4-amino-3-hydroxy-6-methylheptanoic acid, and 2-amino-3-hydroxy-3-methylbutanoic acid; and wherein the histidine analog is selected from the group consisting of β-(1,2,3-triazol-4-yl)-DL-alanine, and 1,2,4-triazole-3-alanine.

2 . The modified polypeptide of claim 1 , wherein the fatty acid is selected from the group consisting of palmitic acid, octanoic acid, hexanoic acid, docosahexaenoic acid, lauric acid, nonanoic acid, valeric acid, decanoic acid, oleic acid, arachidic acid, myristic acid, arachidonic acid, linoleic acid, stearic acid, decosanoic acid, tetracosanoic acid, sapienic acid, elaidic acid, vaccenic acid, eicosapentaenoic acid, and erucic acid.

3 . The modified polypeptide of claim 1 , wherein the fatty acid is attached to the N-terminus of the synthetic peptide.

4 . The modified polypeptide of claim 1 , wherein the fatty acid is attached to the C-terminus of the synthetic peptide.

5 . The modified polypeptide of claim 3 , wherein the synthetic polypeptide comprises a negatively charged C-terminal residue selected from the group consisting of aspartic acid, glutamic acid, methyl aspartic acid, methyl glutamic acid, 2-aminoadipic acid, 2-aminoheptanedioic acid, and iminodiacetic acid.

6 . The modified polypeptide of claim 4 , wherein the synthetic polypeptide comprises an N-terminal residue selected from the group consisting of glycine, lysine, arginine, citrulline, ornithine, and 2-amino-3-guanidinopropionic acid.

7 . The modified polypeptide of claim 1 , wherein the synthetic polypeptide comprises an amino acid sequence selected from the group consisting of:

(SEQ ID NO: 2)

Ala-Cys-Ala-His-Ala;

(SEQ ID NO: 3)

Ala-Ser-Ala-His-Ala;

(SEQ ID NO: 4)

Phe-Cys-Phe-His-Ala;

(SEQ ID NO: 5)

Phe-Ser-Phe-His-Ala;

(SEQ ID NO: 6)

Phe-His-Phe-Cys-Ala;

(SEQ ID NO: 7)

Phe-His-Phe-Ser-Ala;

(SEQ ID NO: 8)

Ala-Cys-Ala-His-Ala-Asp;

(SEQ ID NO: 9)

Ala-Ser-Ala-His-Ala-Asp;

(SEQ ID NO: 10)

Phe-Cys-Phe-His-Ala-Asp;

(SEQ ID NO: 11)

Phe-Ser-Phe-His-Ala-Asp;

(SEQ ID NO: 12)

Asp-Phe-His-Phe-Cys-Ala;

(SEQ ID NO: 13)

Asp-Phe-His-Phe-Ser-Ala;

(SEQ ID NO: 14)

Ala-Ala-Cys-Ala-His-Ala-Asp;

(SEQ ID NO: 15)

Ala-Ala-Ser-Ala-His-Ala-Asp;

(SEQ ID NO: 16)

Ala-Phe-Cys-Phe-His-Ala-Asp;

(SEQ ID NO: 17)

Ala-Phe-Ser-Phe-His-Ala-Asp;

(SEQ ID NO: 18)

Asp-Phe-His-Phe-Cys-Ala-Gly;

(SEQ ID NO: 19)

Asp-Phe-His-Phe-Ser-Ala-Gly;

(SEQ ID NO: 20)

Gly-Ala-Ala-Cys-Ala-His-Ala-Asp;

(SEQ ID NO: 21)

Gly-Ala-Ala-Ser-Ala-His-Ala-Asp;

(SEQ ID NO: 22)

Gly-Ala-Phe-Cys-Phe-His-Ala-Asp;

Assignees

• Univ California
• Z-Field Tech Llc

Inventors

• Heller Michael J
• Takahashi Tsukasa
• Sheldon Iii Edward Lewis

Classifications

• C12Y301/00

  Hydrolases acting on ester bonds (3.1) · CPC title

• C12N9/16

  acting on ester bonds (3.1) · CPC title

• C12N9/20 Primary

  Triglyceride splitting, e.g. by means of lipase · CPC title

• C12Y301/01

  Carboxylic ester hydrolases (3.1.1) · CPC title

• C12N9/18

  Carboxylic ester hydrolases {(3.1.1)} · CPC title