Enzyme and application thereof
AssigneeShanghai Jiao Tong University
InventorsPing XU, Hongzhi TANG, Kunzhi ZHANG, Geng WU, Fei TAO
Life of this patent
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Abstract
Provides an artificial enzyme obtained by improving upon a sequence of a natural nicotine dehydrogenase, wherein the improvement comprises replacing at least one amino acid hindering product release with an amino acid with smaller side chains, thereby improving a catalytic rate.
Claims26
An artificial enzyme, manufactured by modifying a sequence of a natural enzyme, the natural enzyme having the activity for catalyzing the conversion reaction from compound I to compound II as shown by the following formula,
- or, the natural enzyme being nicotine dehydrogenase;
- wherein the modification comprises:
- substituting at least one of the amino acids that block product release with amino acids with smaller side chains.
An enzyme, having one or more of the following features:
- 1) the amino acid sequence of the enzyme includes amino acid sequence with more than 85%, or more than 90%, or more than 95%, or more than 98%, or more than 99% homology to the amino acid sequence shown in SEQ ID NO:2;
- or the amino acid sequence of the enzyme includes the amino acid sequence shown in SEQ ID NO:2;
- or the amino acid sequence of the enzyme is as shown in SEQ ID NO:2;
- 2) the enzyme is encoded by nucleic acid hybridized with complementary chain of nucleic acid encoding protein with an amino acid sequence shown by SEQ ID NO:2 under a high stringent condition;
- and
- 3) the amino acid sequence of the enzyme includes the amino acid sequence in which the amino acid sequence as shown in SEQ ID NO:2 is conservatively substituted;
- wherein, according to the relative locations, at least one of the amino acids at positions 353, 355 and 364 of the enzyme is a first small-molecular-weight amino acid;
- at least one of the amino acids at positions 163, 214, 218, 242, 246 and 249 of the enzyme is a second small-molecular-weight amino acid, the first small-molecular-weight amino acid has the molecular weight less than 140, and the second small-molecular-weight amino acid has the molecular weight less than 110.
An enzyme, the enzyme having the activity for catalyzing the conversion reaction from compound I to compound II as shown by the following formula:
- or, the enzyme being nicotine dehydrogenase;
- and
- the enzyme having a product release channel for product release, wherein the amino acid with side chains located at the narrowest site of product release channel has the molecular weight of less than 140.
(canceled)
-23. (canceled)
A method for modifying enzymes of monoamine oxidase family, wherein the method comprises:
- substituting at least one of the amino acids that block product release with amino acids with smaller side chains.
Dependencies are inferred from the claim text.