EP-4737475-A1 - PROTEIN VARIANT AND METHOD FOR PRODUCING L-ISOLEUCINE BY USING SAME

Abstract

The present disclosure relates to: a protein variant; a polynucleotide encoding the protein variant; a microorganism of the genus Corynebacterium, comprising at least one selected from the group consisting of the protein variant and the polynucleotide; and a method for producing L-isoleucine by using the microorganism.

Inventors

• YOON, Jeong-Hye
• KIM, Gyuree
• JUNG, MOO YOUNG
• SHIM, JIHYUN
• SEO, CHANG IL

Assignees

• CJ Cheiljedang Corporation

Dates

Publication Date

20260506

Application Date

20240724

Claims (11)

1. A protein variant comprising an amino acid sequence in which the amino acid corresponding to the 27th position from the N-terminus in the amino acid sequence of SEQ ID NO:116 is substituted with aspartic acid, an amino acid sequence in which the amino acid corresponding to the 461st position from the N-terminus in the amino acid sequence of SEQ ID NO:117 is substituted with glutamic acid, or an amino acid sequence in which the amino acid corresponding to the 75th position from the N-terminus in the amino acid sequence of SEQ ID NO:118 is substituted with valine.
2. The protein variant according to claim 1, wherein the protein variant has, respectively, an amino acid sequence identity of 95% or more to less than 100% to the amino acid sequence of SEQ ID NO:116, SEQ ID NO:117, or SEQ ID NO:118.
3. The protein variant according to claim 1, wherein the protein variant comprises an amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 8, or SEQ ID NO: 14.
4. A polynucleotide encoding the protein variant of claim 1.
5. A microorganism of the genus of Corynebacterium comprising at least one selected from the group consisting of the protein variant according to claim 1 and the polynucleotide encoding the protein variant.
6. The microorganism according to claim 5, wherein the microorganism of the genus of Corynebacterium is Corynebacterium glutamicum.
7. The microorganism according to claim 5, wherein the microorganism has increased L-isoleucine productivity, compared to a microorganism of the genus of Corynebacterium which does not comprise any one selected from the group consisting of the protein variant and the polynucleotide encoding the protein variant.
8. A method of producing L-isoleucine, comprising culturing the microorganism according to claim 5 in a medium.
9. The method of producing L-isoleucine according to claim 8, wherein the method further comprises recovering L-isoleucine from the cultured medium or microorganism.
10. A use for producing L-isoleucine, of the microorganism according to any one claim of claim 5 to claim 7.
11. A composition, method, product, process, or use characterized by at least one element disclosed in the present disclosure.

Description

[TECHNICAL FIELD] Cross-reference to related application(s) The present disclosure claims the benefit of the priority based on Korean Patent Application No. 10-2023-0096087 filed on July 24, 2023, and the entire contents disclosed in the document of the corresponding Korean patent application are incorporated as a part of the present disclosure. Throughout the present disclosure, numerous papers and patent documents are referenced and citations thereof are indicated. The disclosures of the cited papers and patent documents are incorporated into the present disclosure in their entirety by reference to more clearly describe the level of the technical field to which the present invention pertains and the contents of the present invention. The present disclosure relates to a protein variant, a microorganism of the genus of Corynebacterium comprising the protein variant, and a method of producing L-isoleucine using the microorganism. [BACKGROUND ART] L-isoleucine (lle, I) is classified as an essential amino acid and is one of the 20 essential amino acids. It belongs to the group of branched-chain amino acids (BCAAs) and is used in the production of various products such as animal feed, food additives, and pharmaceuticals. Because L-isoleucine contributes to energy generation after metabolism, hemoglobin synthesis, blood sugar regulation, and muscle building and maintenance, its applications are expanding not only in infusion solutions, nutritional supplements, and sports nutrition but also in the field of animal feed. Based on this trend, various attempts have been made to enhance the production capacity of L-isoleucine using microorganisms, including those belonging to the genus Corynebacterium (US6072083A). Despite these efforts, there remains a continuing need to develop technologies that can further improve the productivity of L-isoleucine. [DISCLOSURE] [TECHNICAL PROBLEM] One object of the present disclosure is to provide a protein variant. Another object of the present disclosure is to provide a polypeptide encoding the protein variant. Other object of the present disclosure is to provide a microorganism of the genus of Corynebacterium, comprising at least one selected from the group consisting of the protein variant and the polynucleotides encoding the same. Other object of the present disclosure is to provide a method of producing L-isoleucine, comprising culturing the microorganism in a medium. Other object of the present disclosure is to provide a use for producing L-isoleucine of the microorganism. [TECHNICAL SOLUTION] Description thereof in detail is as follows. On the other hand, each description, aspect and embodiment disclosed in the present disclosure can also be applied to each other description, aspect and embodiment. In other words, all combinations of various elements disclosed in the present disclosure belong to the scope of the present disclosure. In addition, the scope of the present disclosure is not to be considered limited by the specific description described below. Furthermore, those skilled in the art can recognize or confirm numerous equivalents to the specific aspects of the present application disclosed in the present application using only common experiments. In addition, these equivalents are intended to be included in the present application. One aspect of the present disclosure provides a protein variant comprising an amino acid sequence in which the amino acid corresponding to the 27th position from the N-terminus in the amino acid sequence of SEQ ID NO:116 is substituted with an amino acid different from the original amino acid, an amino acid sequence in which the amino acid corresponding to the 461st position from the N-terminus in the amino acid sequence of SEQ ID NO:117 is substituted with an amino acid different from the original amino acid, or an amino acid sequence in which the amino acid corresponding to the 75th position from the N-terminus in the amino acid sequence of SEQ ID NO:118 is substituted with an amino acid different from the original amino acid. In the present disclosure, the sequence of SEQ ID NOs: 116 to 118 are shown in table 1 below. [Table 1]Protein name (gene ID, gene name)Sequence (N-terminus → C-terminus)SEQ ID NOflavoprotein disulfide reductase (NCgI0658, IpdA)116ATP synthase F0F1 subunit beta (NCgI1165, atpD)117transcriptional regulator of sugar metabolism (NCgI1859, fruR)118 In one embodiment, the afore-mentioned protein variant of the present disclosure may be essentially consisting of an amino acid sequence in which the amino acid corresponding to the 27th position from the N-terminus in the amino acid sequence of SEQ ID NO:116 is substituted with an amino acid different from the original amino acid, an amino acid sequence in which the amino acid corresponding to the 461st position from the N-terminus in the amino acid sequence of SEQ ID NO:117 is substituted with an amino acid different from the original amino acid, or an amino acid sequence